فهرست مطالب mohammadhossein nasr-esfahani

Ovarian granulosa cells (GCs) are essential for follicular development. Ovarian advanced glycation end-products (AGEs) accumulation is related to GCs dysfunction. Alpha-lipoic acid (ALA) illustrates therapeutic capabilities for infertility-related disorders. Therefore, this study assessed the effects of ALA on AGEs-induced GCs hormonal dysfunction.

The study was conducted from October 2021 to September 2022 at the Department of Medical Genetics, Shiraz University of Medical Sciences. Isolated GCs (n=50) were divided into control, human glycated albumin (HGA), HGA+ALA, and ALA treatments. Steroidogenic enzymes and AGE receptor (RAGE) genes were assessed by qRT-PCR. Steroid hormones and RAGE protein were evaluated using ELISA and Western blotting. Data were analyzed using GraphPad Prism software (ver. 9), and P<0.05 was considered significant.

Our findings showed that HGA treatment significantly (P=0.0001) increased RAGE (by 140.66%), STAR (by 117.65%), 3β-HSD (by 165.68%), and 17β-HSD (by 122.15%) expression, while it decreased CYP19A1 (by 68.37%) expression. RAGE protein level (by 267.10%) was also increased in HGA-treated GCs. A significant decrease in estradiol (by 59.66%) and a slight and sharp elevation in progesterone (by 30.40%) and total testosterone (by 158.24%) levels was also observed. ALA treatment ameliorated the HGA-induced changes in steroidogenic enzyme mRNA levels (P=0.001) and steroid hormone secretion (P=0.010).

This work shows that ALA therapy likely corrects hormonal dysfunctions caused by AGEs in luteinized GCs. This effect is probably achieved by decreased RAGE expression. Clinical research is needed to understand how AGEs and ALA interact in the ovary, which might lead to a more targeted ovarian dysfunction therapy.

Advanced glycation end products (AGEs) that accompany many metabolic disorders including diabetes, obesity, and a wide range of dyslipidemia conditions, are strongly associated with adverse effects on cell and tissue homeostasis. Accordingly, our objective was to investigate the impact of AGE-promoting diets on mouse models, considering both scenarios with and without methylglyoxal (MGO) as a primary precursor of AGEs.

In this experimental study, 5-week-old C57BL/6 mice were split into four groups as a control group (n=5), AGE (n=5), MGO (n=8), and AGE-MGO-diets (n=8). After five weeks the level of fasting blood sugar (FBS), body weight, food intake, sperm parameters, and functional tests were evaluated. Furthermore, testicular superoxide dismutase (SOD) activity, malondialdehyde, and total antioxidant capacity (TAC) were assessed.

After five weeks, AGE, AGE-MGO, and MGO groups showed the highest level of body weight and FBS in comparison to the control group. Mean sperm concentration, sperm malondialdehyde, testicular lipid peroxidation, and TAC did not differ significantly among the study groups. While, AGE, MGO, and AGE-MGO groups showed a significant reduction in sperm motility and progressive motility compared to the control group (P<0.05). The greatest increases in abnormal sperm morphology and intracytoplasmic reactive oxygen species (ROS) were observed in the MGO and AGE-MGO groups than in the control group (P<0.05). Sperm protamine deficiency and residual histone were significantly increased in the three treatment groups compared to the control group (P<0.05). Regarding the DNA damage, the AGE and AGE-MGO groups showed the most severe damage. The lowest amount of testicular superoxide dismutases (SOD, P<0.001) was observed in the AGE-MGO group.

AGEs and MGO have a negative influence on sperm function and reproductive potential. These effects could be possibly attributed to both increased oxidative stress (OS) and inflammation.

In this article published in Cell J, Vol 26, No 1, 2024, on pages 81-90, the authors found that the affiliation of authorsin address 1 and also the two corresponding authors had accidentally missed during the formatting of the paper.Therefore, we corrected them.The authors would like to apologize for any inconvenience.

Separation of healthy sperm (motile and with the best DNA structure) is traditionally done by density gradient centrifugation or upward swimming methods. In recent years, microfluidic methods have been developed to separate motile sperm with the best morphology and DNA structure and will replace traditional methods in close future. Microfluidic systems require a smaller sample volume, which, in addition to higher purity, has advantages such as cost reduction and test time reduction.. In this research, by using thigmotaxis phenomenon in which sperms follow the channel wall and also positive dielectrophoresis phenomenon in which traps the particles based on applying non uniform electric field, a microfluidic actuator is presented. By using a large number of microchannel parallel to each other, the motile sperm, follow the walls and move to the end of channels. Then, by applying 1 MHz in 3 V, the positive dielectrophoresis force traps the sperms with low motility, but the sperms with high motility have the ability to escape from the electric field and reach the outlet of the channel. In order to check the electric field and also the fluid flow in the channels, Comsol software was used. Based on the design and simulation, the sperm separation actuator was manufactured and tested. The clinical tests results showed that compared to the inlet of the device, the motility was increased from 54% to 81% and the viability was increased from 56% to 75% in the outlet of the manufactured system.

