bromochloroacetic-acid and Diabetes-Mellitus--Type-2

Impaired wound healing is a major medical problem in diabetes. The objective of this study was to determine the possible application of an insoluble fraction of fur-derived keratin biomaterial as a wound dressing in a full thickness surgical skin wound model in mice ( n = 20) with iatrogenically induced diabetes. The obtained keratin dressing was examined in vitro and in vivo. In vitro study showed the keratin dressing is tissue biocompatible and non-toxic for murine fibroblasts. Antimicrobial examination revealed the keratin dressing inhibited the growth of S. aureus and E. coli. In vivo studies showed the obtained dressing significantly ( p < 0.05) accelerated healing during the first week after surgery compared to control wounds. Keratin dressings were incorporated naturally into granulation and regenerating tissue without any visible signs of inflammatory response, which was confirmed by clinical and histopathological analysis. It is one of the first studies to show application of insoluble keratin proteins and its properties as a wound dressing. The obtained keratin dressing accelerated wound healing in mice with iatrogenically induced diabetes. Therefore, it can be considered as a safe and efficient wound dressing. Although future studies are needed to explain the molecular mechanism behind fur-derived keratin effect during the multilayer wound healing process, our findings may open the way for a new class of insoluble fur keratin dressings in chronic difficult to heal wounds treatment.

Topics: Animals; Anti-Bacterial Agents; Bandages; Biocompatible Materials; Cell Survival; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Escherichia coli; Fibroblasts; Keratins; Male; Mice; Mice, Inbred C57BL; NIH 3T3 Cells; Skin; Staphylococcus aureus; Streptozocin; Wound Healing; Wounds and Injuries

Diabetes mellitus is chronic metabolic disorder, resulting from insulin deficiency, characterized by hyperglycemia altered metabolism of carbohydrates, proteins and lipids and an increased risk of vascular complications. There are different classes of anti-diabetic drugs in allopathic system of medicine. Metformin (dimethyl biguanide) is a blood glucose lowering agent used in the treatment of non-insulin dependent diabetes mellitus. Almost in all diseases the blood serves as the primary metabolic transport system in the body. Its composition is the preferred indicator with respect to the pathophysiological condition of the patient. Instead of analyzing blood to diagnose diabetes, hair could be used to detect diabetes using FTIR-ATR technique. The most important components of hair are fibrous proteins (keratins), melanins, glycogen, and lipids. Hair follicles are located 3-4mm below the surface of the skin and are surrounded by rich blood capillary system. In the present study, ten diabetic subjects were considered to evaluate the efficacy of metformin hydrochloride for the treatment of diabetes mellitus using FTIR-ATR spectroscopy. The spectra of diabetic hair fibre samples have been recorded in the mid infrared region of 4000-450cm

Topics: Biomarkers; Data Interpretation, Statistical; Diabetes Mellitus, Type 2; Female; Glucose; Glycogen; Hair; Humans; Hypoglycemic Agents; Keratins; Lipoproteins, HDL; Lipoproteins, LDL; Male; Metformin; Middle Aged; Spectroscopy, Fourier Transform Infrared; Treatment Outcome

