bromochloroacetic-acid and Diabetes-Mellitus

The growth of Diabetes Mellitus (DM) is a serious public health issue which is more prevalent in developing countries. The main problems related to DM are the gradual changes in the structural and functional integrity of tissues caused by hyperglycemia, which calls for early diagnosis and periodic monitoring exams. Recent studies suggest that the quality of the nail plate has great potential to assess the secondary complications of DM. Hence, this study aimed to determine the biochemical characteristics of the nails of individuals with DM2 by Raman confocal spectroscopy (CRS).. We collected fragments from the distal region of the fingernails of 30 healthy volunteers and 30 volunteers with DM2. The samples were analyzed by CRS (Xplora - Horiba) coupled to a 785 nm laser.. Alterations in different biochemical components, such as proteins, lipids, amino acids, and final agents of advanced glycation, and alterations in the disulfide bridges, which are important in stabilizing keratin in nails were identified.. The spectral signatures and new DM2 markers in nails were identified. Therefore, the possibility of acquiring biochemical information by evaluating the nails of diabetics, a simple and easily acquired material associated with the CRS technique, may allow health complications to be detected quickly.

Topics: Biomarkers; Diabetes Mellitus; Humans; Keratins; Nails; Spectrum Analysis, Raman

Topics: Anti-Bacterial Agents; Diabetes Mellitus; Glucans; Humans; Hydrogels; Keratins; Oxygen

Glycated keratin allows the monitoring of average tissue glucose exposure over previous weeks. In the present study, we wanted to explore if near-infrared (NIR) spectroscopy could be used as a non-invasive diagnostic tool for assessing glycation in diabetes mellitus.. A total of 52 patients with diabetes mellitus and 107 healthy subjects were enrolled in this study. A limited number (n=21) of nails of healthy subjects were glycated in vitro with 0.278 mol/L, 0.556 mol/L and 0.833 mol/L glucose solution to study the effect of glucose on the nail spectrum. Consequently, the nail clippings of the patients were analyzed using a Thermo Fisher Antaris II Near-IR Analyzer Spectrometer and near infrared (NIR) chemical imaging. Spectral classification (patients with diabetes mellitus vs. healthy subjects) was performed using partial least square discriminant analysis (PLS-DA).. In vitro glycation resulted in peak sharpening between 4300 and 4400 cm-1 and spectral variations at 5270 cm-1 and between 6600 and 7500 cm-1. Similar regions encountered spectral deviations during analysis of the patients' nails. Optimization of the spectral collection parameters was necessary in order to distinguish a large dataset. Spectra had to be collected at 16 cm-1, 128 scans, region 4000-7500 cm-1. Using standard normal variate, Savitsky-Golay smoothing (7 points) and first derivative preprocessing allowed for the prediction of the test set with 100% correct assignments utilizing a PLS-DA model.. Analysis of protein glycation in human fingernail clippings with NIR spectroscopy could be an alternative affordable technique for the diagnosis of diabetes mellitus.

Topics: Adult; Diabetes Mellitus; Discriminant Analysis; Female; Glycation End Products, Advanced; Glycosylation; Humans; Keratins; Least-Squares Analysis; Male; Middle Aged; Nails; Spectroscopy, Near-Infrared

Limbal epithelial stem cells (LESC) maintenance requires communication between stem cells and neighboring stromal keratocytes. Extracellular vesicles (EVs) are important for intercellular communication in various stem cell niches. We explored the regulatory roles of limbal stromal cell (LSC)-derived exosomes (Exos), an EV sub-population, in limbal epithelial cells (LEC) in normal and diabetic limbal niche and determined differences in Exo cargos from normal and diabetic LSC. Wound healing and proliferation rates in primary normal LEC were significantly enhanced upon treatment by normal Exos (N-Exos), but not by diabetic Exos (DM-Exos). Western analysis showed increased Akt phosphorylation in wounded LECs and organ-cultured corneas treated with N-Exos, compared to untreated wounded cells and DM-Exos treated fellow corneas, respectively. N-Exos treated organ-cultured corneas showed upregulation of putative LESC markers, keratin 15 (K15) and Frizzled-7, compared to the DM-Exos treated fellow corneas. By next generation sequencing, we identified differentially expressed small RNAs including microRNAs in DM-Exos vs. N-Exos. Overall, N-Exos have greater effect on LEC proliferation and wound healing than DM-Exos, likely by activating Akt signaling. The small RNA differences in Exos from diabetic vs. normal LSC could contribute to the disease state. Our study suggests that exosomes may serve as novel therapeutic tools for diabetic cornea.

