Biosensors; a novel concept in real-time detection of autophagy.

Autophagy is an early-stage response with self-degradation properties against several insulting conditions. To date, the critical role of autophagy has been well-documented in physiological and pathological conditions. This process involves various signaling and functional biomolecules, which are involved in different steps of the autophagic response. During recent decades, a range of biochemical analyses, chemical assays, and varied imaging techniques have been used for monitoring this pathway. Due to the complexity and dynamic aspects of autophagy, the application of the conventional methodology for following autophagic progression is frequently associated with a mistake in discrimination between a complete and incomplete autophagic response. Biosensors provide a de novo platform for precise and accurate analysis of target molecules in different biological settings. It has been suggested that these devices are applicable for real-time monitoring and highly sensitive detection of autophagy effectors. In this review article, we focus on cutting-edge biosensing technologies associated with autophagy detection.

Prolonged stress-induced depression-like behaviors in aged rats are mediated by endoplasmic reticulum stress and apoptosis in the hippocampus.

Structural and functional recovery from stress-induced depression is impaired in the context of aging brain. Since investigating the molecular substrates that facilitate behavioral recovery may have important implications for understanding brain plasticity and resilience of individuals, we studied depressive-like behaviors in young and aged rats 6 weeks after chronic stress exposure as a recovery period and examined the levels of TNF-α and IL-6 inflammatory cytokines, NADH oxidase activity, NADPH oxidase, endoplasmic reticulum (ER) stress markers, and apoptosis in the hippocampus. Young (3 months old) and aged (22 months old) male Wistar rats were divided into four groups; young control (Young), depression model of young rats that received chronic stress procedure followed by a 6-week recovery period (Young+S), aged control (Aged), and depression model of aged rats that received chronic stress procedure followed by a 6-week recovery period (Aged+S). After the recovery period, aged but not young rats showed depression-like behaviors, evaluated by the sucrose preference test (SPT) and forced swimming test (FST), coincided with the altered levels of TNF-α, IL-6, NADH oxidase activity, NADPH oxidase, GRP78, CHOP, and cleaved caspase-12 in the hippocampus of these animals. These data suggested that oxidative and ER stress-induced apoptosis in the aging hippocampus may affect the recovery-related outcomes after the stress paradigm.

Application of Pt@ZIF-8 nanocomposite-based electrochemical biosensor for sensitive diagnosis of tau protein in Alzheimer's disease patients.

Alzheimer's disease (AD) is a progressive brain disorder characterized by the ongoing decline of brain functions. Studies have revealed the detrimental effects of hyperphosphorylated tau (p-tau) protein fibrils in AD pathogenesis, highlighting the importance of this factor in the early-stage detection of AD conditions. We designed an electrochemical immunosensor for quantitative detection of the cis conformation of the p-tau protein (cis-p-tau) employing platinum nanoparticles (Pt NPs) supported on zeolitic imidazolate frameworks (ZIF) for modifying the glassy carbon electrode (GCE) surface. Under optimum conditions, the immunosensor selectively and sensitively detected cis-p-tau within the broad linear range of 1 fg mL-1 to 10 ng mL-1 and the low limit of detection (LOD) of 1 fg mL-1 with desired reproducibility and stability. Furthermore, the fabricated immunosensor's performance was examined for the cis-p-tau analysis in the serum of AD patients, indicating its accuracy and feasibility for real-sample analysis. Notably, this is the first application of Pt@ZIF-8 nanocomposite in fabricating a valid immunosensor for selective cis-p-tau detection, even in the presence of trans-p-tau. It is worth mentioning that the enzyme-linked immunosorbent assay (ELISA) reference technique is not able to evaluate pico- or femtomolar concentrations of cis-p-tau, making the fabricated immunosensor superior for early-stage measurement and screening of AD.

Lived experience caring for a child with Covid-19 disease: A phenomenological study.

