An Urgent Industrial Scheme Both for Total Synthesis, and for Pharmaceutical Analytical Analysis of Umifenovir as an Anti-viral API for Treatment of COVID-19.

BACKGROUND: This paper aims to reveal an urgent industrial scheme for a fast and facile total synthesis of umifenovir (arbidol) (by one-pot stages) as an antiviral agent for treating the 2019-nCoV virus via inhibiting its viral replication in the human cells. As COVID-19 takes thousands of lives all around the world, it seems that the medicinal resources would not be enough to supply billions of peoples currently living on the planet. Thus, this pandemic and its subsequent impacts on the natural order of our life would be one of the most important threats against the entire human race. METHODS: In this project, we have made attempts to find an operative approach for synthesizing this compound as an active pharmaceutical ingredient (API), which showed it could be effective in inhibiting the newly emerged coronavirus.. RESULTS: The designed scheme uses relatively cheap precursors and contains one pot stage instead of seven time-consuming and more costly linear steps. Moreover, safe and cheap solvents have been used like water and ethanol, instead of toxic ones like methanol and pyridine which could cause rejection of the API in the organic volatile impurities (OVI) test of pharmacopeia analysis, as well as increase the concern of inflammability, explosivity, and carcinogenic properties of those common solvents. CONCLUSION: The most important pharmaceutical analytical methods containing OVI test (mainly ethanol (about 171 ppm) much lower than the limits, by gas chromatography-Flame Ionization Detector (GC-FID) instrument), assay content (about 99.6% by potentiometric titration), and related purity analysis (by high-performance liquid chromatography-Ultraviolet Detector (HPLCUV)) (about 99.8%) were performed and described to give a more clear industrial scheme.

α-Lipoic acid prevents senescence, cell cycle arrest, and inflammatory cues in fibroblasts by inhibiting oxidative stress.

Senescence is a process characterized by an irreversible growth arrest in cells and induced by oxidative stress. In the current study, anti-aging potential of a well-known antioxidant, α-lipoic acid (α-LA), in rat embryonic fibroblast (REF) cells was assessed. In this regard, oxidative stress, inflammation, and apoptosis pathways were investigated on REF cells exposed to H2O2 as a senescence inducer and α-LA as a protective compound. In cells treated with α-LA and H2O2, level of ß-galactosidase, as an aging marker, and oxidative stress biomarkers, were significantly lower than those exposed to H2O2 only. Furthermore, flow cytometry assay showed that α-LA caused a significant reduction in the number of apoptotic cells via the caspase-dependent pathway. In addition, it could neutralize the inflammatory effects of H2O2 and attenuated the concentration of inflammatory cytokines. In comparison to H2O2 group, a significant increase in G0/G1 arrest was observed during cell cycle analysis in cells exposed to H2O2 and α-LA. The results of this study show that α-LA has beneficial effects on H2O2-induced cellular senescence. α-LA works by attenuating the reactive oxygen species, subsiding inflammation, and affecting cell division.

Regulation of aging and oxidative stress pathways in aged pancreatic islets using alpha-lipoic acid.

Oxidative stress has been involved in the aging process and the pathogenesis of type-2 diabetes, which is a serious health problem worldwide. This study investigates the anti-aging, anti-apoptotic, and antioxidant properties of alpha-lipoic acid (ALA), aiming to improve aged rat pancreatic cells. In this regard, half maximal effective concentration (EC50) of ALA based on the survival of aged pancreatic islet cells was determined as 100 µM. Following this, p38 and p53 genes expression as key factors in aging, oxidative stress biomarkers, insulin secretion, and Pdx1 protein expression were evaluated using real-time PCR, ELISA reader, and fluorescence microscope. It was revealed that ALA reduces and controls the effects of aging on beta cells mainly by suppressing p38 and p53 at the gene level (P < 0.001 and P < 0.01), respectively, reducing reactive oxygen species (P < 0.001) and enhancing levels of thiols (P < 0.05) compared with the aged islets. Furthermore, both qualitative and quantitative investigations of insulin secretion have shown that ALA can improve aged cells' function and increase insulin secretion specially in the stimulating concentration of glucose. Also, the expression of Pdx1 was considerably increased by ALA in comparison to the aged pancreatic islets (P < 0.001). As far as the authors of the present study are concerned, this is the first study, which evaluated aging associated with p38 and p53 pathways, oxidative stress parameters, and the expression of insulin in beta cells of an aged rat and reaffirmed the fact that ALA has a significant antioxidant role in reducing the aging process.