Epithelial-mesenchymal transition (EMT) is a critical process for cancer metastasis and recurrence. Metformin, an effective oral antidiabetic drug, has been associated with decreased cancer risk and mortality. In this pilot study, we started to evaluate the effect of metformin on EMT in vivo and in vitro in endometrial cancer (EC).. Endometrial cancer cell lines and freshly isolated EC tumor specimens were used to assess EMT after metformin treatment. Cell lines were subjected to wound healing and AlamarBlue assays to determine cell migration and cell proliferation; messenger RNA levels were measured by real-time reverse transcriptase (RT) quantitative polymerase chain reaction (PCR), and protein levels were measured by Western blots to detect EMT marker expression.. Protein expression and messenger RNA of E-cadherin was found to be increased (P = 0.02 and 0.04, respectively) in 30 EC tumor specimens of diabetic patients treated with metformin compared with 20 EC tumor specimens of diabetic patients treated with other antidiabetic agents. In vitro, metformin reduced cell migration at 5 mM for 48 hours, as determined by the wound healing assay in EC cell lines (Ishikawa, 45% reduction; HEC50, 40% reduction), whereas more than 90% of the cells remained viable on the AlamarBlue assay. Metformin reduced EMT in the cell lines and regulated the expression of the EMT-related epithelial markers, E-cadherin and Pan-keratin; the mesenchymal markers, N-cadherin, fibronectin, and vimentin; and the EMT drivers, Twist-1, snail-1, and ZEB-1.. Tumors of patients on metformin have increased E-cadherin expression, and metformin decreases EMT in EC cell lines in vitro, suggesting clinical biological relevance of metformin in women with EC.

Topics: Adult; Aged; Aged, 80 and over; Cadherins; Carcinoma; Cell Line, Tumor; Cell Movement; Diabetes Mellitus, Type 2; Drug Screening Assays, Antitumor; Endometrial Neoplasms; Epithelial-Mesenchymal Transition; Female; Humans; Hypoglycemic Agents; Keratins; Metformin; Middle Aged; Pilot Projects; Retrospective Studies

Although assessment of glycated nail proteins may be a useful marker for monitoring of diabetes, their nature and formation are still poorly understood. Besides a detailed anatomical analysis of keratin glycation, the usefulness of glycated nail protein assessment for monitoring diabetic complications was investigated.. 216 patients (94 males, 122 females; mean age ± standard deviation: 75.0 ± 8.7 years) were enrolled. Glycation of nail and eye lens proteins was assessed using a photometric nitroblue tetrazolium-based assay. Following chromatographic separation of extracted nail proteins, binding and nonbinding fractions were analyzed using one-dimensional gel electrophoresis. Using a hand piece containing a latch-type-bur, a meticulous cutting of the nail plate into superficial and deep layers was performed, followed by a differential analysis of fructosamine.. Using SDS PAGE, four and two bands were identified among the nonglycated and glycated nail fraction respectively. Significantly lower fructosamine concentrations were found in the superficial nail layer (mean: 2.16 ± 1.37 μmol/g nails) in comparison with the deep layer (mean: 4.36 ± 2.55 μmol/g nails) (P<0.05). A significant higher amount of glycated eye lens proteins was found in diabetes mellitus patients (mean: 3.80 ± 1.57 μmol/g eye lens) in comparison with nondiabetics (mean: 3.35 ± 1.34 μmol/g eye lens) (P<0.05). A marked correlation was found between glycated nail and glycated eye lens proteins [y (glycated nail proteins) = 0.39 + 0.99 x (eye lens glycated proteins); r2 = 0.58, P<0.001]. The concentration of glycated eye lens proteins and the HbA1c level were found to be predictors of the concentration of glycated nail proteins.. Glycation of nail proteins takes place in the deep layer of finger nails, which is in close contact with blood vessels and interstitial fluid. Glycation of nail proteins can be regarded as a representative marker for diabetic glycation-associated target organ damage.

Topics: Aged; Aged, 80 and over; Biomarkers; Case-Control Studies; Crystallins; Diabetes Complications; Diabetes Mellitus, Type 2; Female; Glycation End Products, Advanced; Humans; Keratins; Lens, Crystalline; Male; Nails

A 58-year-old patient who smoked and had uncontrolled type 2 diabetes mellitus was referred to our clinic. The patient had a suspicious asymptomatic lesion that was diagnosed as B-cell non-Hodgkin lymphoma (NHL). Immunohistochemistry revealed intense and diffuse expression of CD20, CD10, BCL-6, and Ki-67. A positron emission tomography/computed tomography (PET/CT) scan showed focal pathological uptake of F-18-fluorodeoxyglucose only in the subcutaneous tissue anterior to the left maxillary sinus. After lesion excision and five courses of chemotherapy, PET/CT scans demonstrated complete resolution of the lesion. Smoking, uncontrolled diabetes mellitus, and periodontal disease might be predisposing factors for oral NHL.