Background: Caregivers of COVID-19 patients, including mothers with an infected child, are affected by adverse physical and psychological effects. Since mothers perform an important role in caring for a child with Covid-19, explaining their challenges and experiences positively affects the health conditions of children and society, in general. Aim: This study aimed to explain mothers' experiences with children diagnosed with COVID-19, who were referred to Bandar Abbas Children's Hospital. Method: The present study was qualitative and used the Van-Manen method. The statistical population included 13 mothers with children hospitalized for coronary heart disease and COVID-19, who were selected by purposive sampling. Semi-structured interviews were implemented to collect data until saturation. Data analysis was performed using MAXQDA 10 software. Lincoln and Guba's criteria were used to evaluate the data's accuracy and strength. Results: The mean age of mothers participating in the study was 37.92 ± 4.87. On the other hand, the mean age of children was 5.15 ± 2.07 years. Three main themes and 13 sub-themes were extracted from the data analysis. The main themes were "Inability to Caring", "Mental and Physical drain" and "Conflict of Roles and Responsibilities". In this study, when a child develops COVID-19 disease, the mother experiences a series of negative emotions, which lead to feelings of helplessness and inability to care for the child. Conclusion: Mental and physical manifestations strains were the most important issues that the mothers experienced. The majority of the participants need psychological support to cope with the disease. Therefore, it is recommended to make a proper design to psychologically and socially support these mothers, while alleviating the physical manifestations of their children's disease.

Melatonin: a promising neuroprotective agent for cerebral ischemia-reperfusion injury.

Cerebral ischemia-reperfusion (CIR) injury is initiated by the generation of reactive oxygen species (ROS), which leads to the oxidation of cellular proteins, DNA, and lipids as an initial event. The reperfusion process impairs critical cascades that support cell survival, including mitochondrial biogenesis and antioxidant enzyme activity. Failure to activate prosurvival signals may result in increased neuronal cell death and exacerbation of CIR damage. Melatonin, a hormone produced naturally in the body, has high concentrations in both the cerebrospinal fluid and the brain. However, melatonin production declines significantly with age, which may contribute to the development of age-related neurological disorders due to reduced levels. By activating various signaling pathways, melatonin can affect multiple aspects of human health due to its diverse range of activities. Therefore, understanding the underlying intracellular and molecular mechanisms is crucial before investigating the neuroprotective effects of melatonin in cerebral ischemia-reperfusion injury.

Electrochemical Biosensors as a Novel Platform in the Identification of Listeriosis Infection.

Listeria monocytogenes (L.M.) is a gram-positive bacillus with wide distribution in the environment. This bacterium contaminates water sources and food products and can be transmitted to the human population. The infection caused by L.M. is called listeriosis and is common in pregnant women, immune-deficient patients, and older adults. Based on the released statistics, listeriosis has a high rate of hospitalization and mortality; thus, rapid and timely detection of food contamination and listeriosis cases is necessary. During the last few decades, biosensors have been used for the detection and monitoring of varied bacteria species. These devices are detection platforms with great sensitivity and low detection limits. Among different types of biosensors, electrochemical biosensors have a high capability to circumvent several drawbacks associated with the application of conventional laboratory techniques. In this review article, different electrochemical biosensor types used for the detection of listeriosis were discussed in terms of actuators, bioreceptors, specific working electrodes, and signal amplification. We hope that this review will facilitate researchers to access a complete and comprehensive template for pathogen detection based on the different formats of electrochemical biosensors.

Electrochemical biosensors in exosome analysis; a short journey to the present and future trends in early-stage evaluation of cancers.

The tumor microenvironment consists of a multiplicity of cells such as cancer cells, fibroblasts, endothelial cells, and immune cells within the specific parenchyma. It has been indicated that cancer cells can educate other cells within the tumor niche in a paracrine manner by the release of nano-sized extracellular vesicles namely exosomes (Exo), resulting in accelerated tumor mass growth. It is suggested that exosomal cargo with remarkable information can reflect any changes in metabolic and proteomic profiles in parent tumor cells. Therefore, exosomes can be touted as prognostic, diagnostic, and therapeutic elements with specific biomarkers in patients with different tumor types. Despite the advantages, conventional exosome separation and purification protocols are time-consuming and laborious with low abnormal morphology and purity rate. During the last decades, biosensor-based modalities, as emerging instruments, have been used to detect and analyze Exo in biofluids. Due to suitable specificity, sensitivity, and real-time readout, biosensors became promising approaches for the analysis of Exo in in vitro and in vivo settings. The inherent advantages and superiority of electrochemical biosensors in the determination of tumor grade based on exosomal cargo and profile were also debated. Present and future challenges were also discussed related to the application of electrochemical biosensors in the clinical setting. In this review, the early detection of several cancer types associated with ovaries, breast, brain, colon, lungs, T and B lymphocytes, liver and rare types of cancers were debated in association with released exosomes.

