bromochloroacetic-acid and Inflammation

Intermediate filament (IF) proteins make up the largest family of cytoskeletal proteins in metazoans, and are traditionally known for their roles in fostering structural integrity in cells and tissues. Remarkably, individual IF genes are tightly regulated in a fashion that reflects the type of tissue, its developmental and differentiation stages, and biological context. In cancer, IF proteins serve as diagnostic markers, as tumor cells partially retain their original signature expression of IF proteins. However, there are also characteristic alterations in IF gene expression and protein regulation. The use of high throughput analytics suggests that tumor-associated alterations in IF gene expression have prognostic value. Parallel research is also showing that IF proteins directly and significantly impact several key cellular properties, including proliferation, death, migration, and invasiveness, with a demonstrated impact on the development, progression, and characteristics of various tumors. In this review, we draw from recent studies focused on three IF proteins most associated with cancer (keratins, vimentin, and nestin) to highlight how several "hallmarks of cancer" described by Hanahan and Weinberg are impacted by IF proteins. The evidence already in hand establishes that IF proteins function beyond their classical roles as markers and serve as effectors of tumorigenesis.

Topics: Animals; Carcinogenesis; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Immunity, Innate; Inflammation; Intermediate Filaments; Keratins; Mice; Neoplasm Metastasis; Neovascularization, Pathologic; Nestin; Vimentin

Topics: Humans; Inflammation; Keratins; Skin Diseases

Hereditary keratin disorders of the skin and its appendages comprise a large group of clinically heterogeneous disfiguring blistering and ichthyotic diseases, primarily characterized by the loss of tissue integrity, blistering and hyperkeratosis in severely affected tissues. Pathogenic mutations in keratins cause these afflictions. Typically, these mutations in concert with characteristic features have formed the basis for improved disease diagnosis, prognosis and most recently therapy development. Examples include epidermolysis bullosa simplex, keratinopathic ichthyosis, pachyonychia congenita and several other tissue-specific hereditary keratinopathies. Understanding the molecular and genetic events underlying skin dysfunction has initiated alternative treatment approaches that may provide novel therapeutic opportunities for affected patients. Animal and in vitro disease modelling studies have shed more light on molecular pathogenesis, further defining the role of keratins in disease processes and promoting the translational development of new gene and pharmacological therapeutic strategies. Given that the molecular basis for these monogenic disorders is well established, gene therapy and drug discovery targeting pharmacological compounds with the ability to reinforce the compromised cytoskeleton may lead to promising new therapeutic strategies for treating hereditary keratinopathies. In this review, we will summarize and discuss recent advances in the preclinical and clinical modelling and development of gene, natural product, pharmacological and protein-based therapies for these disorders, highlighting the feasibility of new approaches for translational clinical therapy.

Topics: Animals; Botulinum Toxins, Type A; Genetic Therapy; Humans; Induced Pluripotent Stem Cells; Inflammation; Keratins; MAP Kinase Signaling System; Molecular Chaperones; Mutation; Nanoparticles; Proteasome Endopeptidase Complex; Retinoids; Skin Diseases, Genetic; Ubiquitin

To review recently published research into the use of dietary cysteine and/or its derivatives as functional food supplements that will enhance antioxidant status and improve outcome in certain diseases.. L-cysteine is now widely recognized as a conditionally essential or (indispensible) sulphur amino acid. It plays a key role in the metabolic pathways involving methionine, taurine and glutathione (GSH), and may help fight chronic inflammation by boosting antioxidant status. In stressed and inflammatory states, sulphur amino acid metabolism adapts to meet the increased requirements for cysteine as a rate-limiting substrate for GSH. Critically ill patients receiving enteral or parenteral nutrition, enriched with cysteine, exhibit decreased cysteine catabolism and improved GSH synthesis. The naturally occurring cysteine-rich proteins, whey or keratin, have the potential to be manufactured into high quality, high cysteine-containing functional foods for clinical investigation.. Cysteine-rich proteins, such as keratin, may have advantages over the simple amino acid or its derivatives, as nutraceuticals, to safely and beneficially improve antioxidant status in health and disease.

Topics: Antioxidants; Cysteine; Dietary Supplements; Functional Food; Glutathione; Humans; Inflammation; Keratins; Methionine; Nutritional Support; Oxidation-Reduction; Sulfur; Taurine

The plasma kinin-forming cascade can be activated by contact with negatively charged macromolecules leading to binding and autoactivation of factor XII, activation of prekallikrein to kallikrein by factor XIIa, and cleavage of high molecular weight kininogen (HK) by kallikrein to release the vasoactive peptide bradykinin. Once kallikrein formation begins, there is rapid cleavage of unactivated factor XII to factor XIIa, and this positive feedback is favored kinetically over factor XII autoactivation. Examples of surface initiators that can function in this fashion are endotoxin, sulfated mucopolysaccharides, and aggregated Abeta protein. Physiological activation appears to occur along the surface of endothelial cells both by the aforementioned contact-initiated reactions as well as bypass pathways that are independent of factor XII. Factor XII binds primarily to cell surface u-PAR (urokinase plasminogen activator receptor); HK binds to gC1qR via its light chain (domain 5) and to cytokeratin 1 by its heavy chain (domain 3) and, to a lesser degree, by its light chain. Prekallikrein circulates bound to HK (as does coagulation factor XI), and prekallikrein is thereby brought to the surface as HK binds. All cell-binding reactions are dependent on zinc ion. Endothelial cells (HUVECs) have bimolecular complexes of u-PAR-cytokeratin 1 and gC1qR-cytokeratin 1 at the cell surface plus free gC1qR, which is present in substantial molar excess. Factor XII appears to interact primarily with the u-PAR-cytokeratin 1 complex, whereas HK binds primarily to the gC1qR-cytokeratin 1 complex and to free gC1qR. Release of endothelial cell heat shock protein 90 (Hsp90) or the enzyme prolylcarboxypeptidase leads to activation of the bradykinin-forming cascade by activating the prekallikrein-HK complex. In contrast to factor XIIa, neither will activate prekallikrein in the absence of HK, both reactions require zinc ion, and the stoichiometry suggests interaction of one molecule of Hsp90 (for example) with one molecule of prekallikrein-HK complex. The presence of factor XII, however, leads to a marked augmentation in reaction rate via the kallikrein feedback as well as to a change to classic enzyme-substrate kinetics. The circumstances in which activation is initiated by factor XII autoactivation or by these factor XII bypasses are yet to be defined. The pathologic conditions in which bradykinin generation appears important include hereditary and acquired C1 inhibitor deficiency, c

Topics: Amino Acid Sequence; Animals; Bradykinin; Endothelial Cells; Humans; Immunity, Innate; Inflammation; Keratins; Vasodilator Agents

This review indicates that the patient-to-patient uniqueness commonly seen in chronic laminitis represents the variable presence of the digital pathologies. Although some degree of mechanical failure is always present, the secondary metabolic and growth dysplasias, vascular pathologies, and sepsis may or may not be evident. The presence and severity of these pathologies appear to have a more significant impact on the prognosis of individual cases than does the displacement of the distal phalanx. It should be reiterated that it is often the combined presence of these individual pathologies that gives rise to the patient that is totally refractory to treatment. In the absence of these pathologies, many horses with significant displacement of the distal phalanx are not in pain and are not in need of treatment. It thus follows that a key to the improved rehabilitation of difficult patients is focusing research on the physiopathology and diagnosis of these nonmechanical problems.

Topics: Animals; Chronic Disease; Foot Diseases; Hoof and Claw; Horse Diseases; Horses; Inflammation; Keratins

The last decade has witnessed a remarkable expansion in the understanding of the skin as an immunologic organ. A prime example of this is the discovery that keratinocytes can synthesize and secrete immunoregulatory and proinflammatory glycoproteins termed cytokines. Initial studies focused on interleukin 1-like molecules produced by keratinocytes, initially termed epidermal cell-derived thymocyte activating factor (ETAF). It is now clear that the functional activities described by the term ETAF are composed mainly of interleukin 1 alpha and interleukin 1 beta, although other epidermal cytokines may contribute to the biologic activity. The biologic properties of interleukin 1 are the basis of this review.

Topics: Animals; Epidermis; Humans; Inflammation; Interleukin-1; Keratins; Lymphocytes; Molecular Structure; Tumor Necrosis Factor-alpha

Topics: Animals; Biological Factors; Cytokines; Epidermal Cells; Epidermis; Gene Expression Regulation; Hematopoiesis; Humans; Inflammation; Keratins; Lymphoid Tissue; Mice; RNA, Messenger

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Epidermal Cells; Humans; Inflammation; Keratins; Melanins; Melanocytes; Monophenol Monooxygenase; Phorbol Esters; Photosensitivity Disorders; Platelet Activating Factor; Prostaglandins; Swine

The retinoids are synthetic derivatives of vitamin A. Isotretinoin (13-cis-retinoic acid) is now being widely used in the United States for severe acne and etretinate is available in Europe and other countries for psoriasis. These drugs are also effective for a number of other skin diseases. This is an attempt to review basic knowledge of retinoids with which the practicing dermatologist should be familiar, to review the current status of studies, and to speculate on the present and future roles of these drugs in dermatology.

Topics: Acne Vulgaris; Etretinate; Humans; Inflammation; Isotretinoin; Keratins; Psoriasis; Retinoids; Sebum; Skin Diseases; Skin Neoplasms; Sweat Gland Diseases; Tretinoin; Vitamin A

Nineteen patients with carcinoma cuniculatum are presented. Of these, 17 were male and two were female. The age range was from 26 to 73 years with a mean of approximately 54 years. Sixteen tumours were located on the foot, the other three were situated on the knee, wrist and finger respectively. The pathological features of carcinoma cuniculatum are described and the aetiology of the tumour is discussed.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cytoplasmic Granules; Female; Foot Diseases; Humans; Inflammation; Keratins; Male; Middle Aged; Mitosis; Neoplasm Invasiveness; Skin Neoplasms

Topics: Acid Phosphatase; Animals; Cathepsins; Chediak-Higashi Syndrome; Fabry Disease; Inflammation; Keratins; Langerhans Cells; Light; Lysosomes; Melanophores; Microscopy, Electron; Peptide Hydrolases; Phagocytosis; Pigmentation; Protease Inhibitors; Sebaceous Glands; Skin; Skin Diseases; Skin Neoplasms; Vitamin A

Topics: Epithelium; Gingiva; Gonadal Steroid Hormones; Humans; Hydrogen-Ion Concentration; Inflammation; Keratins; Mouth; Vitamin A

Topics: Age Factors; Carcinoma, Squamous Cell; Cell Membrane; Cell Nucleus; Cervix Mucus; Cervix Uteri; Chromatin; Cytoplasm; Female; Herpes Simplex; Humans; Hyperplasia; Inflammation; Keratins; Metaplasia; Pregnancy; Trichomonas Infections

Dandruff is a common scalp condition characterized by flakes, pruritus and sometimes mild erythema. These symptoms reflect underlying histopathologic and biochemical events that must be reversed if treatment is to be effective.. This study aimed to better characterize the state of the epidermis in dandruff and to determine how a defined set of skin surface biomarkers of this state change during a successful course of treatment with a potentiated zinc pyrithione (ZPT) shampoo.. A population of dandruff sufferers was treated for 3 weeks with a commercial ZPT shampoo or a non-medicated product, and the effect of treatment on adherent scalp flake (ASF) scores was evaluated. Biopsies were taken from lesional sites at baseline and at the end of the study for histomorphometric and histopathologic analysis. Stratum corneum (SC) samples were likewise obtained for evaluation of biochemical markers of inflammation (IL-1α, IL-1RA, IL-8) and barrier integrity (keratin 1, 10, 11; involucrin; SC lipids; human serum albumin). The biomarker profile was evaluated first by comparison with that in non-dandruff subjects at baseline, and then to determine whether any treatment-induced changes were correlated with reductions in flaking in dandruff sufferers.. Taken together, our studies showed that treatment with the ZPT shampoo led to an improvement in the overall scalp condition as assessed by the resolution of flaking, reduction in epidermal thickness and inflammatory biomarkers, and a dramatic improvement in biomarkers of epidermal barrier integrity.. The combination of biomarkers examined appears to be a good overall descriptor of the health of the scalp in dandruff, and changes in these biomarkers track with tissue-level events that underlie clinical efficacy in the treatment of dandruff by ZPT shampoo. For the first time, we demonstrate a set of tools that extend beyond flaking scores to provide insight into specific biological changes occurring on the scalp to enable an objective assessment of scalp health.

Topics: Adolescent; Adult; Aged; Biomarkers; Biopsy; Dermatitis, Seborrheic; Epidermis; Female; Hair Preparations; Humans; Inflammation; Interleukins; Keratins; Male; Middle Aged; Organometallic Compounds; Pyridines; Scalp; Treatment Outcome; Young Adult

To determine whether inflamed and uninflamed epidermoid cysts differ in the number and/or type of bacteria inhabiting them.. A controlled study. We obtained aerobic and anaerobic bacterial culture specimens from 25 inflamed and 25 uninflamed epidermoid cysts.. A university medical center.. Nonimmunocompromised adults without recent systemic use of antibiotics.. The 2 groups did not differ significantly with respect to number of bacterial isolates, "no growth" cultures, and aerobic, anaerobic, or potential pathogens cultured.. The microbiological milieu of inflamed epidermoid cysts is similar to that of uninflamed cysts. Possible mechanisms for inflammation are discussed.

Topics: Adult; Aged; Bacteria, Aerobic; Bacteria, Anaerobic; Colony Count, Microbial; Cysts; Erythema; Female; Gram-Positive Bacterial Infections; Humans; Inflammation; Keratins; Male; Middle Aged; Peptostreptococcus; Skin Diseases; Staphylococcal Infections; Staphylococcus aureus; Suppuration

The clinical efficacy and tolerability of the vitamin D3 analogues calcitriol, calcipotriol and 1 alpha, 24 dihydroxyvitamin D3 in the treatment of psoriasis have been assessed in various clinical studies. In vitro and in vivo investigations have shown interference of these compounds with epidermal growth, keratinisation and inflammation. In this study we quantified the in vivo cell biological effects during treatment of psoriatic plaques with 1 alpha, 24 dihydroxyvitamin D3. By using a flow cytometric triple labelling procedure, we could discriminate different epidermal subpopulations, permitting precise assessment of epidermal cell cycle kinetics. Twenty patients with plaque-type psoriasis were treated in a double-blind placebo-controlled left-right comparative study with 1 alpha, 24 dihydroxyvitamin D3 ointment (4 micrograms/g applied once daily) for 8 weeks. Epidermal cell suspensions prepared from keratotome biopsies taken before and after treatment were stained with TO-PRO-3 iodide (a new DNA fluorochrome) and monoclonal antibodies against keratin 10 (as a marker for differentiation) and vimentin (as a marker for inflammation), simultaneously. The flow cytometric analyses showed a significant decrease of proliferating basal keratinocytes in verum-treated lesions, whereas such a decrease was not observed in placebo-treated lesions. The amount of keratin 10-positive keratinocytes increased and the presence of vimentin-positive cells decreased in cell suspensions derived from both verum- and placebo-treated lesions, but these effects were not significant. We conclude that multiparameter flow cytometry promises to be an adequate approach to assess the interference of antipsoriatic treatments with cutaneous inflammation, epidermal proliferation and keratinisation. Topical 1 alpha, 24 dihydroxyvitamin D3 seems to exert its in vivo antipsoriatic effect mainly through an inhibition of epidermal growth.

Topics: Administration, Cutaneous; Adult; Aged; Biopsy; Cell Cycle; Cell Differentiation; Cell Division; Cells, Cultured; Dermatologic Agents; Dihydroxycholecalciferols; DNA; Double-Blind Method; Epidermis; Female; Flow Cytometry; Fluorescent Dyes; Humans; Inflammation; Keratins; Male; Middle Aged; Placebos; Psoriasis; Vimentin

Corticosteroids are important in the treatment of inflammatory dermatoses, such as psoriasis. They have anti-inflammatory, anti-proliferative and immunosuppressive effects. In this study, the effect of budesonide on proliferation, inflammatory cells and cytokines in psoriasis was investigated. In order to elucidate the time course of the different effects of corticosteroid treatment in psoriasis, six patients were treated for 3 weeks with budesonide 0.025% ointment (Preferid), and biopsies were studied immunohistochemically, before treatment and after 1 and 3 weeks of treatment. Clinical scores together with staining with antibodies indicating proliferation, keratin 16, keratin 10, T-lymphocytes, monocytes, polymorphonuclear leukocytes, Langerhans cells, interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), and intercellular adhesion molecule-1 (ICAM-1) were performed. 'Psoriasis area' and 'severity index' (PASI) scores were significantly reduced after 1 week and 3 weeks of treatment. Epidermal hyperproliferation (Ki-67 binding) and suprabasal keratin 16 (Ks8.12) expression decreased within 1 week, while keratin 10 (RKSE60) expression did not change. Five out of 6 patients showed cytokine levels (IL-1alpha, IL-6, IL-8, and TNF-alpha; detected immunohistochemically) in the normal range, while 1 patient had highly increased cytokine levels. In this patient, cytokine levels decreased during treatment. In 4 patients, showing high dermal ICAM-1 expression before treatment, a consistent reduction of ICAM-1 on endothelial cells was observed. The inflammatory infiltrate (T-lymphocytes (T11), monocytes/macrophages (WT14), polymorphonuclear leukocytes (PMN, anti-elastase)) was reduced to some extent after 3 weeks. The number of Langerhans cells (OKT6) did not change. These results indicate that the psoriatic lesions, although clinically comparable, show interindividual differences in cytokine expression. Corticosteroid treatment for 1-3 weeks improves clinical scores and hyperproliferation. Cytokine levels are reduced during steroid treatment in the patient who showed high levels before treatment. To suppress the infiltrate entirely, longer steroid treatment is probably necessary. This may explain the relapse seen after short term corticosteroid therapy.

Topics: Administration, Topical; Adult; Aged; Anti-Inflammatory Agents; Biomarkers; Budesonide; Cell Division; Cytokines; Female; Glucocorticoids; Humans; Immunohistochemistry; Inflammation; Inflammation Mediators; Keratins; Male; Middle Aged; Pregnenediones; Psoriasis; Time Factors

The aim of the present study was to discover to what extent 1,24(OH)2D3 ointment (tacalcitol; 4 micrograms/g) can modulate epidermal proliferation and keratinization, and several aspects of inflammation. Ten patients with psoriasis vulgaris were included in a placebo-controlled, double-blind study, using 1,24(OH)2D3 ointment (4 micrograms/g). Before, and after 8 weeks of treatment, punch biopsies were taken from lesions treated with the active agent and placebo-treated lesions. An immunohistochemical study was carried out using monoclonal antibodies against the hyperproliferation-associated keratin 16, against cycling nuclei, filaggrin, involucrin, T lymphocytes, Langerhans cells, CD14 and polymorphonuclear leucocytes (PMN). The Wilcoxon test for matched pairs was used for statistical analysis of results. The biopsies from the lesions treated with the active agent showed a statistically significant change towards normalization of all aspects of inflammation studied, and of epidermal proliferation and keratinization, but there did not appear to be any effect on Langerhans cells. The only parameter which showed a significant alteration in the placebo-treated lesions was the number of cycling nuclei in the epidermis (P < or = 0.02). However, the biopsies from the plaques treated with the active agent showed a greater decrease of cycling cells (decrease: Mactive = 70, Mplacebo = 53) and a lower P-value (< or = 0.01). We therefore conclude that at the cell biological level 1,24(OH)2D3 ointment (4 micrograms/g) has a substantial effect on several cell types, with regard to inflammation, epidermal proliferation and keratinization, with the exception of Langerhans cells.

