Correction: Probiotic and prebiotic supplementation ameliorates chronic restraint stress-induced male reproductive dysfunction.

Correction for 'Probiotic and prebiotic supplementation ameliorates chronic restraint stress-induced male reproductive dysfunction' by Mohd Akram et al., Food Funct., 2023, 14, 8558-8574, https://doi.org/10.1039/D3FO03153E.

Probiotic and prebiotic supplementation ameliorates chronic restraint stress-induced male reproductive dysfunction.

Restraint stress (RS) can induce male reproductive deficits by activating the hypothalamic-pituitary-adrenal (HPA) axis and causing oxidative stress. Previous studies have shown that probiotics can alleviate neurological and metabolic disorders induced by stress. However, the effects of probiotics on RS-induced reproductive deficits have not been fully elucidated. This study aimed to investigate whether Lactobacillus rhamnosus NCDC-610 (Probiotic-1) and Lactobacillus fermentum NCDC-400 (Probiotic-2) with prebiotic (fructooligosaccharides (FOS)) could prevent RS-induced reproductive deficits. C57BL6/J mice were subjected to RS for four hours daily before oral administration of probiotics (4 × 109 CFU per mice) either separately or concurrently with FOS. The results showed that oral administration of Probiotic-1 and Probiotic-2 protected against RS-induced sperm deficits, including sperm count, motility, morphology, and histopathology of testes, and improved intestinal health. Furthermore, Probiotic-1 and Probiotic-2 prevented RS-induced changes in testosterone levels by up-regulating the expressions of steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage enzyme (P450scc), and 17ß-hydroxysteroid dehydrogenase (17ßHSD) in the testes. Additionally, Probiotic-1 and Probiotic-2 increased the activities of catalase and superoxide dismutase and reduced the fold change of interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-α), indicating a protective effect against RS-induced oxidative stress. Oral administration of Probiotic-1 and Probiotic-2, either separately or concurrently with FOS (probiotic dose of 4 × 109 CFU per mice and prebiotic 5% w/v), prevented RS-induced activation of the HPA axis and improved male fertility. These findings suggest that L. rhamnosus NCDC-610 and L. fermentum NCDC-400 are safe and effective probiotics for mitigating stress-induced male reproductive deficits.

Protective effect of probiotic and prebiotic fermented milk containing Lactobacillus fermentum against obesity-induced hepatic steatosis and inflammation.

Obesity has reached epidemic proportions, with major economic and health implications. The complex pathophysiology of obesity explains the difficulty provided to health policy for its clinical management. Increasing data show that obesity and metabolic abnormalities are intimately connected to differences in consumption of probiotics, its relevance to gut microbiota activity and composition. The goal of this investigation was to assess the effect of oral delivery of indigenous probiotic Lactobacillus fermentum NCDC 400 and prebiotic fructo-oligosaccharide (FOS) on obesity-associated hepatic steatosis and inflammation produced by a high-fat diet (HFD). C57BL/6 mice treated with L. fermentum NCDC 400 either independently or in conjunction with FOS demonstrated reduced body weight and abdominal obesity after 24 weeks of treatment. Also, the anti-oxidative enzyme activity went down, and the inflammatory profile got better, with less fat getting into the hepatocytes. The lipid profile changed, with HDL cholesterol going up and LDL cholesterol and triglyceride levels going down. Further, L. fermentum NCDC 400 and FOS combinations decreased fasting glucose, gHbA1c, gastric inhibitory peptide, and insulin levels in mice fed with HFD, thus improving glucose homeostasis. Overall, consumption of L. fermentum NCDC 400 alone or its combinational effects had a protective role on obesity-associated hepatic steatosis. PRACTICAL APPLICATIONS: The potential indigenous probiotic Lactobacillus fermentum NCDC 400 and prebiotic FOS had a preventive role in obesity-induced hepatic steatosis and improves anti-oxidant and anti-inflammatory properties in HFD-fed obese mice. Our finding would be helpful to prevent obesity-associated hepatic steatosis and inflammation upon supplementation of pre- and pro-biotics (synbiotics).

Dietary intake of probiotic fermented milk benefits the gut and reproductive health in mice fed with an obesogenic diet.

