Methylation Status of cAMP-responsive Element Modulator (CREM) Gene in Infertile Men and Its Association with Sperm Parameters.

The methylation pattern of non-imprinting genes was little studied, although it is widely known that the abnormal methylation levels of imprinting genes are associated with different forms of male infertility. The purpose of this research was to assess the CREM gene's methylation status and seminal characteristics in infertile individuals who were potential intracytoplasmic sperm injection (ICSI) candidates. A total of 45 semen samples (15 normospermia, 15 asthenospermia, and 15 oligoasthenoteratospermia) were examined. Using aniline blue (AB) staining, we carried out conventional semen analysis, chromatin quality, and sperm maturity testing. DNA was taken from semen samples, and all isolated DNA was assessed using Nanodrop and gel electrophoresis. A quantitative methylation-specific polymerase chain reaction (Q-MSP) approach was used to quantify the methylation at the DMRs of the CREM gene. According to our findings, sperm count (P=0.012), concentration (P= 0.019), motility (P=0.006), progression (P=0.006), and normal morphology (P=0.004) were all inversely correlated with abnormal sperm chromatin condensation. Additionally, we noted that the methylation level of the CREM gene was considerably more significant in the oligoasthenoteratospermia group compared to the asthenospermia and normospermia groups (P<0.05). Additionally, sperm count (P=0.043), progression (P=0.026), and normal morphology (P=0.024) were all inversely linked with CREM methylation. Overall, the abnormal CREM methylation patterns have a negative impact on sperm parameters. Additionally, the CREM gene's DNA methylation status may serve as an epigenetic indicator of male infertility.

Can Rosuvastatin Reduce the Risk of Thrombosis in Patients with Hypercholesterolemia with its Effect on Coagulation Factors and Homocysteine Levels?

BACKGROUND AND OBJECTIVE: Hypercholesterolemia is one of the main risk factors for vascular thrombosis in individuals. Therefore, the use of statins is very effective in reducing cholesterol and can reduce the risk of thrombosis in these patients. Rosuvastatin, a member of the statin family which, inhibits cholesterol synthesis. Very few studies have been done in relation to how rosuvastatin can affect thrombosis. So, this research has been tried whether rosuvastatin can have an effect on coagulation factors and homocysteine as risk factors for thrombosis in hypercholesterolemia? METHODS: In this experimental study, 60 patients (30 men and 30 women with a mean age of 40- 70 years) diagnosed with hypercholesterolemia (cholesterol >250 mg/dl) participated in this research. 30 patients were prescribed rosuvastatin (20 mg/day), and 30 patients were simultaneously taken placebo for three months. All parameters, including FVIII, FV, Fibrinogen, D-Dimer, plasma homocysteine level and lipid profile, were measured before and after treatment. All the results were statistically compared between the two groups. RESULTS: In patients who took rosuvastatin, the drug was able to significantly reduce the concentrations of total cholesterol, triglycerides, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) (P <0.001). Also, rosuvastatin was able to reduce the concentrations of homocysteine significantly, D-Dimer (P <0.001), coagulation factor VIII and factor V (P <0.05). In patients with hypercholesterolemia who took the placebo, did not affect the mentioned variables (P >0.05). CONCLUSION: According to the results, it seems that rosuvastatin may be able to reduce the risk of thrombosis in patients by affecting coagulation factors and homocysteine levels.

Impact of DNA methylation of the human mesoderm-specific transcript (MEST) on male infertility.