Topics: Antineoplastic Combined Chemotherapy Protocols; Biopsy; Combined Modality Therapy; Diabetes Mellitus, Type 2; Fluorodeoxyglucose F18; Gingival Neoplasms; Humans; Immunohistochemistry; Keratins; Lymphoma, Non-Hodgkin; Male; Middle Aged; Neoplasm Staging; Oral Surgical Procedures; Positron Emission Tomography Computed Tomography; Radiography, Panoramic; Radiopharmaceuticals

Diabetes mellitus (DM) has become a global problem. By monitoring the health status of these individuals, diabetic complications can be prevented. We aimed to analyze alterations in the morphology and cytomorphometry of buccal epithelial cells of type 2 DM patients using oral exfoliative cytology technique and determine its importance in public health screening, diagnosis and monitoring of diabetes mellitus.. The study was carried out in 100 type 2 DM patients and 30 healthy individuals. Smears were taken from the right buccal mucosa and stained by the Papanicolaou technique. Staining with Acridine orange was carried out to view qualitative changes with confocal laser scanning microscope (LSM-510 Meta). The cytomorphometry was evaluated using IMAGE PRO PLUS 5.5 software with Evolution LC camera. All findings were statistically analyzed.. The results showed that with increase in fasting plasma glucose levels, there is significant increase in nuclear area, decrease in cytoplasmic area, and increase in nuclear cytoplasmic ratio (p < 0.05) when compared to the control group. Various qualitative changes were noted, such as cell degeneration, micronuclei, binucleation, intracytoplasmic inclusion, candida and keratinization.. In the present study, we found significant alterations in the cytomorphometry and cytomorphology of buccal epithelial cells of type 2 DM patients. This study supports and extends the view that these cellular changes can alert the clinician to the possibility of diabetes and aid in monitoring of diabetes throughout the lifetime of the patient.

Topics: Acridine Orange; Blood Glucose; Candida; Cell Death; Cell Nucleus; Cytodiagnosis; Cytoplasm; Diabetes Mellitus, Type 2; Epithelial Cells; Fasting; Fluorescent Dyes; Humans; Image Processing, Computer-Assisted; Inclusion Bodies; Keratins; Mass Screening; Micronucleus Tests; Microscopy; Microscopy, Confocal; Mouth Mucosa; Neutrophils; Papanicolaou Test

Hair analysis is an area of increasing interest in the fields of medical and forensic sciences. Human scalp hair has attractive features in clinical studies because hair can be sampled easily and noninvasively from human subjects, and unlike blood and urine samples, it contains a chronological record of medication use. Keratin protein is the major component of scalp hair shaft material and it is composed of 21 amino acids. The method used herein for the amino acid determination in hair included keratin protein acid hydrolysis using 6 M hydrochloric acid (HCl), followed by amino acids derivatization using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), and the determination of derivatized amino acids by gas chromatography/mass spectrometry (GC/MS). Amino acid profiles of scalp hair of 27 Jordanian subjects (15 diabetes mellitus (DM) type 2 patients and 12 control subjects) were analyzed. A fuzzy rule-building expert system (FuRES) classified the amino acid profiles into diabetic and control groups based on multivariate analyses of the abundance of 14 amino acids. The sensitivity and specificity were 100% for diabetes detection using leave-one-individual-out cross-validation. The areas under the receiver operative characteristics (ROC) curves were 1.0, which represents a highly sensitive and specific diabetes test. The nonessential amino acids Gly and Glu, and the essential amino acid Ile were more abundant in the scalp hair of diabetic patients compared to the hair of control subjects. The associations between the abundance of amino acids of human hair and health status may have clinical applications in providing diagnostic indicator or predicting other chronic or acute diseases.