Celiac disease is a common chronic inflammatory condition of the small intestine caused by permanent intolerance to gluten/gliadin. It has been demonstrated that oxidative stress is one of the mechanisms that is involved in gliadin toxicity, and there is a correlation between oxidative damage with this disease. Similarly, increased oxidative stress was repeatedly reported in infertile men which led to low-quality of sperm function. Therefore, we aimed to assess sperm parameters and chromatin status in men with Celiac disease.

In this case-control study, semen samples were collected from 11 fertile men without Celiac and 10 men with diagnostic Celiac disease. Basic semen analyses were performed according to the World Health Organization (WHO) 2010 protocol. The percentage of sperm with persistence histones, protamine deficiency, DNA fragmentation, malondialdehyde (MDA), and intracellular reactive oxygen species (ROS) were assessed using aniline blue, chromomycin A3, sperm chromatin structure assay, thiobarbituric acid reactive substances (TBARS) assay, and diacetyldichlorofluorescein staining, respectively.

Unlike the sperm parameters, which did not show significant differences between men with Celiac disease and fertile individuals, sperm chromatin maturation (persistence histones and protamine deficiency) and sperm DNA damage in men with Celiac disease were significantly higher compared to fertile individuals (P<0.05). In addition, the percentage of sperm viability in these individuals was significantly lower than that in the fertile individuals (P<0.05). We did not observe any significant differences in sperm lipid peroxidation and intracellular ROS levels between the two study groups (P>0.05).

Celiac disease affects sperm chromatin maturation and DNA fragmentation, emphasizing its impact on reproductive health.

Sperm motility is a fundamental factor for sperm penetration into the oocyte and fertilization. According to the World Health Organization (WHO) 2021, less than 42 percent of motility is called asthenozoospermia, which is one of the most common causes of male infertility. This retrospective cohort study aims to investigate sperm DNA damage with TUNEL and SCSA methods and its relationship with sperm parameters, age, and body mass index (BMI) in severe asthenospermia (<2% sperm motility) and normozoospermia.

The study parameters between 111 subjects with severe asthenozoospermia and 113 subjects with normozoospermia were investigated. The 2010 World Health Organization guidelines were used to evaluate sperm parameters, and TUNEL and SCSA methods were used to assess sperm DNA damage. The statistical analysis was done using the t-test of two independent samples. Pearson's correlation coefficient was used to investigate the relationship between sperm DNA damage and sperm parameters, age, and BMI P<0.05 was considered significant.

The mean seminal volume, sperm concentration and count, and the percentage of sperm total motility and progressive motility were significantly lower in severe asthenospermic subjects compared to normozoospermic subjects (P<0.001). In addition, the mean percentage of sperm DNA damage in severe asthenozoospermic individuals was significantly higher than in normozoospermic individuals (P<0.001). There was also a positive and significant relationship between sperm DNA damage and the age of severely asthenozoospermic men.

Sperm motility, sperm DNA damage, and the father's age are essential in the successful conception, development, and health of the embryo. Therefore, the evaluation of sperm DNA damage is recommended in examining male fertility and selecting the appropriate treatment approach for men with severe asthenozoospermia.

Practical Implications:

Considering that the sperm genome constitutes 50 percent of the genetic material of the next generation, evaluating sperm DNA health to determine the appropriate treatment approach in infertile men can be influential in the success of assisted reproductive techniques and in maintaining the next generation’s health.

Antioxidant apigenin (AP) is a natural, non-mutagenic, and less toxic flavonoid with pharmacological anti-cancer, anti-viral, anti-bacterial, anti-apoptotic, and anti-inflammatory activities. This antioxidant is easily received by the cell, binds to sperm DNA, and forms a DNA-AP complex, thereby protecting sperm DNA. The present study was conducted to determine the antioxidant effect of AP on human sperm quality after freezing-thawing.

In this descriptive-analytical study, 10 normozoospermic samples underwent freezing-thawing conditions, and sperm functional tests were investigated in different AP concentrations, including 0.4 mM, 0.2 mM, 0.1 mM, and 0.05 mM.

