Effect of melatonin supplemented at the light or dark period on recovery of sciatic nerve injury in rats

Peripheral nerve injuries can cause disabilities, social or economic problems. Melatonin, the secretory product of the pineal gland has antioxidant and anti-inflammatory actions. The aim of the present study was to investigate the effect of melatonin on the recovery of sciatic nerve after injury, comparing its effect when given in the light or the dark periods. Forty adult male Albino rats were allocated into four groups: control, nerve injury, nerve injury + melatonin given at light and nerve injury + melatonin given at dark. Nerve injury was initiated by clamping the sciatic nerve. Sciatic functional index (SFI) was measured preoperatively and postoperatively. Melatonin was given daily for six weeks. Recovery of the function was analyzed by functional analysis, electrophysiological analysis and biochemical measurement of Superoxide dismutase (SOD), Interleukin 1-beta (IL-1 β), Nerve growth factor (NGF), and bcl-2. Melatonin improved SFI, nerve conduction velocity (NCV) and the force of gastrocnemius muscle contraction as compared to the untreated rats. SOD activity, NGF, and bcl-2 were significantly increased, while IL-1β was significantly decreased after melatonin treatment as compared to the untreated injury group. SFI reached the control level; muscle contraction and IL-1B were significantly improved in the group treated with melatonin in the dark. Melatonin fastened the neural recovery and may be used in the treatment of nerve injury and it induced better nerve regeneration when the rats were treated during the dark period

Effect of undifferentiated versus hepatogenic partially differentiated mesenchymal stem cells on hepatic and cognitive functions in liver cirrhosis

Liver cirrhosis is the outcome of chronic liver injury. The current study aimed to investigate the therapeutic effect of undifferentiated mesenchymal stem cells versus in vitro partially differentiated mesenchymal stem cells on liver cirrhosis and hepatic encephalopathy. 50 adult male albino rats constituted the animal model and were divided into the following groups: control, thioacetamide, undifferentiated mesenchymal stem cells and hepatocyte growth factor-differentiated mesenchymal stem cells groups. Cognitive assessment was achieved by open field test and Y-maze task. We measured serum alanine aminotransferase, albumin and transforming growth factor-beta1, gene expression of α-smooth muscle actin, matrix metalloprotein-2, its tissue inhibitor and apoptotic markers: Bax and Bcl2, brain glial fibrillary acidic protein, synaptophysin, and dopaminergic receptors

Evaluation of Renal Gene Expression of Protein Kinase C (PKC) Isoforms in Diabetic and Nondiabetic Proliferative Glomerular Diseases