Cohort profile: Bandar Kong prospective study of chronic non-communicable diseases.

Chronic non-communicable diseases (NCDs), are the leading causes of death among adults worldwide. It is projected that half of the NCDs could be avoided by preventing measures. Under the prospective epidemiological research studies in Iran (PERSIAN), we established a prospective population-based cohort study in southern Iran. The present study was designed to observe changing pattern of lifestyle transition over time and investigate the incidence and prevalence of regional modifiable risk factors as well as their associations with major NCDs. At baseline, 4063 participants aged 35-70 years were recruited on Oct, 2016and planned to get re-evaluated every 5 years along with annual follow-up. Data using validated electronic questionnaire comprising 55 questions and 482 items including general, medical and nutrition queries was collected. Blood, hair, nails, urine specimens and anthropometric measurements were taken. The response rate was 99%. In the results; male and female participants were 42.5% and 57.5%, respectively. Of note, 30.4% of women and 16.1% of men were obese. The prevalence of hypertension in men and women was 14.6% and 21%; however, diabetic men and women were 17.4% and 12.4%, respectively. Living in rural areas increased the odds of having hypertension by 1.33 (AOR = 1.33, 95% CI:1-09, 1.61). Noteworthy, logistic regression displayed that aging could predispose individuals to be more overweight, hypertensive and diabetic. The prevalence of multimorbidity of 3 or more NCDs were 8% (No. 326) and 6% (No.240), respectively. Intake of fruits, vegetables and dairy was less than two servings per day in 9.2%, 13% and 58.3% of the participants. Lower cardiovascular diseases and serum level of FBS and higher HDL level in sailors/fishermen compared to other job groups were significant (p-value <0.001). The second annual follow-up was completed and now at the end of the third wave. Findings of the present study signified the high prevalence of behavioral risk factors and their associations with respective NCDs. Subsequently, it is essential to keep track lifestyle variations, the modifiable risk factors and NCDs trends by prospective population-based cohort studies.

Swimming exercise improves SIRT1, NF-κB, and IL-1ß protein levels and pancreatic tissue injury in ovariectomized diabetic rats.

OBJECTIVES: In this study, we investigated the beneficial effects of swimming exercise on the SIRT1, NF-κB, IL-1ß protein levels, and pancreatic tissue damage in an ovariectomized diabetic rat model based on the anti-inflammatory effect of exercise. METHODS: Forty mature female Wistar rats were purchased and divided into sham (n=10) and OVX (bilateral ovariectomy) (n=30) groups. The ovariectomized rats were divided into 1-OVX, 2-ovariectomized diabetic (OVX.D), 3-OVX.D + exercise (OVX.D. E). After surgical recovery, animals in the diabetic group received a high-fat diet for one month. Swimming exercise (1 h/day) was performed concurrently with the start of the HFD diet for eight weeks. At the end of the high-fat diet, streptozotocin (30 mg/kg) was injected intraperitoneally. At the end of the second month, pancreatic tissue was collected from the animals after deep anesthesia for molecular evaluation and histology by Western blotting and hematoxylin-eosin, respectively. RESULTS: Swimming exercise significantly decreased inflammatory cytokines and tissue damage, and this decrease in cytokine expression appears to be associated with SIRT1 expression. The increase in SIRT1 by training was associated with decreased NF-κB-p65 and IL-1ß expression and preventing tissue damage. Induction of diabetes in the ovariectomized group (OVX.D) resulted in a significant increase in NF-κB-p65 and IL-1ß proteins and a decrease in the expression of SIRT1 compared with the sham group. However, swimming training significantly reversed these effects compared with the OVX.D group. CONCLUSIONS: Increased inflammation of ß-cells impairs insulin secretion in estrogen insufficiency. Swimming exercise eliminates inflammation in post-menopausal diabetes and supports the potential to prevent pancreatic activity after menopause.

Diabetic neovascularization defects in the retina are improved by genistein supplementation in the ovariectomized rat.