On the mechanism of genotoxicity of ethephon on embryonic fibroblast cells.

Ethephon is one of the most widely used plant growth regulator in agriculture that its application has been increased in recent years. Many reports have raised concern over the safety of this organophosphorus compound. The aim of the current study was to assess the potential genotoxic effect of ethephon on murine embryonic fibroblast (MEF) cell line, using two genotoxicity endpoints: γH2AX expression and comet assay. γH2AX served as an early and sensitive biomarker of genotoxic damage. Oxidative stress biomarkers, including reactive oxygen species (ROS), lipid peroxidation (LPO) and total antioxidant capacity were also examined. The results showed a significant increase in cell proliferation, 24 h post-treatment with 10, 40,160 µg/ml ethephon, while at the higher concentrations cytotoxic effect was observed. The γH2AX expression and γH2AX foci count per cell were significantly increased at non-cytotoxic concentrations of ethephon, accompanied with increased DNA damage as illustrated by comet assay. LPO and ROS levels were elevated only at 160 µg/ml and higher doses. The results interestingly showed that low non-cytotoxic doses of ethephon promoted DNA damage inducing cell proliferation, raising the possibility of ethephon mutagenicity. The genotoxic effect of ethephon at low doses might not relate to oxidative damage and that increased in the level of ROS and LPO generation at higher doses could account for the cytotoxic effect of ethephon. Taken together, our study provides strong in vitro evidence on potential genotoxicity of ethephon at low doses. More precise studies are needed to clarify the mutagenic effect of chronic exposure to ethephon.

Amniotic membrane mesenchymal stem cells can differentiate into germ cells in vitro.

This is the first report on differentiation of mouse amniotic membrane mesenchymal stem cells (AM-MSCs) into male germ cells (GCs). AM-MSCs have the multipotent differentiation capacity and can be differentiated into various cell types. In the present study, AM-MSCs were induced for differentiation into GCs. AM-MSCs were isolated from mouse embryonic membrane by enzymatic digestion. AM-MSCs were characterized with osteogenic and adipogenic differentiation test and flow cytometric analysis of some CD-markers. AM-MSCs were induced to differentiate into GCs using a creative two-step method. Passage-3 AM-MSCs were firstly treated with 25 ng/ml bone morphogenetic protein 4 (BMP4) for 5 d and in continuing with 1 µM retinoic acid (RA) for 12 d (total treatment time was 17 d). At the end of the treatment period, real-time reverse transcription (RT)-PCR was performed to evaluate the expression of GC-specific markers-Itgb1, Dazl, Stra8, Piwil2, Mvh, Oct4, and c-Kit- in the cells. Moreover, flow cytometry and immunofluorescence staining were performed to evaluate the expression of Mvh and Dazl at protein level. Real-time RT-PCR showed that most of the tested markers were upregulated in the treated AM-MSCs. Furthermore, flow cytometric and immunofluorescence analyses both revealed that a considerable part of the treated cells expressed GC-specific markers. The percentage of positive cells for Mvh and Dazl was about 23 and 46%, respectively. Our results indicated that a number of AM-MSCs successfully differentiated into the GCs. Finally, it seems that AM-MSCs would be a potential source of adult pluripotent stem cells for in vitro generation of GCs and cell-based therapies for treatment of infertility.

Growing knowledge of using embryonic stem cells as a novel tool in developmental risk assessment of environmental toxicants.

Developmental toxicology is an important area of novel toxicology. In recent years, there have been big concerns toward the increasing exposure to pharmaceutical agents, food additives, pesticides, occupational toxicants, and environmental pollutants, as well as their possible association with all aspects of male or female-mediated transient or permanent defects in progeny. Therefore, it is of great importance to look for new predictive models to evaluate environmental toxicants before they can harm the human health and embryo development. In this regard, new cell-based in vitro screening models have been developed and validated in predictive toxicology to minimize assay costs and animal usage. Stem cell-based models have been increasingly applied for predicting the toxicity of chemicals. One of the most promising existing in vitro developmental toxicity tests is the validated embryonic stem cell test (EST) which employs marine or human embryonic stem cells to assess the potential of chemicals embryotoxicity. These cells are very suitable for embryotoxicity assessment as they have been demonstrated to specify cellular developmental processes during early embryogenesis and gene expression patterns of differentiation to functionally competent specialized cell types. The present paper aimed at criticizing the human and experimental evidence for developmental toxic effects of environmental toxicants based on ESCs models. Accordingly, pesticides, heavy metals, plasticizers, nanomaterials and some solvents have been considered as the main evaluated environmental toxicants inducing developmental toxicity. At the end, current challenges, pros and cons of using ESCs as an alternative validated in vitro model for specific developmental toxicity screening are discussed.