Topics: Cell Division; Dihydroxycholecalciferols; Double-Blind Method; Epidermis; Filaggrin Proteins; Humans; Immunohistochemistry; Inflammation; Keratins; Psoriasis

Topics: Alveolar Process; Animals; Clinical Trials as Topic; Connective Tissue; Epithelial Cells; Fibroblasts; Freeze Drying; Gingiva; Gingivectomy; Graft vs Host Reaction; Haplorhini; Inflammation; Keratins; Macaca; Mouth Mucosa; Osteoclasts; Postoperative Complications; Skin Transplantation; Transplantation Immunology; Transplantation, Homologous; Wound Healing

Keratin pearls are intraepithelial accumulations of squamous cells and debris that can be an etiology of vulvovaginal irritation in pediatric patients and are often associated with clitoral adhesions. Historically, most cases have been managed with manual or operative lysis of adhesions.. Two prepubertal girls presented to our clinic with chronic clitoral irritation and were found to have clitoral adhesions with keratin pearls. Both were managed with topical estrogen cream, which resulted in resolution of their symptoms.. Keratin pearls can form when the overlying clitoral epithelium becomes blocked by clitoral adhesions. Hypoestrogenism is thought to be implicated in adhesion development; thus, topical estrogen cream is a reasonable option in initial management. Our results demonstrate a noninvasive alternative to the initial treatment of clitoral keratin pearls.

Topics: Administration, Topical; Child; Clitoris; Emollients; Estrogens; Female; Humans; Inflammation; Keratins

Controlling infection-driven inflammation is a major clinical dilemma because of limited therapeutic options and possible adverse effects on microbial clearance. Compounding this difficulty is the continued emergence of drug-resistant bacteria, where experimental strategies aiming to augment inflammatory responses for enhanced microbial killing are not applicable treatment options for infections of vulnerable organs. As with corneal infections, severe or prolonged inflammation jeopardizes corneal transparency, leading to devastating vision loss. We hypothesized that keratin 6a-derived antimicrobial peptides (KAMPs) may be a two-pronged remedy capable of tackling bacterial infection and inflammation at once. We used murine peritoneal neutrophils and macrophages, together with an in vivo model of sterile corneal inflammation, to find that nontoxic and prohealing KAMPs with natural 10- and 18-amino acid sequences suppressed lipoteichoic acid (LTA)- and lipopolysaccharide (LPS)-induced NFκB and IRF3 activation, proinflammatory cytokine production, and phagocyte recruitment independently of their bactericidal function. Mechanistically, KAMPs not only competed with bacterial ligands for cell surface Toll-like receptor (TLR) and co-receptors (MD2, CD14, and TLR2) but also reduced cell surface availability of TLR2 and TLR4 through promotion of receptor endocytosis. Topical KAMP treatment effectively alleviated experimental bacterial keratitis, as evidenced by substantial reductions of corneal opacification, inflammatory cell infiltration, and bacterial burden. These findings reveal the TLR-targeting activities of KAMPs and demonstrate their therapeutic potential as a multifunctional drug for managing infectious inflammatory disease.

Topics: Animals; Carrier Proteins; Inflammation; Keratins; Lipopolysaccharide Receptors; Lipopolysaccharides; Mice; Peptides; Toll-Like Receptor 2

The aim of this research was to investigate the role of the cornified epithelium, the outermost layer of the oral mucosa, engineered to prevent water loss and microorganism invasion, in severe forms of periodontitis (stage III or IV, grade C).. Porphyromonas gingivalis, a major periodontal disease pathogen, can affect cornified epithelial protein expression through chronic activation of signal transducer and activator of transcription 6 (Stat6). We used a mouse model, Stat6VT, that mimics this to determine the effects of barrier defect on P gingivalis-induced inflammation, bone loss, and cornified epithelial protein expression, and compared histologic and immunohistologic findings with tissues obtained from human controls and patients with stage III and IV, grade C disease. Alveolar bone loss in mice was assessed using micro-computerised tomography, and soft tissue morphology was qualitatively and semi-quantitatively assessed by histologic examination for several proteins, including loricrin, filaggrin, cytokeratin 1, cytokeratin 14, a proliferation marker, a pan-leukocyte marker, as well as morphologic signs of inflammation. Relative cytokine levels were measured in mouse plasma by cytokine array.. In the tissues from patients with periodontal disease, there were greater signs of inflammation (rete pegs, clear cells, inflammatory infiltrates) and a decrease and broadening of expression of loricrin and cytokeratin 1. Cytokeratin 14 expression was also broader and decreased in stage IV. P gingivalis-infected Stat6VT mice showed greater alveolar bone loss in 9 out of 16 examined sites, and similar patterns of disruption to human patients in expression of loricrin and cytokeratins 1 and 14. There were also increased numbers of leukocytes, decreased proliferation, and greater signs of inflammation compared with P gingivalis-infected control mice.. Our study provides evidence that changes in epithelial organisation can exacerbate the effects of P gingivalis infection, with similarities to the most severe forms of human periodontitis.

Topics: Alveolar Bone Loss; Animals; Cytokines; Humans; Inflammation; Keratin-14; Keratins; Mice; Periodontitis; Porphyromonas gingivalis

Polycyclic aromatic hydrocarbons like benzo[a]pyrene (BaP) are generated during incomplete combustion of organic materials. Prior research has demonstrated that BaP is a prenatal ovarian toxicant and carcinogen. However, the metabolic pathways active in the embryo and its developing gonads and the mechanisms by which prenatal exposure to BaP predisposes to ovarian tumors later in life remain to be fully elucidated. To address these data gaps, we orally dosed pregnant female mice with BaP from embryonic day (E) 6.5 to E11.5 (0, 0.2, or 2 mg/kg/day) for metabolite measurement or E9.5 to E11.5 (0 or 3.33 mg/kg/day) for embryonic gonad RNA sequencing. Embryos were harvested at E13.5 for both experiments. The sum of BaP metabolite concentrations increased significantly with dose in the embryos and placentas, and concentrations were significantly higher in female than male embryos and in embryos than placentas. RNA sequencing revealed that enzymes involved in metabolic activation of BaP are expressed at moderate to high levels in embryonic gonads and that greater transcriptomic changes occurred in the ovaries in response to BaP than in the testes. We identified 490 differentially expressed genes (DEGs) with false discovery rate P-values < 0.05 when comparing BaP-exposed to control ovaries but no statistically significant DEGs between BaP-exposed and control testes. Genes related to monocyte/macrophage recruitment and activity, prolactin family genes, and several keratin genes were among the most upregulated genes in the BaP-exposed ovaries. Results show that developing ovaries are more sensitive than testes to prenatal BaP exposure, which may be related to higher concentrations of BaP metabolites in female embryos.

Topics: Animals; Benzo(a)pyrene; Chromatography, High Pressure Liquid; Computational Biology; Female; Gonads; Inflammation; Keratins; Macrophages; Male; Mice; Mice, Inbred C57BL; Monocytes; Ovary; Placenta; Pregnancy; Pregnancy, Animal; RNA-Seq; Sex Factors; Testis; Time Factors; Transcriptome

Amniotic membrane (AM) is often applied as a substitute material during ocular surface reconstruction. However, since AM has several disadvantages, alternative materials must be considered for this application. Keratin films made from human hair (KFs) have previously been presented as a promising option; they exhibited suitable characteristics and satisfactory biocompatibility in an in vivo rabbit model. Nevertheless, dexamethasone (DEX) eye drops are necessary after surgery to suppress inflammation. Since eye drops must be administered frequently, this might result in poor patient compliance, and the release of DEX at the transplant site would be clinically beneficial. Therefore, we aimed to incorporate DEX into KFs without hindering the positive film characteristics. Drug-loaded KFs were generated either by suspension technique or by the addition of solubilizing agents. The resulting specimens were analyzed regarding appearance, loading capacity, transparency, mechanical characteristics, swelling behavior and in vitro release. Furthermore, biocompatibility was assessed in vitro by determining the cell viability, seeding efficiency and growth behavior of corneal epithelial cells. The amount of incorporated DEX influenced the transparency and biomechanical properties of the films, but even highly loaded films showed properties similar to those of AM. The suspension technique was identified as the best incorporation approach regarding chemical stability and prolonged DEX release. Moreover, suspended DEX in the films did not negatively impact cell seeding efficiencies, and the cell-growth behaviors on the specimens with moderate DEX loads were satisfactory. This suggest that these films could comprise a suitable alternative material with additional anti-inflammatory activity for ocular surface reconstruction. Graphical abstract.

Topics: Amnion; Animals; Anti-Inflammatory Agents; Dexamethasone; Inflammation; Keratins; Ophthalmology; Rabbits; Tissue Scaffolds

Abnormal endometrial repair after injury results in the formation of intrauterine adhesions (IUA) and a thin endometrium, which are key causes for implantation failure and infertility. Stem cell transplantation offers a potential alternative for some cases of severe Asherman's syndrome that cannot be treated with surgery or hormonal therapy. Umbilical cord-derived mesenchymal stem cells (UCMSCs) have been reported to repair the damaged endometrium. However, there is no report on the effects of UCMSCs previously seeded on human acellular amniotic matrix (AAM) on endometrial injury.. Absolute ethanol was injected into rat uteri to damage the endometrium. UCMSCs previously seeded on AAM were surgically transplanted. Using a variety of methods, the treatment response was assessed by endometrial thickness, endometrial biomarker expression, endometrial receptivity, cell proliferation, and inflammatory factors.. Endometrial thickness was markedly improved after UCMSC-AAM transplantation. The expression of endometrial biomarkers, namely, vimentin, cytokeratin, and integrin. UCMSC transplantation using AAM as the carrier can be applied to treat endometrial injury in rats. The successful preparation of lyophilized AAM provides the possibility of secondary infectious disease screening and amniotic matrix quality detection, followed by retrospective analysis. The UCMSC-AAM complex may promote the better application of UCMSCs on the treatment of injured endometrium.

Topics: Amnion; Animals; Biomarkers; Cell Transplantation; Disease Models, Animal; Endometrium; Female; Humans; Inflammation; Integrins; Keratins; Mesenchymal Stem Cells; Placenta; Pregnancy; Rats; Rats, Sprague-Dawley; Regeneration; Retrospective Studies; Stem Cell Transplantation; Tissue Adhesions; Umbilical Cord; Uterine Diseases; Uterus; Vimentin

Concentration of hyaluronic acid (HA) in the lungs increases in idiopathic pulmonary fibrosis (IPF). HA is involved in the organization of fibrin, fibronectin, and collagen. HA has been proposed to be a biomarker of fibrosis and a potential target for antifibrotic therapy. Hyaluronidase (HD) breaks down HA into fragments, but is a subject of rapid hydrolysis. A conjugate of poloxamer hyaluronidase (pHD) was prepared using protein immobilization with ionizing radiation. In a model of bleomycin-induced pulmonary fibrosis, pHD decreased the level of tissue IL-1β and TGF-β, prevented the infiltration of the lung parenchyma by CD16

Topics: Animals; Cell Differentiation; Collagen Type I; Hyaluronic Acid; Hyaluronoglucosaminidase; Hydroxyproline; Idiopathic Pulmonary Fibrosis; Inflammation; Interleukin-1beta; Keratins; Lung; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Platelet Endothelial Cell Adhesion Molecule-1; Poloxamer; Receptors, IgG; Spiperone; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Keratin is a cytoskeletal scaffolding protein essential for wound healing and tissue recovery. The aim of the study was to evaluate the potential role of insoluble fur keratin-derived powder containing silver nanoparticles (FKDP-AgNP) in the allogenic full-thickness surgical skin wound model in diabetic mice. The scanning electron microscopy image evidenced that the keratin surface is covered by a single layer of silver nanoparticles. Data obtained from dynamic light scattering and micellar electrokinetic chromatography showed three fractions of silver nanoparticles with an average diameter of 130, 22.5, and 5 nm. Microbiologic results revealed that the designed insoluble FKDP-AgNP dressing to some extent inhibit the growth of Escherichia coli and Staphylococcus aureus. In vitro assays showed that the FKDP-AgNP dressing did not inhibit fibroblast growth or induce hemolysis. In vivo studies using a diabetic mice model confirmed biocompatible properties of the insoluble keratin dressings. FKDP-AgNP significantly accelerated wound closure and epithelization at Days 5 and 8 (p < .05) when compared with controls. Histological examination of the inflammatory response documented that FKDP-AgNP-treated wounds contained predominantly macrophages, whereas their untreated variants showed mixed cell infiltrates rich in neutrophils. Wound inflammatory response based on macrophages favors tissue remodeling and healing. In conclusion, the investigated FKDP-AgNP dressing consisting of an insoluble fraction of keratin, which is biocompatible, significantly accelerated wound healing in a diabetic mouse model.

Topics: Animals; Anti-Bacterial Agents; Bandages; Biocompatible Materials; Cell Movement; Cell Proliferation; Cell Survival; Colloids; Cytokines; Diabetes Mellitus, Experimental; Escherichia coli; Inflammation; Keratins; Kinetics; Light; Male; Metal Nanoparticles; Mice; Mice, Inbred C57BL; Microbial Sensitivity Tests; NIH 3T3 Cells; Signal Transduction; Silver; Skin; Staphylococcus aureus; Wound Healing

Topics: Animals; Autoimmunity; Humans; Immunity, Innate; Inflammation; Keratins; Phenotype; Signal Transduction; Skin; Skin Diseases

Topics: Animals; Autoimmune Diseases; Autoimmunity; Humans; Inflammation; Inflammation Mediators; Keratins; Signal Transduction; Skin; Skin Diseases

Endometriosis is a common disorder of unknown etiology, and non-surgical therapies are still a challenge. To understand the pathogenesis and preclinical testing of drugs for endometriosis, animal models are highly desirous. Herein, we carried out longitudinal characterization of a mouse model for endometriosis where uterine tissue was transplanted onto the intestinal mesentery. During the course of lesion development from day 15 to 60 post-induction, the ectopic endometrium became pale, fluid-filled and the animals developed peritoneal adhesions. Most lesions resembled a well-differentiated type of endometriosis and ~13% of animals had mixed type of lesions. There was extensive stromal compaction in the ectopic tissue. During the progression of endometriosis, there was increased proliferation of epithelial and stromal cells as evident by PCNA staining.

Topics: Animals; Aromatase; Cadherins; Cell Proliferation; Choristoma; Disease Models, Animal; Endometriosis; Endometrium; Epithelial Cells; Epithelial-Mesenchymal Transition; Estrogen Receptor alpha; Estrogen Receptor beta; Female; Fibrosis; Gene Expression Regulation; Humans; Inflammation; Interferon-gamma; Interleukin-1beta; Keratins; Mesentery; Mice; Myofibroblasts; Proliferating Cell Nuclear Antigen; Stromal Cells; Transplantation, Autologous; Transplantation, Heterotopic; Tumor Necrosis Factor-alpha

The aim of this study is to determine the physical and functional interplay between fatty acid-binding protein 4 (FABP4) and its membrane receptor-like candidate protein, cytokeratin 1 (CK1), and to determine the effect of hindering CK1-mediated FABP4 cellular uptake on non-disturbed or metabolically stressed endothelial cells.. We monitored the direct interaction between FABP4 and CK1 using surface plasmon resonance, and the effects of blocking exogenous FABP4 (eFABP4) cellular uptake were determined by using specific siRNA to knock down the expression of CK1 in human umbilical vein endothelial cells (HUVECs). The expression and nuclear translocation of transcription factors involved in oxidative stress (NRF2) and inflammation (p65 subunit of NF-ĸB transcription factor) were determined by Western blotting analysis.. Our data showed that FABP4 and CK1 bind to each other and that the putative FABP4 binding domain would be within the. We demonstrated that CK1 facilitates eFABP4 cellular uptake in endothelial cells. Therefore, the CK1-targeted inhibition of exogenous FABP4 cellular uptake might be a potential therapeutic strategy to protect endothelial cells against FABP4-induced activation of inflammation and oxidative stress.

Topics: Biological Transport; Endothelial Cells; Fatty Acid-Binding Proteins; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Keratin-1; Keratins; Oxidative Stress; Receptors, Cell Surface; Signal Transduction

Peritoneal dialysis (PD) is often used to treat patients with end stage renal disease, and its long-term complications include excessive inflammation and oxidative stress. Allograft inflammatory factor 1 (AIF-1), as a cytoplasmic protein, is originally identified from infiltrating macrophages, and it was associated with inflammation in the cells other than macrophages, such as endothelial cells and vascular smooth muscle cells. To clarify the molecular mechanisms of AIF-1-modulated pathological changes in the peritoneum during PD, we first detected the AIF-1 expression in peritoneal tissues from PD mice. Results revealed that the pro-fibrotic stimulation caused AIF-1 upregulation and triggered inflammation in peritoneal tissues, and that AIF-1 co-expressed with pan-cytokeratin (a marker of peritoneal mesothelial cells). We next treated primary mouse peritoneal mesothelial cells (pan-cytokeratin and intercellular adhesion molecule 1 positive cells) with 50 or 100 ng/mL recombinant AIF-1, and evaluated the direct effects of AIF-1 on these cells in vitro. We found that exogenous AIF-1 treatment induced inflammation and oxidative stress in mesothelial cells. Apart from the augmented IL-6 and TNF-α secretion, the level of ROS was upregulated and the activity of anti-oxidative SOD was reduced in cells exposed to AIF-1. Moreover, AIF-1 simulation triggered the activation of NF-κB pathway-enhanced the conversion of IκB to phosphorylated IκB and promoted the translocation of NF-κB p65 from cytoplasm into nucleus. Additionally, AIF-1-evoked inflammation in peritoneal mesothelial cells was attenuated by the addition of NF-κB inhibitor (BAY 11-7082). In brief, this study provides us novel information to understand the molecular regulation mechanisms of AIF-1 in peritoneal fibrosis.

Topics: Allografts; Animals; Calcium-Binding Proteins; Epithelial Cells; Fibrosis; I-kappa B Proteins; Inflammation; Keratins; Macrophages; Male; Mice; Mice, Inbred C57BL; Microfilament Proteins; NF-kappa B; Oxidative Stress; Peritoneal Dialysis; Peritoneal Fibrosis; Peritoneum; Reactive Oxygen Species; Signal Transduction; Transcription Factor RelA

Acne vulgaris is a disease of pilosebaceous units with multifactorial pathogenesis, including hyperkeratinization, increased sebum secretion, and inflammation. Recently, it was suggested that acne subjects may have also impaired skin barrier. We hypothesized that excess unsaturated free fatty acids (UFFA) present in the sebum may cause barrier impairment associated with increased follicular stratum corneum (SC) thickening and inflammation seen in acne. Therefore, epidermal and sebaceous lipid profiles from acne and healthy subjects were analyzed and an in vitro epidermal tissue model was developed to validate this hypothesis. Significantly increased levels of free fatty acids (p < 0.05) were observed in skin lipids of human acne vs. healthy subjects. Exposure of human epidermal equivalents (HEEs) to the UFFA oleic acid (OA), also present in sebum, led to barrier impairment associated with increased SC lipid disorder, increased secretion of interleukin-1α (IL-1α), and excessive SC thickening. Furthermore, the expression of genes encoding for inflammatory cytokines and epidermal differentiation proteins was also increased both in acne lesions and in OA-treated HEEs. Taken together, these data are in agreement with the hypothesis that excess UFFAs in sebum of acne subjects may contribute to impaired skin barrier associated with the increased follicular SC thickness and inflammation seen in acne. Moreover, OA induces similar molecular and phenotypic changes in HEEs as those seen in acne lesions and suggests that an UFFA-treated epidermal tissue model can be used to study the UFFA-mediated pathways involved in the pathogenesis of inflammatory acne and for the development of appropriate therapies.

Topics: Acne Vulgaris; Adolescent; Adult; Fatty Acids, Nonesterified; Female; Humans; Inflammation; Interleukin-8; Keratins; Lipids; Propionibacterium acnes; Sebum; Skin; Young Adult

Psoriasis is a common chronic dermatosis characterized by keratinocyte hyperproliferation accompanied by inflammatory reactions. Pathological changes upset the balance between keratinocyte proliferation, differentiation, and death in psoriatic lesions, suggesting that molecules with topical anti-inflammatory, anti-proliferation and anti-angiogenesis abilities may be useful for its treatment. The flavonoid astilbin is the major active component extracted from the rhizome of Smilax glabra, which has been widely used in China to treat inflammatory and autoimmune diseases. Here, we investigate the potential of astilbin as a treatment for psoriasis. We reveal that astilbin inhibits the growth of HaCaT keratinocytes. Detailed study shows that astilbin leads to S phase arrest of the cell cycle by induction of p53 and p21 and activated-AMPK. Additionally, astilbin induced keratinocyte differentiation correlated with suppression of keratin 5 (KRT5) and KRT14 proteins (the markers of epidermal basal layer) and induction KRT1 and KRT10 proteins (occurring in the upper layers). Moreover, astilbin regulates the expression of VEGF in human HaCaT keratinocytes. These results suggest that astilbin may be a promising agent for psoriasis treatment.