The quality of total sperm parameters and functional tests decreased after freezing compared to before freezing. Among the AP concentrations, only in the 0.2 mM AP concentration, the improvement of the additional histone percentage, protamine deficiency, and sperm DNA health were observed compared to the control; this finding was not statistically significant.

The use of AP with a concentration of 0.2 mM during freezing-thawing culminates in improving sperm functional tests.

Diabetic men suffer an increased risk of infertility associated with signs of oxidative damage and decreased methylation in sperm pointing to a deficit of the one-carbon cycle (1CC). We aimed to investigate this deficit using mice models (type 1 and 2) of streptozotocin-induced diabetes.

In this experimental study, 50 male mice, aged eight weeks, were divided randomly into four groups: sham, control, type 1 diabetes mellitus (DM1), and DM2. The DM1 group was fed a normal diet (ND) for eight weeks, followed by five consecutive days of intraperitoneal administration of Streptozotocin (STZ, 50 mg/kg body weight). The DM2 group was fed a high-fat diet (HFD) for eight weeks, followed by a single intraperitoneal injection of STZ (100 mg/kg). After twelve weeks, all the mice were euthanized, and study parameters assessed. In the sham group, citrate buffer as an STZ solvent was injected.

Both types of diabetic animals had serious impairment of spermatogenesis backed by increased DNA damage (P=0.000) and decreased chromatin methylation (percent: P=0.019; intensity: P=0.001) and maturation (P=0.000). The 1CC was deeply disturbed with increased homocysteine (P=0.000) and decreased availability of carbon units [methionine (P=0.000), serine (P=0.088), folate (P=0.016), B12 (P=0.025)] to feed methylations.

We have observed a distinct impairment of 1CC within the testes of individuals with diabetes. We speculate that this impairment may be linked to inadequate intracellular glucose and diminished carbon unit supply associated with diabetes. As a result, interventions focusing on enhancing glucose uptake into sperm cells and providing supplementary methyl donors have the potential to improve fertility issues in diabetic patients. However, additional clinical testing is required to validate these hypotheses.

Asthenozoospermia is an infertility condition in which a person has reduced sperm motility and so, the chance of the sperm fertilizing the egg in the female reproductive tract is reduced. This study aimed to investigate sperm DNA damage using SCSA and TUNEL tests in a large population of infertile men with asthenozoospermia, comparing them with a population of normozoospermic individuals with healthy sperm samples.

In this observational case-control study, semen parameters were assessed according to the World Health Organization guidelines (2010) guideline in a large population of asthenozoospermic (1383) and normozoospermic (2235) men referring to Isfahan Fertility and Sterility Center. In addition, sperm DNA fragmentation were assessed by TUNEL, and SCSA tests. For comparison of study variations between two groups, independent samples t-test was used, and for correlation between parameters, Pearson's correlation coefficient was carried out. P<0.05 was considered significant.

Despite no significant differences in sperm volume and body mass index, the means of sperm concentration, count, total sperm motility and total progressive were significantly lower in the asthenozoospermic men compared to normozoospermic men (P<0.001). In addition, the means of high DNA stainability as well as sperm DNA fragmentation assessed by SCSA and TUNEL tests were significantly higher in asthenozoospermic men compared to normozoospermic men (P<0.001).

In asthenozoospermic men, sperm DNA is severely damaged, so before using assisted reproductive techniques in these individuals, it is better to use antioxidant treatment and/or novel sperm preparation methods for treatment of them.

Varicocele is a common cause of male infertility, affecting a substantial proportion of infertile men. Recent studies have employed transcriptomic analysis to identify candidate genes that may be implicated in the pathogenesis of this condition. Accordingly, this study sought to leverage rat gene expression profiling, along with protein-protein interaction networks, to identify key regulatory genes, related pathways, and potentially effective drugs for the treatment of varicocele.

In this in-silico study, differentially expressed genes (DEGs) from the testicular tissue of 3 rats were screened using the edgeR package in R software and the results were compared to 3 rats in the control group. Data was obtained from GSE139447. Setting a -11 and P<0.05 as cutoff points for statistical significance, up and down-regulated genes were identified. Based on Cytoscape plugins, protein-protein interaction (PPI) networks were drawn, and hub genes were highlighted. ShinyGO was used for pathway enrichment. Finally, effective drugs were identified from the drug database.