The protein kinase C (PKC) family consists of 13 members categorized as conventional or novel depending on whether diacylglycerol, calcium, or phosphatidylserine is required for activation. High glucose leads to activation of different forms of PKC across tissue types, thus determining the kind of diabetes-induced organ damage. PKC β was reported to have a positive role in B-lymphocyte activity through activation of NF-κB, leading to various immune disorders. We examined renal expression of two PKC isoforms α and β in renal biopsies of patients with diabetic nephropathy, lupus nephritis (LN) (Class 3-4), and mesangioproliferative glomerulonephritis (MPGN) to explore the role of each isoform in different glomerular diseases. PKC α and β gene expression was studied by quantitative real-time reverse transcription-PCR in 20 patients with type 2 diabetes and proteinuria (serum creatinine 2.04 ± 0.85 mg/dl, 24-h urinary protein 3.61 ± 1.75 g, eGFR 37.85 ± 17.89 ml/min/1.73 m2), 20 patients with proliferative LN (serum creatinine 1.67 ± 1.50 mg/dl, 24-h urinary protein 4.46 ± 5.01 g, eGFR 69.62 ± 40.93 ml/min/1.73 m2), and 20 patients with MPGN (serum creatinine 3.32 ± 2.79 mg/dl, 24-h urinary protein 4.65 ± 4.11 g, eGFR 32.62 ± 29.56 ml/min/1.73 m2). Normal tissues from the normal pole of four kidneys removed because of renal tumor served as controls. PKC α gene expression was significantly increased in diabetic kidneys compared to LN and MPGN (316.95 ± 152.94 µg/ml vs. 185.97 ± 32.13 and 195.46 ± 46.45 µg/ml, p < 0.05). PKC β gene expression was significantly increased in the LN and MPGN groups compared to the diabetic nephropathy group (41.01 ± 14.03 and 39.93 ± 16.41 µg/ml, respectively, vs. 18.20 ± 4.91 µg/ml, p < 0.05). Significant correlation was noted between the PKC α gene concentrations and proteinuria in diabetic patients. Renal expression of PKC α and β genes in control tissues were significantly lower compared to diabetic kidneys, LN, and MPGN groups (32.31 ± 0.36 and 4.67 ± 2.41 µg/ml, respectively, p < 0.001). The study revealed enhanced renal gene expression of both PKC isoforms α and β in diabetic kidney tissues, LN, and MPGN, but in different patterns. PKC α gene expression was significantly increased in diabetic patients with chronic kidney disease. The increased expression of the PKC β gene in LN and MPGN highlights its role in regulation of the immune system. This may represent potential therapeutic targets for prevention of progressive kidney injury in diabetic and proliferative glomerular diseases

Upregulation of Anti-Desmocollin 3 Antibodies in Pemphigus Diseases: A Case-control Study

Background: Pemphigus diseases are a subgroup of autoimmune bullous diseases characterized by autoantibodies against desmogleins and occasionally desmocollins. Desmocollin 3 is the main desmocollin isoform that contributes to cell adhesion in the epidermis. Objective: To evaluate the presence and level of anti-desmocollin 3 antibodies in pemphigus diseases, and to investigate whether their presence is associated with a specific type, presentation, or clinical pattern. Methods: Forty patients with pemphigus diseases and forty healthy controls were enrolled. Medical history, clinical examination, and pemphigus disease area index (PDAI) scoring were recorded for all patients. Serum samples were collected from both groups for assessment of anti-desmocollin 3 antibody reactivity by ELISA. Results: The presence of anti-desmocollin 3 antibodies was significant among patients with pemphigus compared with controls (P=0.003). The level of anti-desmocollin 3 antibodies was also significantly higher in patients with pemphigus compared with controls (P=0.01). There was no significant relationship between the presence of antidesmocollin 3 antibodies and any of the clinical presentations of pemphigus (type, severity, duration, activity, presence of annular pattern, or site of affection – mucosal, cutaneous, on the scalp, palmoplantar, or flexural). Conclusion: Anti-desmocollin 3 antibodies are upregulated in pemphigus diseases and can contribute to the pathogenesis of pemphigus. No specific clinical type, presentation, or pattern was found to be associated with the presence of anti-desmocollin 3 antibodie