Topics: Anti-Inflammatory Agents; Cell Differentiation; Cell Line; Cell Proliferation; China; Cyclin-Dependent Kinase Inhibitor p21; Flavonols; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; Rhizome; S Phase; Tumor Suppressor Protein p53

Immune cell infiltrates (ICI) of tumors are scored by pathologists around tumor glands. To obtain a better understanding of the immune infiltrate, individual immune cell types, their activation states and location relative to tumor cells need to be determined. This process requires precise identification of the tumor area and enumeration of immune cell subtypes separately in the stroma and inside tumor nests. Such measurements can be accomplished by a multiplex format using immunohistochemistry (IHC).. We developed a pipeline that combines immunohistochemistry (IHC) and digital image analysis. One slide was stained with pan-cytokeratin and CD45 and the other slide with CD8, CD4 and CD68. The tumor mask generated through pan-cytokeratin staining was transferred from one slide to the other using affine image co-registration. Bland-Altman plots and Pearson correlation were used to investigate differences between densities and counts of immune cell underneath the transferred versus manually annotated tumor masks. One-way ANOVA was used to compare the mask transfer error for tissues with solid and glandular tumor architecture.. The overlap between manual and transferred tumor masks ranged from 20%-90% across all cases. The error of transferring the mask was 2- to 4-fold greater in tumor regions with glandular compared to solid growth pattern (p < 10. In summary, we developed a general method to integrate data from IHC stained slides into a single dataset. Because of the transfer error between slides, we recommend applying the antibody for demarcation of the tumor on the same slide as the ICI antibodies.

Topics: Biomarkers, Tumor; Breast Neoplasms; Cell Count; Cohort Studies; Female; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Inflammation; Keratins; Leukocyte Common Antigens

Intrauterine infection is a primary cause of preterm birth and fetal injury. The pro-inflammatory role of the fetal skin in the setting of intrauterine infection remains poorly characterized. Whether or not inflammation of the fetal skin occurs in primates remains unstudied. Accordingly, we hypothesized that: i) the fetal primate skin would mount a pro-inflammatory response to preterm birth associated pro-inflammatory agents (lipopolysaccharides from Escherichia coli, live Ureaplasma parvum, interleukin-1β) and; ii) that inhibiting interleukin-1 signaling would decrease the skin inflammatory response.. Rhesus macaques with singleton pregnancies received intraamniotic injections of either sterile saline (control) or one of three pro-inflammatory agonists: E. coli lipopolysaccharides, interluekin-1β or live U. parvum under ultrasound guidance. A fourth group of animals received both E. coli lipopolysaccharide and interleukin-1 signaling inhibitor interleukin-1 receptor antagonist (Anakinra) prior to delivery. Animals were surgically delivered at approximately 130 days' gestational age.. Intraamniotic lipopolysaccharide caused an inflammatory skin response characterized by increases in interluekin-1β,-6 and -8 mRNA at 16 hours. There was a modest inflammatory response to U. parvum, but interleukin-1β alone caused no inflammatory response in the fetal skin. Intraamniotic Anakinra treatment of lipopolysaccharide-exposed animals significantly reduced skin inflammation.. Intraamniotic lipopolysaccharide and U. parvum were associated with modest increases in the expression of inflammatory mediators in primate fetal skin. Although administration of Interleukin-1β alone did not elicit an inflammatory response, lipopolysaccharide-driven skin inflammation was decreased following intraamniotic Anakinra therapy. These findings provide support for the role of the fetal skin in the development of the fetal inflammatory response.

Topics: Animals; Chorioamnionitis; Disease Models, Animal; Female; Fetus; Inflammation; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Keratins; Lipopolysaccharides; Macaca mulatta; Male; Polymerase Chain Reaction; Pregnancy; RNA, Messenger; Skin; Ureaplasma

The aim of this study was to optimize the culture system of keratinocytes obtained from patients with oral lichen planus (OLP) and verify whether this model could simulate the local inflammatory environment of OLP.. Keratinocytes were isolated from 48 patients with OLP and cultured in vitro. The ultrastructure of OLP keratinocytes was observed via electron microscopy. The expression of pancytokeratin and vimentin was determined by immunohistochemistry, and the proliferation of OLP keratinocytes was measured by CCK-8 assay. Immunofluorescence staining was used to detect TLR4 and NF-κB p65 expression, and the levels of IL-1β, IL-6, and TNF-α in the supernatant were measured by ELISA.. When seeded in plates precoated with recombinant human type-1 collagen, keratinocytes isolated from patients who received systemic antifungal treatment and were younger than 40 years were more successful to be cultured in vitro. Characteristic pancytokeratin was expressed in almost all OLP keratinocytes. Compared with normal oral keratinocytes, OLP keratinocytes demonstrated higher levels of TLR4/NF-κB p65 and inflammatory cytokines, including IL-1β, IL-6, and TNF-α.. We successfully optimized the culture system of OLP keratinocytes,which mimicked the local inflammatory environment of OLP and may be used as a cell model of OLP.

Topics: Adolescent; Adult; Age Factors; Aged; Antifungal Agents; Case-Control Studies; Cell Proliferation; Cytokines; Female; Humans; Inflammation; Keratinocytes; Keratins; Lichen Planus, Oral; Male; Microscopy, Electron; Middle Aged; Primary Cell Culture; Toll-Like Receptor 4; Transcription Factor RelA; Vimentin; Young Adult

Non-invasive sample collection methods could facilitate clinical research on hair diseases. In an exploratory experimental study on six male volunteers with untreated androgenetic alopecia (AGA), Hamilton-Norwood stage IIIv-IV, skin surface and infundibular protein as well as RNA extracts from plucked hair follicles were analyzed from frontal skin, vertex and clinically unaffected occiput. Slightly increased levels of inflammatory markers were only found in AGA-affected scalp skin and infundibulum, not in RNA from plucked hair follicles. RNA expression profiles point towards differential expression of genes involved in hair cycle regulation, hair keratin production, but also RNA methylation and ion channel regulation.

Topics: Alopecia; Biomarkers; Disease Progression; DNA Methylation; Gene Expression Profiling; Gene Expression Regulation; Hair; Hair Follicle; Humans; Inflammation; Keratins; Male; Methylation; Pilot Projects; RNA; Scalp; Skin

Alopecia areata (AA) is a common inflammatory disease targeting the anagen-stage hair follicle. Different cytokines have been implicated in the disease profile, but their pathogenic role is not yet fully determined. We studied biopsies of pretreatment lesional and non-lesional (NL) scalp and post-treatment (intra-lesional steroid injection) lesional scalp of 6 patchy patients with AA using immunohistochemistry and gene expression analysis. Immunohistochemistry showed increases in CD3(+) , CD8(+) T cells, CD11c(+) dendritic cells and CD1a(+) Langerhans cells within and around hair follicles of pretreatment lesional scalp, which decreased upon treatment. qRT-PCR showed in pretreatment lesional scalp (compared to NL) significant increases (P < 0.05) in expression of inflammatory markers (IL-2, IL-2RA, JAK3, IL-15), Th1 (CXCL10 and CXCL9), Th2 (IL-13, CCL17 and CCL18), IL-12/IL-23p40 and IL-32. Among these, we observed significant downregulation with treatment in IL-12/IL-23p40, CCL18 and IL-32. We also observed significant downregulation of several hair keratins in lesional scalp, with significant upregulation of KRT35, KRT75 and KRT86 in post-treatment lesional scalp. This study shows concurrent activation of Th1 and Th2 immune axes as well as IL-23 and IL-32 cytokine pathways in lesional AA scalp and defined a series of response biomarkers to corticosteroid injection. Clinical trials with selective antagonists coupled with cytokine-pathway biomarkers will be necessary to further dissect pathogenic immunity.

Topics: Adrenal Cortex Hormones; Alopecia Areata; Biomarkers; Biopsy; Cytokines; Female; Gene Expression Profiling; Gene Expression Regulation; Hair Follicle; Humans; Immunohistochemistry; Inflammation; Interleukins; Keratins; Langerhans Cells; Male; Scalp; Th1 Cells; Th2 Cells

Previously, we generated a preclinical mouse prostate tumor model based on PSA-Cre driven inactivation of Pten. In this model homogeneous hyperplastic prostates (4-5m) developed at older age (>10m) into tumors. Here, we describe the molecular and histological characterization of the tumors in order to better understand the processes that are associated with prostate tumorigenesis in this targeted mouse Pten knockout model. The morphologies of the tumors that developed were very heterogeneous. Different histopathological growth patterns could be identified, including intraductal carcinoma (IDC), adenocarcinoma and undifferentiated carcinoma, all strongly positive for the epithelial cell marker Cytokeratin (CK), and carcinosarcomas, which were negative for CK. IDC pattern was already detected in prostates of 7-8 month old mice, indicating that it could be a precursor stage. At more than 10 months IDC and carcinosarcoma were most frequently observed. Gene expression profiling discriminated essentially two molecular subtypes, denoted tumor class 1 (TC1) and tumor class 2 (TC2). TC1 tumors were characterized by high expression of epithelial markers like Cytokeratin 8 and E-Cadherin whereas TC2 tumors showed high expression of mesenchyme/stroma markers such as Snail and Fibronectin. These molecular subtypes corresponded with histological growth patterns: where TC1 tumors mainly represented adenocarcinoma/intraductal carcinoma, in TC2 tumors carcinosarcoma was the dominant growth pattern. Further molecular characterization of the prostate tumors revealed an increased expression of genes associated with the inflammatory response. Moreover, functional markers for senescence, proliferation, angiogenesis and apoptosis were higher expressed in tumors compared to hyperplasia. The highest expression of proliferation and angiogenesis markers was detected in TC2 tumors. Our data clearly showed that in the genetically well-defined PSA-Cre;Pten-loxP/loxP prostate tumor model, histopathological, molecular and biological heterogeneity occurred during later stages of tumor development.

Topics: Adenocarcinoma; Animals; Apoptosis; Biomarkers; Biomarkers, Tumor; Cadherins; Carcinoma; Carcinosarcoma; Cellular Senescence; Disease Progression; Epithelial Cells; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Inflammation; Keratins; Male; Mesoderm; Mice; Mice, Inbred Strains; Mice, Knockout; Neoplasm Proteins; Neovascularization, Pathologic; Prostatic Hyperplasia; Prostatic Neoplasms; PTEN Phosphohydrolase; RNA, Messenger; RNA, Neoplasm; Stromal Cells

Amniotic membrane (AM) transplantation is the clinical standard for ocular surface reconstruction, however recently keratin film (KF) has been proposed as an alternative material. Aim of the current study was to evaluate corneal biocompatibility of KF in a rabbit model. Forty-six New Zealand white rabbits underwent dissection of a corneal intrastromal pocket in which an AM or KF implant was inserted and observed for 10 days and for 4 weeks. Half of animals received topical steroids, while the other half were left without. At the end of the follow-up clinical and histology examinations were performed to evaluate transparency, inflammation and degradation. After 10 days the clinical and the histology results appeared to be comparable in KF implanted eyes treated with and without steroids. After 4 weeks, comparable clinical results were observed in all KF implanted eyes, while the inflammation score was lower in non-steroid compared to steroid treated eyes along with a higher degradation rate of the keratin films. In conclusion, keratin films from human hair show a good biocompatibility and transparency in vivo. The administration of topical steroids seems to slow down implant degradation which might be important for the modulation of tissue integration and matrix regeneration.

Topics: Amnion; Animals; Biocompatible Materials; Dissection; Eye; Female; Humans; Implants, Experimental; Inflammation; Keratins; Materials Testing; Models, Animal; Plastic Surgery Procedures; Rabbits

Scrotal calcinosis is a rare and benign condition characterized by multiple calcific deposits occurring in scrotum and formed as nodules and lumps within scrotal skin with any systemic metabolic disorder. The so-called idiopathic scrotal calcinosis does not appear to be idiopathic, but rather a process of dystrophic calcification of epidermal cysts. Histological examination shows calcium deposites with in the dermis that may be surrounded with histiocytes and an inflammatory giant cell reaction. The aim of this paper was to detect dystrophic calcification of epidermal cysts and to take attention to the incorrect terminology of "idiopathic calsification".. This is a two-centered study of scrotal calcinosis with 17 cases, on which clinical and histopathological examinations were conducted.. The patients we examined all had scrotal epidermoid cysts in varying stages of inflammation coexisted with scrotal calcinosis. Some cyts (52.9%) had intact epithelial walls, others (35.2%) showed rupture of their epithelial walls associated with the presence of keratin fibers and calcium granules in the surrounding dermis and all had naked calcium deposits lying in the dermis.. The spectrum of the changes that we experienced in the histology, coupled with the normal values in the biochemical profile, shore up the theory of dystrophic calcification of epithelial cysts. During the time first these cysts become inflamed than rupture in the and calcium depocytes replase with the cysts.

Topics: Adult; Aged; Aged, 80 and over; Calcinosis; Calcium; Epidermal Cyst; Genital Diseases, Male; Humans; Inflammation; Keratins; Male; Middle Aged; Retrospective Studies; Scrotum; Terminology as Topic; Young Adult

Topics: Epithelial Cells; Female; Hair; Hair Follicle; Hair Removal; Humans; Inflammation; Interleukin-6; Keratinocytes; Keratins; PPAR gamma; Propionates

Diagnostic criteria that have been specified for unicystic ameloblastomas (UAs) are not always helpful to differentiate these cystic tumors from common odontogenic cysts. The aim of this study therefore was to identify additional histopathological features (other than the features considered for the diagnosis of UA at present) that would be helpful to differentiate UA from odontogenic cysts.. One hundred histopathologically confirmed unicystic ameloblastomas and 20 cases each of radicular, inflamed dentigerous and non-inflamed dentigerous cysts were selected. Histopathological features of the UAs that are not used as diagnostic criteria at present were identified.. Hyperplastic arcading epithelial proliferations with stellate-reticulum-like and vacuolated cells were always seen associated with inflammation in odontogenic cysts, while in UA plexiform-like areas were also seen without inflammation (P < 0.001). In addition, a spiky rete pattern was observed in non-inflamed UA while this pattern was observed only in inflamed odontogenic cysts. Furthermore, spiky retes together with subepithelial hyalinization were usually observed in UAs while only subepithelial hyalinization was observed in non-inflamed dentigerous cysts.. Combinations of histopathological features were identified to differentiate non-inflamed UA from common odontogenic cysts. However, presence of inflammatory changes in UA precludes the use of features identified in the present study for diagnostic purposes.

Topics: Ameloblastoma; Ameloblasts; Connective Tissue; Dentigerous Cyst; Diagnosis, Differential; Epithelial Cells; Epithelium; Female; Humans; Hyalin; Hyperplasia; Inflammation; Keratins; Male; Radicular Cyst; Vacuoles; Young Adult

Topics: Hidradenitis Suppurativa; Humans; Hypoxia; Hypoxia-Inducible Factor 1; Inflammation; Keratinocytes; Keratins

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Calbindin 2; Cholecystitis; Cisplatin; Cystitis; Diagnosis, Differential; Female; Glutamates; Guanine; Humans; Inflammation; Keratins; Lung Neoplasms; Male; Mesothelioma; Mesothelioma, Malignant; Middle Aged; Pemetrexed; Peritoneal Neoplasms; Survival Rate; Vimentin

Clear cell acanthoma, firstly described by Degos as "an epidermal tumor with a particular aspect", although quite a rare lesion, raised an important interest because it may be easily confused with other dermatologic lesions, in the absence of a histopathological examination. Its clinical aspect is of a solitary nodule, with a red-brown varying color, with a size of 3 mm to 2 mm, sometimes covered with a thin scall. We present a case of a multiple rare cell acanthoma (seven nodular formations), having a rapid development (about two months) diagnosed in a 71-year-old patient within the lower 1/3 of the right shin.

Topics: Acanthoma; Aged; B-Lymphocytes; Dermis; Female; Glycogen; Humans; Immunohistochemistry; Inflammation; Keratinocytes; Keratins; Leg; Lymphocyte Count; Skin Neoplasms; T-Lymphocytes

Age related macular degeneration (AMD) is one of the leading causes of blindness in Western society. A hallmark of early stage AMD are drusen, extracellular deposits that accumulate in the outer retina. Advanced glycation endproducts (AGE) accumulate with aging and are linked to several age-related diseases such as Alzheimer's disease, osteoarthritis, atherosclerosis and AMD. AGE deposits are found in drusen and in Bruch's membrane of the eye and several studies have suggested its role in promoting oxidative stress, apoptosis and lipofuscin accumulation. Recently, complement activation and chronic inflammation have been implicated in the pathogenesis of AMD. While AGEs have been shown to promote inflammation in other diseases, whether it plays a similar role in AMD is not known. This study investigates the effects of AGE stimulation on pro- and anti-inflammatory pathways in primary culture of human retinal pigment epithelial cells (RPE). Differential gene expression studies revealed a total of 41 up- and 18 down-regulated RPE genes in response to AGE stimulation. These genes fell into three categories as assessed by gene set enrichment analysis (GSEA). The main categories were inflammation (interferon-induced, immune response) and proteasome degradation, followed by caspase signaling. Using suspension array technology, protein levels of secreted cytokines and growth factors were also examined. Anti-inflammatory cytokines including IL10, IL1ra and IL9 were all overexpressed. Pro-inflammatory cytokines including IL4, IL15 and IFN-γ were overexpressed, while other pro-inflammatory cytokines including IL8, MCP1, IP10 were underexpressed after AGE stimulation, suggesting a para-inflammation state of the RPE under these conditions. Levels of mRNA of chemokine, CXCL11, and viperin, RSAD2, were up-regulated and may play a role in driving the inflammatory response via the NF-kB and JAK-STAT pathways. CXCL11 was strongly immunoreactive and associated with drusen in the AMD eye. The pathways and novel genes identified here highlight inflammation as a key response to AGE stimulation in primary culture of human RPE, and identify chemokine CXCL11 as putative novel agent associated with the pathogenesis of AMD.

Topics: Cell Survival; Cells, Cultured; Chemokine CXCL11; Down-Regulation; Gene Expression Profiling; Gene Expression Regulation; Glycation End Products, Advanced; Humans; Inflammation; Keratins; Lysine; Macular Degeneration; Pigment Epithelium of Eye; Postmortem Changes; Reproducibility of Results; Retinal Drusen; Serum Albumin, Bovine; Tissue Donors; Up-Regulation

Several molecular tests have been developed to estimate risk of distant recurrence and help clinical decision-making regarding adjuvant chemotherapy in patients with early stage breast carcinoma. Both Oncotype DX, a 21-gene expression profile, and Mammostrat, an immunohistochemistry-based assay, are validated to stratify patients into groups with low, intermediate and high risk of distant recurrence. However, they have not been compared head-to-head and little data are available regarding their correlation with clinicopathologic tumor features. In this study, we compared the clinicopathologic tumor features with risk estimations by Oncotype DX and Mammostrat in 106 low-grade estrogen receptor (ER)-positive breast carcinomas. Double immunohistochemical stain for pancytokeratin and Ki-67 was performed to assess cell proliferation in cancer vs stromal/inflammatory cells. Tumors showing intermediate/high risk by Oncotype DX, but not by Mammostrat, showed increased stromal cellularity, presence of inflammatory cells and increased proliferation in stromal/inflammatory cells. Discrepant cases showing intermediate/high risk by Oncotype DX but low risk by Mammostrat were associated with increased stromal cellularity, presence of inflammatory cells and increased proliferation in stromal/inflammatory cells, compared with concordant cases showing low risk by both assays. Our results suggest that low-grade ER-positive breast carcinomas with increased stromal/inflammatory cell proliferation may show an apparent increased risk of distant recurrence as assessed by Oncotype DX, which uses RNA extracted from a mixture of tumor and stromal/inflammatory cells in the assay. Mammostrat, which examines cancer cells only, may provide a better estimation of likely tumor behavior in a subgroup of low-grade breast carcinomas.