Among the 1277 DEGs in this study, 677 genes were up-regulated while 600 genes were down-regulated in rats with varicocele compared to the control group. Using protein-protein interaction networks, we identified the top five up-regulated genes and the top five down-regulated genes. Enrichment analysis showed that the up-regulated genes were associated with the cell division cycle pathway, while the down-regulated genes were linked to the ribosome pathway. Notably, our findings suggested that dexamethasone may be a promising therapeutic option for individuals with varicocele.

The current investigation indicates that in varicocele the cell division cycle pathway is up-regulated while the ribosome pathway is down-regulated compared to controls. Based on these findings, dexamethasone could be considered a future candidate drug for the treatment of individuals with varicocele.

Epigenetic modifications such as DNA methylation play a key role in male infertility etiology. This study aimed to explore the global DNA methylation status in testicular spermatogenic cells of varicocele-induced rats and consider their semen quality, with a focus on key epigenetic marks, namely 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC), as well as the mRNA and proteins of ten-eleven translocation (TET) methylcytosine dioxygenases 1-3.

In this experimental study, 24 mature male Wistar rats (8 in each group) were assigned amongst the control, sham, and varicocele groups. Sperm quality was assessed, and DNA methylation patterns of testicular spermatogenic cells were investigated using reverse transcription-polymerase chain reaction (RT-PCR), western blot, and immunofluorescence techniques.

Sperm parameters, chromatin and DNA integrity were significantly lower, and sperm lipid peroxidation significantly increased in varicocele-induced rats in comparison with control rats. During spermatogenesis in rat testis, 5-mC and 5-hmC epigenetic marks, and TET1-3 mRNA and proteins were expressed. In contrast to the 5-mC fluorescent signal which was presented in all testicular cells, the 5-hmC fluorescent signal was presented exclusively in spermatogonia and a few spermatids. In varicocele-induced rats, the 5-mC signal decreased in all cells within the tubules, whereas a strong signal of 5-hmC was detected in seminiferous tubules compared to the control group. As well, the levels of TET2 mRNA and protein expression were significantly upregulated in varicocele-induced rats in comparison with the control group. Also, our results showed that the varicocele-induced animals exhibited strong fluorescent signals of TET1-3 in testicular cells, whereas weak fluorescent signals were identified in the seminiferous tubules of the control animals.

Consequently, we showed TET2 upregulation and the 5-hmC gain at testicular levels are associated with varicocele and sperm quality decline, and therefore they can be exploited as potential biomarkers of spermatogenesis.

The increasing prevalence of obesity has affected millions of people across the globe, leading to various diseases, such as prediabetes, diabetes, cardiovascular diseases, cancer, and muscle disorders. One of the therapeutic interventions in the prevention of diabetes is the use of herbal supplements, such as green coffee and its phenolic compounds, including chlorogenic acid, due to its antioxidant, anti-inflammatory, and insulin-sensitive properties. Therefore, the present study aimed to investigate the effect of green coffee and chlorogenic acid on glycogen content in the muscles of prediabetic mice.

A total of 20 male C57BL/6 mice were randomly assigned to two groups: normal diet mice (5 mice) and high-fat diet mice (15 mice), which were fed for 12 weeks. After the induction of prediabetes by Fasting blood sugar (FBS) and Glucose tolerance test (GTT) tests, this group was divided into three subgroups: prediabetes mice (5 cases), prediabetes mice with diabetes with green coffee consumption (Prediabetes - Green Coffee, 5 cases) and prediabetes with chlorogenic acid consumption (Prediabetes - Chlorogenic Acid, 5 cases) which were treated for 10 weeks. In the end, the mice were sacrificed, and the relative level of expression of genes involved in the glycogen content of the Gastrocnemius muscle, including PI3K, AKT, GSK3, GYS1, as well as the LRRC8A gene, was measured through the Real-Time PCR technique. The data were analyzed using the one-way analysis of variance test by GraphPad Prism software (version 8).

The results pointed to a significant increase in the blood glucose level in the HFD group (129.9±5.4 mg/dL) compared to ND (95±3.3 mg/dL) and a significant decrease in the PD-GC group (103.5±8.16 mg/dL) and PD-CA (99±8 mg/dL) compared to the PD group (129.5±9.4 mg/dL) (P<0.001). The periodic acid-Schiff ( PAS) staining technique demonstrated a significant decrease in the level of glycogen content in the rats of the PD group compared to the ND group. While in the treatment groups, a significant increase was observed in muscle glycogen accumulation compared to the PD group (P<0.001). The expression level of PI3K, AKT, LRRC8A, and GYS1 genes decreased, and GSK3 significantly increased in the PD group compared to the ND group (P<0.001), indicating a disturbance in the signaling pathway of glycogen content in the PD group. Nonetheless, in the treated groups, a significant increase was observed in the expression level of PI3K, AKT, LRRC8A, and GYS1 genes compared to the PD group (P<0.001).