Male infertility accounts for nearly 40%-50% of all infertile cases. One of the most prevalent disorders detected in infertile men is errors in the MEST differentially methylated region (DMR), which has been correlated with poor sperm indexes. The aim of our study was to characterize the methylation pattern of the MEST gene, along with assessing seminal factors and chromatin condensation in sperm samples from both infertile patients and fertile cases, all of whom were candidates for intracytoplasmic sperm injection. We collected forty-five semen specimens from men undergoing routine spermiogram analysis at the Infertility Treatment Center. The specimens consisted of 15 samples of normospermia as the control group, 15 individuals of asthenospermia, and 15 individuals of oligoasthenoteratospermia as the cases group. Standard semen analysis and the chromatin quality and sperm maturity tests using aniline blue staining were performed. The DNA from spermatozoa was extracted and treated with a sodium bisulfite-based procedure. The methylation measure was done quantitatively at the DMRs of the MEST gene by quantitative methylation-specific polymerase chain reaction (qMSP). The mean percentages of total motility, progression, and morphology in normospermia were significantly higher than oligoasthenoteratospermia and asthenospermia, and they were substantially higher in asthenospermia compared to oligoasthenoteratospermia (P ≤ 0.05). The mean percentages of histone transition abnormality and MEST methylation in oligoasthenoteratospermia were significantly higher than asthenospermia and normospermia (P ≤ 0.05). A negative correlation existed between the histone transition abnormality and MEST methylation with sperm parameters. In conclusion, chromatin integrity, sperm maturity, and MEST methylation may be considered important predictors for addressing male factor infertility. Therefore, we suggest that male infertility may be linked to methylation of the imprinted genes.

Differentiation of Human Adipose-derived Stem Cells to Exosome-affected Neural-like Cells Extracted from Human Cerebrospinal Fluid Using Bioprinting Process.

BACKGROUND: Advancement in tissue engineering has provided novel solutions for creating scaffolds as well as applying induction factors in the differentiation of stem cells. The present research aimed to investigate the differentiation of human adipose-derived mesenchymal stem cells to neural-like cells using the novel bioprinting method, as well as the effect of cerebrospinal fluid exosomes. METHODS: In the present study, the extent of neuronal proliferation and differentiation of adipose- derived stem cells were explored using the MTT method, immunocytochemistry, and real-- time PCR in the scaffolds created by the bioprinting process. Furthermore, in order to investigate the veracity of the identity of the CSF (Cerebrospinal fluid) derived exosomes, after the isolation of exosomes, dynamic light scattering (DLS), scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques were used. RESULTS: MTT findings indicated survivability and proliferation of cells in the scaffolds created by the bioprinting process during a 14-day period. The results obtained from real-time PCR showed that the level of MAP2 gene (Microtubule Associated Protein 2) expression increased on days 7 and 14, while the expression of the Nestin gene (intermediate filament protein) significantly decreased compared to the control. The investigation to confirm the identity of exosomes indicated that the CSF-derived exosomes had a spherical shape with a 40-100 nm size. CONCLUSION: CSF-derived exosomes can contribute to the neuronal differentiation of adipose- derived stem cells in alginate hydrogel scaffolds created by the bioprinting process.

Artemisin and human endometrial-derived stem cells improve cognitive function and synaptic plasticity in a rat model of Alzheimer disease and diabetes.

Alzheimer disease (AD) is a common form of dementia associated with loss of memory and disruption of synaptic plasticity. There is a strong correlation between the pathophysiological features of AD and diabetes, including induction of oxidative stress, inflammation, and abnormality in blood vessels. Considering the brain's limited capacity to repair damage and the potential of stem cell-derived neural cells in the repair of neurodegenerative disease, we investigated the effects of artemisinin and TSP­1­human endometrial-derived-derived stem cells (TSP­1­hEDSCs) on the cognitive function and synaptic plasticity in AD-diabetes rats. The authors previously showed that artemisinin and TSP­1­hEDSCs suppressed oxidative stress and inflammation in AD-diabetes rats. Thrombospondins-1 (TSPs-1) is a glycoprotein that inhibits angiogenesis. AD and diabetes were induced using streptozotocin. Synaptic plasticity and learning and memory function were studied using the Morris water maze and electrophysiological test, respectively. Streptozotocin increased traveled swimming distance and escape latency in the morris water maze test, decreased the percent time spent in the target quadrant, inhibited the long-term potentiation (LTP), and increased the blood glucose levels. Simultaneous or separate administration of artemisinin and TSP­1­hEDSCs decreased the blood levels of glucose and improved cognitive tasks and synaptic plasticity by considerably reducing traveled swimming distance and escape latency, increasing the percent time spent in the target quadrant, and retrieval of the LTP; therefore, they could be utilized as an adjunct treatment for AD treatment. These results may be due to a decrease in oxidative stress and inflammation.