Reduction of chlorpyrifos-induced toxicity in human lymphocytes by selected phosphodiesterase inhibitors.

One of the most famous and commonly used compounds from organophosphate (OP) family is chlorpyrifos (CP) which is widespreadly used as a powerful insecticide. Previous studies have shown that OPs induce oxidative stress, inflammation and apoptosis by generating the free radicals. The protective effects of three members of phosphodiesterase inhibitor (PDEI) family, including rolipram (RLP), milrinon (MLR) and pentoxifylline (PTX) were evaluated in the human lymphocytes against CP's toxicity. In this case, the level of oxidative stress biomarkers, the viability of the cells and the rate of apoptosis by flow cytometry were investigated. The results of this study revealed that CP makes a significant increase in the level of inflammatory and oxidative stress markers such as meyloperoxidase (MPO), lipid peroxidation (LPO), total thiol molecules (TTM) and total antioxidant potential (TAP), and also makes an enhancement in the rate of apoptosis process. On the other hand, PDEIs and specifically the combination of them restored the negative effects of CP and significantly prevented the apoptosis and oxidative stress imbalance. It is concluded that these PDEIs have positive effects in attenuation, recovery, and protection of CP-induced toxicity in the human lymphocytes.

Dual targeting of TNF-α and free radical toxic stress as a promising strategy to manage experimental polycystic ovary.

CONTEXT: It is now clear that oxidative stress (OS) and chronic low-grade inflammation are two main pathways involved in polycystic ovary syndrome (PCOS) pathogenesis. Therefore, simultaneous targeting of these pathways by means of carvedilol and Semelil (ANGIPARS™), as established medicines with dual anti-cytokine and anti-oxidant potential may be a therapeutic alternative approach to the current treatments. OBJECTIVE: The objective of this study is to study the protective effects of carvedilol and ANGIPARS™ on inflammatory and oxidative response in hyperandrogenism-induced polycystic ovary (PCO). MATERIALS AND METHODS: The murine model of PCO was induced by letrozole (1 mg/kg/d; orally) and effective doses of carvedilol (10 mg/kg/d; orally) and ANGIPARS™ (2.1 mg/kg/d; orally) were administrated for 21 d in PCO and non-PCO healthy rats. Ovarian folliculogenesis, sex hormones concentrations, OS, inflammatory, and metabolic biomarkers were assessed in serum and ovaries. RESULTS: PCO rats exhibited ovarian cystogenesis which was preserved by the application of carvedilol and ANGIPARS™. In comparison with controls, decreased level of the total antioxidant power (TAP) and higher levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) in serum and ovaries (2.41 ± 0.67 versus 0.72 ± 0.11; and 0.17 ± 0.04 versus 0.05 ± 0.01; 5.48 ± 1.30 versus 10.56 ± 0.77; and 7.06 ± 1.94 versus 17.98 ± 0.98; p < 0.05, respectively) were detected in PCO rats. Moreover, the PCO rats exhibited hyperandrogenism due to a 3.7-fold increase in serum testosterone concentration (35.04 ± 3.17 versus 131.09 ± 13.24; p < 0.05) along with a 2.98-fold decrease in serum progesterone (6.19 ± 0.40 versus 18.50 ± 1.03; p < 0.05) and 5.2-fold decrease in serum estradiol (9.30 ± 0.61 versus 48.3 ± 2.10; p < 0.05) when compared with those of the control group. However, similar to the control group, normal levels of OS markers and sex hormones were detected in ANGIPARS™ and carvedilol co-treated PCO rats. Besides, when compared with controls, increased levels of TNF-α (770.75 ± 42.06 versus 477.14 ± 28.77; p < 0.05) and insulin (1.27 ± 0.10 versus 0.36 ± 0.05; p < 0.05) in PCO rats were significantly inhibited by carvedilol and ANGIPARS™ co-treatment. DISCUSSION AND CONCLUSION: We evidenced the beneficial effects of carvedilol and ANGIPARS™ in PCO, which underpin the new alternative approach in using these kinds of medicines in female reproductive disorders.