Genistein seems to have a protective and therapeutic effect on conditions associated with neovascular growth in the retina. This study investigated the angiogenesis, antioxidant, and anti-inflammatory effect of genistein on the retinas in ovariectomized diabetic rats. In this study, 40 female albino Wistar rats were divided into four groups (n = 8 per group): sham, ovariectomized group (OVX), OVX + diabetes (OVX.D), and OVX.D + genistein (OVX.D.G). OVX induced by removal of bilateral ovaries and then high-fat diet (HFD) and a low dose of streptozotocin (STZ) (1 mg/kg; intraperitoneal (IP) injection) was used for diabetes induction (OVX.D) with 8 weeks of genistein treatment (OVX.D.G). At the end of 8 weeks, the retina was removed under anesthesia. The samples were used to measure extracellular signal-regulated kinase (ERK), matrix metalloproteinase 2 (MMP-2), vascular endothelial growth factor (VEGF), and nuclear factor NF-kappa-B (NF-κB) by western blotting and inflammatory factors ELISA and oxidative stress. Measurements of glutathione (GSH) and malondialdehyde (MDA) showed that OVX and especially OVX.D significantly decreased GSH and increased MDA level in the retina, but genistein reversed these effects in OVX.D.G groups. Also, OVX and OVX.D significantly increased VEGF, MMP-2, p-ERK, NF-κB, interleukin-1beta (IL-1ß), and tumor necrosis factor alpha (TNFα) expression in the retina of OVX and OVX.D groups in comparison to the sham group (p < 0.05). However, a significant reduction of these proteins was observed in the genistein-treated group (p < 0.05). In conclusion, bilateral ovariectomy and subsequently estrogen deficiency caused the development of inflammation, neovascularization, and then retinopathy in STZ-induced diabetic ovariectomized rats. On the basis of the results, genistein administration may be a practical approach for improving symptoms and complications of ovariectomized diabetic retinopathy.

Synergism effect of swimming exercise and genistein on the inflammation, oxidative stress, and VEGF expression in the retina of diabetic-ovariectomized rats.

AIMS: Retinal neovascularization is one of the visual disorders during the postmenopausal period or types two diabetes. Physical activities and also phytoestrogens with powerful antioxidant features have been widely considered to improve nervous system diseases. Therefore, this study investigated the effects of genistein, swimming exercise, and their co-treatment on retina angiogenesis, oxidative stress, and inflammation in diabetic-ovariectomized rats. MAIN METHODS: Wistar rats were randomly divided into six groups (n = 8 per group): sham, ovariectomized group (OVX), OVX + diabetes (OVX.D), OVX.D+ genistein (1 mg/kg, eight weeks; daily SC), OVX.D + exercise (eight weeks), and OVX.D+ genistein+exercise (eight weeks). At the end of 8 weeks, the retina was removed under anesthesia. The assessed effects of treatment were by measuring MiR-146a and miR-132 expression via RT-PCR, the protein levels of ERK, MMP-2, VEGF, and NF-κB via western blotting, inflammation, and oxidative stress markers levels via the Eliza. KEY FINDINGS: The results showed miR-132, miR-146b, and MMP-2, NF-κB, ERK, VEGF, TNF-α, IL-1ß proteins, and MDA factor in the OVX.D group were increased, but glutathione (GSH) was decreased in comparison with the sham and OVX groups. Both exercise and genistein treatment has reversed the disorder caused by diabetes. However, the combination of exercise and genistein was more effective than each treatment alone. SIGNIFICANCE: It can be concluded that the interaction of exercise and genistein on microRNAs and their target protein was affected in the inflammation, stress oxidative, and extracellular matrix metalloproteinase pathways, can leading to a decrease in impairment of retinal neovascularization of the ovariectomized diabetic rats.

Ultrasensitive and label free electrochemical immunosensor for detection of ROR1 as an oncofetal biomarker using gold nanoparticles assisted LDH/rGO nanocomposite.

In the present article, we developed a highly sensitive label-free electrochemical immunosensor based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO)/gold nanoparticles modified glassy carbon electrode for the determination of receptor tyrosine kinase-like orphan receptor (ROR)-1. In this electrochemical immunoassay platform, NiFe-LDH/rGO was used due to great electron mobility, high specific surface area and flexible structures, while Au nanoparticles were prepared and coated on the modified electrodes to improve the detection sensitivity and ROR1 antibody immobilizing (ROR1Ab). The modification procedure was approved by using cyclic voltammetry and differential pulse voltammetry based on the response of peak current to the step by step modifications. Under optimum conditions, the experimental results showed that the immunosensor revealed a sensitive response to ROR1 in the range of 0.01-1 pg mL-1, and with a lower limit of quantification of 10 attogram/mL (10 ag mL-1). Furthermore, the designed immunosensor was applied for the analysis of ROR1 in several serum samples of chronic lymphocytic leukemia suffering patients with acceptable results, and it also exhibited good selectivity, reproducibility and stability.