Topics: Biomarkers, Tumor; Breast Neoplasms; Cell Proliferation; Decision Support Techniques; Female; Gene Expression Profiling; Genetic Testing; Humans; Immunohistochemistry; Inflammation; Keratins; Ki-67 Antigen; Middle Aged; Neoplasm Grading; Neoplasm Invasiveness; Patient Selection; Predictive Value of Tests; Receptors, Estrogen; Risk Assessment; Risk Factors; Stromal Cells

Cells of the epidermis renew constantly from germinal layer stem cells. Although epithelial cell differentiation has been studied in great detail and the role of Wnt signaling in this process is well described, the contribution of epidermal Wnt secretion in epithelial cell homeostasis remains poorly understood. To analyze the role of Wnt proteins in this process, we created a conditional knockout allele of the Wnt cargo receptor Evi/Gpr177/Wntless and studied mice that lacked Evi expression in the epidermis. We found that K14-Cre, Evi-LOF mice lost their hair during the first hair cycle, showing a reddish skin with impaired skin barrier function. Expression profiling of mutant and wild-type skin revealed up-regulation of inflammation-associated genes. Furthermore, we found that Evi expression in psoriatic skin biopsies is down-regulated, suggesting that Evi-deficient mice developed skin lesions that resemble human psoriasis. Immune cell infiltration was detected in Evi-LOF skin. Interestingly, an age-dependent depletion of dendritic epidermal T cells (DETCs) and an infiltration of γδ(low) T cells in Evi mutant epidermis was observed. Collectively, the described inflammatory skin phenotype in Evi-deficient mice revealed an essential role of Wnt secretion in maintaining normal skin homeostasis by enabling a balanced epidermal-dermal cross talk, which affects immune cell recruitment and DETC survival.

Topics: Animals; CD3 Complex; Cell Proliferation; Chronic Disease; Dendritic Cells; Dermatitis; Epidermis; Gene Deletion; Humans; Inflammation; Intracellular Signaling Peptides and Proteins; Keratinocytes; Keratins; Lymphocyte Activation; Mice; Mice, Transgenic; Neutrophil Infiltration; Phenotype; Psoriasis; Receptors, Antigen, T-Cell, gamma-delta; Receptors, G-Protein-Coupled; STAT3 Transcription Factor; T-Lymphocytes; Wnt Proteins

Human skin mainly functions as an effective barrier against unwanted environmental influences. The barrier function strongly relies on the outermost layer of the skin, the stratum corneum (SC), which is composed of corneocytes embedded in an extracellular lipid matrix. The importance of a proper barrier function is shown in various skin disorders such as atopic dermatitis (AD), a complex human skin disorder strongly associated with filaggrin (FLG) null mutations, but their role in barrier function is yet unclear. To study the role of FLG in SC barrier properties in terms of SC lipid organization and lipid composition, we generated an N/TERT-based 3D-skin equivalent (NSE) after knock-down of FLG with shRNA. In these NSEs, we examined epidermal morphogenesis by evaluating the expression of differentiation markers keratin 10, FLG, loricrin and the proliferation marker ki67. Furthermore, the SC was extensively analysed for lipid organization, lipid composition and SC permeability. Our results demonstrate that FLG knock-down (FLG-KD) did not affect epidermal morphogenesis, SC lipid organization, lipid composition and SC permeability for a lipophilic compound in NSEs. Therefore, our findings indicate that FLG-KD alone does not necessarily affect the functionality of a proper barrier function.

Topics: Cell Proliferation; Dermatitis, Atopic; Epidermis; Fibroblasts; Filaggrin Proteins; Gene Knockdown Techniques; Heterozygote; Humans; Inflammation; Intermediate Filament Proteins; Keratin-10; Keratins; Ki-67 Antigen; Lipids; Membrane Proteins; Permeability; Phenotype; Skin; Skin Diseases

The aim of this study is to characterize immunohistochemical profiles of lining epithelia of nasopalatine duct cyst (NPC) as well as to correlate those findings with their clinicopathological features to understand the histopathogenesis of NPC.. Forty-one surgical specimens from NPC were examined for clinical profiles and expression of keratin-7, 13, MUC-1, and P63 by immunohistochemistry, compared to radicular cyst (RC) and maxillary sinusitis.. Nasopalatine duct cyst was clinically characterized by male predominant occurrence: 44% of the cases involved tooth roots, and 70% with inflammatory backgrounds. Lining epithelia of NPCs without daughter cysts were immunohistochemically distinguished into three layers: a keratin 7-positive (+) ciliated cell layer in the surface, a keratin-13+ middle layer, and a MUC-1+/P63+ lower half, indicating that they were not respiratory epithelia, and the same layering pattern was observed in RC. However, those immunolocalization patterns of the main cyst lining with daughter cyst were exactly the same as those of daughter cyst linings as well as duct epithelia of mucous glands.. Two possible histopathogenesis of NPC were clarified: one was inflammatory cyst like RC and the other was salivary duct cyst-like mucocele.

Topics: Adult; Aged; Epithelial Cells; Female; Humans; Inflammation; Keratins; Male; Maxillary Diseases; Maxillary Sinusitis; Membrane Proteins; Middle Aged; Mucins; Mucocele; Nasal Cavity; Nonodontogenic Cysts; Palate, Hard; Radicular Cyst; Sex Ratio; Terminology as Topic; Tooth Root; Young Adult

Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine with pro-inflammatory functions and involved in tumorigenesis. The aim of this study was to evaluate the expression and localization of the macrophage MIF in oral squamous carcinoma (OSC). In addition, the relationship between MIF expression and clinicopathological parameters such as survival data, tobacco use, alcohol habits, TNM stage, tumor graduation, and peritumoral inflammatory infiltrate were evaluated.. Using immunohistochemistry, expression and localization of MIF was detected in 44 specimens of OSC. The absolute number and relative proportions of MIF-positive cells detected were also determined separately for tumor parenchyma vs. stroma. All counts were determined from 10 consecutive high-power fields using an integration graticule. Moreover, some parameters were analyzed separately for lip and intra-oral cancers.. Migration inhibitory factor-positive cells were observed in both the tumor parenchyma and in inflammatory cells of all specimens. In contrast, MIF expression was not detected in tumoral nests associated with poorly differentiated tumors. In specimens of lip cancer, a greater number of MIF-positive stromal immune cells were detected than in intra-oral cancer specimens (Mann-Whitney test, P = 0.049).. Oral squamous carcinoma cells consistently express MIF independent of their location. Lip tumors presented more MIF-positive peritumoral inflammatory cells, similar to control, suggesting that immunological differences in leukocyte activation exist between in lip and intra-oral cancers.

Topics: Alcohol Drinking; Carcinoma, Squamous Cell; Cell Count; Cohort Studies; Epithelium; Female; Humans; Inflammation; Keratins; Leukocytes; Leukoplakia, Oral; Lip Neoplasms; Macrophage Migration-Inhibitory Factors; Male; Middle Aged; Mouth Neoplasms; Neoplasm Grading; Neoplasm Staging; Smoking; Stromal Cells; Survival Rate

In this retrospective study, the aim was to compare individual histopathologic parameters of malignancy between nonmetastatic and metastatic squamous cell carcinoma of the tongue.. Sixty-two cases of squamous cell carcinoma of the tongue were selected and examined according to the system established by Brandwein-Gensler et al (Am J Surg Pathol 29:167, 2005) and included the pattern of invasion (most to least favorable), lymphocytic infiltration, perineural invasion, risk score, keratinization, eosinophilia, perivascular invasion, and tumor thickness.. The least favorable pattern had no association with nodal metastasis (P > .05). The scarcity or density of the lymphocytic infiltration, perineural invasion, and a risk score ≥ 3 were associated with nodal metastasis (P < .05). Keratinization, eosinophilia, perivascular invasion, and tumor thickness had no association with nodal metastasis (P > .05). A significant positive correlation was found between the pattern of invasion and perineural invasion and between the pattern of invasion and tumor thickness (P < .05).. The scarcity or density of the lymphocytic infiltration, perineural invasion, and histopathologic risk score may be helpful as parameters of histologic malignancy for the evaluation of metastatic and nonmetastatic squamous cell carcinoma of the tongue.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Chemotaxis, Leukocyte; Eosinophilia; Female; Follow-Up Studies; Humans; Inflammation; Keratins; Lymphatic Metastasis; Lymphocytes; Male; Microvessels; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Peripheral Nerves; Retrospective Studies; Risk Assessment; Survival Rate; Tongue Neoplasms

This study evaluated the early recovery process of the palatal wounds of dogs using bismuth subgallate. Five healthy adult male dogs underwent eight 5-mm partial-thickness punch biopsies in two paired columns on the palatal mastigatory mucosa. For the haemostasis, one side received moistened gauze pressure (test group 1), and the other received bismuth subgallate (test group 2). A description of the epithelium and connective tissue repair was made at 3, 7, 14 and 21 days. During the first days, a mass of disorganized tissue covered the connective tissue, in which there was intense chronic inflammation, and migration of epithelium cells from the edges towards the central region to close to the wound was seen. The final evaluation demonstrated well organized epithelial and connective tissues in all the samples. Epithelium thickness was measured at 0, 14 and 21 days, from images of the digitalized histological sections. In comparisons between the test groups, the bismuth subgallate group was slightly better than the saline group, but no statistically significant difference was found at 21 days. It was possible to conclude that bismuth subgallate did not interfere in the tissue repair of the palatal mastigatory mucosa in dogs.

Topics: Animals; Biopsy, Needle; Bismuth; Blood Coagulation; Cell Movement; Collagen; Connective Tissue; Dogs; Epithelial Cells; Epithelium; Fibrin; Fibroblasts; Gallic Acid; Granulation Tissue; Hemostatics; Image Processing, Computer-Assisted; Inflammation; Keratins; Lymphocytes; Macrophages; Male; Mouth Mucosa; Neutrophils; Organometallic Compounds; Palate; Pressure; Sodium Chloride; Time Factors; Wound Healing

Intraperitoneal injection of Freund's adjuvant induces acute peritonitis. By the time of the Freund's adjuvant treatment the flat, simple squamous epithelial cells became rounded, cuboidal shaped, many of them have lost their connection with the neighbouring cells and detached from the basement membrane. The macrophage markers' (ED1, OX43 and CD68) expression also increased in the mesothelial cells and more mesothelin and anti-ED1 double-labelled cells were found freely present close to the surface. The cytokeratin expression of the mesothelial cells has gradually decreased. At the 5th day of the inflammation practically there was no cytokeratin labelling present in the mesothelial cells and the mesothelin expression has significantly decreased. Parallel to this mesothelial cells started to express vimentin, a characteristic mesenchymal intermediate filament protein indicating that they gradually lost their epithelial character and gained mesenchymal phenotype. These results strongly suggest that under the effect of Freund's adjuvant treatment (inflammation) mesothelial cells can undergo epithelial-to-mesenchymal transition and differentiate into phagocytotic (macrophage-like) cells. Studying the caveolae/caveolin-1 on the plasma membrane of mesothelial cells we found that the Freund's adjuvant treatment has changed the cellular distribution of caveolin-1: as the inflammation progressed strong caveolin-1 labelling was found inside of the cytoplasm (in perinuclear localization) indicating that inflammation induced the caveolae internalization. These results indicate that caveolae/caveolin-1 might play important regulatory role in signal transduction leading to trasdifferentiation.

Topics: Adjuvants, Immunologic; Animals; Biomarkers; Caveolin 1; Cell Differentiation; Cell Membrane; Cells, Cultured; Cytoplasm; Epithelial-Mesenchymal Transition; Freund's Adjuvant; Immunoenzyme Techniques; Inflammation; Keratins; Macrophages; Male; Rats; Rats, Sprague-Dawley; Vimentin

Gastrointestinal tract xanthomas are non tumor, well demarcated mucosal lesions that consist of foamy histiocytes, most commonly diagnosed in the stomach. The histologic appearance of xanthomas can resemble certain malignant lesions. After retrospective data base search, we have encountered only 2 cases of xanthomas, both in the antral part of the stomach. Lamina propria of the mucosa contained rare, chronic inflammatory infiltrate and clusters of oval and polygonal cells with abundant, foamy cytoplasm. The cytoplasm of described cells did not show the presence of mucin (Periodic acid-Schiff (PAS) and Alcian blue staining). The cells showed distinct cytoplasmic CD68 positivity and CKMNF116 negativity, which confirmed the diagnosis of xanthoma. Given the frequent association of xanthomas and known precancerous lesions of gastric mucosa, and occasional coexistence of malignant change, we need to pay attention to its diagnosis, and it is advisable to use both histochemical and immunohistochemical methods.

Topics: Adult; Aged; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biopsy; Coloring Agents; Cytoplasm; Diagnosis, Differential; Female; Gastric Mucosa; Humans; Immunohistochemistry; Inflammation; Keratins; Male; Pyloric Antrum; Retrospective Studies; Stomach Diseases; Xanthomatosis

Topics: Actins; Adult; Anaplastic Lymphoma Kinase; Diagnosis, Differential; Female; Fibroma; Fibrosarcoma; Follow-Up Studies; Humans; Hysterectomy; Inflammation; Keratins; Leiomyoma; Neoplasm Recurrence, Local; Neoplasms, Muscle Tissue; Receptor Protein-Tyrosine Kinases; Uterine Neoplasms; Young Adult

Objectives were to (1) determine the feasibility of performing hoof biopsies without impairing locomotion; (2) evaluate the feasibility of using biopsied tissue for quantitative PCR; and (3) compare relative gene expression among claws for several target genes. Biopsies were performed on 6 Holstein cows, yielding 4 tissue specimens per cow from front leg, right limb, and medial claw (claw position 3); rear leg, left limb, and lateral claw (claw position 5); and rear leg, right limb, medial claw (claw position 7). Cows were monitored for lameness daily for 7 d post-biopsy and then weekly for 8 wk. Histopathological analysis confirmed that tissue collected was from between the stratum corneum and dermis. Biopsied tissue was used for RNA extraction, including evaluation of yield and purity. The profile by claw position of 19 genes with key functions in cell differentiation, proliferation, inflammation, and keratin formation was assessed via quantitative reverse transcription-PCR. Other than transient disturbances in locomotion score in some cows during 2 to 4 d post-biopsy, no signs of pain, locomotion impairment, or clinical lameness were observed post-biopsy. Total RNA yields averaged 259.7±100, 447.8±288, and 496.4±118 μg/mg of tissue for claw positions 3, 5, and 7, respectively. The biopsy procedure was successful for obtaining corium for gene expression. Among 5 keratin proteins analyzed, only keratin 5 was expressed. Transcripts related to inflammation and oxidative stress (STAT3, MYD88, SOD2, and TLR4) were among the more abundant in corium tissue, but expression did not differ between claws. Biotinidase (BTD) expression was greater in claw 3 versus claw 5, whereas the ligand-activated nuclear receptor retinoic acid receptor-α (RXRA) was greater in claws 3 + 5 compared with claw 7. Overall, results from this pilot study revealed modest differences at the transcriptome level, suggesting that biotin availability and lipid metabolism differ between claw positions, whereas inflammation and oxidative stress seem to play an important role across claws. More comprehensive studies of the hoof transcriptome are required to improve our understanding of the mechanisms that link environmental and dietary factors to development of lameness.

Topics: Animals; Biopsy; Cattle; Cattle Diseases; Dermis; Female; Foot Diseases; Gene Expression Regulation; Genes; Hoof and Claw; Inflammation; Keratin-5; Keratins; Lameness, Animal; Locomotion; Oxidative Stress; Reverse Transcriptase Polymerase Chain Reaction

Humanized mouse models offer a challenging possibility to study human cell function in vivo. In the huPBL-SCID-huSkin allograft model human skin is transplanted onto immunodeficient mice and allowed to heal. Thereafter allogeneic human peripheral blood mononuclear cells are infused intra peritoneally to induce T cell mediated inflammation and microvessel destruction of the human skin. This model has great potential for in vivo study of human immune cells in (skin) inflammatory processes and for preclinical screening of systemically administered immunomodulating agents. Here we studied the inflammatory skin response of human keratinocytes and human T cells and the concomitant systemic human T cell response.As new findings in the inflamed human skin of the huPBL-SCID-huSkin model we here identified: 1. Parameters of dermal pathology that enable precise quantification of the local skin inflammatory response exemplified by acanthosis, increased expression of human β-defensin-2, Elafin, K16, Ki67 and reduced expression of K10 by microscopy and immunohistochemistry. 2. Induction of human cytokines and chemokines using quantitative real-time PCR. 3. Influx of inflammation associated IL-17A-producing human CD4+ and CD8+ T cells as well as immunoregulatory CD4+Foxp3+ cells using immunohistochemistry and -fluorescence, suggesting that active immune regulation is taking place locally in the inflamed skin. 4. Systemic responses that revealed activated and proliferating human CD4+ and CD8+ T cells that acquired homing marker expression of CD62L and CLA. Finally, we demonstrated the value of the newly identified parameters by showing significant changes upon systemic treatment with the T cell inhibitory agents cyclosporine-A and rapamycin. In summary, here we equipped the huPBL-SCID-huSkin humanized mouse model with relevant tools not only to quantify the inflammatory dermal response, but also to monitor the peripheral immune status. This combined approach will gain our understanding of the dermal immunopathology in humans and benefit the development of novel therapeutics for controlling inflammatory skin diseases.

Topics: Animals; Antigens, Differentiation, T-Lymphocyte; beta-Defensins; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Differentiation; Cyclosporine; Disease Models, Animal; Elafin; Gene Expression Regulation; Humans; Inflammation; Injections, Intraperitoneal; Interleukin-17; Keratinocytes; Keratins; Ki-67 Antigen; L-Selectin; Membrane Glycoproteins; Mice; Mice, SCID; Sirolimus; Skin; Skin Transplantation; Transplantation, Heterologous

To evaluate the immunohistochemical expression of MMP-1, -2, -7, -9 and -26 in oral squamous cell carcinomas (SCCs) according to tumour site and histological grade of malignancy.. Fifteen cases of SCC of the lower lip and 15 cases of tongue SCC were selected and divided into low grade malignancy (n = 17) and high grade malignancy (n = 13).. Higher immunohistochemical expression of MMPs by neoplastic cells was observed in tongue SCCs, with a statistically significant difference for MMP-9 (P < 0.05). High-grade SCCs showed a higher expression of MMPs, except for MMP-2, with a statistically significant difference for MMP-7 (P < 0.05) and MMP-26 (P < 0.05). In addition, a direct association was observed between morphological scores of malignancy and MMP immunoreactivity, with the association being significant for MMP-7 and MMP-26.. The present results demonstrate the important role of MMPs in the development of SCCs of the lower lip and tongue.

Topics: Carcinoma, Squamous Cell; Cell Nucleus; Disease Progression; Humans; Immunohistochemistry; Inflammation; Keratins; Lip Neoplasms; Matrix Metalloproteinase 1; Matrix Metalloproteinase 2; Matrix Metalloproteinase 7; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Matrix Metalloproteinases, Secreted; Neoplasm Invasiveness; Stromal Cells; Tongue Neoplasms

Sulfur mustard (SM, bis(2-chloroethyl)sulfide) is a bifunctional alkylating agent that causes dermal inflammation, edema and blistering. To investigate the pathogenesis of SM-induced injury, we used a vapor cup model which provides an occlusive environment in which SM is in constant contact with the skin. The dorsal skin of SKH-1 hairless mice was exposed to saturated SM vapor or air control. Histopathological changes, inflammatory markers and DNA damage were analyzed 1-14 days later. After 1 day, SM caused epidermal thinning, stratum corneum shedding, basal cell karyolysis, hemorrhage and macrophage and neutrophil accumulation in the dermis. Cleaved caspase-3 and phosphorylated histone 2A.X (phospho-H2A.X), markers of apoptosis and DNA damage, respectively, were increased whereas proliferating cell nuclear antigen (PCNA) was down-regulated after SM exposure. By 3 days, epithelial cell hypertrophy, edema, parakeratosis and loss of epidermal structures were noted. Enzymes generating pro-inflammatory mediators including myeloperoxidase and cyclooxygenase-2 were upregulated. After 7 days, keratin-10, a differentiation marker, was evident in the stratum corneum. This was associated with an underlying eschar, as neoepidermis began to migrate at the wound edges. Trichrome staining revealed increased collagen deposition in the dermis. PCNA expression in the epidermis was correlated with hyperplasia, hyperkeratosis, and parakeratosis. By 14 days, there was epidermal regeneration with extensive hyperplasia, and reduced expression of cleaved caspase-3, cyclooxygenase-2 and phospho-H2A.X. These findings are consistent with the pathophysiology of SM-induced skin injury in humans suggesting that the hairless mouse can be used to investigate the dermatoxicity of vesicants and the potential efficacy of countermeasures.