A systematic review on the role of environmental toxicants in stem cells aging.

Stem cells are an important target for environmental toxicants. As they are the main source for replenishing of organs in the body, any changes in their normal function could affect the regenerative potential of organs, leading to the appearance of age-related disease and acceleration of the aging process. Environmental toxicants could exert their adverse effect on stem cell function via multiple cellular and molecular mechanisms, resulting in changes in the stem cell differentiation fate and cell transformation, and reduced self-renewal capacity, as well as induction of stress-induced cellular senescence. The present review focuses on the effect of environmental toxicants on stem cell function associated with the aging process. We categorized environmental toxicants according to their preferred molecular mechanism of action on stem cells, including changes in genomic, epigenomic, and proteomic levels and enhancing oxidative stress. Pesticides, tobacco smoke, radiation and heavy metals are well-studied toxicants that cause stem cell dysfunction via induction of oxidative stress. Transgenerational epigenetic changes are the most important effects of a variety of toxicants on germ cells and embryos that are heritable and could affect health in the next several generations. A better understanding of the underlying mechanisms of toxicant-induced stem cell aging will help us to develop therapeutic intervention strategies against environmental aging. Meanwhile, more efforts are required to find the direct in vivo relationship between adverse effect of environmental toxicants and stem cell aging, leading to organismal aging.

Wound Healing Activity of a Traditionally Used Poly Herbal Product in a Burn Wound Model in Rats.

BACKGROUND: Burns are known as one of the most common and destructive forms of injury with a vast spectrum of consequences. Despite the discovery of various antibacterial and antiseptic agents, burn wound healing still has remained a challenge to modern medicine. Plants, with a valuable traditional support, have been considered as potential agents for prevention and treatment of disorders in recent years. However, modern scientific methods should be applied to validate the claims about the therapeutic effects of the herbal products. OBJECTIVES: This study was conducted to evaluate the wound-healing activity of a poly herbal cream (PHC), retrieved from Iranian Traditional Medicine (ITM), in a rat burn wound model in Iran. MATERIALS AND METHODS: In this experimental study, PHC containing aqueous extracts of Malva sylvestris and Solanum nigrum leaves and oily extract of Rosa damascena petals was used. Second-degree burn wounds were induced in four groups of five rats each. Group 1 received no treatment while groups 2, 3 and 4 were given cream base, silver sulfadiazine (SS) 1% and PHC, respectively to compare the efficacy of PHC with the negative and positive control groups. The percentage of wound healing on days 2, 6, 10 and 14 and histopathological parameters of healed wounds on the 14th day were assessed. Antioxidant and antimicrobial activities of PHC were evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and micro-dilution methods, respectively. RESULTS: There was a significant improvement in healing percentage of PHC-treated rats in comparison to the other groups at the end of the treatment period (87.0% ± 2.1% for PHC in comparison to 32.2% ± 1.6%, 57.0% ± 5.3% and 70.8% ± 3.5% for the control, cream base and SS groups, respectively). Moreover, the healed wounds in PHC-treated animals contained less inflammatory cells and had desirable re-epithelialization with remarkable neovascularization. In addition to the antioxidant activity, PHC exhibited antibacterial effect against Staphylococcus aureus. CONCLUSIONS: Poly herbal cream experimentally and histopathologically revealed a burn wound healing activity probably due to the antioxidant, anti-inflammatory and antimicrobial activities of its phytochemical contents. Therefore, this study confirms the use of M. sylvestris, S. nigrum and R. damascena in burn prescriptions in ITM.

On The Protection by The Combination of CeO2 Nanoparticles and Sodium Selenite on Human Lymphocytes against Chlorpyrifos-Induced Apoptosis In Vitro.