Considering the complications caused by prediabetes disease, in addition to early diagnosis of prediabetes, it seems to control weight and increase insulin sensitivity, the consumption of green coffee and chlorogenic acid can improve the prediabetic condition caused by the signaling pathway of glycogen synthesis in the muscle tissue of prediabetic rats.

One of the main spermatogenesis events is the replacement of histones with small proteins called protamines, which leads to chromatin's condensation in the sperm nucleus and protects it against possible damage. Today, tests such as aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining are used based on different characteristics to evaluate sperm chromatin compaction. For the assessment of DNA fragmentation in sperm, several tests such as 8-hydroxy-2-deoxyguanosine (8-OHdG), TUNEL, Comet, sperm chromatin structure assay (SCSA), sperm chromatic dispersion (SCD), and acridine orange have been introduced that directly and indirectly assay DNA damage. The articles in PubMed and Google Scholar, as well as related books, from 2007 to 2022, were collected and reviewed based on keywords 8-OHdG, TUNEL, Comet, SCD, and acridine orange. So far, many studies have been conducted at the treatment level and on sperm chromatin tests, but the number of cases published so far is limited. Various sperm samples have been used in different studies, with different threshold limits in the tests. The sixth edition of the World Health Organization (WHO) book notes that each laboratory has its threshold limit. Therefore, in this review study, common methods of evaluating chromatin packaging and DNA damage are introduced, and the advantages and disadvantages of each test are discussed based on the latest achievements related to infertility.

Objectices:

skeletal muscles play an important role in the homeostasis of the whole body.Any dysfunction of skeletal muscles can cause problems such as prediabetes and type 2 diabetes.MondoA is a transcription factor that is activated under normal conditions by increasing the amount of glucose inside the cell and transcribes its target genes(TXNIP/ARRDC4).The purpose of this study is to investigate the effect of aerobic training on the level of MondoA transcription factor activity and the expression of its target genes in the skeletal muscle of prediabetic model rats.

Methods & Materials:

In order to induce prediabetes15 male C57BL/6 mice,(aged four weeks and weighing between 12 and 14 g)were fed with a high-fat diet for 12 weeks.After induction of prediabetes mice were divided into two groups of high fat diet(5 mice/no exercise) and aerobic training group (5 mice/10 weeks of aerobic training)The aerobic exercise protocol was performed in the form of running on a treadmill 5 days a week and 45 minutes a day.The velocity also increased from 15 to 23 m/min with moderate intensity.

The results showed a significant decrease(p<0.001)in blood glucose level and insulin resistance in the PD-Ex group compared to PD. Also exercise training decreased the relative level of expression of TXNIP(p<0.01) and ARRDC4(p<0.05)genes, the presence of transcription factor MondoA in the nucleus, and increased the expression of Glut4 protein in the cell membrane(p<0.01).

In general, aerobic exercise can be effective in lowering blood sugar by decreasing the activity of MondoA in the nucleus and increasing the presence of Glut4 in the cell membrane

Role of Endoplasmic Reticulum Stress in The Male Reproductive SystemTesticular dysfunction, whether linked to varicocele, obesity, diabetes, aging, inflammation, or lifestyle or environmental issues, is frequently accompanied by an accumulation of unfolded or misfolded proteins, indicating impaired endoplasmic reticulum (ER) function. In this review, we examined the Google Scholar, Scopus and PubMed databases (from 2011 to 2022) to support the association of ER stress with defective spermatogenesis in animal models and humans. ER stress, whether in its pro-survival or pro-apoptotic aspect, appears to be closely linked to each studied situation. Several studies have demonstrated a significant increase in oxidative stress (OS) levels in infertile men compared to fertile individuals, which is associated with poor spermatogenesis quality. OS is likely the result of the interplay between ER stress and spermatogenesis defects. These findings suggest that therapeutic strategies aimed at mitigating both ER stress and OS could be of interest in restoring male reproductive function.

Oligozoospermia or reduced sperm concentration is often associated with abnormal motility and morphology, reflecting abnormal spermatogenesis in the testes. The sperm DNA integrity plays an important role in embryo development, and fertility. Therefore, we aimed to assess sperm DNA integrity in a population of oligozoospermic and normozoospermic men.