Fatty Acids in Subcutaneous Adipose Tissue of Pregnant Women with and without Polycystic Ovary Syndrome Are Associated with Genes Related to Steroidogenesis: A Case-Control Study.

BACKGROUND: The qualitative analysis of adipose tissue (AT) is an exciting area for research and clinical applications in several diseases and it is emerging along with the quantitative approach to research on overweight and obese people. While the importance of steroid metabolism in women with polycystic ovary syndrome (PCOS) has been reported, limited data exists on the effective roles of AT in pregnant women suffering from PCOS. The aim of this study was to determine association of fatty acid (FA) profiles with expression of 14 steroid genes in abdominal subcutaneous AT of PCOS vs. non-PCOS pregnant women. MATERIALS AND METHODS: In this case-control study, the AT samples of 36 non-PCOS pregnant women and 12 pregnant women with PCOS (3:1 ratio control: case) who underwent cesarean section were collected. Relationship of expressing gene targets and different features were performed using Pearson correlation analysis on the R 3.6.2 software. The ggplot2 package in R tool was used to draw the plots. RESULTS: Age (31.4 and 31.5 years, P=0.99), body mass index (BMI) (prior pregnancy 26 and 26.5 kg.m-2, P=0.62) and at delivery day (30.1 and 31, P=0.94), gestational period (264 and 267 days, P=0.70) and parity (1.4 and 1.4, P=0.42) of non-PCOS and PCOS pregnant women were similar. Expression of steroidogenic acute regulator (STAR) and 11ß-Hydroxysteroid dehydrogenase (11BHSD2) in non-PCOS pregnant women showed the highest association with eicosapentaenoic acid (EPA, C20:5 n-3, r=0.59, P=0.001) and (r=0.66, P=0.001), respectively. In the all participants, STAR mRNA level showed the greatest association with the EPA fatty acid concentration (P=0.001, r=0.51). CONCLUSION: Our results showed a link between the genes involved in steroid metabolism and fatty acids in AT of pregnant women, especially for omega-3 FA and the gene involved in the first step of steroidogenesis in subcutaneous AT. These findings warrant further studies.

Spectroscopic analysis of recombinant human growth hormone in the presence of sucrose and trehalose.

Recombinant human growth hormone (rhGH) is a therapeutic protein, associated with various human diseases, such as growth hormone deficiency. One of the interesting issues in the formulation of therapeutic proteins is excipients like disaccharides. In the current study, we try to compare the effect of sucrose and trehalose on the structure of rhGH in the liquid state at 25°C and 55°C. We use spectroscopic techniques including intrinsic and extrinsic fluorescence, Fourier-transform infrared (FTIR), circular dichroism (CD), dynamic light scattering (DLS), and time-resolved fluorescence. FTIR shows a slight change in the secondary structure of rhGH in presence of the sugars as sucrose is more effective than trehalose. Fluorescence investigations also confirm the enhancements of folding of rhGH and fluorescein isothiocyanate (FITC)-rhGH in presence of sucrose (1.5-fold more than trehalose). Also, we studied sucrose's effect on the rete of aggregation of rhGH using spectroscopy of Congo red, and fluorescence imaging of thioflavin T (ThT)-treated samples. It can be suggested that sucrose facilitates the amyloid formation of rhGH during 20 days of incubation at 37°C. This study will help to understand the growth hormone structural behavior in the liquid state in the presence of sucrose and trehalose in vitro.

Effect of eugenol on lipid profile, oxidative stress, sex hormone, liver injury, ovarian failure, and expression of COX-2 and PPAR-α genes in a rat model of diabetes.