OBJECTIVE: Chlorpyrifos (CP) as an organophosphorus pesticide is thought to induce oxidative stress in human cells via producing reactive oxygen species (ROS) that leads to the presence of pathologic conditions due to apoptosis along with acetylcholinesterase (AChE) inhibition.This study aimed to evaluate the apoptotic effects of CP and to assess the protective potential of CeO2nanoparticle (CNP) and sodium selenite (SSe) by measuring cascades of apoptosis, oxidative stress, inflammation, and AChE inhibition in human isolated lymphocytes. MATERIALS AND METHODS: In the present experimental study, we examined the anti-oxidative and AChE activating potential of CNP and SSe in CP-treated human lymphocytes. Therefore, the lymphocytes were isolated and exposed to CP, CP+CNP, CP+SSe, and CP+CNP+SSe after a three-day incubation. Then tumor necrosis factor-alpha (TNF-α) release, myeloperoxidase (MPO) activity, thiobarbituric acid-reactive substances (TBARS) levels as inflammatory/oxidative stress indices along with AChE activity were assessed. In addition, the apoptotic process was measured by flow cytometry. RESULTS: Results showed a significant reduction in the mortality rate, TNF-α, MPO activity, TBARS, and apoptosis rate in cells treated with CNP, SSe and their combination. Interestingly, both CNP and SSe were able to activate AChE which is inhibited by CP. The results supported the synergistic effect of CNP/SSe combination in the prevention of apoptosis along with oxidative stress and inflammatory cascade. CONCLUSION: CP induces apoptosis in isolated human lymphocytes via oxidative stress and inflammatory mediators. CP firstly produces ROS, which leads to membrane phospholipid damage. The beneficial effects of CNP and SSe in reduction of CP-induced apoptosis and restoring AChE inhibition relate to their anti-oxidative potentials.

In vitro protection of human lymphocytes from toxic effects of chlorpyrifos by selenium-enriched medicines.

OBJECTIVES: Chlorpyrifos (CP) is a broad-spectrum organophosphorus pesticide used extensively in agricultural and domestic pest control, accounting for 50% of the global insecticidal use. In the present study, protective effects of two selenium-enriched strong antioxidative medicines IMOD and Angipars were examined in human lymphocytes treated with CP in vitro. MATERIALS AND METHODS: Isolated lymphocytes were exposed to 12 µg/ml CP either alone or in combination with effective doses (ED50) of IMOD (0.2 µg/ml) and Angipars (1 µg/ml). After 3 days incubation, the viability and oxidative stress markers including cellular lipid peroxidation (LPO), myeloperoxidase (MPO), total thiol molecules (TTM), and total antioxidant power (TAP) were evaluated. Also, the levels of tumor necrosis factor-α (TNF-α), as inflammatory index along with acetylcholinesterase (AChE) activity and cell apoptosis were assessed by flow cytometry. RESULTS: Results indicated that effective doses of IMOD and Angipars reduced CP-exposed lymphocyte mortality rate along with oxidative stress. Both agents restored CP-induced elevation of TNF-α and protected the lymphocytes from CP-induced apoptosis and necrosis. CONCLUSION: Overall, results confirm that IMOD and Angipars reduce the toxic effects associated with CP through free radical scavenging and protection from apoptosis and necrosis.

A comprehensive review of pesticides and the immune dysregulation: mechanisms, evidence and consequences.

Nowadays, in many communities, there is a growing concern about possible adverse effects of pesticides on human health. Reports indicate that during environmental or occupational exposure, pesticides can exert some intense adverse effects on human health through transient or permanent alteration of the immune system. There is evidence on the relation between pesticide-induced immune alteration and prevalence of diseases associated with alterations of the immune response. In the present study, direct immunotoxicity, endocrine disruption and antigenicity have been introduced as the main mechanisms working with pesticides-induced immune dysregulation. Moreover, the evidence on the relationship between pesticide exposure, dysregulation of the immune system and predisposition to different types of psychiatric disorders, cancers, allergies, autoimmune and infectious diseases are criticized.

A Systematic Review of the Molecular Mechanisms of Uranium -Induced Reproductive Toxicity.

Uranium is the heaviest metal known as nuclear fuel, and employed in the production of glass tinting compounds, ceramic glazes, gyroscope wheels, chemical catalysts and X-ray tube targets. Inhalation and ingestion are two of the most usual ways of exposure. Uranium may be released into drinking water through the mining leading to contamination. Uranium is able to damage the DNA by generation of free radicals and acting as a catalyst in the Fenton reactions causing oxidative stress. In fact, reproductive system contains high amount of polyunsaturated fatty acids, and therefore it is highly vulnerable to reactive oxygen species (ROS) and sensitive to uranium toxicity. Toxic effects of uranium are generally reported through different mechanisms of action including inflammation, degeneration of testis, vacuolization of Leydig cells, spermatocytes necrosis, and oocyte dysmorphism. The present article provides a comprehensive review of the recent findings mostly about the molecular and biochemical toxicity of uranium on the reproductive system.