Topics: Animals; Apoptosis; Biomarkers; Caspase 3; Cell Degranulation; Cell Differentiation; Cyclooxygenase 2; DNA Damage; Histones; Inflammation; Keratinocytes; Keratins; Male; Mast Cells; Mice; Mice, Hairless; Mustard Gas; Peroxidase; Proliferating Cell Nuclear Antigen; Skin; Staining and Labeling; Wound Healing

Respiratory epithelium is the target of therapies, such as gene therapy, for cystic fibrosis (CF) lung disease. To determine the usefulness of the nasal epithelium as a pre-screen for lung-directed therapies, we profiled gene expression in CF and non-CF nasal and bronchial epithelium samples using Illumina HumanRef-8 Expression BeadChips. 863 genes were differentially expressed between CF and non-CF bronchial epithelium but only 15 were differentially expressed between CF and non-CF nasal epithelium (≥1.5-fold, P≤0.05). The most enriched pathway in CF bronchial epithelium was inflammatory response, whereas in CF nasal epithelium it was amino acid metabolism. We also compared nasal and bronchial epithelium in each group and identified differential expression of cellular movement genes in CF patients and cellular growth genes in non-CF subjects. We conclude that CF and non-CF nasal and bronchial epithelium are transcriptionally distinct and CF nasal epithelium is not a good surrogate for the lung respiratory epithelium.

Topics: Adolescent; Adult; Bronchi; Case-Control Studies; Child; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Gene Expression Regulation; Humans; Immunohistochemistry; Inflammation; Keratins; Male; Nasal Mucosa; Oligonucleotide Array Sequence Analysis; Young Adult

Cardiac dysfunction following acute myocardial infarction is a major cause of advanced cardiomyopathy. Conventional pharmacological therapies rely on prompt reperfusion and prevention of repetitive maladaptive pathways. Keratin biomaterials can be manufactured in an autologous fashion and are effective in various models of tissue regeneration. However, its potential application in cardiac regeneration has not been tested. Keratin biomaterials were derived from human hair and its structure morphology, carryover of beneficial factors, biocompatibility with cardiomyocytes, and in vivo degradation profile were characterized. After delivery into infarcted rat hearts, the keratin scaffolds were efficiently infiltrated by cardiomyocytes and endothelial cells. Injection of keratin biomaterials promotes angiogenesis but does not exacerbate inflammation in the post-MI hearts. Compared to control-injected animals, keratin biomaterials-injected animals exhibited preservation of cardiac function and attenuation of adverse ventricular remodeling over the 8 week following time course. Tissue western blot analysis revealed up-regulation of beneficial factors (BMP4, NGF, TGF-beta) in the keratin-injected hearts. The salient functional benefits, the simplicity of manufacturing and the potentially autologous nature of this biomaterial provide impetus for further translation to the clinic.

Topics: Animals; Biocompatible Materials; Blotting, Western; Disease Models, Animal; Female; Hair; Heart; Heart Function Tests; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate; Inflammation; Injections; Keratins; Mechanical Phenomena; Myocardial Infarction; Myocardium; Myocytes, Cardiac; Neovascularization, Pathologic; Paracrine Communication; Rats; Rats, Sprague-Dawley; Tissue Scaffolds; Ventricular Remodeling

Tibia fracture in rats results in chronic vascular and nociceptive changes in the injured limb resembling complex regional pain syndrome (CRPS) and up-regulates expression of interleukin 1β (IL-1β), interleukin IL-6 (IL-6), tumor necrosis factor-α (TNF-α), and nerve growth factor-β (NGF-β) in the hindpaw skin. When fractured rats are treated with cytokine or NGF inhibitors nociceptive sensitization is blocked. Because there is no leukocyte infiltration in the hindpaw skin we postulated that resident skin cells produce the inflammatory mediators causing nociceptive sensitization after fracture. To test this hypothesis rats underwent distal tibia fracture and hindlimb casting for 4 weeks, then the hindpaw skin was harvested and immunostained for keratin, cytokines and NGF. BrdU staining was used to evaluate cell proliferation. Hindpaw nociceptive thresholds, edema, and temperature were tested before and up to 96h after intraplantar injections of IL-6 and TNF-α. Tibia fracture caused keratinocyte activation, proliferation, and up-regulated IL-1β, IL-6, TNF-α and NGF-β protein expression in the hindpaw keratinocytes. Local injections of IL-6 and TNF-α induced hindpaw mechanical allodynia lasting for several days and modest increases in temperature and edema. These data indicate that activated keratinocytes proliferate and express IL-1β, IL-6, TNF-α, and NGF-β after fracture and that excess amounts of inflammatory mediators in the skin cause sustained nociceptive sensitization. This is the first study demonstrating in vivo keratinocyte expression of IL-6, TNF-α and NGF-β in a CRPS model and we postulate that the keratinocyte is the primary cellular source for the inflammatory signals mediating cutaneous nociceptive sensitization in early CRPS.

Topics: Analysis of Variance; Animals; Body Temperature; Cell Proliferation; Cytokines; Dose-Response Relationship, Drug; Immunohistochemistry; Inflammation; Inflammation Mediators; Keratinocytes; Keratins; Male; Nerve Growth Factor; Pain; Rats; Rats, Sprague-Dawley; Tibial Fractures

Medullary carcinoma and serrated adenocarcinoma are two variants of colon cancer which are associated to particular pathways. Medullary carcinoma is invariably associated with MSI while serrated adenocarcinoma is characterized by excess of methylation. TNM classification and the tumoral grade are still the most important prognostic factors. Several parameters including morphological criteria, molecular features or immunohistochemical markers seem to be relevant but none of them are today used in clinical practice. A more accurate approach of the evaluation of these additional parameters should improve their prognosis values. double dagger.

Topics: Adenocarcinoma; Carcinoma, Medullary; Colorectal Neoplasms; DNA Mismatch Repair; Humans; Inflammation; Keratins; Neoplasm Invasiveness; Prognosis

Heme oxygenase-1 (HO-1), a cytoprotective, pro-angiogenic and anti-inflammatory enzyme, is strongly induced in injured tissues. Our aim was to clarify its role in cutaneous wound healing. In wild type mice, maximal expression of HO-1 in the skin was observed on the 2(nd) and 3(rd) days after wounding. Inhibition of HO-1 by tin protoporphyrin-IX resulted in retardation of wound closure. Healing was also delayed in HO-1 deficient mice, where lack of HO-1 could lead to complete suppression of reepithelialization and to formation of extensive skin lesions, accompanied by impaired neovascularization. Experiments performed in transgenic mice bearing HO-1 under control of keratin 14 promoter showed that increased level of HO-1 in keratinocytes is enough to improve the neovascularization and hasten the closure of wounds. Importantly, induction of HO-1 in wounded skin was relatively weak and delayed in diabetic (db/db) mice, in which also angiogenesis and wound closure were impaired. In such animals local delivery of HO-1 transgene using adenoviral vectors accelerated the wound healing and increased the vascularization. In summary, induction of HO-1 is necessary for efficient wound closure and neovascularization. Impaired wound healing in diabetic mice may be associated with delayed HO-1 upregulation and can be improved by HO-1 gene transfer.

Topics: Adenoviridae; Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents; Diabetes Mellitus, Experimental; Gene Transfer Techniques; Heme Oxygenase-1; Humans; Inflammation; Keratins; Mice; Mice, Inbred C57BL; Promoter Regions, Genetic; Transgenes; Wound Healing

Four different modifications of the keratin polymer were made as small rods and inserted into the subcutaneous tissue of the hind limbs of adult female sheep. Material 1 was porous, whereas materials 2, 3, and 4 had little or no internal porosity. The tissue response to each material was determined with regard to the extent of the inflammatory reaction and formation of a fibrous capsule around the implant, and the integrity and morphological appearance of the implant was assessed. An inflammatory cell infiltrate and fibrous capsule formed at an early stage around implants of material 1. Subsequently the inflammation decreased, the fibrous capsule became more mature, and the implants became cavitated and invaded by mono- and multi-nucleated macrophages, fibroblasts and blood vessels, with breakdown of the implant material occurring at its periphery. Similar changes were observed for implants made of materials 2 and 4. Implants of material 3 were remarkable in that, while surrounded by an inflammatory infiltrate and a fibrous capsule, they did not show any disturbance even at 24 weeks. The fibrous capsule around material 3 was thinner than that around material 1 at 6 to 24 weeks (both materials prepared using ammonium thioglycollate). No difference in capsule thickness was found for materials 2 and 4 (both materials prepared using thioglycollic acid).

Topics: Animals; Biocompatible Materials; Female; Granulation Tissue; Implants, Experimental; Inflammation; Keratins; Materials Testing; Polymers; Porosity; Sheep; Subcutaneous Tissue

GPR125 belongs to the family of Adhesion G protein-coupled receptors (GPCRs). A single copy of GPR125 was found in many vertebrate genomes. We also identified a Drosophila sequence, DmCG15744, which shares a common ancestor with the entire Group III of Adhesion GPCRs, and also contains Ig, LRR and HBD domains which were observed in mammalian GPR125.. We found specific expression of GPR125 in cells of the choroid plexus using in situ hybridization and protein-specific antibodies and combined in situ/immunohistochemistry co-localization using cytokeratin, a marker specific for epithelial cells. Induction of inflammation by LPS did not change GPR125 expression. However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.. These findings suggest that GPR125 plays a functional role in choroidal and hippocampal response to injury.

Topics: Animals; Base Sequence; Brain Injuries; Choroid Plexus; Gene Expression Profiling; Hippocampus; Humans; Immunohistochemistry; In Situ Hybridization; Inflammation; Keratins; Lipopolysaccharides; Male; Membrane Glycoproteins; Membrane Proteins; Mice; Platelet Glycoprotein GPIb-IX Complex; Rats; Receptors, G-Protein-Coupled; RNA, Messenger; Time Factors; Up-Regulation

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Hamartoma; Humans; Inflammation; Keratins; Nasal Mucosa; Nasopharyngeal Neoplasms; Papilloma, Inverted; Paranasal Sinus Neoplasms; Polyps; Teratoma

Exhaled breath condensate (EBC) is increasingly studied as a noninvasive research method of sampling the lungs, measuring several biomarkers. The exact site of origin of substances measured in EBC is unknown, as is the clinical applicability of the technique. Special techniques might be needed to measure EBC biomarkers.. To assess biomarker concentrations in clinical disease and investigate the site of origin of EBC, we compared EBC and bronchoalveolar lavage (BAL) biomarkers in 49 patients undergoing bronchoscopy for clinical indications.. We measured exhaled nitric oxide, 8-isoprostane, hydrogen peroxide, total nitrogen oxides, pH, total protein, and phospholipid (n = 33) and keratin (n = 15) to assess alveolar and mucinous compartments, respectively. EBC was collected over 10 min using a refrigerated condenser according to European Respiratory Society/American Thoracic Society recommendations, and BAL performed immediately thereafter.. 8-Isoprostane, nitrogen oxides, and pH were significantly higher in EBC than in BAL (3.845 vs. 0.027 ng/ml, 28.4 vs. 3.8 microM, and 7.35 vs. 6.4, respectively; p < 0.001). Hydrogen peroxide showed no difference between EBC and BAL (17.5 vs. 20.6 microM, p = not significant), whereas protein was significantly higher in BAL (33.8 vs. 183.2 microg/ml, p < 0.001). Total phospholipid was also higher in EBC, but keratin showed no difference. No significant correlation was found between EBC and BAL for any of the biomarkers evaluated either before or after correction for dilution.. In clinical disease, markers of inflammation and oxidative stress are easily measurable in EBC using standard laboratory techniques and EBC is readily obtained. However, EBC and BAL markers do not correlate.

Topics: Biomarkers; Breath Tests; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Dinoprost; Exhalation; Female; Humans; Hydrogen Peroxide; Inflammation; Keratins; Lung Diseases; Male; Middle Aged; Nitric Oxide; Phospholipids

Keratin 8 (K8) and keratin-18 (K18) are the major intermediate filament proteins in the intestinal epithelia. The regulation and function of keratin in the intestinal epithelia is largely unknown. In this study we addressed the role and regulation of K8 and K18 expression by interleukin 6 (IL-6). Caco2-BBE cell line and IL-6 null mice were used to study the effect of IL-6 on keratin expression. Keratin expression was studied by Northern blot, Western blot, and confocal microscopy. Paracellular permeability was assessed by apical-to-basal transport of a fluorescein isothiocyanate dextran probe (FD-4). K8 was silenced using the small interfering RNA approach. IL-6 significantly up-regulated mRNA and protein levels of K8 and K18. Confocal microscopy showed a reticular pattern of intracellular keratin localized to the subapical region after IL-6 treatment. IL-6 also induced serine phosphorylation of K8. IL-6 decreased paracellular flux of FD-4 compared with vehicle-treated monolayers. K8 silencing abolished the decrease in paracellular permeability induced by IL-6. Administration of dextran sodium sulfate (DSS) significantly increased intestinal permeability in IL-6-/- mice compared with wild type mice given DSS. Collectively, our data demonstrate that IL-6 regulates the colonic expression of K8 and K18, and K8/K18 mediates barrier protection by IL-6 under conditions where intestinal barrier is compromised. Thus, our data uncover a novel function of these abundant cytoskeletal proteins, which may have implications in intestinal disorders such as inflammatory bowel disease wherein barrier dysfunction underlies the inflammatory response.

Topics: Animals; Caco-2 Cells; Colon; Epithelial Cells; Fluorescent Dyes; Humans; Inflammation; Interleukin-6; Intestines; Keratin-8; Keratins; Mice; Mice, Transgenic; Microscopy, Confocal; Permeability

Little is known about the behavior of the ovarian surface epithelium (OSE), which plays a central role in ovarian cancer etiology. It has been suggested that incessant ovulation causes OSE changes leading to transformation and that high gonadotropin levels during postmenopause activate OSE receptors, inducing proliferation. We examined the chronology of OSE changes, including tumor appearance, in a mouse model where ovulation never occurs due to deletion of follitropin receptor. Changes in epithelial cells were marked by pan-cytokeratin (CK) staining. Histologic changes and CK staining in the OSE increased from postnatal day 2. CK staining was observed inside the ovary by 24 days and increased thereafter in tumor-bearing animals. Ovaries from a third of aged (1 year) mutant mice showed CK deep inside, indicating cell migration. These tumors resembled serous papillary adenoma of human ovaries. Weak expression of GATA-4 and elevation of PCNA, cyclooxygenase-1, cyclooxygenase-2, and platelet-derived growth factor receptors alpha and beta in mutants indicated differences in cell proliferation, differentiation, and inflammation. Thus, we report that OSE changes occur long before epithelial tumors appear in FORKO mice. Our results suggest that neither incessant ovulation nor follicle-stimulating hormone receptor presence in the OSE is required for inducing ovarian tumors; thus, other mechanisms must contribute to ovarian tumorigenesis.

Topics: Adenoma; Animals; Cell Differentiation; Cell Proliferation; Cyclooxygenase 1; Cyclooxygenase 2; Cystadenoma, Serous; Epithelial Cells; Female; Fluorescent Antibody Technique; GATA4 Transcription Factor; Humans; Immunoenzyme Techniques; Inflammation; Keratins; Mice; Ovarian Neoplasms; Ovary; Proliferating Cell Nuclear Antigen; Receptors, FSH; Receptors, Platelet-Derived Growth Factor

Affymetrix gene-expression analysis was performed on mRNAs from involved and noninvolved skin biopsies from three volunteers with Lentigo senilis. Of the 42,000 transcripts scanned, 17 were downregulated (<1.4 times below the control level) and 23 were upregulated (>1.9 times above the control level). A serine peptidase gene was downregulated in keeping with the suggestion that age spots are associated with impaired melanin degradation. Three genes involved in the keratinization and synthesis and the organization of fibers in the basement membrane were downregulated, two metalloproteinase genes were upregulated, as were six genes associated with the inflammatory response, in keeping with the postulate that the visible aspects of aged skin are causally linked with a microinflammatory response. The regulation of five genes associated with the Wnt family was altered. Antiapoptotic genes were downregulated, and six genes associated with transmembrane transport were upregulated.

Topics: Aging; Basement Membrane; Female; Gene Expression Profiling; Gene Expression Regulation; Humans; Inflammation; Keratins; Lentigo; Male; Models, Biological; Oligonucleotide Array Sequence Analysis; Skin

We previously showed that the EP2 knockout mice were resistant to chemically induced skin carcinogenesis. The purpose of this study was to investigate the role of the overexpression of the EP2 receptor in mouse skin carcinogenesis. To determine the effect of overexpression of EP2, we used EP2 transgenic (TG) mice and wild-type (WT) mice in a DMBA (7,12-dimethylbenz[alpha]anthracene)/TPA (12-O-tetradecanoylphorbol-13-acetate) two-stage carcinogenesis protocol. EP2 TG mice developed significantly more tumors compared with WT mice. Overexpression of the EP2 receptor increased TPA-induced keratinocyte proliferation both in vivo and in vitro. In addition, the epidermis of EP2 TG mice 48 h after topical TPA treatment was significantly thicker compared to that of WT mice. EP2 TG mice showed significantly increased cyclic adenosine monophosphate levels in the epidermis after prostaglandin E2 (PGE2) treatment. The inflammatory response to TPA was increased in EP2 TG mice, as demonstrated by an increased number of macrophages in the dermis. Tumors and 7 x TPA-treated and DMBA-TPA-treated (6 weeks) skins from EP2 TG mice produced more blood vessels than those of WT mice as determined by CD-31 immunostaining. Vascular endothelial growth factor (VEGF) protein expression was significantly increased in squamous cell carcinoma (SCC) samples from EP2 TG mice compared that of WT mice. There was, however, no difference in the number of apoptotic cells in tumors from WT and EP2 TG mice. Together, our results suggest that the overexpression of the EP2 receptor plays a significant role in the protumorigenic action of PGE2 in mouse skin.

Topics: Animals; Animals, Newborn; Bromodeoxyuridine; Carcinoma, Squamous Cell; Cattle; Cell Culture Techniques; Cell Proliferation; Female; Humans; Hyperplasia; Inflammation; Keratinocytes; Keratins; Mice; Mice, Transgenic; Neovascularization, Pathologic; Polymerase Chain Reaction; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP2 Subtype; Skin Neoplasms; Tetradecanoylphorbol Acetate; Up-Regulation

Several studies have suggested the involvement of an autoimmune mechanism in aspirin (ASA)-intolerant asthma. To test this hypothesis, we measured the levels of circulating autoantibodies, such as IgG and IgA to tissue transglutaminase (TGase), IgG to cytokeratins (CKs) 8, 18, and 19, Clq-binding immune complex (CIC), and antinuclear antibody (ANA), in the sera of 79 patients with ASA-intolerant asthma (Group I) and those of two control groups, consisting of 61 patients with ASA-tolerant asthma (Group II) and 88 healthy control subjects (Group III) by means of ELISA. Significantly higher prevalences of IgG antibodies to CK18 (13.9%) and CK19 (17.7%) were noted in Group I, as compared with Group III (p<0.05 for all) not with Group II. Regarding the prevalences of other autoantibodies, the levels of ANA (1.3%), IgG to TGase (3.8%), and CIC (24.7%) in Group I were not significantly different from those in Groups II and III. Significant correlations were found between positivities for the anti-CK18 and anti-CK19 autoantibodies and the PC(20) methacholine values in the analysis of asthma Groups I and II vs. normal controls, (p=0.001 and p=0.003, respectively). Further studies are needed to explore the potential involvement of an autoantibody-mediated mechanism in the clinical manifestation of bronchial asthma.

Topics: Aged; Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Asthma; Autoantibodies; Bronchi; Case-Control Studies; Child; Drug Resistance; Female; Humans; Inflammation; Keratins; Male; Middle Aged

Our knowledge of the etiology and pathogenesis of skin diseases characterized by abnormal growth is relatively limited. Even more important, still very little is known of how epidermopoeisis is controlled in normal epidermis. There is no cure for skin diseases caused by abnormal growth control, such as psoriasis. Mechanisms are complex, additional models for epidermal growth and differentiation, and specific techniques to analyze these processes, are needed. Therefore, we have developed flow cytometric techniques to study epidermal growth over the past two decades. A prerequisite for accurate and reliable flow cytometric analysis is a high quality of epidermal single-cell suspensions. In this chapter, protocols are described for preparation of single-cell suspensions and protocols for the multiparameter flow cytometric analysis of growth behavior in normal and hyperproliferative epidermis.