Topics: Adolescent; Adult; Adult Stem Cells; Aged; Aged, 80 and over; Cell Movement; Cell Proliferation; Cells, Cultured; Corneal Keratocytes; Diabetes Mellitus; Epithelial Cells; Epithelium, Corneal; Exosomes; Female; Frizzled Receptors; Humans; Keratins; Male; Middle Aged; Proto-Oncogene Proteins c-akt; Signal Transduction

The particular combination of polydeoxyribonucleotides, l-carnitine, calcium ions, proteolytic enzyme and other ingredients acts in a synergetic way in the regeneration of skin and connective tissues. This new formulation of active principles was tested in vitro as a cell and tissue culture medium and in vivo for various preparations in support of tissue regeneration. In vitro, the new blend allowed the maintenance of skin biopsies for more than 1 year in eutrophic conditions. Immunocytochemical analyses of fibroblasts isolated from these biopsies confirmed a significant increase of the epidermal and connective wound-healing markers such as collagen type I, collagen type IV, cytokeratin 1 (CK1), CK5, CK10 and CK14 versus controls. To examine the effects of the new compound in vivo, we studied impaired wound healing in genetically diabetic db/db mice. At day 18, diabetic mice treated with the new composition showed 100% closure of wounds and faster healing than mice treated with the other solutions. This complex of vital continuity factors or life-keeping factors could be used as a tissue-preserving solution or a cosmetic/drug/medical device to accelerate wound healing in the treatment of patients with deficient wound repair to promote the regeneration of cutaneous and connective tissues (injuries-wound, dermatitis) and prevent the recurrent relapses.

Topics: Administration, Topical; Animals; Biopsy; Body Weight; Cell Survival; Cells, Cultured; Chemistry, Pharmaceutical; Collagen Type I; Collagen Type II; Connective Tissue; Culture Media; Diabetes Mellitus; Fibroblasts; Humans; Immunohistochemistry; Keratins; Male; Mice; Polydeoxyribonucleotides; Skin; Staining and Labeling; Wound Healing

To explore the biological significance of the lymphatics in the autoimmune process, the thymus from non-obese diabetic (NOD) mice was evaluated by histochemistry and western blot analysis. Thymic lymphatic endothelial cells showed suggestive expression patterns of the functional molecules lymphatic vascular endothelial hyaluronan receptor (LYVE)-1, CCL21, CD31 and podoplanin. With increasing age, the expression of CCL21 was reduced in the medullary epithelial cells and lymphatics. Of note, LYVE-1-expressing lymphatics, filled with a cluster of thymocytes, increased in number and size and extended from the corticomedullary boundary into the medulla as the insulitis progressed. The development of lymphatic compartments was occasionally accompanied by a regional disappearance between the cortex and medulla. The CD4- and CD8-positive T cells frequently penetrated through the slender lymphatic walls. The epithelial reticular cell layer lining the perivascular spaces was extensively stained with cytokeratin, but the expression of cytokeratin showed an age-dependent decrease. These findings indicate that the occurrence of LYVE-1-expressing lymphatic compartments and the alteration of CCL21 expression in the lymphatics may be involved in defective thymocyte differentiation and migration, and play a significant role in insulitic and diabetic processes.

Topics: Aging; Animals; Autoimmune Diseases; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Chemokine CCL21; Chemokines, CC; Diabetes Mellitus; Disease Models, Animal; Epithelial Cells; Glycoproteins; Immunohistochemistry; Islets of Langerhans; Keratins; Lymphatic Vessels; Lymphocyte Activation; Membrane Glycoproteins; Membrane Transport Proteins; Mice; Mice, Inbred NOD; Platelet Endothelial Cell Adhesion Molecule-1; T-Lymphocytes; Thymus Gland

Histopathological changes of biopsied pancreata were examined in three diabetic patients with autoimmune chronic pancreatitis (ACP). We found intra- and periinsular mononuclear cell (MNC)-mainly CD8+ T cell-infiltration as well as MNC infiltration around the ductal cells. Islet beta cell volume decreased significantly. Some ductal cells expressed insulin and insulin promoter factor-1 concomitantly in the affected pancreas. These findings provide new insight toward understanding the pathological mechanisms of corticosteroid treatment-responsive diabetes associated with ACP.