Probiotics have been suggested as alternatives to pharmacological drugs in the treatment of a variety of medical problems, including obesity management, which is often linked to low sperm production. Also, probiotic fermented products are known to boost host immune response, immunosenescence, infection tolerance, and redox homeostasis, but their direct role in male fertility has been less investigated. This study assessed the effect of two probiotic strains, L. fermentum NCDC 400 and L. rhamnosus NCDC 610, and fructooligosaccharide (FOS) fermented milk supplementation. We identified the significantly reduced oxidative stress markers in the plasma and liver of HF diet-fed animals. We determined the role of key testicular enzymes of steroidogenic pathway genes StAR, P450scc, and 17ßHSD in maintaining the testosterone concentration and restoring testicular structures. In conclusion, the present work illustrated the ability of both probiotics L. fermentum NCDC 400 and L. rhamnosus NCDC 610 as regulatory agents with beneficial effects on weight loss and endogenous testosterone with substantially improved sperm motility in male diet-induced obesity (DIO) models. Our findings indicate that fermented milk supplementation may be an alternative treatment for preventing obesity and other related metabolic syndromes.

Novel and known miRNAs in zebu (Tharparkar) and crossbred (Karan-Fries) cattle under heat stress.

MicroRNAs (miRNAs) are small single-stranded non-coding RNAs that act as the master regulator of animal growth and development. RNA-RNA interaction is an important mechanism of gene regulation during biotic and abiotic stress. Heat stress (HS) is one of the most important abiotic factors which affect the growth, milk yield and reproductive health of the dairy animals. In the present investigation, we identified 387 known and 77 novel miRNAs from Tharparkar (TH) and Karan Fries (KF) cattle under HS condition. Family distribution analysis showed the identified miRNAs belong to more than 15 different families in which miR-2284 was the most abundant. We identified 42,350 targets for the known miRNAs reported in cattle. Pathway analysis of the identified targets showed most of the target genes were involved in cancer, mitogen-activated protein kinase (MAPK) signaling, calcium signaling, Ras signaling, and cAMP signaling pathways. Differential gene expression showed more than 344 miRNAs changed their expression significantly between control and HS condition. Heat map was generated for the top 20 most up and downregulated miRNAs. Ten miRNAs were validated using qRT-PCR to be heat responsive, based on read count value and differential gene expression. These novel miRNAs are new addition to the miRNA database of cattle. This study provides an overview of miRNA profile and their interaction with the target genes which leads to further understanding in deciphering the thermotolerance mechanism in cattle.

MSN, MWCNT and ZnO nanoparticle-induced CHO-K1 cell polarisation is linked to cytoskeleton ablation.

BACKGROUND: The cellular response to nanoparticles (NPs) for the mechanical clue and biochemical changes are unexplored. Here, we provide the comprehensive analysis of the Chinese Hamster Ovary (CHO-K1) cell line to study cell behaviour following the exposure of mesoporous silica nanoparticle (MSN), multiwall carbon nanotubes (MWCNTs), and zinc oxide (ZnO) NPs. RESULTS: Through the high-throughput proteomic study, we observed that the effect of NPs is alone not restricted to cell viability but also on cell polarisation. In the case of MSN, no drastic changes were observed in cellular morphology, but it upregulated chaperons that might prevent protein aggregation. However, MWCNT showed elongated cell appearance with numerous cytoplasmic vacuoles, and induce lamellipodia formation through actin polymerisation. The cytoskeleton remodelling was accompanied by the increased expression of Dlc-1, cofilin and Rac1 proteins. While ZnO NPs resulted in the rounded cell morphology along with nuclear abnormalities. The proteome analysis revealed that UBXN11 control cell roundness and DOCK3 leads to actin stress fibre formation and finally, loss of cell adhesion. It enhances the expression of catastrophic DNA damage and apoptotic proteins, which was unrecoverable even after 72 h, as confirmed by the colony formation assay. All three NPs trigger over-expression of the endocytic pathway, ubiquitination, and proteasomal complex proteins. The data indicate that ZnO and MSN entered into the cells through clathrin-mediated pathways; whereas, MWCNT invades through ER-mediated phagocytosis. CONCLUSIONS: Based on the incubation and concentration of NPs, our work provides evidence for the activation of Rac-Rho signalling pathway to alter cytoskeleton dynamics. Our results assist as a sensitive early molecular readout for nanosafety assessment.

Exploration of immunomodulatory and protective effect of Withania somnifera on trace metal oxide (zinc oxide nanoparticles) induced toxicity in Balb/c mice.

