sodium-dodecyl-sulfate and Necrosis

Resistance and resilience constitute the two complementary aspects of epithelial host defenses in Drosophila. Epithelial cell homeostasis is necessary for the recovery of damages caused by stress or infections. However, the genes responsible for gut epithelial homeostasis remain poorly understood. Here, we show that rgn(G4035) mutant flies have higher mortality than wild-type flies after ingestion of sodium dodecyl sulfate (SDS). Excessive melanization and increased necrotic cells in the gut contribute to the reduced survival of rgn(G4035) mutant flies following SDS ingestion. rgn mutant flies have a defect in the replenishment of intestinal stem cells (ISCs) following gut damage. The antimicrobial peptide (AMP) expression is affected in rgn(G4035) mutant fly guts. Together, our study provides evidence that rgn gene is essential for gut cell homeostasis following damage in Drosophila.

Topics: Animals; Antimicrobial Cationic Peptides; Apoptosis; Drosophila melanogaster; Drosophila Proteins; Epithelial Cells; Female; Gene Expression Regulation, Developmental; Intestinal Mucosa; Intestines; Male; Melanins; Mutation; Necrosis; Sodium Dodecyl Sulfate; Stem Cells

The effect of a single time exposure of SLS to the buccal mucosa of mice was compared to one application of the hapten OXA (oxazolone), evaluated by routine histology, immunohistochemistry and ELISA quantifications of cytokines. The SLS concentrations (2%, 4% and 8%) resulted in epithelial surface necrosis at 1-6 h, after 2-6 h accumulation of intra-epithelial neutrophils and at 24 h the main inflammatory cells were mononuclear. Increased concentrations of SLS gave more severe damage. CD4(+) T cells were found at 6 h and increased slightly up to 24 h and were most frequently seen at the lowest SLS dose. The CD8(+) T cells were kept at a low number during the whole 24 h observation period, but increased proportionally to the CD4(+) T cells. One application of 1% OXA did not raise the number of cells of either phenotype (2-24 h). Neither IL-2 nor IFN-γ demonstrated increased levels during the week of observation at any concentration of SLS, contrary to one application of OXA which caused increased IL-2 levels both at the local application site and in the regional and distant lymph nodes. Regardless of SLS concentration, a minor increase in regional lymph node weight was observed 8-12 h after substance application, quickly to subside whilst one OXA application gave a maximal weight increase at 48-72 h. We conclude that oral mucosa irritant SLS reactions gave early surface necrosis and neutrophil infiltrations and later mononuclear cell infiltrations dominated by CD4(+) T cells. The cytokines IL-2 and IFN-γ and lymphocyte proliferation in the regional lymph nodes was not observed after SLS application, contrary to hapten application.

Topics: Animals; Enzyme-Linked Immunosorbent Assay; Immunoenzyme Techniques; Inflammation; Interleukin-2; Mice; Mouth Mucosa; Necrosis; Oxazolone; Sodium Dodecyl Sulfate; T-Lymphocytes; Tumor Necrosis Factor-alpha

Contact allergens induce the augmentation of cell surface molecules on and release of cytokines from Langerhans cells (LC) in skin sensitization. THP-1 and U937 cell lines, surrogates of LC, were used as analytical tools of this phenomenon recently. In THP-1 cells, contact allergens are reported to induce the phenotypic alteration including the production of pro-inflammatory cytokines and augmentation of cell surface molecules especially at sub-toxic doses. However, the relationship between phenotypic alteration and cytotoxicity is not clear yet. The purpose of this study is to understand the relationship between the protein expression and cytotoxicity induced by contact allergens. First, we observed that the cytotoxicity induced by contact allergens is caused by both apoptosis and necrosis. Apoptosis was preferentially confirmed in stimulation with contact allergens, but non-allergen sodium lauryl sulfate (SLS) hardly induced apoptosis. Moreover, there was no effect to augmentation of protein expression when apoptosis induction pathways were inhibited. Based on these findings, we proposed that the protein expression and cytotoxicity were controlled independently. Next, oxidative stress was found to be generated by contact allergens at the early phase, and this regulated the protein expression and cytotoxicity at least partially. Finally, the humoral factors from dead cells induced by dinitrochlorobenzene (DNCB) were exposed to fresh THP-1 cells to confirm whether protein expression depended on cytotoxicity. The protein expression was not induced. Altogether, these results suggest that cytotoxicity induced by contact allergens may result in apoptosis and may also be stimulated in parallel with protein expression through an intracellular signal or signals.