Topics: Acetamides; Amino Acids; Asian People; Case-Control Studies; Diabetes Mellitus, Type 2; Gas Chromatography-Mass Spectrometry; Hair; Humans; Hydrochloric Acid; Jordan; Keratins; Pilot Projects; Principal Component Analysis; ROC Curve; Scalp; Trimethylsilyl Compounds

Sitagliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor-based incretin therapy intended for the treatment of type 2 diabetes mellitus (T2DM), has not been linked to adverse effects on the pancreas in prospective clinical trials or in nonclinical toxicology studies. To further assess potential pancreatic effects, sitagliptin was studied in the male Zucker diabetic fatty (ZDF) rat model of T2DM. Following 3 months of oral dosing with vehicle, or sitagliptin at doses 3- to 19-fold above the clinically therapeutic plasma concentration, which increased active plasma glucagon-like peptide-1 levels up to approximately 3-fold, or following 3 months of oral dosing with metformin, a non-incretin-based reference T2DM treatment, the pancreas of male ZDF rats was evaluated using qualitative and quantitative histopathology techniques. In the quantitative evaluation, proliferative index was calculated in exocrine pancreatic ducts and ductules using computer-based image analysis on sections stained by immunohistochemistry for cytokeratin (a cytoplasmic epithelial cell marker) and Ki-67 (a nuclear marker of recent cell division). Relative to controls, sitagliptin treatment did not alter disease progression based on detailed clinical signs and clinical pathology assessments. Sitagliptin treatment did not result in pancreatitis or any adverse effect on the pancreas based on a qualitative histopathology evaluation. Proliferative index did not increase with sitagliptin treatment based on quantitative assessment of more than 5000 sections of pancreas, where control group means ranged from 0.698-0.845% and sitagliptin-treated group means ranged from 0.679-0.701% (P = .874). Metformin treatment was similarly evaluated and found not to have adverse effects on pancreas.

Topics: Administration, Oral; Animals; Blood Glucose; Body Weight; Cell Proliferation; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Hypoglycemic Agents; Keratins; Ki-67 Antigen; Male; Metformin; Pancreas, Exocrine; Pyrazines; Rats; Rats, Zucker; Sitagliptin Phosphate; Triazoles

Topics: Alleles; Animals; Crohn Disease; Diabetes Mellitus, Type 2; Disease; DNA; Evolution, Molecular; Female; Genome, Human; Humans; Interleukin-18; Keratins; Liver Cirrhosis, Biliary; Male; Neanderthals; Optic Disk; Sequence Analysis, DNA; Smoking

In this study, we analyzed the effect of water and frequency on the dielectric properties of the nail plate in healthy individuals and in diabetic patients. The temperature dependencies of the loss tangent for both healthy and diabetic nail represent the relation between the electrical energy lost and stored in keratin-glucose-water complexes of this tissue. The differences between these materials concern the temperature ranges in which there appears the decomposition of loosely bound water. The effect of glycation on the loss tangent of the wet and the dry nail is supported by higher values of this parameter than in those describing the healthy samples in the whole temperature range and above 100°C, respectively. In addition, glycation lead to distinct increase in the dielectric relaxation decrement in permittivity and dielectric loss between the frequency of 500 Hz and 100 kHz for the wet and the dry nail. The results of this study indicate that the dielectric spectroscopy may be useful in observing the decomposition of water in the diabetic nail plate.

Topics: Diabetes Mellitus, Type 2; Electricity; Glucose; Humans; Keratins; Middle Aged; Nails; Water