BACKGROUND: Diabetes is among the leading causes of reproductive system failure and infertility in both women and men. Inflammation and oxidative stress have a main role in the development of diabetes. Eugenol or clove oil is a phenolic monoterpenoid with antioxidant and anti-inflammatory properties. Here, the effects of eugenol on diabetes features and ovarian function were investigated. METHODS AND RESULTS: Streptozotocin-induced diabetes rats were treated with 12 and 24 mg/kg of eugenol for 4 weeks. The biochemical and histological assay was done to evaluate the effects of eugenol on ovary and pancreas function, liver injury, oxidative status, sex hormones, lipid profile, and mRNA levels of cyclooxygenase-2 (COX-2) and peroxisome proliferator-activated receptor alpha (PPAR-α) genes. Streptozotocin increased levels of serum glucose, total cholesterol, triglyceride, low-density lipoprotein, aspartate transaminase, alanine transaminase, alkaline phosphatase, malondialdehyde, pancreas necrosis and inflammation, COX-2 expression, ovarian cystic, and anovulation. It decreased the levels of insulin, high-density lipoprotein, Superoxide dismutase, estradiol, progesterone, testosterone, luteinizing hormone, follicle-stimulating hormone, and PPAR-α expression. Eugenol administration ameliorated diabetes features through the improvement of lipid profile, oxidative status, insulin and glucose levels, sex hormone levels, liver markers, COX-2 and PPAR-α expression, and pancreas histology. It had no effect on ovarian cystic and follicular development. CONCLUSIONS: Therefore, eugenol may be useful for ameliorating some adverse features of diabetes and used as an adjunct treatment or protective agent accompany by other chemicals in diabetes patients.

Effects of Rosa damascena on reproductive improvement, metabolic parameters, liver function and insulin-like growth factor-1 gene expression in estradiol valerate induced polycystic ovarian syndrome in Wistar rats.

BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in reproductive-age women. The present study aimed to evaluate the effects of Rosa damascena (RD) extract in estradiol valerate (EV) induced polycystic ovary syndrome rats. METHODS: Adult female Wistar rats were divided into control (n = 12) and PCOS groups (n = 36). The PCOS model was induced using EV (4 mg/kg/day), which was confirmed in 6 rats in each control and PCOS group by observation of irregular estrous cycles in vaginal smears and ovarian multiple cystic. Then, the rest of the control group (n = 6) and PCOS rats (n = 30 in 5 divided groups) were treated orally for 28 days with metformin (MET) as a positive control (200 mg/kg/day) and RD extract (400, 800, and 1200 mg/kg/day, respectively). Body and ovary weights, biochemical and histological parameters, and expression of the IGF-1 gene were measured. RESULTS: Compared to the PCOS group, metformin and higher doses of RD extract (800 and 1200 mg/kg/day) significantly reduced BW, HOMA-IR, FBS, FINS, TG, LDL, TT, E2, LH, TC, and liver enzymes, and increased HDL and FSH levels. In addition, ovarian weight and CFs decreased, and the findings showed an increment in PFs, CLs, PAFs, AFs, and GFs. IGF-1 gene expression levels were significantly decreased (p < 0.001). CONCLUSION: RD extract seems to have the potential therapeutic effect of alleviating PCOS complications, and IGF-1 signaling may be involved in the beneficial effects of RD on PCOS.

The effects of Ibuprofen and 1, 8- cineol on anxiety and spatial memory in hyperammonemic rats.

In hepatic encephalopathy, hyperammonemia (HA) causes cognitive impairment and anxiety by causing neuroinflammation. Ibuprofen and 1,8- cineol have anti-inflammatory and antioxidant properties, respectively. The aim of this study was to evaluate the effects of ibuprofen alone and in combination with 1,8- cineol on anxiety and oxidative stress in a HA rat animal model. For this purpose, 36 rats were divided into six groups (n = 6) including the HA (received intraperitoneally (IP) ammonium acetate 2.5 mg/kg for four week), ibuprofen (induced HA rats that received 15 mg/kg, IP), cineol (induced HA rats that received 5 and 10 mg/kg, IP), Ib + cineol (induced HA rats that received 15 and 10 mg/kg, respectively, IP), and the control groups (received normal saline, IP). Except the HA group, all other groups received the aforementioned treatment for two weeks.. The Morris water maze and elevated plus maze were used to assess cognitive function and anxiety in the animals, respectively. Superoxide dismutase (SOD) activity was measured to evaluate oxidative stress. The mRNA expression levels of interleukin (IL)-6 and IL-1ß was assessed by real-time PCR in the animal's brain. The results showed a significant improvement in spatial memory and anxiety of the Ib group compared to the HA group (P < 0.01), but no significant change was observed in SOD activity (P > 0.05). There was a significant improvement in spatial memory and anxiety as well as a significant increase in SOD activity in the Ib + cineol group (P < 0.01) compared to the HA group. These results indicate that the Ib + cineol, not only improve cognitive function and reduce anxiety, also reduce oxidative stress, therefore, the simultaneous use of these two compounds may be useful in improving HA-induced cognitive disorders and anxiety.