Dyslipidemia and its associated factors in southern Iranian women, Bandare-Kong Cohort study, a cross-sectional survey.

Dyslipidemia, a major risk factor for cardiovascular diseases, has become a global issue. Due to the variations in the prevalence of dyslipidemia, this study aimed to evaluate dyslipidemia and its associated factors in women of the Bandare-Kong Cohort Study (BKNCD). This study was conducted on women from the population-based BKNCD, as part of the Prospective Epidemiological Research Studies in IrAN (PERSIAN). Sociodemographic data, medical history, and anthropometric indices were collected. Dyslipidemia was defined as any lipid abnormality including low-density lipoprotein (LDL) ≥ 160, total cholesterol (TC) ≥ 240, high-density lipoprotein (HDL) < 40, or triglyceride > 200 mg/dl. From the 2223 women in this study (mean age: 48.28 ± 9.26 years), dyslipidemia was observed in 851 (38.3%). High TC was the most common lipid abnormality (18.5%) followed by high LDL (17.7%). Dyslipidemia was most prevalent among women aged 55-70 years, the married, those with < 6 years of education, the unemployed, the overweight or obese, with low socioeconomic status, diabetes, hypertension, and high waist circumference, those using the hookah and living in urban areas. Logistic regression revealed that women with high waist-to-hip ratio (WHR) (OR = 2.22, 95% CI 1.60-3.08), those aged 45-54 years (OR = 1.34, 95% CI 1.07-1.68) and 55-70 years (OR = 1.33, 95% CI 1.03-1.72), and those living in urban areas (OR = 1.35, 95% CI 1.05-1.73) were at significantly increased risk of dyslipidemia. In addition, the results were confirmed using deep neural network models. Dyslipidemia was highly prevalent in Iranian women in the southern coastal region. Central obesity, age over 45 years, and living in urban areas appear to be relatively significant risk factors for dyslipidemia among women.

Recent progress in nanomaterial-based electrochemical biosensors for pathogenic bacteria.

This review (with 118 refs.) discusses the progress made in electroanalytical methods based on the use of organic and inorganic nanomaterials for the determination of bacteria, specifically of E. coli, Salmonella, Staphylococcus, Mycobacterium, Listeria and Klebsiella species. We also discuss advantages and limitations of electrochemical methods. Strategies based on the use of aptamers, DNA and antibodies are covered. Following an introduction into electrochemical biosensing, a first large section covers methods for pathogen detection using metal nanoparticles, with subsections on silver nanoparticles, gold nanoparticles, magnetic nanoparticles and carbon-based nanomaterials. A second large section covers methods based on the use of organic nanocomposites, graphene and its derivatives. Other nanoparticles are treated in a final section. Several tables are presented that give an overview on the wealth of methods and materials. A concluding section summarizes the current status, addresses challenges, and gives an outlook on potential future trends. Graphical abstract This review demonstrates the progress made in electroanalytical methods based on the use of organic and inorganic nanomaterials for the detection and determination of pathogenic bacteria. We also discuss advantages and limitations of electrochemical methods. Strategies based on the use of aptamers, DNA and antibodies are covered.

Effects of genistein and swimming exercise on spatial memory and expression of microRNA 132, BDNF, and IGF-1 genes in the hippocampus of ovariectomized rats.