The current study was undertaken to investigate the immunomodulatory and protective effects of Withania somnifera (WS) extract and Withaferin A (WA) supplementation on zinc oxide nanoparticles mediated toxicity in Balb/c mice. The animals were exposed to ZnO NPs along with WS and WA for 28 days and various parameters like body weight, organ coefficient, cytotoxicity, nitric oxide (NO), total serum protein, phagocytosis, and the gene expression levels of TLR6 and ARG genes were determined. In vivo study showed that, dose-dependent reduction in phagocytosis, an increase in the levels of NO production along with up-regulation of TLR6, arginase gene was significant (P < 0.05) when ZnO NPs were given. However toxicity of ZnO NP was reduced in presence of WS and WA with decreased TLR6 over expression and restoration of phagocytic activities. Our results provided a valuable insight into the underlying mechanism for the protective effects of WS. Mechanism of toxicity induced by Zinc oxide nanoparticles ZnO NPs and immunomodulatory protective effects of Withania somnifera extract (WS) and Withaferin A (WA), in Balb/c mice modal of peritoneal macrophages. Red arrows: effect of ZnO NPs independently leads to ROS production which attenuated the phagocytosis of yeast by macrophages through, up-regulation of TLR6 and down-regulation of arginase gene expression. Green arrows: co-treatment, Impact of Withania somnifera extract with zinc oxide nanoparticles (WS + ZnO NPs), Withaferin A along with zinc oxide nanoparticles (WA + ZnO NPs)-enhance phagocytic activity by counteracting mechanism of ZnO NPs toxicity. Black arrows: increasing or decreasing effects. Per oral (P.O).

Comparative evaluation of half-maximum inhibitory concentration and cytotoxicity of silver nanoparticles and multiwalled carbon nanotubes using buffalo bull spermatozoa as a cell model.

There is a dearth of information regarding the safety of silver nanoparticles (Ag NPs) and multiwalled carbon nanotubes (MWCNTs) with respect to their impact on human/animal health and the environment. This study aimed to determine the half-maximum inhibitory concentration (IC50) of Ag NPs and MWCNTs by employing different doses and time interval combinations in buffalo bull spermatozoa. Semen samples containing 100 million spermatozoa each were incubated with 1, 10, 25, 50, 75 and 100 µg/mL of Ag NPs and MWCNTs at 37°C for 30, 60 and 120 min. Sperm viability was monitored by the MTT assay and eosin-nigrosin staining followed by estimation of IC50 values using correlation-regression analysis. Spermatozoa treated with IC50 doses of Ag NPs and MWCNTs were also assessed for different sperm functionality parameters including oxidative stress and membrane integrity. These parameters were observed to be significantly affected in treated spermatozoa compared with the controls. We concluded that both nanomaterials showed cytotoxicity, mediated principally via oxidative stress. This work has provided valuable toxicological information that will serve as a benchmark for future studies aimed at safe use of nanomaterials.

Mesoporous silica nanoparticle is comparatively safer than zinc oxide nanoparticle which can cause profound steroidogenic effects on pregnant mice and male offspring exposed in utero.

The increasing use of nanomaterials has naturally caused heightened concerns about their potential risks to human and animal health. We investigated the effect of zinc oxide nanoparticles (ZnO NPs) and mesoporous silica nanoparticles (MSN) on steroidogenesis in the corpus luteum (CL) of pregnant mice and testis of male offspring. Pregnant albino mice were exposed to ZnO NPs and MSN for 2 days on alternate days, gestation days 15-19. Hepatic injury marker enzymes increased in the higher concentration of NM-exposed mother mice, but histological examination revealed no changes in the placenta of pregnant mice, whereas testis of male offspring showed gross pathological changes. The expression pattern of progesterone biosynthesis-related genes was also altered in the CL of NP-exposed pregnant mice. In utero exposure of ZnO NPs increased the relative expression of StAR in 100 mg/kg body weight (BW) ZnO NP-treated and bulk ZnO-treated groups and P450 side-chain cleavage enzyme (P450scc) in 50 mg/kg BW ZnO NP-treated and 100 mg/kg of bulk ZnO-treated male offspring. Serum testosterone concentration significantly increased in the 100 mg/kg of bulk ZnO-treated group and decreased in the 250 mg/kg of MSN-treated group and a single dose of 300 mg/Kg BW of ZnO NPs caused miscarriages and adversely affected the developing foetus in mice.