Effect of metformin on Sirtuin-1 disorders associated with diabetes in male rats

Background: Type 2 diabetes mellitus (T2DM) is characterized by insulin resistance, hyperinsulinaemia and hyperglycaemia. Increased glucose production through abnormally elevated hepatic gluconeogenesis is central to the manifestation of hyperglycaemia in T2DM. Metformin corrects hyperglycaemia mainly through inhibition of gluconeogenesis. Sirtuin 1 (SIRT1) has been identified as regulator of gluconeogenic gene expression. The present study aimed to evaluate the effect of metformin on SIRT1 level and activity in liver and pancreas of diabetic rats. Further, the possible role of SIRT1 on metabolic disorders associated with diabetes mellitus, including serum levels of glucose, insulin, triglyceride (TG) and high density lipoprotiens (HDL), will be explored.Methods: Thirty-two male albino rats were divided into control group (GpI), diabetic (DM) group (GpII), (metformin + DM) group (GpIII) administered 120 mg/kg metformin daily for 1 month before induction of diabetes, (DM + metformin) group (GpIV) administered 250 mg/kg metformin daily for 1 month after induction of diabetes. At the end of the study, BMI%, serum levels of glucose, insulin, TG and HDL, HOMA, SIRT1 level and activity in liver and pancreas and pancreatic DNA ladder were assessed.Results: Our results showed significant decrease in serum glucose, insulin and TG levels and HOMA; significant increase in HDL level and SIRT1 level and activity in liver and pancreas beside the marked disappearance of pancreatic apoptosis in GpIII &amp; IV relative to GpII. Regarding BMI%, it showed no significant changes in GpIV relative to GpII. No significant change was recorded between GpIII and GpIV regarding all studied parameters except on serum TG.Conclusion: Lowered SIRT1 in diabetes was improved by the administration of Metformin. Consequently, the pathophysiological disorders associated with T2DM were improved.Keywords: DM, Metformin, SIRT1, Pancreatic apoptosi

Application of the Methylated Markers (Spectrin Beta and DEAD-Box Protein) for Definitive Differentiation Between Fresh and Aged Semen by evaluating Their Role in Identifying Semen From Mixed Body Fluids

Background: Semen identification is assumed a crucial proof of sexual assault. Moreover, body fluids at the crime scene of a human being, such as blood, semen, and saliva, are often mixed.Methods: Hence, in our study, we aimed to use methylation analysis targeting DNA epigenetic markers Spectrin beta chain (B_SPTB_03) and DEAD-box protein (DDX4) to differentiate between fresh semen (less than 4 hours) and aged semen (after 24 hours) as well as to differentiate between semen alone and semen mixed with other body fluids (blood and saliva) in the fresh and dried state.Results: Our findings showed statistically significant differences in the methylation patterns of the SPTB and DDX4 loci to distinguish semen from mixed body fluids in fresh and old samples. We were able to obtain two novel cutoff values to differentiate between fresh and aged semen, which are (52.25) with the SPTB marker and (70.75) with the DDX4 marker. Conclusion: It is concluded that the methylation approach based on the epigenetic markers of Spectrin beta chain and DEAD-box protein (B_SPTB_03 and DDX4) successfully identified fresh from aged semen and semen-derived alleles from mixed stains, hence it is recommended to be employed in forensic practice

The Effect Of Temperature On Age Estimation Of Semen Stains On Porous Versus Non-Porous Surfaces Using Messenger Ribonucleic Acid Measurement

Background: While mRNA can be used to identify the type of a body fluid, its degradation can also give some indication of the time interval since it was deposited. This study was conducted to evaluate the effect of temperature on the age estimation of human semen stains using mRNA deposited on porous versus non-porous surfaces at different time intervals. Methods: Ten semen samples were applied on two different media (glass and cotton) and exposed to three different temperatures (4°C, room temperature, 40°C) and examined at three time intervals (0, 45, and 90 days). The semen-specific mRNA markers PRM1 and PRM2 were quantitatively assessed along with a reference gene, beta-actin, using reverse transcription-quantitative polymerase chain reaction. Results: Mean Cq values of mRNA markers (PRM1 and PRM2) and the reference gene (beta-actin) increased with time of storage at different temperatures on both examined media. The mean quantification cycle (Cq) values of PRM2 were lower than PRM1, indicating that the levels of PRM2 marker in semen stain were higher than those of PRM1 marker. However, the mean Cq values of PRM2 at each time interval were not significantly different between temperatures, while PRM1 showed statistically significant differences in mean Cq values between temperatures at day 45 on both media. Conclusion: These results indicate that PRM2 can serve as a reliable mRNA marker to estimate the time since deposition of semen stain at different temperatures on two different media

Homing and reparative effect of intra-articular injection of autologus mesenchymal stem cells in osteoarthritic animal model