Effects of empagliflozin on the expression of kisspeptin gene and reproductive system function in streptozotocin-induced diabetic male rats.

One of the main health concerns of diabetes is testicular dysfunction and impairment of reproductive function and sperm quality which can cause male infertility. kisspeptin is a hypothalamic neuropeptide hormone that is involved in the regulation of energy metabolism, gonadotrophin-releasing hormone (GnRH), and reproductive function. In the present study, the therapeutic effects of empagliflozin (sodium-glucose co-transporter 2 inhibitors) on kisspeptin expression along with reproductive function were investigated in diabetic male Wistar rats. Diabetes was induced by a single dose injection of 60 mg/kg streptozotocin. Empagliflozin in doses of 10 and 25 mg/kg body weight was used for 8 weeks. Serum samples, testis, epididymis, and pancreas tissues were collected at the end of the experiments. Lipid profiles, oxidative stress markers, blood hormones, expression of kisspeptin along with pathological alterations of the testis were assayed using real-time PCR, biochemical, and histological technics. Data have shown that empagliflozin improved hyperglycemia, reproductive impairment, oxidative stress condition, and histopathological alterations of pancreatic and testis tissues in diabetic animals. It improved the serum levels of sex hormones, insulin, leptin, and the expression of kisspeptin in the testes tissues. Spermatogenesis is also improved in treated animals. Data indicated that the administration of empagliflozin can ameliorate symptoms of diabetes. It probably has promising antidiabetic potential and may improve the male infertility of diabetic subjects. To our knowledge, this is the first experimental evidence for the potential impact of empagliflozin on kisspeptin expression in diabetic male rats.

Neuroprotective effects of nerolidol against Alzheimer's disease in Wistar rats.

Alzheimer's disease (AD) is the most common type of cognitive disorder in an elderly population associated with the accumulation of amyloid plaques and neurofibrillary tangles. Nerolidol is assumed to have neuroprotection effects. This study aimed to investigate the therapeutic effects of nerolidol on the Aß-induced model of AD in rats. Hippocampal injection of Aß was used to induce AD. Animals were randomly divided into control, sham (received PBS as Aß solvent), AD, DNPZ (AD + donepezil, 4 weeks); NRD-50 (AD + nerolidol, 50 mg/kg, 4 weeks), NRD-100 (AD + nerolidol, 100 mg/kg, 4 weeks; Prot (rats which received 100 mg/kg nerolidol for two weeks before Aß administration), and Solv (AD + sunflower oil as nerolidol solvent, 4 weeks) groups. All rats were subjected to a memory behavioral passive avoidance test by shuttle box. Thioflavin-S staining was performed to confirm Aß plaque formation and measured using ImageJ analyzing program. BDNF and CREB-1 expressions were analyzed by immunohistochemistry assay. Aß induced AD by Aß plaques formation and increasing step-through latency time. It reduced the expression of BDNF and CREB-1 protein. Administration of nerolidol or donepezil improved these features by decreasing Aß and increasing BDNF and CREB-1 expression and latency time. Nerolidol is likely to provide protection against AD. It may prevent dementia through the mediation of BDNF-CREB-1 expression and cholinergic nerve cells restoring. It seems that the administration of nerolidol before the onset of the disease will be more effective than after.

A Survey of Gasoline Ameliorator, Methyl-Tert-Butyl Ether (MTBE) on Bovine Serum Albumin: A Spectroscopy and Molecular Dynamic Simulation Study.