Topics: Adrenal Cortex Hormones; Aged; Autoimmune Diseases; Biopsy; CD8-Positive T-Lymphocytes; Diabetes Complications; Diabetes Mellitus; HLA Antigens; Humans; Immunohistochemistry; Insulin; Islets of Langerhans; Keratins; Male; Middle Aged; Pancreatitis

In this study, 55 diabetic patients and 45 non-diabetic control subjects were examined to determine oral candidal carriage state. The influence of some local aetiologic and systemic factors such as: salivary flow rate and pH, heredity, alcohol drinking, smoking habits, antimicrobial therapy, wearing of denture, burning sensation, dry mouth, taste alteration and tooth brushing habit on candidal carriage rate were investigated. Imprint culture, cytological smears and biochemical tests were used. Oral carrier rate and density of Candida species were non-significantly higher in the diabetic patients than in the non-diabetic control subjects. This increase was confirmed cytologically too. In both groups, Candida albicans was found to be a predominant species on tongue dorsum. Cigarette and alcohol habits of men were higher while tooth brushing habit was less than in women in diabetic and control groups. Salivary flow rate and pH values of diabetic patients were significantly lower while serum glucose values were significantly higher than of non-diabetic controls. The rate of diabetic patients suffering from dry mouth and having diabetic heredity in the family were significantly higher than control subjects. The candidal colonization was higher and keratinization was lower while diabetic treatment tended from diet and oral antidiabetic towards insulin. The decrease in salivary pH, the increase in serum glucose and wearing denture were correlated with the increased rate and density of C. albicans in both groups. Keratinization was also accompanied with the increase in leucocytes. In diabetic group, positive correlations were found between antimicrobial therapy and C. glabrata carriage; the increase in leucocytes and C. albicans carriage; the increase in keratinization and alcohol habit; serum glucose and smoking habit; dry mouth complaint and antimicrobial therapy. There was a negative correlation between salivary flow rate and C. albicans carriage. In control group a positive correlation was found between antimicrobial therapy and keratinization.

Topics: Adolescent; Adult; Aged; Alcohol Drinking; Anti-Infective Agents; Blood Glucose; Candida; Candida albicans; Candidiasis, Oral; Chi-Square Distribution; Colony Count, Microbial; Dentures; Diabetes Mellitus; Female; Humans; Hydrogen-Ion Concentration; Keratins; Leukocytes; Male; Middle Aged; Mouth; Saliva; Secretory Rate; Sex Factors; Smoking; Statistics as Topic; Toothbrushing; Xerostomia

We propose a new indicator for diabetic control that shows the extent of glycation of hair protein (keratin), the glycation index (A(390)/A(412)) which is based on the ratio of glycated protein- to cystine-induced coloration, where A(390) and A(412) represent each absorbance in the color reactions of glycated protein and cystine in the hair protein. Samples can be quickly and non-invasively collected and easily stored. This index for the back and scalp hairs from hypercholesterolemic mice with hyperglycemia, diabetic rats and diabetic patients gave significantly higher values (2.0-6.0-fold) than those of normal subjects (p<0.01). The glycation indices (mean + or - S.D.) of hairs from diabetic and non-diabetic subjects were 3.00 + or - 0.96 (n = 21) and 1.51 + or - 0.45 (n = 30), respectively. These indices (y) correlated well with the levels of glycohemoglobin (HbA(1c), chi) in diabetic and non-diabetic subjects: y = 0.69 chi- 2.03 (r = 0.82, n = 31, p<0.01). Within-run precision (reproducibility, CV) for the assay of the glycation indices of hairs from the three groups was 6.7-9.4% (n = 10 each). The proposed glycation index of hair gave reasonable results for animals and humans with normo- and hyperglycemia, suggesting that it is reliable and can be diagnostically useful.