Topics: Acetaldehyde; Allergens; Amino Acid Chloromethyl Ketones; Apoptosis; B7-2 Antigen; Caspase Inhibitors; Cell Line; Cell Survival; Cysteine Proteinase Inhibitors; Dinitrochlorobenzene; Eugenol; Humans; Intercellular Adhesion Molecule-1; Lactic Acid; Macrophages; Necrosis; Oxidative Stress; Propyl Gallate; Sodium Dodecyl Sulfate

The microbiota of root caries lesions of different grades of severity were studied. Fourteen lesions were examined. The experimental design of the study allowed correlation of histopathologically distinguishable stages with specific and distinct microbial populations. Dentin samples were ground in a sterile mortar and cultured anaerobically on nonselective Columbia blood agar plates supplemented with 5% hemolyzed human blood and on media selective for Lactobacillus spp. and streptococci. The cultivable microbiota were quantitatively speciated using Rapid ID 32A, Rapid ID 32 Strep, API 20 Strep, APIZYM, and API50 CH tests and SDS-PAG electrophoresis. In initial as well as in advanced lesions gram-positive bacteria accounted for approximately 90% of the CFUt. The proportion of Actinomyces, and in particular A. naeslundii was significantly higher (p < 0.05) in initial lesions than in advanced lesions. In contrast, the percentage of Streptococcus and especially S. mutans was higher (p < 0.05) in advanced than in initial lesions. Surprisingly low (0.8% of the CFUt) was the percentage of lactobacilli in advanced lesions. Gram-negative bacteria formed a minor part of the microbiota in both initial and advanced lesions. Among the gram-negative isolates, Prevotella, Selenomonas, and Bacteroides spp. were most noticeable. In advanced lesions, only the outermost layer of 0.5 mm thickness was populated by a high number of bacteria; the following segments harbored a negligible number of bacteria only. It is concluded that root caries is a continuous destruction process which is restricted to a subsurface zone of limited depth. The necrotic dentin is successively worn away, leading to a saucer-shaped cavitation which is repopulated by plaque. The creation of cavitations favors an aciduric flora. This might explain the succession of bacterial populations observed during the destruction process.

Topics: Actinomyces; Adult; Agar; Aged; Bacteria; Bacteroidaceae; Bacteroides; Blood; Colony Count, Microbial; Culture Media; Dental Plaque; Dentin; Disease Progression; Electrophoresis, Polyacrylamide Gel; Gram-Positive Bacteria; Humans; Lactobacillus; Middle Aged; Necrosis; Prevotella; Root Caries; Sodium Dodecyl Sulfate; Streptococcus; Streptococcus mutans

An ELISA assay with monoclonal antibody (Mab 2F4) raised against human ventricular myosin heavy chains was developed and used to investigate human sera after myocardial infarction. The monoclonal antibody 2F4 was selected for its high affinity to soluble fragments of myosin heavy chains (subfragment-1) and for its appropriate tissue specificity. By including Mab 2F4 in a simple and rapid dot immunobinding assay sera from patients with acute chest pain and of persons without a history of heart disease were tested. Myosin was detected only in the sera of the patients with myocardial necrosis, confirmed by electrocardiographic data. Negative reactions in all control cases were found. The serum myosin fragments reactive with Mab 2F4 were characterized by immunoblot experiments and protein bands in the region about 43 kDa were found. It was concluded that the myocardial infarction can be demonstrated by detection of cardiac myosin heavy chain fragments in the patients' sera.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Heart Ventricles; Humans; Hybridomas; Immunization; Immunoblotting; Male; Mice; Mice, Inbred BALB C; Middle Aged; Myocardial Infarction; Myocardium; Myosins; Necrosis; Peptide Fragments; Sodium Dodecyl Sulfate; Spleen

HgCl2, CuSO4, SnCl2, SnCl4 or sodium lauryl sulphate (SLS) were openly applied to rat oral mucosa for 1 min, followed 6 h later by histologic examination of the tissue response. Granulocytes were the predominant inflammatory cells and no lymphocytic infiltration could be seen with any of the substances tested. Irritant threshold levels were defined histologically for each of the substances. CuSO4 was found to be non-irritant at all concentrations. The addition of non-irritant concentrations of SLS lowered the threshold levels for HgCl2 and SnCl2, but CuSO4/SLS was non-irritant at all concentrations tested. Preapplication to the mucosa of SLS at non-irritant concentrations gave results with HgCl2, SnCl2 and CuSO4 similar to those with SLS added to the metal salt solutions. Lesions of allergic contact type could not be induced in the oral mucosa to any of the metal salt preparations.

Topics: Animals; Copper; Granulocytes; Mercury; Mouth Mucosa; Necrosis; Rats; Rats, Inbred Strains; Sodium Dodecyl Sulfate; Stomatitis; Tin

Topics: Animals; Copper; Endometrium; Female; Formaldehyde; Granulation Tissue; Irritants; Necrosis; Quinacrine; Rabbits; Regeneration; Sodium Dodecyl Sulfate; Sulfates; Talc; Tissue Adhesions; Uterine Diseases; Wound Healing