In this cross-sectional study, 967 oligozoospermic samples (sperm count lower than 39 million per ejaculation), and 967 normozoospermic samples according to World Health Organization criteria were included. Sperm DNA damage was assessed by SCSA and TUNEL assays. Statistical analyzes were performed using SPSS version 22 software and a significance level of less than 0.05 was considered.

Mean sperm DNA damage and also DNA stainability were significantly higher in oligozoospermic individuals than them in normozoospermic individuals (Ps<0.001). Unlike semen volume and sperm count which was significantly lower (P<0.001), the mean of sperm abnormal morphology was significantly higher in oligozoospermic men compared to normozoospermic men (P<0.001).

In individuals with oligozoospermia, in addition to the count, morphology and motility of sperm can also be abnormal according to the WHO threshold. Also, sperm DNA damage is significantly high, which can indicate abnormal spermatogenesis and epididymal immaturity. Therefore, depending on the severity of the damage, it is preferable to manage therapeutic interventions, such as drug therapies or assisted reproductive techniques, in order to make the best decisions regarding treatment.

Spermatogenesis is a process that leads to the production and differentiation of sperm. During this process, various molecular and metabolic pathways are involved. "1-carbon cycle" is known as one of the most important metabolic cycles, which includes two cycles of folate and methionine and the transsulfuration pathway. This cycle has essential and vital roles in DNA and RNA methylation, DNA condensation and maturation, and maintaining the antioxidant balance in sperm. In this review, the importance of the 1-carbon cycle in the process of spermatogenesis has been considered.

Using the keywords of the study, related studies were reviewed from PubMed, Google Scholar and Science Direct databases between 1993 and 2021 in English, and the information of 69 selected articles was extracted.

Disturbance in the 1-carbon cycle metabolism, such as the transsulfuration pathway, the re-methylation process, folate or cobalamin deficiency, and the presence of single nucleotide polymorphism of the MTHFR gene variant C677T, can affect DNA and RNA methylation and sperm DNA integrity. An increase in homocysteine concentration, especially in individuals with folate deficiency, is associated with an increase in oxidative stress and a decrease in antioxidants in the cell, which can affect sperm function and fertility.

Strengthening the 1-carbon cycle with folate and other micronutrients may lead to the improvement of sperm parameters and fertility potential by activating the enzymes involved in the transsulfuration pathway and reducing homocysteine concentration.

Neural stem cells (NSCs) are multipotent stem cells residing in the central nervous system that is capable of self-renewal to support ongoing requirements for neurogenesis in the adult brain. Since NSCs are considered potential candidate cells for neuro-regenerative medicine, applying safe induction methods for them is very important. Synthetic modified-mRNA (mmRNA) as an alternative to traditional DNA- or protein-based methods, is regarded as a powerful tool for inducing short-term gene expression in cells with no genetic manipulation.

Here, we aimed to develop an optimized condition for mmRNA transfection in primary NSCs. In vitro-transcribed EGFP mmRNA (mmRNAEGFP) was delivered to human embryonic kidney cells (HEK293T) and mouse NSCs by using two commercial agents, Lipofectamine-2000 (LF2000) and TransIT. Also, a plasmid DNA was used to transfect cells considered EGFP-expressing positive control. In addition, the poly(A) tail (poly adenosine tail) elongation and chloroquine (CQ) treatment were performed to improve transfection efficiency. Finally, flow cytometry, fluorescence microscopy, and MTT assays were performed to assess the cells.

In comparison with HEK293T, NSCs were very sensitive to transfection, the efficacy of transfection using DNA/LF2000 was higher in HEK293T cells, but mmRNAEGFP/ TransIT showed better transfection efficacy in NSCs. Poly(A) tail elongation; also, treating the cells with CQ before transfection significantly improved its efficacy.

The mmRNA poly(A) tail elongation and the use of specific transfection agents in combination with TLR inhibitors can lead to a more effective transfection in NSCs.

Stress may have an important role in the origin and progress of depression and can impair metabolichomeostasis. The one-carbon cycle (1-CC) metabolism and amino acid (AA) profile are some of the consequencesrelated to stress. In this study, we investigated the Paroxetine treatment effect on the plasma metabolite alterationsinduced by forced swim stress-induced depression in mice.

In this experimental study that was carried out in 2021, thirty male NMRI mice (6-8 weeksage, 30 ± 5 g) were divided into five groups: control, sham, paroxetine treatment only (7 mg/kg BW/day), depressioninduction, and Paroxetine+depression. Mice were subjected to a forced swim test (FST) to induce depression and thenwere treated with Paroxetine, for 35 consecutive days. The swimming and immobility times were recorded during theinterventions. Then, animals were sacrificed, plasma was prepared and the concentration of 1-CC factors and twentyAAs was measured by spectrophotometry and high-performance liquid chromatography system (HPLC) techniques.Data were analyzed by SPSS, using One-Way ANOVA and Pearson Correlation, and P<0.05 was considered significant.