Background: Methyl-Tert-Butyl Ether (MTBE) as a gasoline modifier is frequently added to fuels and used in plenty of worldwide applications. MTBE biodegradation in groundwater occurs slowly and produces water miscibility; therefore, it causes diverse environmental and human health concerns. Objectives: The interaction of MTBE with bovine serum albumin (BSA) as a model protein at physiological conditions is investigated to illustrate the possible interactions of MTBE with the body's proteins. Materials and Methods: Uv-visible, fluorescence, circular dichroism (CD) spectroscopy methods, and molecular modeling were used to analyze the MTBE's effect on BSA structure and dynamics. The constant protein concentration and various MTBE contents were used for possible interactions. Results: The protein structural analysis shows that MTBE binds to BSA via positive enthalpy and entropy via hydrophobic interactions. Molecular docking shows the participation of several amino acids in the MTBE-BSA interaction. The CD spectroscopy results show that the BSA structure was not changed in the MTBE concentrations utilized in the study. Molecular dynamics (MD) simulation results suggest that MTBE can slightly change protein structure in the last 50ns. Conclusion: Comparing experimental and MD simulation results demonstrated that the BSA secondary structure was maintained in the low concentration of the MTBE. The entropy and enthalpy parameters asserted the hydrophobic interaction was the major force in the interaction between the BSA and MTBE.

Effects of vitamin D supplementation on depression and some selected pro-inflammatory biomarkers: a double-blind randomized clinical trial.

BACKGROUND: Both augmented inflammatory reaction and low vitamin D status are associated with depression but the magnitude of their relationships is unclear. This study was, therefore, conducted to evaluate the effects of vitamin D supplementation on serum 25(OH)D concentration, depression severity and some pro-inflammatory biomarkers in patients with mild to moderate depression. METHODS: An 8-week double-blind randomized clinical trial (RCT) was performed on 56 (18-60 yrs) patients with mild to moderate depression, randomly assigned to intervention (50,000 IU cholecalciferol 2wks-1) and control (placebo) groups. Serum 25(OH)D, intact parathyroid hormone (iPTH), interlukin (IL)-1ß, IL-6, high-sensitivity C-reactive protein (hs-CRP) and depression severity (Beck Depression Inventory-II) (BDI-II)) were initially and finally assessed. RESULTS: At the end point, statistically significant changes were observed only in intervention group as compared with controls including increased 25(OH)D concentration (+ 40.83 ± 28.57 vs. + 5.14 ± 23.44 nmol L-1, P < 0.001) and decreased depression severity (-11.75 ± 6.40 vs. -3.61 ± 10.40, P = 0.003). No significant within- or between group differences were observed in serum IL-1ß, IL-6 and hs-CRP concentrations. CONCLUSION: Increased circulating 25(OH)D concentrations following 8-week vitamin D supplementation (50,000 IU 2wks-1) resulted in a significant decrease in BDI-II scores in patients with mild to moderate depression. However, this effect was independent of the serum concentrations of the studied inflammatory biomarkers. TRIAL REGISTRATION: The clinical trial registration code was obtained from the Iranian Registry of Clinical Trials (date of registration: 17/09/2018, registration number: IRCT20170926036425N1) and ClinicalTrials.gov (date of registration: 04/12/2018, registration number: NCT03766074).

Therapeutic Effects of Eugenol in Polycystic Ovarian Rats Induced by Estradiol Valerate: A Histopathological and A Biochemical Study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common type of endocrinopathy in women which is accompanied by androgens elevation, insulin resistance, and metabolic dysfunction. Eugenol is a phenolic component of clove oil that has an antioxidant, anti-inflammatory, and anti-diabetic activity. The present study aimed to evaluate the therapeutic effects of eugenol on the PCOS models of rats. MATERIALS AND METHODS: In this experimental study, thirty adults female Wistar rats weighing between 180 and 200 g were used. Estradiol valerate-induced PCOS rats (4 mg/rat) were treated with eugenol (12 and 24 mg/kg) for 28 days. The effects of eugenol were studied on levels of glucose, lipid profile, liver enzymes, reproductive hormones, oxidative stress, and the expression of cyclooxygenase-2 (Cox-2) and peroxisome proliferator-activated receptor alpha (Ppar-α) genes, using biochemical analysis of blood and histopathological evaluation of ovaries.
Results: Estradiol valerate-induced PCOS resulted in the formation of cystic follicles in the ovaries, hyperinsulinemia, hyperglycemia, hyperlipidemia, hyperandrogenism, and anovulation. It altered the Cox-2 and Ppar-α gene expression and increased oxidative stress and activities of liver enzymes. Eugenol treatment improved the PCOS-associated endocrine and metabolic disorder and histopathological alterations, mostly through antioxidant, anti-diabetic, anti hyperlipidemic, and anti-androgenic properties. It showed beneficial effects on serum glucose, serum insulin, fat profile, reproductive hormones, liver activity, oxidative stress, expression of Cox-2 and Ppar-α genes, as well as restoration of normal ovulation in the PCOS animals.
Conclusion: Eugenol could represent a promising natural product to prevent PCOS or reduce its symptoms.