Topics: Cell Differentiation; Cell Division; Flow Cytometry; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; Skin

How the inflammatory response is initiated has been well defined but relatively little is known about how such responses are resolved. Here we show that the D6 chemokine receptor is involved in the post-inflammatory clearance of beta-chemokines from cutaneous sites. After induction of inflammation by phorbol esters, wild-type mice showed a transient inflammatory response. However, in D6-deficient mice, an excess concentration of residual chemokines caused a notable inflammatory pathology with similarities to human psoriasis. These results suggest that D6 is involved in the resolution of the cutaneous inflammatory response.

Topics: Animals; Chemokine CXCL2; Chemokine Receptor D6; Chemokines; Chemokines, CC; Dermatitis; Female; Histocytochemistry; Inflammation; Keratins; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Proliferating Cell Nuclear Antigen; Psoriasis; Receptors, CCR10; Receptors, Chemokine; Skin; von Willebrand Factor

The inducible form of cyclooxygenase (COX), COX-2, is up-regulated in many epithelial cancers and its prostaglandin products increase proliferation, enhance angiogenesis, and inhibit apoptosis in several tissues. Pharmacologic inhibition and genetic deletion studies showed a marked reduction of tumor development in colon and skin. COX-2 has also been strongly implicated in urinary bladder cancer primarily by studies with nonselective COX- and COX-2-selective inhibitors. We now show that forced expression of COX-2, under the control of a keratin 5 promoter, is sufficient to cause transitional cell hyperplasia (TCH) in 17% and 75% of the heterozygous and homozygous transgenic lines, respectively, in an age-dependent manner. TCH was strongly associated with inflammation, primarily nodules of B lymphocytes; some T cells and macrophage infiltration were also observed. Additionally, transitional cell carcinoma was observed in approximately 10% of the K5.COX-2 transgenic mice; no TCH or transitional cell carcinoma was observed in wild-type bladders. Immunohistochemistry for vascular proliferation and vascular endothelial growth factor showed significant increases above that in wild-type urinary bladders. Our results suggest that overexpression of COX-2 is sufficient to cause hyperplasia and carcinomas in the urinary bladder. Therefore, inhibition of COX-2 should continue to be pursued as a potential chemopreventive and therapeutic strategy.

Topics: Animals; B-Lymphocytes; Carcinoma, Transitional Cell; Cell Proliferation; Cyclooxygenase 2; Gene Expression Regulation, Enzymologic; Humans; Hyperplasia; Inflammation; Keratin-15; Keratin-5; Keratins; Macrophages; Membrane Proteins; Mice; Mice, Transgenic; Neoplasm Staging; Promoter Regions, Genetic; Prostaglandin-Endoperoxide Synthases; T-Lymphocytes; Transcription, Genetic; Urinary Bladder; Urinary Bladder Neoplasms; Vascular Endothelial Growth Factor A

Keratin 8 (K8) is the major intermediate filament protein present in intestinal epithelia. Depending on the mouse genetic background, absence of K8 causes embryonic lethality or colonic hyperplasia and colitis. We studied disease progression, the inflammatory responses, and role of luminal bacteria in K8-null mice in order to characterize the intestinal pathology of K8-associated colitis. Colon lymphocytes were isolated for analysis of their phenotype and cytokine production, and vascular and lymphocyte adhesion molecule expression in K8-/- mice of varying ages. K8-/- mice had a marked increase in TCR(beta)-positive/CD4-positive T cells infiltrating the colon lamina propria, in association with enhanced Th2 cytokine (IL-4, IL-5 and IL-13) production. K8-/- mice show early signs of inflammation even prior to weaning, that increases with age, and their epithelial cells overexpress MHC class II antigens. The chronic colitis is related to increased CD4-positive infiltrating T cells displaying memory and naive phenotypes, and an altered vascular endothelium with aberrant expression of peripheral node addressin. Analysis of normal gut-specific homing molecules, reveals an increased number of alpha(4)beta(7)-positive cells and vascular mucosal addressin cell adhesion molecule-1 in K8-null colons. Antibiotic treatment markedly decreased colon inflammation and ion transporter AE1/2 mistargeting, indicating that luminal bacteria play an important role in the observed phenotype. Therefore, K8-null mice develop chronic spontaneous Th2-type colitis due to a primary epithelial rather than immune cell defect, which is amenable to antibiotic therapy. These mice provide a model to investigate epithelial-leukocyte and epithelial-microbial cross-talk.

Topics: Animals; Anti-Bacterial Agents; Antigens, Surface; CD4 Antigens; CD4-Positive T-Lymphocytes; CD8 Antigens; Cell Adhesion Molecules; Cell Membrane; Colitis; Colon; Cytokines; Disease Progression; Endothelium, Vascular; Epithelial Cells; Flow Cytometry; Immunoglobulins; Immunohistochemistry; Inflammation; Integrins; Interleukin-13; Interleukin-4; Interleukin-5; Ions; Keratins; Lymphocytes; Membrane Proteins; Mice; Mice, Transgenic; Microscopy, Fluorescence; Models, Biological; Mucoproteins; Phenotype; Protein Structure, Tertiary; Receptors, Antigen, T-Cell; Th2 Cells; Time Factors

The function of intestinal keratins is unknown, although keratin 8 (K8)-null mice develop colitis, hyperplasia, diarrhea, and mistarget jejunal apical markers. We quantified the diarrhea in K8-null stool and examined its physiologic basis. Isolated crypt-units from K8-null and wild-type mice have similar viability. K8-null distal colon has normal tight junction permeability and paracellular transport but shows decreased short circuit current and net Na absorption associated with net Cl secretion, blunted intracellular Cl/HCO3-dependent pH regulation, hyperproliferation and enlarged goblet cells, partial loss of the membrane-proximal markers H,K-ATPase-beta and F-actin, increased and redistributed basolateral anion exchanger AE1/2 protein, and redistributed Na-transporter ENaC-gamma. Diarrhea and protein mistargeting are observed 1-2 d after birth while hyperproliferation/inflammation occurs later. The AE1/2 changes and altered intracellular pH regulation likely account, at least in part, for the ion transport defects and hyperproliferation. Therefore, colonic keratins have a novel function in regulating electrolyte transport, likely by targeting ion transporters to their cellular compartments.

Topics: Age Factors; Animals; Bicarbonates; Biomarkers; Bumetanide; Chlorides; Colon; Cyclic AMP; Diuretics; Electrolytes; Electrophysiology; Female; Hydrogen-Ion Concentration; Inflammation; Intestinal Mucosa; Ion Transport; Keratin-8; Keratins; Male; Mice; Mice, Knockout; Nitrobenzoates; Phenotype; Sodium

We have identified miss-sense mutations in keratin 8 in a subset of patients with inflammatory bowel disease (Crohn disease and ulcerative colitis). Inflammatory bowel diseases are a group of disorders that are polygenic in origin and involve intestinal epithelial breakdown. We investigated the possibility that these keratin mutations might contribute to the course of the disease by adversely affecting the keratin filament network that provides mechanical support to cells in epithelia. The mutations (Gly62 to Cys, Ile63 to Val and Lys464 to Asn) all lie outside the major mutation hotspots associated with severe disease in epidermal keratins, but using a combination of in vitro and cell culture assays we show that they all have detrimental effects on K8/K18 filament assembly in vitro and in cultured cells. The G62C mutation also gives rise to homodimer formation on oxidative stress to cultured intestinal epithelial cells, and homodimers are known to be polymerization incompetent. Impaired keratin assembly resulting from the K8 mutations found in some inflammatory bowel disease patients would be predicted to affect the maintenance and re-establishment of mechanical resilience in vivo, as required during keratin cytoskeleton remodeling in cell division and differentiation, which may lead to epithelial fragility in the gut. Simple epithelial keratins may thus be considered as candidates for genes contributing to a risk of inflammatory bowel disease.

Topics: Actin Cytoskeleton; Animals; Antibodies, Monoclonal; Base Sequence; Cell Differentiation; Chromosomes, Human, Pair 12; Colitis, Ulcerative; Crohn Disease; Dimerization; Electrophoresis, Polyacrylamide Gel; Humans; Inflammation; Inflammatory Bowel Diseases; Keratin-8; Keratins; Mice; Models, Genetic; Molecular Sequence Data; Mutation; Oxidative Stress; Polymers; Protein Binding; Protein Conformation; Sequence Analysis, DNA; Time Factors; Transfection; Xenopus

MAIL (molecule-possessing ankyrin repeats induced by lipopolysaccharide) is a nuclear IkappaB protein that is also termed interleukin-1-inducible nuclear ankyrin repeat protein or inhibitor of nuclear factor kappaB (IkappaB) zeta. In this study, we generated Mail-/- mice to investigate the roles of MAIL in whole organisms. Mail-/- mice grew normally until 4-8 weeks after birth, when they began to develop lesions in the skin of the periocular region, face, and neck. MAIL mRNA and protein were constitutively expressed in the skin of wild type controls, especially in the keratinocytes. Serum IgE was higher in Mail-/- mice than in normal. Histopathological analysis indicated that the Mail-/- skin lesions appeared to be atopic dermatitis (AD) eczema with inflammatory cell infiltration. In addition, markedly elevated expression of some chemokines such as thymus and activation-regulated chemokine was detected in the Mail-/- skin lesions, similar to that observed in the skin of patients with AD. In Mail-/- mice, MAIL-deficient keratinocytes might be activated to produce chemokines and induce intraepidermal filtration of inflammatory cells, resulting in the onset of the AD-like disease. These findings suggest that MAIL is an essential molecule for homeostatic regulation of skin immunity. The Mail-/- mouse is a valuable new animal model for research on AD.

Topics: Adaptor Proteins, Signal Transducing; Alleles; Animals; Ankyrins; Chemokines; Dermatitis, Atopic; Disease Models, Animal; Genetic Vectors; Genome; I-kappa B Proteins; Immunoglobulin E; Immunohistochemistry; Inflammation; Islets of Langerhans; Keratinocytes; Keratins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Models, Genetic; Nuclear Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transgenes

Within the human prostate epithelium four cell populations can be discriminated based on their expression of keratins (K). Basal cells express high levels of K5 and K14, as well as p63, whereas they have very low levels of androgen receptor, prostate-specific antigen (PSA), K8, and K18. Luminal secretory cells lack p63, K5, and K14 but express high levels of K8, K18, androgen receptor, and PSA. Additionally, cells have been identified with a keratin phenotype intermediate between basal and luminal cells that co-express high levels of K5 and K18 (K5/18) as well as hepatocyte growth factor receptor c-MET. Although intermediate cells have been proposed as precursor cells of prostate cancer, their biology is ill defined. Epithelial cells in proliferative inflammatory atrophy (PIA) appear to be cycling rapidly as indicated by expression of Ki-67, and morphological transitions have been identified between PIA and high-grade prostate intraepithelial neoplasia. Many of the atrophic epithelial luminal cells in PIA are candidates for intermediate cells based in part on weak expression of PSA and androgen receptor, high levels of K8/18, and lack of p63. The objective of this study was to further clarify the phenotype of the proposed intermediate cells in PIA and to quantitatively determine the level in which these intermediate cells preferentially occur in PIA lesions. Intermediate cells were immunohistochemically demonstrated using antibodies to K5, K14, K18, and c-MET. Using radical prostatectomy specimens (n = 15) the area fraction of intermediate cells in normally differentiated prostate epithelium and PIA were quantified by a grid point counting method. Atrophic luminal cells of PIA lesions expressed K5 in 39.2 +/- 7.4% of cells compared to 2.4 +/- 2.3% in normal epithelium (P < 0.00001). By contrast, K14 was only expressed in 3.0 +/- 3.2% of the luminal cells. Previous studies have shown that virtually 100% of these atrophic luminal cells are strongly positive for K8/18. c-MET was present in 44.1 +/- 14.1% of luminal cells in PIA but only in 2.1 +/- 2.8% of luminal cells in normal epithelium (P < 0.00001). To unambiguously determine whether intermediate luminal cells in PIA show increased proliferative activity and decreased p27(kip1) expression, double-staining immunofluorescence of Ki-67 and K5, as well as p27(Kip1) and K5 was performed. Luminal cells in PIA often co-expressed K5 and Ki-67. Although p27(Kip1) was strongly expressed in K5-negative differentia

Topics: Cell Division; Epithelial Cells; Humans; Inflammation; Keratin-14; Keratin-5; Keratins; Male; Prostate; Prostate-Specific Antigen; Prostatectomy; Prostatic Hyperplasia; Proto-Oncogene Proteins c-met

Efforts have been made to deliver transgenes to the airway epithelia of laboratory animals and humans to develop gene therapy for cystic fibrosis. These investigations have been disappointing due to combinations of transient and low-level gene expression, acute toxicity, and inflammation. We have developed new helper-dependent adenoviral vectors to deliver an epithelial cell-specific keratin 18 expression cassette driving the beta-galactosidase (beta-gal) or human alpha-fetoprotein (AFP) reporter genes. Following intranasal administration to mice, we found that the reporter genes were widely expressed in airway epithelial and submucosal cells, and secreted human AFP was also detectable in serum. In contrast to a first-generation adenoviral vector, inflammation was negligible at doses providing efficient transduction, and expression lasted longer than typically reported-up to 28 days with beta-gal and up to 15 weeks with human AFP. These results suggest that delivery to the airway of helper-dependent adenoviral vectors utilizing a tissue-specific promoter could be a significant advance in the development of gene therapy for cystic fibrosis.

Topics: Adenoviridae; alpha-Fetoproteins; Animals; beta-Galactosidase; Cystic Fibrosis; Epithelial Cells; Female; Gene Expression; Genetic Therapy; Genetic Vectors; Helper Viruses; Humans; Inflammation; Keratins; Mice; Mice, Inbred C57BL; Mucous Membrane; Respiratory Mucosa

p40, the common subunit of the proinflammatory cytokines IL-12 and IL-23, is produced by resident skin cells. Whereas the in vivo effects of IL-12 are well established, little is known about the role of IL-23 in cutaneous immune responses. In this study we show that p40 transgenic (TG) mice constitutively produce IL-23 (p19/p40), but not IL-12 (p35/p40), in basal keratinocytes by cosecretion of TG p40 with endogenous p19. Repeated injections of rIL-23 in littermate (LM) mice result in an inflammatory skin disease similar to that of p40 TG mice, confirming the proinflammatory activity of IL-23. Furthermore, IL-23 secretion by p40 TG keratinocytes induces elevated numbers of Langerhans cells (LC) with a marked up-regulation of costimulatory molecules, indicating advanced maturation of keratin 14 (K14)/p40 LC when compared with LM LC. At the functional level, freshly isolated K14/p40 LC greatly exceeded LC from LM animals in their capacity to stimulate allogeneic T cell proliferation. To assess whether IL-23 regulates cutaneous immune responses in vivo, we used an allogeneic skin transplantation model. Full thickness skin grafts from K14/p40 donors (H-2(q)) transplanted across a MHC class I and class II barrier onto BALB/c (H-2(d)) recipients were rejected in a significantly accelerated fashion (mean survival time: 8.8 days) when compared with skin grafts from non-TG LM (H-2(q)) (mean survival time: 10.7 days, p < 0.01). Based on these results we propose that IL-23-induced changes of LC may be an important mechanism in directing the outcome of cutaneous immune responses.

Topics: Animals; Cell Count; Cell Movement; Cells, Cultured; Female; Graft Rejection; Immunity, Cellular; Immunophenotyping; Inflammation; Injections, Subcutaneous; Interleukin-12; Interleukin-12 Subunit p40; Interleukin-23; Interleukin-23 Subunit p19; Interleukins; Interphase; Keratin-14; Keratins; Langerhans Cells; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Transgenic; Organ Culture Techniques; Protein Subunits; RNA, Messenger; Skin; Skin Diseases; Skin Transplantation

Atopic dermatitis (AD) is a pruritic inflammatory skin disease. Because IL-18 directly stimulates T cells and mast cells to release AD-associated molecules, Th2 cytokines, and histamine, we investigated the capacity of IL-18 to induce AD-like inflammatory skin disease by analyzing KIL-18Tg and KCASP1Tg, which skin-specifically overexpress IL-18 and caspase-1, respectively. They spontaneously developed relapsing dermatitis with mastocytosis and Th2 cytokine accumulation accompanied by systemic elevation of IgE and histamine. Stat6-deficient KCASP1Tg displayed undetectable levels of IgE but manifested the same degree of cutaneous changes, whereas IL-18-deficient KCASP1Tg evaded the dermatitis, suggesting that IL-18 causes the skin changes in the absence of IgE/stat6. KIL-18Tg and IL-1-deficient KCASP1Tg took longer to display the lesion than KCASP1Tg. Thus, AD-like inflammation is initiated by overrelease of IL-18 and accelerated by IL-1. Our present study might provide insight into understanding the pathogenesis of and establishing therapeutics for chronic inflammatory skin diseases including AD.

Topics: Animals; Dermatitis, Atopic; Humans; Immunoglobulin E; Inflammation; Interleukin-18; Keratins; Mice; Mice, Transgenic; Promoter Regions, Genetic; Pruritus; Rabbits; Signal Transduction; Specific Pathogen-Free Organisms; STAT6 Transcription Factor; T-Lymphocytes; Th2 Cells; Trans-Activators

Tissue repair is determined by many signals provided in the local environment. Central to this process is the commitment of the parenchymal cell to undergo apoptosis, survive, or proliferate following inflammation. We hypothesize that lung epithelial cell apoptosis is influenced by exposure to cytokines released into the alveolar microenvironment during the inflammatory process. In this investigation we demonstrate that interferon (IFN)-gamma and interleukin (IL)-1beta have opposing effects on Fas-mediated apoptosis in A549 cells, a human lung epithelial cell line. Exposure to IFN-gamma before Fas activation significantly increased caspase activity, caspase processing of CK-18, a key cytoskeletal protein in epithelial cells, and increased the appearance of apoptotic nuclei. Induction of Fas-mediated death by IFN-gamma was 3-fold higher than with Fas activation alone. In contrast, pretreatment with IL-1beta before Fas activation completely inhibited apoptosis. Furthermore, our results demonstrate that IFN-gamma and IL-1beta induce opposite effects at multiple checkpoints during Fas-mediated apoptosis. Most striking, IL-1beta prevented the activation of caspases involved in Fas-mediated death by inducing an anti-apoptotic effect proximal to or at the point of caspase-8 activation. Finally, our investigation demonstrates that the differential impact of IL-1beta and IFN-gamma on Fas-mediated apoptosis are in part dependent on modulation of the PI 3-K/Akt survival pathway.

Topics: Apoptosis; bcl-X Protein; Blotting, Western; Caspase 8; Caspase 9; Caspase Inhibitors; Caspases; Cell Nucleus; Cell Survival; Coumarins; Cytokines; Cytoplasm; Enzyme Activation; Enzyme Inhibitors; Epithelial Cells; fas Receptor; Flow Cytometry; Humans; Immunohistochemistry; Inflammation; Interferon-gamma; Interleukin-1; Keratins; Lung; Microscopy, Fluorescence; Phosphatidylinositol 3-Kinases; Poly(ADP-ribose) Polymerases; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Tumor Cells, Cultured

Transgenic mice (K5-PKC alpha) in which the keratin 5 promoter directs the expression of protein kinase C-alpha (PKC alpha) to epidermal keratinocytes display a 10-fold increase in PKC alpha protein in their epidermis and alterations in phorbol ester-induced cutaneous inflammation [J Cell Science 1999;112:3497-3506]. In the current study, we have used these K5-PKC alpha mice to examine the role of PKC alpha in keratinocyte phospholipid metabolism/eicosanoid production and cutaneous inflammation. Primary keratinocytes from wild-type and transgenic mice were prelabeled in culture with [(3)H]arachidonic acid (AA) and subsequently treated with TPA. Compared with wild-type keratinocytes, K5-PKC alpha keratinocytes displayed a 2-fold increase in AA release. TPA treatment resulted in the phosphorylation of cPLA(2). PKC inhibitors GF-109203X or H7, but not mitogen-activated protein/extracellular signal-regulated protein kinase (MEK) inhibitor PD 98059, could inhibit phosphorylation and AA release. Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment of K5-PKC alpha mice resulted in a 5-fold increase in epidermal COX-2 induction and a 2- to 3-fold increase in prostaglandin (PG) E(2) levels above that observed in TPA-treated wild-type mice. PD 98059, GF-109203X, or H7 could block cyclooxygenase-2 (COX-2) induction by TPA. Because C/EBP beta, a basic leucine zipper transcription factor, can be activated via a PKC alpha/mitogen-activated protein kinase pathway and can influence COX-2 expression, we examined whether C/EBP beta is involved in TPA-induced epidermal COX-2 expression. TPA-induced COX-2 expression was similar in C/EBP beta nullizygous and wild-type mice. In summary, our results indicate that epidermal PKC alpha coordinately regulates cPLA(2) activity and COX-2 expression resulting in increased levels of AA and PGE(2). Furthermore, PKC alpha-induced AA release and cPLA(2) phosphorylation are independent of MEK, whereas PKC alpha-induced COX-2 expression and PGE(2) production are MEK-dependent and C/EBP beta-independent events.