Although chronic wounds are common, treatment for these disabling conditions remains limited and largely ineffective. In this study, we examined the benefit of bone marrow-derived mesenchymal stem cells (BM-MSCs) in wound healing. Using an excisional wound splinting model, we showed that injection around the wound and application to the wound bed of green fluorescence protein (GFP)(+) allogeneic BM-MSCs significantly enhanced wound healing in normal and diabetic mice compared with that of allogeneic neonatal dermal fibroblasts or vehicle control medium. Fluorescence-activated cell sorting analysis of cells derived from the wound for GFP-expressing BM-MSCs indicated engraftments of 27% at 7 days, 7.6% at 14 days, and 2.5% at 28 days of total BM-MSCs administered. BM-MSC-treated wounds exhibited significantly accelerated wound closure, with increased re-epithelialization, cellularity, and angiogenesis. Notably, BM-MSCs, but not CD34(+) bone marrow cells in the wound, expressed the keratinocyte-specific protein keratin and formed glandular structures, suggesting a direct contribution of BM-MSCs to cutaneous regeneration. Moreover, BM-MSC-conditioned medium promoted endothelial cell tube formation. Real-time polymerase chain reaction and Western blot analysis revealed high levels of vascular endothelial growth factor and angiopoietin-1 in BM-MSCs and significantly greater amounts of the proteins in BM-MSC-treated wounds. Thus, our data suggest that BM-MSCs promote wound healing through differentiation and release of proangiogenic factors. Disclosure of potential conflicts of interest is found at the end of this article.

Topics: Angiogenic Proteins; Animals; Cell Differentiation; Diabetes Mellitus, Type 2; Female; Fibroblasts; Genes, Reporter; Green Fluorescent Proteins; Keratins; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred Strains; Mice, Mutant Strains; Mice, Transgenic; Neovascularization, Physiologic; Random Allocation; Skin; Transplantation, Homologous; Wound Healing; Wounds and Injuries

The case of a 62-year-old woman with severe post-menopausal hirsutism is described. Her clinical history revealed regular menstrual periods until menopause at the age of 50, hysterectomy for fibromatosis at 58 years, non-insulin dependent diabetes mellitus, hypertension, obesity, severe hirsutism, which had developed in the previous 3 years, with a deeping of the voice. Examination showed android obesity, hypertension and severe hirsutism involving the face and the trunk. Endocrine evaluation pointed out regular adrenal function, serum total and free-testosterone in the adult male range, with normal androstenedione, DHEAS and 17OHP levels. Estradiol was slightly increased and LH and FSH were inappropriately low for her post-menopausal age. Computed tomography of the abdomen showed regular adrenal glands, and a radio-labeled cholesterol scan was negative. A further pelvic transvaginal ultrasonography revealed a small cystic formation near the right ovary and a slight increase in the size of the left ovary. The patient underwent bilateral ovariectomy. Histological examination showed a lipoid cell tumor within the left ovary. Immunohistochemical studies were positive for inhibin and cytokeratin. After surgery, serum testosterone fell to normal levels, gonadotropins increased to menopausal levels, confirming that the tumor was able to produce both LH, and FSH-inhibiting factors, and hirsutism greatly improved. Periodic hormonal tests remained normal and CT of the abdomen and pelvic ultrasonography did not show alterations at a 3 years follow-up.

Topics: Androgens; Biomarkers, Tumor; Diabetes Mellitus, Type 2; Estradiol; Female; Follicle Stimulating Hormone; Hirsutism; Humans; Hypertension; Inhibins; Keratins; Luteinizing Hormone; Middle Aged; Neoplasms, Gonadal Tissue; Obesity; Ovarian Neoplasms; Ovariectomy; Virilism