Ovarian aging-like phenotype in the hyperandrogenism-induced murine model of polycystic ovary.

There are prominently similar symptoms, effectors, and commonalities in the majority of characteristics between ovarian aging and polycystic ovarian syndrome (PCOS). Despite the approved role of oxidative stress in the pathogenesis of PCOS and aging, to our knowledge, the link between the PCO(S) and aging has not been investigated yet. In this study we investigated the possible exhibition of ovarian aging phenotype in murine model of PCO induced by daily oral administration of letrozole (1 mg/kg body weight) for 21 consecutive days in the female Wistar rats. Hyperandrogenization showed irregular cycles and histopathological characteristics of PCO which was associated with a significant increase in lipid peroxidation (LPO) and reactive oxygen species (ROS) and decrease in total antioxidant capacity (TAC) in serum and ovary. Moreover, serum testosterone, insulin and tumor necrosis factor-alpha (TNF-α) levels, and ovarian matrix metalloproteinase-2 (MMP-2) were increased in PCO rats compared with healthy controls, while estradiol and progesterone diminished. Almost all of these findings are interestingly found to be common with the characteristics identified with (ovarian) aging showing that hyperandrogenism-induced PCO in rat is associated with ovarian aging-like phenotypes. To our knowledge, this is the first report that provides evidence regarding the phenomenon of aging in PCO.

On the benefit of galls of Quercus brantii Lindl. in murine colitis: the role of free gallic acid.

INTRODUCTION: In this study we investigated the effect of gall of Quercus brantii Lindl., a traditional Iranian medicine, in a murine model of experimental colitis induced in male rats by rectal administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS). MATERIAL AND METHODS: Quantification of the main active components was done for estimation of total phenolic content and free gallic acid. Gall of Quercus brantii Lindl. in two forms (gall powder and gall hydro alcoholic extract) was gavaged for 10 days (500 mg/kg). Ten days after induction of colitis, colonic status was examined by macroscopic, microscopic and biochemical analyses. Colonic tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) were analyzed as biomarkers of inflammatory condition. To determine the role of oxidative stress (OS) in colitis, the levels of cellular lipid peroxidation (LPO), total antioxidant power (TAP) and myeloperoxidase (MPO) were measured in colon tissues. RESULTS: TNBS-induced colitis exhibited a significant increase in colon MPO activity and concentrations of cellular LPO, TNF-α and IL-1ß, while TAP was significantly reduced. Microscopic evaluations of the colonic damage in the colitis group revealed multifocal degenerative changes in the epithelial lining and areas of necrosis, extensive mucosal and sub-mucosal damage with congested blood vessels, edema and hemorrhages along with extensive infiltration of inflammatory cells. Parameters including macroscopic and microscopic scores, TNF-α, IL-1ß, LPO, TAP and MPO improved by both gall extract and gall powder of Quercus brantii Lindl. and reached close to normal levels. The level of total phenols (GAE/100 g of sample) and free gallic acid were estimated to be 88.43 ±7.23 (mean ± SD) and 3.74% of dry weight, respectively. CONCLUSIONS: The present study indicates that the gall of Quercus brantii Lindl. is able to exert antioxidative and anti-inflammatory effects on the biochemical and pathological parameters of colitis.

Mesenchymal stem cell derived microvesicles: trophic shuttles for enhancement of sperm quality parameters.

Diminishing sperm quality during cryopreservation process ends up in a complete or partial loss of sperm's fertilizing potential. Rehabilitation of such affected sperm is crucial to improve their fertilizing potential. A variety of evidence has indicated that secreted microvesicles (MVs) from mesenchymal stem cells (MSCs) are involved in regeneration of injured endogenous cells via shuttling MSC trophic molecules. Sperm obtained from cauda epididymides of adult male Wistar rats were equally assigned to four separate groups. Following suspension in cryoprotectant extender, sperm were untreated or treated with increasing concentrations of MSC-derived MVs (25, 50 and 100µg). After incubation in successive steps, sperm were cryopreserved. The frozen-thawed sperm were assessed for viability, motility and antioxidant capacity parameters. Consequently, expression levels of surface adhesion molecules (CD29, CD44, ICAM-I and VCAM-I) involved in sperm fusogenic and signaling properties, were assessed by flow cytometry. Results showed an enhanced quality parameters and adhesive properties of cryopreserved sperm following treatment with MSC-derived MVs.