<p>Abstract</p> <p>Background</p> <p>This work aimed to study the homing evidence and the reparative effect of mesenchymal stem cells (MSCs) in the healing process of induced osteoarthritis in experimental animal model (donkeys).</p> <p>Methods</p> <p>Twenty-seven donkeys were equally divided into 3 groups based on the observation period after induction of arthritis (3, 6 and 9 weeks) to achieve different degrees of osteoarthritis. Each group was subdivided into three subgroups of three animals each based on the follow-up period (1, 2 and 6 months) after treatment. The induction was done through intra-articular (IA) injection of 2 ml of Amphotericin-B in both carpal joints. MSCs were harvested in a separate procedure, labeled with green fluorescent protein (GFP) using monster GFP vector and suspended in hyaluronic acid for IA injection. Treatment approaches consisted of cell-treatment using MSCs suspended in 3 ml of hyaluronic acid (HA) for the right carpal joint; and using the same amount of (HA) but without MSCs for the left contralateral carpal joint to serve as a control. Animals were assessed clinically and radiologically before and after treatment. Synovial fluid was also evaluated. Histopathologically; articular cartilage structural changes, reduction of articular cartilage matrix staining, osteophyte formation, and subchondral bone plate thickening were graded. Data was summarized using median and percentile for scores of histopathologic grading. Comparison between groups was done using non-parametric Mann Whitney test.</p> <p>Results</p> <p>The reparative effect of MSCs was significant both clinically and radiologically in all treated groups (P < 0.05) compared to the control groups. Fluorescence microscopy of sections of the cell-treated joints of all animals indicated that the GFP-transduced injected cells have participated effectively in the reparative process of the damaged articular surface and have integrated within the existing articular cartilage. The cells were associated with the surface of the cartilage and, were also detected in the interior.</p> <p>Conclusions</p> <p>Homing was confirmed by the incorporation of injected GFP-labeled MSCs within the repaired newly formed cartilage. Significant recovery proves that the use of IA injection of autologous MSCs is a viable and a practical option for treating different degrees of osteoarthritis.</p

CHOLESTATIC LIVER FIBROSIS IN A RAT MODEL OF BILE DUCT LIGATION: EVALUATING BIOCHEMICAL VERSUS HISTOPATHOLOGICAL CHANGES

Objective: Bile duct ligation (BDL), chronic liver injury model, was extensively used in studying mechanisms of fibrogenesis and antifibrotic agents. Considering the liver regenerative capacity and the diverse results from BDL, the present study aimed to evaluate the biochemical and histopathological changes over 10 weeks following BDL assessing if BDL-induced changes remain in a deterioration state or improve at a certain stage.Methods: Sham operation and BDL were conducted in Male Wistar rats. Serum AST, ALT, total bilirubin and albumin and hepatic hydroxyproline (HYP), reduced glutathione (GSH) and malondialdehyde (MDA) were measured in sham-operated (n=3) and BDL-rats (n=6) at 0, 1, 2, 4, 6, 8 and 10 weeks following operation. Liver tissue was also processed for histopathological analysis (H&amp;E and Sirus red staining).Results: Progressive liver injury (H&amp;E) and collagen deposition (Sirus red and HYP) in BDL-rats were observed starting from the first week post-operation and reached their maximum with early signs of cirrhosis on the 10th week of BDL. Severe and sustained cholestatic injury appeared in 2 weeks (increased ALT, AST, bilirubin along with decreased albumin (P&lt;0.001) compared to sham-operated rats). AST peaked on first week, however, bilirubin, ALT and MDA peaked on the 4th week (P&lt;0.001) then gradually decreased compared to their peaks.Conclusion: The relative improvement in liver function/cholestasis following their peaks in BDL model despite progression of fibrosis and hepatic injury require investigators using this model to consider not only biochemical, but also histopathological findings to guarantee an accurate interpretation of their results.Â