OBJECTIVES: The aim of the present study was to investigate the effects of genistein and exercise on the spatial memory and expression of microRNA-132, BDNF, and IGF-1 in the hippocampus of ovariectomized rats. MATERIALS AND METHODS: Sixty animals were divided into six groups of control, sham, ovariectomy (OVX), ovariectomized with 8 weeks of genistein administration (OVX.G), with 8 weeks of swimming training (OVX.E), and with 8 weeks of both of them (OVX.G.E). The effect of genistein and/or exercise was evaluated by measuring microRNA-132, BDNF, and IGF-1 expression levels in the hippocampus tissue. Grafts were analyzed using Real-time polymerase chain reaction for microRNA-132, BDNF, IGF-1, and spatial memory via a Morris water maze (MWM). RESULTS: Our findings showed that ovariectomy decreased the expression of microRNA-132, BDNF, and IGF-1 in the hippocampus (P<0.05) in comparison with the sham group as well as performance in the water maze (P<0.05). Also according to results ovariectomized groups that were treated with genistein/exercise or both of them showed significant difference in expression of microRNA-132, BDNF, and IGF-1 in the hippocampus (P<0.05) and decreased latency in MWM (P<0.05) compared with the OVX group but combination treatment was more effective in the OVX.G.E group in comparison with OVX.E and OVX.G groups. CONCLUSION: Overall our results emphasized that combination treatment with genistein and exercise could improve microRNA-132, BDNF, and IGF-1 expression in the hippocampus as well as the spatial memory of ovariectomized rats. These effects may have beneficial impacts on the menopausal period.

Therapeutic potential of genistein in ovariectomy-induced pancreatic injury in diabetic rats: The regulation of MAPK pathway and apoptosis.

OBJECTIVES: Genistein, as a phytoestrogen found in legumes, has several biological activities in general and anti-diabetic activity particularly. In this study, we investigated the effect of genistein on proteins involved in ß-cell proliferation, survival and apoptosis to further reveal its anti-diabetic potential in the ovariectomized diabetic rat. MATERIALS AND METHODS: We used three-month-old female Wistar rats that either underwent ovariectomy (OVX) or received a sham surgery (Sham). In a subsequent series of experiments, OVX rats received high-fat diet and low dose STZ to induce diabetes (OVX.D) and genistein treatment (OVX.D.G). Western blot analysis was used for the assessment of phosphorylation of ERK1/2 and AKT and expression of Bcl-2 and caspase-3 in pancreas tissue. Hematoxylin-Eosin (H&E) staining was used for histopathological assessment. RESULTS: Genistein induced AKT and ERK1/2 phosphorylation protein expression of Bcl-2 in the pancreas. In addition, genistein suppressed protein level of caspase-3. Administration of genistein significantly improved hyperglycemia in ovariectomized diabetic rat, concomitant with improved islet ß-cell morphology and mass. CONCLUSION: These findings suggest that the beneficial antidiabetic effect of genistein partially mediated by directly modulating pancreatic ß-cell function via activation of the AKT, ERK1/2, and Bcl-2, as cell survival and anti-apoptotic factors, and decreasing of proapoptotic caspase-3.

Effect of genistein on expression of pancreatic SIRT1, inflammatory cytokines and histological changes in ovariectomized diabetic rat.

OBJECTIVES: Genistein is reported to have anti-diabetic and anti-inflammatory functions, in particular, direct effects on ß-cell proliferation and insulin secretion. In this study, we investigated the anti-inflammatory effect of genistein on the pancreatic ß-cells in ovariectomized diabetic rat. MATERIALS AND METHODS: Forty female rats were divided into four groups: sham, bilateral ovariectomy (OVX), OVX.D (OVX+diabetes) and OVX.D.G (OVX.D+genistein). After bilateral ovariectomy, rats in the diabetic groups were fed high-fat diet (HFD), ad libitum for 4 weeks, and then a low dose of streptozotocin (STZ) (30 mg/kg) injected intraperitoneally. Genistein (1 mg/kg/day; SC) was administrated for 8 weeks. At the end of 8 weeks, pancreas tissue was removed and used for western blotting and Hematoxylin-Eosin staining. RESULTS: Treatment with genistein declined inflammation and tissue injury, and this decline was correlated with the expression of SIRT1. OVX and OVX.D significantly increased Nf-κB and IL-1ß expression and decreased SIRT1 levels compared to sham group (P<0.05). Significant reduction of Nf-κB and IL-1ß, and increasing of SIRT1 were observed during genistein treatment (P<0.05). CONCLUSION: Estrogen deficiency alone or with HFD increased pancreatic inflammation. However, subcutaneous administration of gtenistein prevented from these inflammatory changes in the pancreas of a surgery animal model of ovariectomy with or without diabetes. Our results support the potential preventing effect of genistein from pancreatic injury.