Plasma concentrations of phenylalanine, glutamate, aspartate, arginine, ornithine, citrulline, threonine,histidine, and alanine were significantly reduced in the depression group in comparison with the control group.The Homocysteine (Hcy) plasma level was increased in the Paroxetine group which can be associated withhyperhomocysteinemia. Moreover, vitamin B12, phenylalanine, glutamate, ornithine, citrulline, and glycine plasmalevels were significantly reduced in the depression group after Paroxetine treatment.

This study has demonstrated an impairment in the plasma metabolites’ homeostasis in depression andnormal conditions after Paroxetine treatment, although, further studies are required.

The intracytoplasmic sperm injection (ICSI) has significantly improved male factor infertility treatment; however, complete fertilization failure still occurs in 1-5% of ICSI treatment cycles mainly due to oocyte activation failure. It is estimated that around 40-70% of oocyte activation failure is associated with sperm factors after ICSI. Assisted oocyte activation (AOA) as an effective approach to avoid total fertilization failure (TFF) has been proposed following ICSI. In the literature, several procedures have been described to overcome failed oocyte activation. These include mechanical, electrical, or chemical stimuli initiating artificial Ca2+ rises in the cytoplasm of oocytes. AOA in couples with previous failed fertilization and those with globozoospermia has resulted in varying degrees of success. The aim of this review is to examine the available literature on AOA in teratozoospermic men undergoing ICSI-AOA and determine whether the ICSI-AOA should be considered as an adjunct fertility procedure for these patients.

Increased sperm DNA damage is known as one of the causes of recurrent pregnancy loss(RPL) which can be due to increased levels of oxidative stress. In this study, the effect of alpha-lipoic acid (ALA) as an antioxidant soluble in water and fat was investigated on sperm parameters and sperm function tests in couples with a history of RPL.

In this preliminary study, a total of 37 patients (n=12 and n=25 for placebo and ALA groups, respectively) were considered. Men with high sperm DNA damage were treated with ALA (600 mg/day) or placebo for 80 days. Semen samples were acquired from the participants before initiation and after completion of the medication course and assessed regarding conventional sperm parameters, DNA damage/integrity, intracellular oxidative stress, lipid peroxidation, and seminal antioxidant characteristics. Individuals were further followed up for twelve months for pregnancy occurrence and outcomes. Finally, after excluding patients with no history of RPL, the data was analyzed.

No statistically significant difference was observed between the baseline measures except for seminal volume. However, after the intervention, the mean sperm DNA damage (TUNEL), nuclear protamine deficiency, and persisted histones were significantly lower in the ALA group than placebo receivers (p<0.05). We noticed a decrease in the mean levels of seminal total antioxidant capacity (p=0.03), malondialdehyde (p=0.02), and SCSA-assayed sperm DNA damage (p=0.004) as well as an increase in mean sperm total motility (p=0.04) after treatment with ALA. In addition, the mean of nuclear protamine deficiency and remnant histone content were declined post-ALA therapy (p=0.003 and 0.002, respectively). Regarding post-medication pregnancy loss, no statistically significant difference was observed between the two groups (p=0.15).

ALA-therapy attenuates sperm DNA damage and lipid peroxidation while enhancing sperm total motility and chromatin compaction in the male partner of couples with RPL.

In infertility clinics, preserving high-quality spermatozoa for a long time is a necessity. Pentoxifylline (PT) and L-carnitine (LC) are effective in improving sperm motility as well as protecting the sperm membrane. The present study aimed to investigate the protective impacts of PT and LC on the quality of the normal sperm motility, protamine content, and viability on prolonged storage for 12 days at 4-6°C.

The present experimental work included 26 samples, which were first prepared based on the swim-up technique, of normozoospermic men. They were divided into three aliquots as untreated control, LC-treated, and PT-treated groups and incubated for up to 12 days at 4-6°C. Thereafter, chromatin maturity, sperm viability, and motility were assessed on 0, 1, 2, 5, 7, and 12 days. Data were analyzed using a one-way analysis of variance.

The obtained data revealed that PT supplementation increased the percentage of motile spermatozoa in comparison with control and LC-treated specimens. On the other hand, LC supplementation increased the percentage of viable spermatozoa in comparison with the PT-treated and control samples. During the 12-day storage, the percentage of spermatozoa with a normal protamine content was nearly unchanged in the three groups (P>0.05).