Atrial natriuretic peptide (ANP) has been recently described as an endogenous inhibitor of the synthesis and angiogenic action of vascular endothelial growth factor (VEGF). Given VEGF's key role in promoting neovascularization in proliferative diabetic retinopathy (PDR), this study was designed to evaluate the possibility that ANP could be involved in the neovascular and fibrotic complications of PDR.. We determined ANP by radioimmunoassay in plasma and vitreous humor samples collected from diabetic patients with and without PDR and from non-diabetic subjects. ANP was also immunohistochemically localized in the epiretinal membranes of patients with PDR.. Vitreous ANP concentrations were significantly higher in patients with active PDR compared to patients with quiescent PDR, diabetes without PDR or controls <0.05. Significant differences were also observed between vitreous ANP levels in diabetic patients without PDR and control subjects. There was no significant correlation between serum and vitreous ANP levels in any of the patient groups. ANP was detected in the fibrovascular epiretinal tissue of patients with PDR.. Diabetic patients with active neovascularization have significantly higher levels of ANP in the vitreous humor than those without active PDR. Diabetic patients without PDR were also found to have significantly higher vitreous ANP levels than non-diabetic patients. Since plasma and vitreous ANP concentrations were found to be unrelated, we suggest intraocular ANP synthesis and/or an increase in the release of ANP into the vitreous, as opposed to diffusion from the blood, as the main factors contributing to the high vitreous ANP levels observed in diabetic patients. In the fibrovascular epiretinal tissue of these patients, ANP was found to be localized in vascular, glial, fibroblast-like and retinal pigment epithelium cells. Our findings suggest a role for ANP in PDR.

Topics: Adult; Aged; Atrial Natriuretic Factor; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Epiretinal Membrane; Female; Glial Fibrillary Acidic Protein; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Radioimmunoassay; Retinal Neovascularization; Vitrectomy; Vitreous Body

Topics: Adult; Aged; Amyloid; Cell Differentiation; Diabetes Mellitus, Type 2; Humans; Immunohistochemistry; Insulin; Islet Amyloid Polypeptide; Islets of Langerhans; Keratins; Middle Aged; Pancreatic Ducts

It has been reported that cytokeratin 19 fragment (CYFRA 21-1) is superior to tissue polypeptide antigen (TPA) as a tumor marker, although there is a high correlation between CYFRA 21-1 and TPA levels in patients with lung cancer. We investigated correlations between these tumor markers in patients with non-malignant diseases. Marked correlations were found between CYFRA 21-1 and TPA levels in healthy subjects (n = 31), non-insulin-dependent diabetes mellitus (n = 160) and hemodialysis patients (n = 83) (range of r-value = 0.90-0.93, P < 0.0001). However in liver cirrhosis patients (n = 36), only a weak correlation was found (r = 0.39, P < 0.0001) and there were correlations between only TPA and both aspartate aminotransferase and alanine aminotransferase levels (r2 = 0.48 and 0.36, P < 0.0001). The elevated TPA levels in liver cirrhosis patients may be related to the decreased specificity of TPA as a tumor marker.

Topics: Alanine Transaminase; Antigens, Neoplasm; Aspartate Aminotransferases; Biomarkers, Tumor; Diabetes Mellitus, Type 2; Humans; Keratin-19; Keratins; Kidney Failure, Chronic; Liver Cirrhosis; Renal Dialysis; Tissue Polypeptide Antigen

We examined serum and urinary cytokeratin 19 fragment (CYFRA 21-1) levels in patients with diabetic nephropathy as a model of chronic renal failure, to investigate the mechanism of increased serum CYFRA 21-1 levels in chronic renal failure. Serum and urinary CYFRA 21-1 levels in non-insulin-dependent diabetes mellitus (NIDDM) patients with abnormal urinary immunoglobulin G (IgG) levels (>1.1 mg/g x Cr, n=126) were higher than those with normal urinary IgG levels. In NIDDM patients with normal urinary IgG levels (n=81); the urinary albumin or transferrin levels were not related to serum or urinary CYFRA 21-1 levels. We speculate that the increased serum CYFRA 21-1 levels contribute to metabolic abnormality in the kidney itself rather than the decreased urinary excretion per se, and that increased urinary CYFRA 21-1 levels are found in advanced cases of diabetic nephropathy with destruction of the size barrier.

Topics: Albuminuria; Antigens, Neoplasm; Biomarkers, Tumor; Creatinine; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Female; Humans; Immunoglobulin G; Immunoradiometric Assay; Keratin-19; Keratins; Kidney Failure, Chronic; Male; Middle Aged; Prognosis; Severity of Illness Index; Transferrin