Topics: Animals; Arachidonic Acid; CCAAT-Enhancer-Binding Protein-beta; Cells, Cultured; Cyclooxygenase 2; Cytosol; Dinoprostone; Enzyme Inhibitors; Inflammation; Isoenzymes; Keratinocytes; Keratins; Mice; Mice, Transgenic; Mitogen-Activated Protein Kinase Kinases; Phospholipases A; Phospholipids; Phosphorylation; Promoter Regions, Genetic; Prostaglandin-Endoperoxide Synthases; Protein Isoforms; Protein Kinase C; Protein Kinase C-alpha; Signal Transduction; Skin; Tetradecanoylphorbol Acetate

Glucocorticoids (GCs) are potent inhibitors of epidermal proliferation and effective anti-inflammatory compounds, which make them the drug of choice for a wide range of inflammatory and hyperproliferative skin disorders. GC action is mediated via the glucocorticoid receptor (GR). To study the role of GR in skin development and the molecular mechanisms underlying its action, we generated transgenic mice overexpressing GR in epidermis and other stratified epithelia, under the control of the keratin K5 promoter. Newborn mice show altered skin development, manifested as variable-sized skin lesions that range from epidermal hypoplasia and underdeveloped dysplastic hair follicles to a complete absence of this tissue. In the most affected individuals, skin was absent at the cranial and umbilical regions, and the vibrissae and eyebrows appear scarce, short, and curly. In addition, as a consequence of transgene expression in other ectodermally derived epithelia, K5-GR mice exhibited further abnormalities that strikingly resemble the clinical findings in patients with ectodermal dysplasia, which includes aplasia cutis congenita. In adult transgenic skin, topical application of the tumor promoter TPA did not elicit hyperplasia or transcriptional induction of several proinflammatory cytokines. This anti-inflammatory role of GR was due at least in part to interference with NF-kB, leading to a strong reduction in the kB-binding activity without altering the transcriptional levels of the inhibitor IkBa.

Topics: Animals; Cell Division; Ectoderm; Gene Expression; Inflammation; Keratin-15; Keratin-5; Keratins; Mice; Mice, Transgenic; Promoter Regions, Genetic; Receptors, Glucocorticoid; Skin; Tetradecanoylphorbol Acetate

We implanted keratin and poly(methyl methacrylate) (PMMA) particles to the surface of mouse calvariae to produce a quantitative, localized, inflammatory bone remodeling similar to that seen in cholesteatoma. Both types of particles resulted in increased osteoclast density compared with controls. Osteoclasts infiltrated from marrow and vascular spaces and were active at the periphery of these spaces leading to significant bone remodeling, as demonstrated by the incorporation of bone-labelling fluorophores. Osteoclasts were rarely found on the surface of the calvariae, and mineral apposition rate at the ventral surface was not altered in keratin-implanted animals compared with nonoperated controls. While not useful for the study of the root cause of cholesteatoma, this model will allow the study ofpathologic bone remodeling related to cholesteatoma in a genetically defined animal.

Topics: Animals; Bone and Bones; Bone Diseases; Bone Remodeling; Calcification, Physiologic; Cholesteatoma; Disease Models, Animal; Inflammation; Keratins; Mice; Mice, Inbred C57BL; Osteoclasts; Osteolysis; Peptide Fragments; Skull

Epidermal keratinocytes respond to injury by becoming activated, i.e. hyperproliferative, migratory, and proinflammatory. These processes are regulated by growth factors and cytokines. One of the markers of activated keratinocytes is keratin K6. We used a novel organ culture system to show that tumor necrosis factor alpha (TNFalpha) induces the expression of K6 protein and mRNA in human skin. Multiple isoforms of K6 are encoded by distinct genes and have distinct patterns of expression. By having shown previously that proliferative signals, such as epidermal growth factor (EGF), induce expression of the cytoskeletal protein keratin K6b, we here demonstrate that the same isoform, K6b, is also induced by TNFalpha, a proinflammatory cytokine. Specifically, TNFalpha induces the transcription of the K6b gene promoter. By using co-transfection, specific inhibitors, and antisense oligonucleotides, we have identified NFkappaB and C/EBPbeta as the transcription factors that convey the TNFalpha signal. Both transcription factors are necessary for the induction of K6b by TNFalpha and act as a complex, although only C/EBPbeta binds the K6b promoter DNA. By using transfection, site-directed mutagenesis, and footprinting, we have mapped the site that responds to TNFalpha, NFkappaB, and C/EBPbeta. This site is separate from the one responsive to EGF and AP1. Our results show that the proinflammatory (TNFalpha) and the proliferative (EGF) signals in epidermis separately and independently regulate the expression of the same K6b keratin isoform. Thus, the cytoskeletal responses in epidermal cells can be precisely tuned by separate proliferative and inflammatory signals to fit the nature of the injuries that caused them.

Topics: Base Sequence; Binding Sites; CCAAT-Enhancer-Binding Protein-beta; Cell Division; DNA Footprinting; Epidermal Cells; Epidermal Growth Factor; Epidermis; Fluorescent Antibody Technique; HeLa Cells; Humans; Inflammation; Keratinocytes; Keratins; Molecular Sequence Data; Mutation; NF-kappa B; Oligonucleotides, Antisense; Promoter Regions, Genetic; Protein Isoforms; Response Elements; Transcription Factor AP-1; Transcriptional Activation; Transfection; Tumor Necrosis Factor-alpha

The immunohistochemical expression of PCNA and Ki-67 proteins and the histochemical expression of AgNORs were studied in 20 odontogenic keratocysts in order to assess the relationship between epithelial cell proliferation and inflammation within the capsule. Immunostained cells were quantified by conventional methods, and both quantitative and morphometric analyses of AgNORs were performed by TV image analysis. Non-inflamed odontogenic keratocysts showed a typical epithelial lining and inflamed odontogenic keratocysts were lined also by hyperplastic non-keratinized stratified squamous epithelium. A statistically significant increase of PCNA+ and Ki-67+ cells and of AgNOR numbers was detected in the linings of inflamed odontogenic keratocysts compared to non-inflamed lesions. The results suggest the existence of greater proliferative activity in the epithelial cells of inflamed odontogenic keratocysts, which may be associated with the disruption of the typical structure of odontogenic keratocyst linings.

Topics: Biomarkers; Cell Division; Epithelial Cells; Humans; Immunoenzyme Techniques; Inflammation; Keratins; Ki-67 Antigen; Nucleolus Organizer Region; Odontogenic Cysts; Proliferating Cell Nuclear Antigen; Silver Staining

The purpose of this study was to evaluate how well intraorally transferred skin flaps endure their new surroundings.. Biopsy specimens were taken from 20 patients who had undergone microsurgical reconstruction and as pretransferred skin from 5 of these patients at the time of surgery. The study used immunohistochemistry for immunocompetent cells, differentiation markers for the epidermis and desmosomal proteins, and immunoelectron microscopy for desmosomal protein, in addition to routine histologic examination, including Sudan IV, periodic acid Schiff, and Grocott stains. We also measured the thickness of the epidermis and stratum corneum. Oral swabs from the skin flaps were examined for the presence of yeasts, particularly Candida albicans, by means of a culture method.. According to the results of periodic acid-Schiff and Grocott staining, 20 cases were divided into 2 groups: fungal element-positive cases (n = 15) and fungal element-negative cases (n = 5). All swabs from the former were positive for Candida albicans. In these fungus-positive cases, histopathologic evaluation revealed marked diminution of stratum corneum and pronounced epidermal hyperplasia. Immunohistochemistry demonstrated the dermal infiltration of numerous immunocompetent cells-CD4+, CD8+, CD20+, CD68+, neutrophil elastase+, and HLA-DR+ cells-and the scarce infiltration of IgA+ and IgG+ cells. There were scattered CD1a+, CD4+, CD8+, and HLA-DR+ cells and elastase+ neutrophils in the epidermis. Expression of cytokeratin subtypes (10, 14, 16, and 19), involucrin, and tenascin showed the characteristic features of epidermal proliferation. Enumeration of Ki-67+ keratinocytes showed an increase, indicating epidermal proliferation. Expression of desmoglein 1 and desmocollin 1 in the epidermal keratinocytes was decreased in comparison with that in the pretransferred skin. Immunoelectron microscopy for desmoglein 1 confirmed the reduced immunoreactive deposits along the desmosomal plaques. In the fungus-negative cases, all such changes were a great deal milder.. Taken together, our results demonstrate that most intraorally transferred flaps are affected by an inflammatory process that is induced by the influence of the wet oral environment. They present psoriasiform tissue reactions characterized by epidermal hyperproliferation that are mostly due to Candida albicans infection.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Differentiation; Cadherins; Candida albicans; Candidiasis, Oral; Desmoglein 1; Female; Forearm; Humans; Immunoenzyme Techniques; Inflammation; Keratinocytes; Keratins; Lymphocytes; Male; Microscopy, Immunoelectron; Middle Aged; Mouth; Skin; Skin Transplantation; Surgical Flaps

Protein kinase Calpha (PKCalpha) is one of six PKC isoforms expressed in keratinocytes of mouse epidermis. To gain an understanding of the role of epidermal PKCalpha, we have localized its expression to specific cells of normal mouse skin and examined the effect of keratin 5 (K5) promoter directed expression of PKCalpha in transgenic mice. In normal mouse skin, PKCalpha was extensively expressed in the outer root sheath (ORS) keratinocytes of the anagen hair follicle and weakly expressed in keratinocytes of interfollicular epidermis. K5-targeted expression of PKCalpha to epidermal basal keratinocytes and follicular ORS keratinocytes resulted in a tenfold increase in epidermal PKCalpha. K5-PKCalpha mice exhibited no abnormalities in keratinocyte growth and differentiation in the epidermis. However, a single topical treatment with the PKC activator, 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a striking inflammatory response characterized by edema and extensive epidermal infiltration of neutrophils that formed intraepidermal microabscesses in the epidermis. Compared to TPA-treated wild-type mice, the epidermis of TPA-treated K5-PKCalpha mice displayed increased expression of cyclooxygenase-2 (COX-2), the neutrophil chemotactic factor macrophage inflammatory protein-2 (MIP-2) mRNA and the proinflammatory cytokine TNFalpha mRNA but not IL-6 or IL-1alpha mRNA. To determine if K5-PKCalpha mice display an altered response to TPA-promotion, 7, 12-dimethylbenz[a]anthracene-initiated K5-PKCalpha mice and wild-type mice were promoted with TPA. No differences in papilloma incidence or multiplicity were observed between K5-PKCalpha mice and wild-type littermates. These results demonstrate that the overexpression of PKCalpha in epidermis increases the expression of specific proinflammatory mediators and induces cutaneous inflammation but has little to no effect on epidermal differentiation, proliferation or TPA tumor promotion.

Topics: Abscess; Animals; Cattle; Chemokine CXCL2; Chemotactic Factors; Cyclooxygenase 2; Edema; Enzyme Activation; Epidermis; Gene Expression Regulation; Hair; Inflammation; Isoenzymes; Keratinocytes; Keratins; Mice; Mice, Transgenic; Monokines; Neutrophils; Promoter Regions, Genetic; Prostaglandin-Endoperoxide Synthases; Protein Kinase C; Protein Kinase C-alpha; RNA, Messenger; Skin; Skin Diseases; Tetradecanoylphorbol Acetate; Transcription, Genetic; Tumor Necrosis Factor-alpha

By introducing certain irritants into the middle ear it is possible to produce cholesteatoma. Propylene glycol, the main agent used for this purpose, produces a long-standing inflammation that causes hyperplasia and migration of the epithelium through an intact tympanic membrane. In the present investigation topical prednisolone was used in order to inhibit the production of cholesteatoma. The results indicate that there is a marked decrease in inflammation and hence experimental cholesteatoma production when prednisolone is administered into the middle ear.

Topics: Animals; Anti-Inflammatory Agents; Cell Movement; Cholesteatoma, Middle Ear; Ear, Middle; Epithelium; Fibrosis; Glucocorticoids; Granulation Tissue; Hyperplasia; Inflammation; Injections; Irritants; Keratins; Otitis Media; Prednisolone; Propylene Glycol; Rats; Rats, Wistar; Temporal Bone; Tympanic Membrane

An experimental model of proliferative vitreoretinopathy was developed in the rabbit eye by injecting a solution of human platelet-rich plasma. In this model we evaluated the progression with time of intraocular inflammation and the rate and origin of cell proliferation. A sterile solution adjusted to 107 platelets was injected into the right eye of a total of 46 pigmented and 14 albino rabbits. Animals were sequentially sacrificed at days 7, 14, 21 and 1 month after injection. Clinical evaluation of vitreoretinal proliferation, using a classification in six grades, and of anterior segment inflammation assessed by a Laser Flare Meter, were done for 1 month after injection, before histopathological analysis. Eighty percent of eyes developed tractional retinal detachment in 1 month. Histopathology showed intense cell migration and proliferation in the area of the ciliary body, as early as the seventh day, then further increasing rapidly. Infiltrates were composed of cytokeratin- and vimentin-expressing cells. Abnormal expression of vimentin was also found in ciliary and retinal epithelia and in M¿ller cells. Inflammation measured by the Laser Flare Meter was maximal at day 11 and then reached a plateau at significantly higher levels than controls. Albino rabbits showed significantly lower grades of proliferation, as compared to pigmented rabbits. This study thus clarified some characteristics of experimental vitreoretinal proliferations that that proved similar to those in human diseases, such as the involvement of ciliary body and retinal pigment epithelium, the existence of inflammatory reactions preceding cell proliferation and strong changes in intermediate filaments. This may provide a simple and valuable model for antiproliferative assays and shed some light on the pathogenesis of intraocular proliferative disorders.

Topics: Albinism; Animals; Cell Division; Ciliary Body; Immunohistochemistry; Inflammation; Keratins; Lasers; Male; Rabbits; Retina; Vimentin; Vitreoretinopathy, Proliferative; Vitreous Body

Several reverse transcriptase polymerase chain reaction (RT-PCR) assays have been described for the detection of circulating tumour cells in blood and bone marrow. Target mRNA sequences for this purpose are the cytokeratins (CK) 19 and 20, the carcinoembryonic antigen (CEA), and the prostate-specific antigen messages. In this study, we investigated biological factors influencing the specificity of the CK19 and CEA RT-PCR assays. Bone marrow, granulocyte colony-stimulating factor (G-CSF)-mobilized blood stem cells and peripheral blood samples obtained from healthy volunteers (n = 15; CEA n = 7), from patients with epithelial (n = 29) and haematological (n = 23) cancer and from patients with chronic inflammatory diseases (n = 16) were examined. Neither CEA nor cytokeratin 19 messages could be amplified from bone marrow samples from healthy subjects and from patients with haematological malignancies. In contrast, specimens from patients with inflammatory diseases scored positive up to 60%. To investigate the influence of inflammation on target mRNA expression, haemopoietic cells were cultured with and without cytokine stimulation in vitro. CK19 messages could be easily detected in cultured marrow cells without further stimulation, CEA messages only after gamma-interferon (gamma-INF) stimulation. In contrast, G-CSF-mobilized peripheral blood stem cells were positive for CK19 messages only after stem cell factor (SCF) or interleukin stimulation. We conclude that transcription of so-called tissue-specific genes is inductible in haemopoietic tissues under certain conditions. These factors have to be considered in future applications of RT-PCR for the detection of minimal residual disease.

Topics: Abdominal Neoplasms; Bone Marrow; Carcinoembryonic Antigen; Cytokines; Granulocyte Colony-Stimulating Factor; Hematopoietic Stem Cell Mobilization; Hematopoietic Stem Cells; Humans; Inflammation; Keratins; Leukapheresis; Neoplastic Cells, Circulating; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Stimulation, Chemical; Stromal Cells

The aim of this study was to investigate prospectively the epithelialization process in the healing temporalis myofascial flap (TMF). Eight cats underwent maxillectomy and immediate reconstruction with TMF. They were killed at the determined time and the reconstructed maxillae were processed for examination by light microscopy and scanning electron microscopy. Results revealed that epithelialization of the healing TMF was initiated by hyperplastic changes followed by active migration of epithelial cells deriving from the wound margins. The partial maxillectomy wound was completely covered by a smooth oral mucosa at postoperative week 24. The mucosa had histological and ultrastructural features different from normal palatal mucosa.

Topics: Animals; Cats; Cell Movement; Collagen; Connective Tissue; Edema; Elastin; Epithelium; Fascia; Granulation Tissue; Hyperplasia; Inflammation; Keratins; Lymphocytes; Macrophages; Maxilla; Microscopy, Electron, Scanning; Mouth; Mouth Mucosa; Plasma Cells; Prospective Studies; Regeneration; Surgical Flaps; Temporal Muscle; Wound Healing

The purpose of this study was to observe the epithelialization process of the muscle-only flap used for reconstruction of the oral mucosal defects.. Forty-three male adult Japanese rabbits were used. A superiorly based cleidomastoid muscle flap was designed after vascular assessment. The flap was transferred into the oral cavity to cover a mucoperiosteal defect made in the mandibular alveolus. Epithelialization of the flap was histologically evaluated at designated intervals.. The flaps survived without ischemic necrosis. By 8 days postoperation, the flap was infiltrated by acute inflammatory cells and being replaced by granulation tissue originating from the adjacent tissues. The oral epithelial cells advanced onto this granulating muscle flap, with eventual coverage by 21 days. The granulation tissue matured to fibrous tissue with significant contraction by 2 months. At 6 months postoperation, abnormally hyperkeratinized epithelium was seen on the flap. This differed from the surrounding parakeratinized oral epithelium.. The muscle-only flap in the oral cavity epithelializes after the granulation process.

Topics: Actins; Animals; Cell Movement; Connective Tissue; Epithelial Cells; Epithelium; Follow-Up Studies; Graft Survival; Granulation Tissue; Inflammation; Ischemia; Keratins; Male; Mandible; Mandibular Diseases; Mouth; Mouth Mucosa; Muscle, Skeletal; Necrosis; Periosteum; Rabbits; Surgical Flaps; Wound Healing

The prognostic weight of histological and biological factors was compared with that of known clinical prognostic factors in a population of 108 consecutive previously untreated patients with head and neck squamous cell carcinoma. Parameters studied were: tumour vascularisation, mitotic index, histological differentiation, nuclear grade, keratinisation, desmoplasia, growth pattern, inflammation, tumour emboli in peripheral vessels, keratins 6, 13, 19 immunohistochemical expression, cytofluorometric ploidy and S-phase. In multivariate analysis (Cox), only age and nodal status had a significant impact on the overall survival, whereas T stage was the only significant factor associated with locoregional failure. The cumulative incidence of metastases was correlated not only with age, T and N stage, but also with histological differentiation. All the other histological and biological factors studied failed to provide further prognostic information. These findings may help to select patients with high metastatic risk.