Therapeutic Effect of P-Cymene on Lipid Profile, Liver Enzyme, and Akt/Mtor Pathway in Streptozotocin-Induced Diabetes Mellitus in Wistar Rats.

Diabetes is a serious public health problem in low- and middle-income countries. There is a strong link between hyperglycemia, oxidative stress, inflammation, and the development of diabetes mellitus. PI3K/Akt/mTOR is the main signaling pathway of insulin for controlling lipid and glucose metabolism. P-cymene is an aromatic monoterpene with a widespread range of therapeutic properties including antioxidant and anti-inflammatory activity. In the present study, the antidiabetic effects of p-cymene were investigated. Diabetes was induced using streptozotocin in male Wistar rats. The effects of p-cymene and metformin were studied on levels of glucose (Glu), lipid profile, liver enzymes, oxidative stress, and the expression of Akt, phospho-Akt, and mTOR (mammalian target of rapamycin) proteins, using biochemical, histological, and immunohistochemical analysis. Data have shown that p-cymene can improve serum levels of Glu, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and the expression of mTOR, Akt, and phospho-Akt protein in diabetic animals. These results suggest that p-cymene has hypoglycemia, hypolipidemia, and antioxidant properties. It can regulate Akt/mTOR pathway and reduce hepatic and pancreas injury. It can be suggested for diabetes management alone or simultaneously with metformin.

Local Insulin-Derived Amyloidosis Model Confronted with Silymarin: Histological Insights and Gene Expression of MMP, TNF-α, and IL-6.

Amyloidosis is a heterogeneous group of protein deposition diseases associated with the presence of amyloid fibrils in tissues. Analogs of insulin that are used for treating diabetic patients (including regular insulin) can form amyloid fibrils, both in vitro and in vivo as reported in patients. The main purpose of this study was the induction of localized insulin-generated amyloidosis and the observation of silymarin effects on this process. In order to obtain amyloid structures, regular insulin was incubated at 37 °C for 24 h. Congo red absorbance and transmission electron microscopy images validated the formation of amyloid fibrils. Those fibrils were then injected subcutaneously into rats once per day for 6, 12 or 18 consecutive days in the presence or absence of silymarin, and caused development of firm waxy masses. These masses were excised and stained with Hematoxylin and Eosin, Congo red and Thioflavin S. Histological examination showed adipose cells and connective tissue in which amyloid deposition was visible. Amyloids decreased in the presence of silymarin, and the same effect was observed when silymarin was added to normal insulin and injected subsequently. Furthermore, plasma concentrations of MMP2, TNF-α, and IL-6 inflammatory factors were measured, and their gene expression was locally assessed in the masses by immunohistochemistry. All three factors increased in the amyloidosis state, while silymarin had an attenuating effect on their plasma levels and gene expression. In conclusion, we believe that silymarin could be effective in counteracting insulin-generated local amyloidosis.

The Protective Effects of Indole-Acetic Acid on the Renal Ischemia-Reperfusion Injury via Antioxidant and Antiapoptotic Properties in A Rat Model.