Protection of cisplatin-induced spermatotoxicity, DNA damage and chromatin abnormality by selenium nano-particles.

Cisplatin (CIS), an anticancer alkylating agent, induces DNA adducts and effectively cross links the DNA strands and so affects spermatozoa as a male reproductive toxicant. The present study investigated the cellular/biochemical mechanisms underlying possible protective effect of selenium nano-particles (Nano-Se) as an established strong antioxidant with more bioavailability and less toxicity, on reproductive toxicity of CIS by assessment of sperm characteristics, sperm DNA integrity, chromatin quality and spermatogenic disorders. To determine the role of oxidative stress (OS) in the pathogenesis of CIS gonadotoxicity, the level of lipid peroxidation (LPO), antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) and peroxynitrite (ONOO) as a marker of nitrosative stress (NS) and testosterone (T) concentration as a biomarker of testicular function were measured in the blood and testes. Thirty-two male Wistar rats were equally divided into four groups. A single IP dose of CIS (7 mg/kg) and protective dose of Nano-Se (2 mg/kg/day) were administered alone or in combination. The CIS-exposed rats showed a significant increase in testicular and serum LPO and ONOO level, along with a significant decrease in enzymatic antioxidants levels, diminished serum T concentration and abnormal histologic findings with impaired sperm quality associated with increased DNA damage and decreased chromatin quality. Coadministration of Nano-Se significantly improved the serum T, sperm quality, and spermatogenesis and reduced CIS-induced free radical toxic stress and spermatic DNA damage. In conclusion, the current study demonstrated that Nano-Se may be useful to prevent CIS-induced gonadotoxicity through its antioxidant potential.

ADMET considerations for phosphodiesterase-5 inhibitors.

INTRODUCTION: ADMET (absorption, distribution, metabolism, excretion, and toxicity) profiling is an important aspect of all drug developments. The pharmaceutical industry must always consider ADMET properties in order to optimize drug candidates and to introduce new formulations against existing marketed drugs. Consequently, candidate drug development may be halted early in the discovery phase or during the more costly drug development process because of their poor ADMET properties. AREAS COVERED: The main focus of this article is ADMET profiling, pharmacokinetic (PK) drug interactions, mechanisms and possible adverse drug reactions (ADRs) for approved phosphodiesterase-5 inhibitors (PDE5Is). The authors also look at the efficacy and non-erectogenic benefits of current PDE5Is, which are widely used by patients with erectile dysfunction (ED). The authors also discuss other unapproved PDE5Is such as aildenafil and udenafil, which are currently in use in clinical trials. EXPERT OPINION: The authors believe that the enhancing effect of PDE5Is on the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway means that PDE5Is could be used to treat various conditions. An important issue in their development is 'cross-talk' between PDE5 and other PDEs and thus their specificity for other PDEs. But while it might be difficult to achieve the ideal ADMET profile, it should not necessarily prevent further development of a lead PDE5I. The risk assessment of PDE5Is, with respect to their ADMET properties, is therefore very important for predicting drug-drug interactions, possible side effects, ADRs and its future clinical applications.

Benefit of Satureja khuzestanica in subchronically rat model of cyclophosphamide-induced hemorrhagic cystitis.

Cyclophosphamide (CP) as a widely used antineoplastic drug causes hemorrhagic cystitis (HC) mainly via induction of oxidative stress. Regarding established antioxidant potential of Satureja khuzestanica (Lamiaceae) essential oil (SKEO), we aimed to investigate its protective effects in a subchronic rat model of CP-induced HC. CP (6mg/kg/day) and SKEO (225mg/kg/day) were administered alone or in combination by gavage for 28 days. Histopathological changes were investigated by light microscopy. Plasma samples were assayed for lipid peroxidation and total antioxidant power as biomarkers of toxic stress. In the CP-treated animals, irregular mucus layer, severe hemorrhage and edema, infiltration of inflammatory cells, and accumulation of mast cells were observed. In the CP+SKEO group, a relatively normal urothelial topography with decreased number of mucosal mast cells and inflammatory cells were observed. Increased lipid peroxidation along with decreased total antioxidant capacity resulting from CP treatment was significantly recovered by SKEO co-treatment. It is concluded that SKEO protects rats from CP-induced HC by reduction of free radical-induced toxic stress. It is strongly recommended to examine SKEO in the clinic to approve its benefit in patients undertaking CP.