Topics: Age Factors; Aged; Analysis of Variance; Biopsy; Carcinoma, Squamous Cell; Confidence Intervals; Disease-Free Survival; Female; Flow Cytometry; Follow-Up Studies; Head and Neck Neoplasms; Humans; Inflammation; Keratins; Male; Middle Aged; Mitotic Index; Multivariate Analysis; Ploidies; Predictive Value of Tests; Prognosis; Prospective Studies; Retrospective Studies; S Phase; Survival Rate; Time Factors; Treatment Outcome

Psoriasis is characterized by immune activation, increased proliferation and abnormal differentiation of keratinocytes. The reported anti-psoriatic mechanisms of action in vivo of vitamin D analogues include reduction of keratinocyte proliferation and induction of keratinocyte terminal differentiation. We investigated whether the anti-psoriatic effect of the natural active vatamin D analogue, calcitriol, applied topically, is due to direct effects on keratinocytes alone or also due to immunoregulatory effects of calcitriol. Psoriasis patients were treated with topical calcitriol (0.005%) and a vehicle control for 8 weeks. Disease activity was assessed by a severity index and quantitative histopathological markers. In vitro studies of lymphocyte proliferation and gamma interferon secretion and of keratinocyte proliferation complemented the clinicohistopathologic studies. A heterogeneous response to calcitriol treatment could be segregated based upon elimination of K-16 keratin expression. Calcitriol treatment decreased keratinocyte proliferation, normalized keratinocyte differentiation and decreased immune activation in plaques. The histologic response to vitamin D treatment of psoriasis includes suppression of both immune and keratinocyte activation in situ. These studies provide a basis for rational combination of anti-psoriatic treatments and for the design of new vitamin D analogues to treat psoriasis.

Topics: Administration, Topical; Calcitriol; Cell Differentiation; Chemotaxis, Leukocyte; Epithelium; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; T-Lymphocytes

The odontogenic keratocyst (OKC) is a locally aggressive neoplasm with rates of recurrence reported as high as 60%. Correlation between histopathology and the likelihood of recurrence remains a subject of controversy. In this review of the authors' experience in treating 50 patients with OKC between 1977 and 1993, 58 specimens were studied to correlate the likelihood of recurrence with the presence of the following histologic features--parakeratosis, orthokeratosis, satellite cysts, epithelial rests, or epithelial proliferation. Orthokeratinized cysts were associated with a higher recurrence rate than in previously reported studies. Disruption of the epithelial lining in the resected specimen was found to be a primary determinant of recurrence.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cell Division; Child; Epithelium; Female; Follow-Up Studies; Humans; Inflammation; Keratins; Keratosis; Male; Mandibular Diseases; Maxillary Diseases; Middle Aged; Odontogenic Cysts; Radiography; Recurrence

Many theories concerning the development of chronic pilonidal sinus have been proposed. A histologic study of primary pilonidal sinus in 53 patients is presented. Subcutaneous tissue contained sinuses surrounded by chronic inflammation. Hair in sinuses was found in three quarters of the specimens examined. Examination showed that hair entered via one of the sinus openings created. Pits (defined as darker spots of varying width in the midline of the internatal cleft) were found to be indentations of the skin containing keratin plugs and debris. They may be isolated or connected with hair follicles. Pilonidal sinuses are chronic inflammatory processes of the skin caused by keratin plugs and debris clinically observed as pits, having penetrated the dermis.

Topics: Adolescent; Adult; Chronic Disease; Female; Hair; Humans; Inflammation; Keratins; Male; Middle Aged; Pilonidal Sinus

Keratinocytes at sites of cutaneous inflammation have increased expression of intercellular adhesion molecule 1 (ICAM-1), a cytokine-inducible adhesion molecule which binds the leukocyte integrins LFA-1 and Mac-1. Transgenic mice were prepared in which the expression of mouse ICAM-1 was targeted to basal keratinocytes by using the human K14 keratin promoter. The level of constitutive expression attained in the transgenic mice exceeded the peak level of ICAM-1 expression induced on nontransgenic mouse keratinocytes in vitro by optimal combinations of interferon gamma and tumor necrosis factor alpha or in vivo by proinflammatory stimuli such as phorbol 12-myristate 13-acetate. In vitro adhesion assays demonstrated that cultured transgenic keratinocytes were superior to normal keratinocytes as a substrate for the LFA-1-dependent binding of mouse T cells, confirming that the transgene-encoded ICAM-1 was expressed in a functional form. However, the high level of constitutive ICAM-1 expression achieved on keratinocytes in vivo in these transgenic mice did not result in additional recruitment of CD45+ leukocytes into transgenic epidermis, nor did it elicit dermal inflammation. Keratinocyte ICAM-1 expression also did not potentiate contact-hypersensitivity reactions to epicutaneous application of haptens. The absence of a spontaneous phenotype in these transgenic mice was not the result of increased levels of soluble ICAM-1, since serum levels of soluble ICAM-1 were equal in transgenic mice and controls. We conclude that elevated ICAM-1 expression on keratinocytes cannot act independently to influence leukocyte trafficking and elicit cutaneous inflammation.

Topics: Animals; Base Sequence; DNA Primers; Endotoxins; Enzyme-Linked Immunosorbent Assay; Exons; Gene Expression; Growth Hormone; Humans; Inflammation; Intercellular Adhesion Molecule-1; Introns; Keratinocytes; Keratins; Mice; Mice, Transgenic; Molecular Sequence Data; Polymerase Chain Reaction; Promoter Regions, Genetic; Restriction Mapping; Skin

The purpose of this study was to test the hypothesis that odontogenic cysts can be induced by periapical infection. Pulp extirpation and reaming beyond the root apices were performed in 53 lower first molars in 27 Sprague-Dawley rats. The cavities were left open to allow continuous contamination by oral bacteria. Animals were killed at 6 and more than 8 months after operation. Odontogenic cysts were found in association with 8/53 teeth in 6 animals. Histologically, cysts were observed around the lower incisors below the first molars. The cyst wall consisted of fibrous connective tissue with inflammation and was lined with keratinized squamous epithelium. The cyst cavity contained a mass of keratin and necrotic debris. These results support the hypothesis that inflammatory stimulation from the apices can cause cystic changes in the enamel epithelium of underlying teeth.

Topics: Animals; Bacterial Infections; Connective Tissue; Dental Pulp Cavity; Dental Pulp Exposure; Disease Models, Animal; Epithelium; Female; Inflammation; Keratins; Male; Mandibular Diseases; Necrosis; Odontogenic Cysts; Periapical Diseases; Periapical Periodontitis; Pulpectomy; Rats; Rats, Sprague-Dawley; Root Canal Therapy; Time Factors

Discrimination of benign from malignant is fundamental to accurate histologic diagnosis. This article describes a series of nine patients with cutaneous squamous cell carcinoma who were noted to have in the vicinity of their neoplasm seemingly benign but biologically significant dermal granulomatous inflammation to cornified cells (GRC).. Two patients were reported as having this finding alone on incisional biopsy specimens only to have subsequent biopsy specimens demonstrate bona fide malignant neoplasms. On Mohs' frozen sections, four patients were noted to have GRC without concomitant tumor and four patients had GRC admixed with tumor. Two patients who demonstrated GRC but no neoplasm on final Mohs' sections had development of recurrent neoplasm after the initial procedure.. In the setting of suspected or proved cutaneous squamous cell carcinoma, GRC signifies the presence of viable neoplasm and warrants additional tissue resection.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cells; Female; Granuloma, Foreign-Body; Humans; Inflammation; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Skin Neoplasms

Interleukin 6 (IL-6) is a cytokine that mediates a wide range of inflammatory and immune responses. Its expression is elevated in inflammatory or immunodeficient diseases, including psoriasis, rheumatoid arthritis, and AIDS. To explore the role of IL-6 in skin, we utilized a human keratin 14 (K14) promoter to express IL-6 in the basal cells of stratified squamous epithelia of transgenic mice. Mice expressing the K14-IL-6 transgene were smaller than normal and exhibited retarded hair growth. Surprisingly, IL-6 expression did not lead to enhanced epidermal proliferation, but it did result in a thicker stratum corneum, with an otherwise seemingly normal program of differentiation. IL-6 expression did not lead to leukocytic infiltration, making it unlikely that it has direct proinflammatory activity in skin. Based on this study, one role of IL-6 relevant to host defense may be to enhance the stratum corneum, thereby providing increased protection from injurious stimuli or infection. If IL-6 plays additional roles in the skin, it is likely to act synergistically with factors that IL-6 alone cannot induce.

Topics: Animals; Base Sequence; Epithelium; Gene Expression; Inflammation; Interleukin-6; Keratinocytes; Keratins; Leukocytes; Mice; Mice, Transgenic; Molecular Sequence Data; Nucleic Acid Hybridization; Oligodeoxyribonucleotides; Phenotype; Polymerase Chain Reaction; RNA, Messenger; Skin; Skin Physiological Phenomena

Six cases are described of benign thymic cysts of the anterior mediastinum showing focal pseudoepitheliomatous hyperplasia of the lining epithelium. The patients' ages ranged from 11 to 54 years; five cysts occurred in males and one in a female. Histologically, the lesions were characterized by exuberant proliferation of the cyst lining epithelium that grew as sheets and tongues of atypical squamous cells with large, hyperchromatic nuclei, prominent nucleoli, and scattered mitotic figures. The walls of the cyst adjacent to the areas of epithelial proliferation showed abundant hemorrhage, necrosis, and severe inflammatory changes. All cases were treated by local surgical excision. There was no evidence of recurrence or metastases over a follow-up period of up to 8 years (average follow-up, 4 years). It is proposed that pseudoepitheliomatous hyperplasia may develop in thymic cysts as an expression of regeneration of the lining epithelium in response to the inflammatory, hemorrhagic, and necrotizing changes which often accompany these lesions. This should not be mistaken for malignancy, and should be distinguished from the exceptional cases of true thymic neoplasms seen in association with thymic cysts.

Topics: Adult; Child; Epithelium; Female; Follow-Up Studies; Humans; Hyperplasia; Immunoenzyme Techniques; Inflammation; Keratins; Male; Mediastinal Cyst; Middle Aged

A system of histological grading based on retrospective analysis of 239 patients with squamous cell carcinoma of the penis is presented. A new scoring system with 4 histological grades was used. The results of this study confirm previous reports that penile cancer is usually a highly (50%) or moderately (29%) differentiated squamous cell carcinoma. Poorly differentiated carcinomas and cancers of other types are very rare. The new grading system was found to be practical and a correlation between histological grade, clinical findings and prognosis was established. Patients with grade 1 tumours had an exceptionally favourable prognosis, with more than 80% being long-term survivors; for these patients, treatment with delayed side effects should be avoided and new forms of treatment should be explored.

Topics: Adult; Age Factors; Aged; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Humans; Inflammation; Keratins; Male; Methods; Middle Aged; Penile Neoplasms; Prognosis; Retrospective Studies

Cytoskeletal proteins (i.e., cytokeratins, vimentin, actin, and desmin) are normally present in the placental chorionic villi and are related to the maintenance of the villous shape, and to the ability of the villi to contract and permit a normal blood flow. In areas of villitis of unestablished etiology, the normal reactivity of these proteins in the villous core around fetal stem vessels disappears, and an increased number of cytokeratin positive cells is identified. The vascular damage in stem villi with villitis could develop ischemia in the villous tree promoting cytotrophoblast proliferation. Increased numbers of these areas could be related with the appearance of abnormal pregnancies.

Topics: Chorionic Villi; Cytoskeletal Proteins; Desmin; Female; Fluorescent Antibody Technique; Humans; Inflammation; Keratins; Pregnancy; Vimentin

Extravascular coagulation and fibrinolysis are intimately involved in and modulate cancer cell growth, invasion and metastasis. Samples from resection specimens of patients with primary lung cancer (adenocarcinomas) were tested with monoclonal (MAb) and polyclonal (PAb) antibodies against various factors of the coagulation or fibrinolysis systems, or against antigens of inflammatory or proliferating cells. MAb Ki-67 specific to nuclear antigens of proliferating cells showed a distinct but variable staining of cell nuclei throughout the tumor tissue. Nests of tumor tissue stained with cytokeratin-specific antibodies (PKK1), whereas other parts were negative. Fibrin(ogen) and fibronectin were found throughout the tumor tissue stroma and in the alveolar lining, and the most densely stained areas were at the transition zone between normal and tumor tissue. Fibrinolytic system components like tissue plasminogen activators (t-PA), and urokinase (u-PA), and their inhibitors PAI-1 and PAI-2 were all studied. All specimens were negative for t-PA (except endothelial linings), whereas urokinase-specific antibodies stained loosely packed tumor cells and macrophages within the tumor stromal tissue and alveolar septa. Both PAI-1 and PAI-2 were most prominently expressed within interstitial and alveolar macrophages. A weaker staining of tumor tissue cells was demonstrated. Inflammatory cells like macrophages and T lymphocytes were located in aggregates or diffusely spread within tumor stromal tissue. The inflammatory reaction was most intense at the border between normal lung and tumor tissue.

Topics: Adenocarcinoma; Antibodies; Antibodies, Monoclonal; Biomarkers; Blood Coagulation; Fibrin; Fibrinogen; Fibrinolysis; Fibronectins; Humans; Immunohistochemistry; Inflammation; Keratins; Ki-67 Antigen; Lung Neoplasms; Macrophages; Nuclear Proteins; Plasminogen Inactivators; T-Lymphocytes; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

Topics: Animals; Cell Division; Cell Line; Cells, Cultured; Epidermis; Humans; Inflammation; Interleukin-6; Interleukins; Keratins; Macrophages; Male; Skin

The studies presented in this report indicate that the mechanisms responsible for both ultraviolet radiation (UVR)- and lipopolysaccharide (LPS)-induced desensitization are different from one another and appear to be regulated at the site(s) of administration of the inflammatory agent. Furthermore, desensitization to either UVR or LPS is not due to the inability of interleukin 1 (IL 1)-sensitive target cells within these animals to respond to this endogenous mediator of inflammation. These conclusions were based on the demonstrated ability of UVR-desensitized mice to undergo an acute-phase response after exposure to a systemically administered inflammatory agent (LPS). In a reciprocal manner, LPS-desensitized mice were found to elicit a normal acute-phase response after a single UVR exposure. In addition, both UVR- and LPS-desensitized mice were found to respond normally to the systemic administration of an exogenous source of semi-purified IL 1. Desensitization to the inflammatory properties of either UVR or LPS appears to be controlled at the site of interaction between the tissues capable of producing epidermal-derived thymocyte-activating factor (ETAF)/IL 1 (epidermal keratinocytes or reticuloendothelial cells, respectively) and the exogenous inflammatory stimulus. Peritoneal macrophages obtained from LPS-desensitized mice were found to have a markedly reduced capacity to secrete ETAF/IL 1 in vitro when compared to peritoneal exudate cells (PEC) obtained from normal mice. In parallel with this decreased secretory potential by PEC was the appearance of membrane-associated forms of this mediator. Membrane-associated IL 1 was not found to be present on PEC obtained from normal mice. Keratinocytes obtained from the skin of normal mice or keratinocytes isolated from the irradiated skin site of UVR-desensitized mice were both found to secrete high levels of ETAF/IL 1 constitutively in vitro. Furthermore, both sources of keratinocytes also expressed membrane-associated forms of ETAF/IL 1 constitutively. Therefore, unlike LPS desensitization, the phenomenon of UVR desensitization does not appear to induce changes in the ability of keratinocytes to secrete soluble forms or to express membrane forms of ETAF/IL 1. UVR desensitization may be a result of the inability of ETAF/IL 1 generated within the skin to reach the various IL 1-responsive target cells throughout the body, or may result from the impaired ability of UVR to stimulate ETAF/IL 1 production due to c

Topics: Animals; Desensitization, Immunologic; Endotoxins; Epidermis; Immune Tolerance; Inflammation; Interleukin-1; Keratins; Lipopolysaccharides; Macrophage Activation; Macrophages; Mice; Mice, Inbred C3H; Serum Amyloid A Protein; Time Factors; Ultraviolet Rays

Topics: Animals; Cyanoacrylates; Gingiva; Inflammation; Keratins; Rats; Root Canal Therapy

A clinical and histopathologic analysis of 464 oral squamous cell papillomas is presented. Data on age, sex, race, location, clinical appearance, duration, recurrence, and clinical diagnosis are reviewed. One hundred seventy-six of the 464 specimens were examined for hyperkeratosis, character and amount of inflammatory infiltrate, and evidence of cellular atypia. The trends seen in this study support claims made by previous authors regarding incidence and inflammatory involvement. The data support a slightly higher occurrence rate in males than in females and in white as opposed to black patients. Papillomas were most abundant on the palatal complex, dorsum and lateral tongue borders, and lower lips, respectively. Confusion of papilloma for fibroma in the clinical diagnosis was less common than expected. Recurrence rate and incidence of multiple papillomas were low. Histologic study revealed a tendency for hyperkeratotic lesions to arise from nonkeratinized oral sites. Cellular atypia was found, but it is still unclear whether these changes are preneoplastic or due to an increased growth rate.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Female; Humans; Hyperplasia; Inflammation; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Recurrence, Local; Papilloma; Time Factors

In a study of the behavior of the keratin layer of the drumhead epidermis in some otologic conditions, a marker dye was applied to the tympanic membrane, and migration and/or desquamation of the dotted spots observed. Decelerated migration with desquamation in situ was observed during the quiescent phase of recurrent otitis externa, with or without myringitis. The same findings were observed in most cases of chronic secretory otitis media and its sequelas (retraction pockets, atrophic membranes, and scarring). The remnants of drumhead with keratotic and hyperplastic changes in the middle ear cleared themselves of the dye very slowly.

Topics: Adolescent; Adult; Aged; Child; Ear Diseases; Humans; Inflammation; Keratins; Middle Aged; Otitis Externa; Otitis Media; Skin; Tympanic Membrane

Cytochemical studies of glycogen of oral mucosa cells have been made on the smears by freeze-drying and PAS staining. The specimens were obtained from different areas of oral cavity of 77 human subjects and an attempt was made to find some interrelation amoung glycogen deposition, keratinization and inflammation. The largest glycogen deposition was found in the mucosa cells from mouth floor and cheek, a little in those from gingiva and quite a small or no glycogen in those from mucosa of hard palate and tongue. In gingiva the cells showing much more keratinization were less in glycogen contents, and vice versa. In inflammation some increase in glycogen contents were found in the gingivitis and the highest glycogen content in the cases of denture irritation of the palate as far as the present observation is concerned.

Topics: Adolescent; Adult; Aged; Cheek; Denture, Complete; Female; Gingiva; Gingivitis; Glycogen; Humans; Inflammation; Keratins; Male; Middle Aged; Mouth Diseases; Mouth Floor; Mouth Mucosa; Palate; Stomatitis, Aphthous; Tongue

Topics: Animals; Catechol Oxidase; Female; Guinea Pigs; Hair; Hydroquinones; Inflammation; Keratins; Langerhans Cells; Lymphocytes; Male; Melanins; Melanocytes; Microscopy, Electron; Organoids; Pigmentation; Skin; Tyrosine

Topics: Abdominal Muscles; Animals; Bladder Exstrophy; Cystitis; Disease Models, Animal; Epithelial Cells; Female; Inflammation; Keratins; Metaplasia; Methods; Rats; Urinary Bladder

The smoking habits of 345 Danish and 184 Hungarian leukoplakia patients were analysed against the histopathology of the leukoplakias, i.e. type of keratinization, epithelial thickness, epithelial dysplasia and inflammation. In spite of the reasonable size of the numbers forming the basis for the analysis, no statistically significant differences were found between smokers and non-smokers. However, it was found that the frequency of epithelial dysplasia is not higher among smokers than among non-smokers.

Topics: Atrophy; Denmark; Epithelium; Female; Humans; Hungary; Hyperplasia; Inflammation; Keratins; Keratosis; Leukoplakia, Oral; Male; Smoking

Topics: Abscess; Aluminum; Animals; Hydrogen-Ion Concentration; Hyperplasia; Inflammation; Keratins; Mice; Permeability; Phospholipids; Protein Binding; Protein Denaturation; Skin; Skin Diseases; Skin Ulcer; Sulfhydryl Compounds; Swine

Topics: Cell Membrane; Cell Nucleus; Cheek; Cytoplasmic Granules; Epithelium; Glycogen; Histocytochemistry; Humans; Inflammation; Keratins; Keratosis; Leukoplakia; Leukoplakia, Oral; Lichen Planus; Microscopy, Electron; Mouth Mucosa; Mouth Neoplasms; Smoking

Topics: Animals; Atrophy; Epithelium; Inflammation; Keratins; Male; Mitosis; Parotid Gland; Rats; Salivary Glands; Sublingual Gland; Submandibular Gland; Tongue; Vitamin A Deficiency

Topics: Adult; Aged; Female; Glycogen; Histocytochemistry; Humans; Inflammation; Keratins; Leukoplakia; Male; Middle Aged; Mouth Mucosa; Staining and Labeling; Vitamin A