Apoptosis and Histopathology of the Heart after Renal Ischemia-Reperfusion in Male Rat Running title: Ischemia-Reperfusion Injury

ABSTRACT Ischemia-reperfusion injury was seen in strokes, myocardial infarctions, acute kidney injury, mesenteric ischemia, liver and systemic shock. Renal ischemia-reperfusion is more importance in the setting of kidney transplantation that affects distant organs. In this study forty Male Albino Wistar rats (200-250g) were randomly divided in four group (n=10) including control, sham operation group, nephrectomy and IRI group. All rats anesthetized with intraperitoneal injection of ketamine (50 mg/kg) and xylazine (10 mg/kg) and maintained the core body temperature at approximately 37°C. For inducing IRI group, it was performed right nephrectomy, and in continuing, the left kidney pedicle occluded to 45 min via nontraumatic microvascular clamp for making ischemia that followed 24 hours reperfusion. TUNEL assay was used to detect the cardiac apoptotic cells. Hematoxylin-Eosin staining and periodic acid-Schiff (PAS) procedure was used to histopathological assessment and glycogen accumulation respectively. There was more heart damage at 24 h reperfusion in IRI group. Renal IRI group showed myocardial degeneration, necrosis and increasing connective tissue in myofibril. There were apparent hypertrophy and swelling of myofibril, fragmentation and vacuolization of sarcoplasm. In addition, it was shown elevated apoptotic cell at 24 hours reperfusion in renal IRI group than sham group. There were increases of glycogen accumulation in cardimyocyte of renal IRI group. Our findings suggest that renal IRI-induced cardiac damage, accompanied by an accumulation of glycogen granules, induced apoptosis and histological changes in cardiomyocytes.

Involvement of microRNA-133 and -29 in cardiac disturbances in diabetic ovariectomized rats.

OBJECTIVES: Menopause and diabetes obviously increase the risk of cardiovascular disease in women. The aims of the present study were to evaluate the effects of ovariectomy in type 2 diabetes on the histology and expression of miRNA-29, miRNA-133, IGF-1 and Bcl-2 genes and Bcl-2 protein and caspase 3 activity in the hearts of female rats. MATERIALS AND METHODS: Forty Female Wistar rats were divided into four groups: control, sham, ovariectomized (OVX), and ovariectomized with type 2 diabetes (OVX.D). After the 8-week experiment, the histological evaluation of the heart tissue was performed using H&E staining and PAS analysis, and cardiac expression of miRNA-29, miRNA-133, IGF-1, and Bcl-2 were evaluated using real-time PCR, and Bcl-2 protein and caspase 3 activity were evaluated using Western blot and ELISA. RESULTS: Ovariectomy significantly decreased miRNA-29, miRNA-133, IGF-1, and BCL-2 expression and Bcl-2 protein and increased caspase 3 activity in the heart compared to sham animals group (P<0.05). Type 2 diabetes in ovariectomized rats markedly decreased expression of miRNA-29, miRNA-133, IGF-1, BCL-2 genes, and Bcl-2 protein, and increased caspase 3 activity and reduced collagen and fibroblast tissue and glycogen granule deposition in relation to OVX group (P<0.05). CONCLUSION: Our findings suggest that type 2 diabetes and menopause synergically could enhance the cardiac fibrosis through dysregulation of miRNA-29, miRNA-133, IGF-1, and Bcl-2 genes expression and Bcl-2 protein and upregulation of caspase 3 activity.

Expression of the Mir-133 and Bcl-2 could be affected by swimming training in the heart of ovariectomized rats.

OBJECTIVES: The beneficial and more potent role of exercise to prevent heart apoptosis in ovariectomized rats has been known. The aim of this study was to examine the effects of swimming training on cardiac expression of Bcl-2, and Mir-133 levels and glycogen changes in the myocyte. MATERIALS AND METHODS: Forty animals were separated into four groups as control, sham, ovariectomy (OVX) and ovariectomized group with 8 weeks swimming training (OVX.E). Training effects were evaluated by measuring lipid profiles, Bcl-2 and Mir-133 expression levels in the cardiac tissue. Grafts were analyzed by reverse transcription-polymerase chain reaction for Bcl-2 mRNA and Mir-133 and by Western blot for Bcl-2 protein. RESULTS: Ovariectomy down-regulated Bcl-2 and Mir-133 expression levels in the cardiac tissue, and swimming training up-regulated their expression significantly (P<0.05). CONCLUSION: Our results showed that regular exercise as a physical replacement therapy could prevent and improve the effects of estrogen deficiency in the cardia.