Although LC supplementation can be considered a better alternative than PT for preserving sperm viability, PT could better preserve sperm motility compared to LC during 12 days at 4-6°C.

Pre-diabetic stressful conditions are status before the onset of type 2 diabetes, in which the blood sugar level is higher than normal and below the diagnostic criteria for type 2 diabetes. Therefore, the purpose of this study was to investigate the effect of endurance training on muscle glycogen storage in pre-diabetic mice. For this purpose, Fifteen male C57BL / 6 mice were randomly divided into two groups: mice fed a normal diet (ND, n= 5) and a high-fat diet (HFD, n=10) and then fed for 12 weeks. After diagnosing prediabetes induction by diagnostic tests in the HFD group, this group was into two groups: the pre-diabetic group (PD) and pre-diabetic mice along with endurance training (PD-Ex). The results showed a significant decrease in blood glucose levels and insulin resistance of HFD-Ex group compared to PD. The PAS technique showed an observational decrease in glycogen storage in the HFD group compared to the ND group, while endurance training increased it in the skeletal muscle of mice. The relative gene expression of the factors involved in glycogen storage such as AKT, PI3K, Gys1, and GSK3 as well as the gene LRRC8A showed significant changes (p <0.05) between the ND group compared to the HFD group, showing the glycogen storage signaling pathway is disturbed. However, endurance training (PD-Ex) has resulted in a significant improvement in glycogen synthase gene levels. In general, Endurance training seems to improve the negative effects of pre-diabetic conditions due to a high-fat diet.

According to the World Health Organization, infertility refers to a couple's inability to conceive following at least one year of regular unprotected sexual intercourse. Male infertility accounts for about half of the various causes of couples' infertility. This review aimed to investigate the cellular and molecular differentiation of spermatozoa, focusing on the structure of the cytoskeleton, microtubules, actin filaments, motor, and non-motor proteins, to study the known genes associated with their formation and function, as well as the proteins involved in intracellular cargo transport, and ultimately investigate their essential role in maintaining sperm morphology and motility and subsequent male reproduction and infertility. The importance of microtubules in the critical processes of sperm production, transformation, maturation, and fertility of spermatozoa is such that the term "microtubule" has been recently used to denote the microtubule and all microtubule-related components in the sperm cell. The cellular process of sperm evolution and differentiation was discussed first, followed by a description of the cytoskeletal system of the acroframosome-acroplaxome-manchette axis, which is involved in acrosome formation and development, sperm head and flagellum shaping mechanisms, in response to the current and future demands of infertility researchers and clinicians in this emerging field of research. The significance of the aberrant function of different components of the sperm cytoskeleton in creating major disorders associated with male infertility was next inspected to clarify the ambiguous aspects of this complex process.

Assessment of the cytotoxicity of novel calcium silicate-based cement is imperative in endodontics. This experimental study aimed to assess the cytotoxicity and odontogenic/osteogenic differentiation potential of a new calcium silicate/pectin cement called Nano-dentine against stem cells from the apical papilla (SCAPs).

In this experimental study, the cement powder was synthesized by the sol-gel technique. Zirconium oxide was added as opacifier and Pectin, a plant-based polymer, and calcium chloride as the liquid to prepare the nano-based dental cement. Thirty-six root canal dentin blocks of human extracted single-canal premolars with 2 mm height, flared with #1, 2 and 3 Gates-Glidden drills were used to prepare the cement specimens. The cement, namely mineral trioxide aggregate (MTA), Biodentine, and the Nano-dentine were mixed according to the manufacturers’ instructions and applied to the roots of canal dentin blocks. The cytotoxicity and odontogenic/osteogenic potential of the cement were evaluated by using SCAPs.

SCAPs were characterized by the expression of routine mesenchymal cell markers and differentiation potential to adipocytes, osteoblasts, and chondrocytes. Cement displayed no significant differences in cytotoxicity or calcified nodules formation. Gene expression analysis showed that all three types of cement induced significant downregulation of COLA1; however, the new cement induced significant up-regulation of RUNX2 and SPP1 compared to the control group and MTA. The new cement also induced significant up-regulation of TGFB1 and inducible nitric oxide synthase (iNOS) compared with Biodentine and MTA.

The new Nano-dentin cement has higher odontogenic/osteogenic potential compared to Biodentine and MTA for differentiation of SCAPs to adipocytes, osteoblasts, and chondrocytes.