INTRODUCTION: The significant role of oxidative stress in the occurrence and development of a variety of diseases, including renal ischemia-reperfusion injury, has been thoroughly studied in this research. In this study, the protective role of indole-acetic acid on antioxidant, apoptotic and histopathological parameters in a rat model of renal ischemia-reperfusion (IR) injury were investigated. METHODS: We divided 40 rats into the following four groups (n = 10 per group): healthy control, IR control, IR + indole-acetic acid 40 mg/kg, and IR + indole-acetic acid 60 mg/kg. After two weeks, the rats were anesthetized and their kidneys were removed. The effects of indole-acetic acid on biochemical parameters [glutathione peroxidase (GPx) and catalase (CAT) were measured by spectrophotometry and expression of apoptotic genes (BAX and Bcl2) using real-time RT-PCR. Tubular necrosis was evaluated using a histopathological study. RESULTS: There were significant improvements in biochemical parameters (GPx), expression of the apoptotic genes (BAX) and tubular necrosis in rats treated with indole-acetic acid. CONCLUSION: Indole-acetic acid could reduce the effects of factors involved in the pathogenesis of IR, including oxidative stress, apoptosis and tubular necrosis. It can be recommended that, indoleacetic acid may be useful for amelioration of damages caused by IR.  DOI: 10.52547/ijkd.6894.

Phenylalanine and indole effects on the pathogenicity of human lysozyme amorphous aggregates.

Protein aggregation can affect the stability and function of proteins, and may lead to developing diseases, but reports on the in vivo effect of aggregates are scarce. In the current study, the effect of phenylalanine (Phe) and indole presence was first investigated on the structure and stability of human lysozyme (HLZ) and its aggregation under in vitro condition. Tm measurements, circular dichroism and spectrofluorimetric spectra, as well as and transmission electron microscopy (TEM) were performed in this stage. In the next step, pathogenicity of HLZ amorphous aggregates formed in presence or absence of the additives was investigated in vivo, by subcutaneous injection to adult male Wistar rats. Resulting inflamed tissues were studied by hematoxylin and eosin (HE), Congo red and Sudan black staining. Serum levels of liver enzymes (Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST)), specific inflammatory cytokines (Tumor Necrosis Factor Alpha (TNF-α) and Interleukin 6 (IL-6)) as well as glucose, cholesterol, and triglyceride levels were measured. Amorphous aggregates of HLZ caused inflammation and affected the number of fat cells, macrophages, cytokines, liver enzymes and glucose. Indole, that increases amorphous aggregates amount as shown with CD, fluorescence, and TEM experiments, leads into more severe inflammation. In presence of Phe, (which stabilizes HLZ structure) a markedly milder inflammatory state is observed in histological results and no increase could be detected in the inflammation-related parameters. In conclusion, amorphous aggregates of HLZ may be pathogenic in vivo, and presence of anti-aggregation compounds (such as Phe) can be effective in diminishing their deleterious manifestations.

High-density lipoprotein lipid peroxidation as a diagnostics biomarker in coronary artery disease.

This study aimed at evaluating the serum High-density lipoprotein lipid peroxidation (HDLox) levels and their association with coronary artery disease (CAD). This case-control study comprised 572 patients with stable CAD and 281 healthy subjects with no history of cardiovascular disease (control group). Based on the results of coronary angiography, the patient group was divided into two groups: CAD- and CAD+. HDLox was measured using a fluorimetric method. The ability of HDLox and serum high-density lipoprotein cholesterol (HDL-C) to detect CAD and coronary artery stenosis ≥50% was also compared using the receiver operating characteristic (ROC) curve analysis. The CAD patients showed significantly higher serum HDLox levels, compared to the control group [1.15 (1.01-1.31) vs. 0.85 (0.62-1.06), no units, p < 0.001]. Moreover, serum HDLox levels were significantly lower in CAD- patients, compared to the CAD+ patients [1.05 (0.92-1.22) vs. 1.24 (1.12-1.35), no units, p < 0.001]. According to the results of univariate and multivariate logistic regression, the HDLox showed association with the presence of CAD (odds ratio [OR]: 1.754; 95% confidence interval [CI]: 1.564-1.968; p < 0.001) and coronary artery stenosis ≥50% (OR: 1.729; 95% CI: 1.534-1.949; p < 0.001). The results obtained from the area under the ROC curve revealed that the HDLox could better detect the risk of CAD and coronary artery stenosis ≥50% compared to serum HDL-C level. The oxidation of HDL leads to a reduction in its antioxidant function and it has a crucial role in the development of atherosclerosis. HDLox is suggested as a diagnostics biomarker for CAD.