Topics: Animals; Blood Glucose; Cystine; Diabetes Mellitus; Glycated Hemoglobin; Glycation End Products, Advanced; Hair; Humans; Hyperglycemia; Keratins; Mice; Rats

In this study, we compared the clinical and endocrinological characteristics, neuroimaging findings, surgical outcome, and conventional histological findings (including immunohistochemistry) with the electron microscopic appearance of 31 growth hormone (GH)-producing adenomas. By electron microscopy, these 31 tumors were divided into 23 densely granulated somatotroph adenomas (DG adenomas) and 8 sparsely granulated somatotroph adenomas (SG adenomas). SG adenomas more frequently affected younger women, but no significant correlation was found between the adenoma type and the characteristic signs and symptoms of acromegaly, the incidence of diabetes mellitus or hypertension, or the basal serum GH and insulin-like growth factor I levels. A distinct response of GH to thyrotropin-releasing hormone, bromocriptine, or GH-releasing hormone was significantly more common in patients with DG adenomas than in those with SG adenomas, whereas the incidence of a response to gonadotropin-releasing hormone or oral glucose was not significantly different between the two groups. An analysis of neuroimaging findings and surgical results indicated that SG adenomas were more likely to be macroadenomas with suprasellar extension or invasive tumors and had a lower surgical cure rate. However, postoperative radiotherapy seemed to be similarly effective in both types of adenoma to prevent a tumor recurrence and to reduce postoperative GH basal level in serum. Light microscopy showed that DG adenomas were mainly acidophilic and were immunopositive not only for GH but also for prolactin (43%), the beta subunit of thyroid-stimulating hormone (26%), and the alpha subunit of glycoprotein hormone (87%), whereas SG adenomas were almost all chromophobic and only revealed immunopositivity for GH.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenoma; Adenoma, Chromophobe; Adult; Biomarkers, Tumor; Bromocriptine; Cytoplasmic Granules; Diabetes Mellitus; Female; Gonadotropin-Releasing Hormone; Growth Hormone; Humans; Hypertension; Keratins; Middle Aged; Neoplasm Proteins; Pituitary Neoplasms; Prolactin; Thyrotropin-Releasing Hormone; Treatment Outcome

Non-enzymatic glycosylation of keratin from the stratum corneum of the sole of the foot was measured by the thiobarbituric acid technique in thirty diabetic and thirty control subjects. A significant increase in the level of glycosylation was demonstrated in the diabetic subjects (P less than 0.001). HbA1 levels were measured in a further eighteen subjects at the same time as keratin was removed, and in this group a significant association between non-enzymatic glycosylation of that protein and diabetic control was demonstrated (P less than 0.01). In vitro incubation of keratin in the presence of free glucose produced increased non-enzymatic glycosylation (P less than 0.01) and this effect was blocked by incubation in the presence of increasing concentrations of aspirin (P less than 0.01). Measurement of non-enzymatic glycosylation of keratin in a further group of twenty diabetics with neuropathic ulceration showed a significant increase in levels when compared with a group of diabetics without ulcers (P less than 0.05). As keratin is the principle structural protein of the stratum corneum of the sole of the foot, it is possible that changes in this protein associated with non-enzymatic glycosylation may contribute to abnormalities seen in the skin of the feet of diabetics.

Topics: Adult; Aged; Aspirin; Diabetes Mellitus; Epidermis; Foot; Glucose; Glycated Hemoglobin; Humans; In Vitro Techniques; Keratins; Middle Aged; Skin Ulcer

Colorimetric techniques such as the thiobarbituric acid assay are widely used for the determination of nonenzymatic glycosylation of proteins. One of the major problems associated with this technique is the high nonspecific background absorbance which, due to its variability, results in loss of sensitivity. This report describes a method of removing the nonspecific absorbance by extracting the final chromogen into cyclohexanone. Using this method for the determination of nonenzymatic glycosylation of hair and epidermal keratin, at least 90% of the background absorbance is removed, increasing the sensitivity of the technique. This allows more effective discrimination of the level of glycolsylation of proteins from control and diabetic patients, by reducing the degree of overlap. The requirement for individual borohydride reduced samples is also avoided, thus simplifying the technique. Cyclohexanone extraction provides a simple addition to the standard thiobarbituric acid technique with significant improvement in results.

Topics: Aged; Cyclohexanes; Cyclohexanones; Diabetes Mellitus; Epidermis; Hair; Humans; Keratins; Methods; Middle Aged; Proteins; Thiobarbiturates