Enhanced steroidogenic and altered antioxidant response by ZnO nanoparticles in mouse testis Leydig cells.

Zinc oxide nanoparticles (ZnO NPs) are important nanomaterials with myriad applications and in widespread use. The main aim of this study was to evaluate the direct effect of ZnO NPs on steroidogenesis by considering mouse testicular Leydig cells (TM3) as an in vitro model system. The uptake, intracellular behaviour, cytotoxicity and morphological changes induced by ZnO NPs (0-200 µg/ml) in a time-dependent manner in the TM3 were assessed. A significant ( p < 0.05) decrease in TM3 viability was observed at 2 µg/ml ZnO NP after a 1-h incubation time period. Increased antioxidant enzyme activity, namely, superoxide dismutase (SOD) and catalase, was regularly observed. Not surprisingly, apoptosis also increased significantly after a 4-h exposure period. Transmission electron micrographs illustrated that ZnO NPs were taken up by Leydig cells and resulted in the formation of autophagosomes, autolysosomes and autophagic vacuoles. Concomitant real-time data indicated that ZnO NPs significantly increased the expression of steroidogenesis-related genes (steroidogenic acute regulatory protein and cytochrome P450 side-chain cleavage enzyme) and significantly ( p < 0.05) decreased antioxidant enzyme gene (SOD) expression after a 4-h incubation period. Moreover, ZnO NPs exposure significantly increased testosterone production at 2 µg/ml concentration after a 12-h incubation period. Our findings confirm the adverse effects of ZnO NPs by being cytotoxic, enhancing apoptosis, causing steroidogenic effect in Leydig cells and increasing autophagic vacuole formation possibly via alteration of antioxidant enzyme activity in TM3 cells.

Comparative in vivo toxicity assessment places multiwalled carbon nanotubes at a higher level than mesoporous silica nanoparticles.

In the present work, we took two nanomaterials (NMs), mesoporous silica nanoparticles (MSNs) and multiwalled carbon nanotubes (MWCNTs), and compared their in vivo toxicity taking albino mice as a test animal model. Presently, conflicting data persist regarding behavior of these NMs with macromolecules like protein and lipid at the cellular level in cell lines as well as in animal models and this generated the interest to study them. The mice were treated orally with a single dose of 50 ppm MWCNTs and intraperitoneally with 10, 25, and 50 mg kg-1 body weight (BW) of MSNs and 1.5, 2.0, and 2.5 mg kg-1 BW of MWCNTs. Liver enzyme markers serum aspartate aminotransferase (AST), alanine aminotransferase, and alkaline phosphatase along with total protein (TP) levels were evaluated 7 days postexposure. No significant differences in organ weight indices or enzyme levels were observed between different treatment doses but there were significant differences between the treatment groups and the controls. Of the three enzymes assayed, AST displayed a peculiar pattern, especially in the MWCNTs intraperitoneally treated group. TP level was significantly increased in the orally treated MWCNTs group. The results showed that MWCNTs even at much smaller doses than MSNs displayed similar toxicity levels, suggesting that toxicity of MWCNTs is greater than MSNs.

Health hazards associated with nanomaterials.

Nanotechnology is a major scientific and economic growth area and presents a variety of hazards for human health and environment. It is widely believed that engineered nanomaterials will be increasingly used in biomedical applications (as therapeutics and as diagnostic tools). However, before these novel materials can be safely applied in a clinical setting, their toxicity needs to be carefully assessed. Nanoscale materials often behave different from the materials with a larger structure, even when the basic material is same. Many mammals get exposed to these nanomaterials, which can reach almost every cell of the mammalian body, causing the cells to respond against nanoparticles (NPs) resulting in cytotoxicity and/or genotoxicity. The important key to understand the toxicity of nanomaterials is that their minute size, smaller than cellular organelles, allows them to penetrate the basic biological structures, disrupting their normal function. There is a wealth of evidence for the noxious and harmful effects of engineered NPs as well as other nanomaterials. The rapid commercialization of nanotechnology field requires thoughtful, attentive environmental, animal and human health safety research and should be an open discussion for broader societal impacts and urgent toxicological oversight action. While 'nanotoxicity' is a relatively new concept to science, this comprehensive review focuses on the nanomaterials exposure through the skin, respiratory tract, and gastrointestinal tract and their mechanism of toxicity and effect on various organs of the body.

Toxicity of titanium oxide nanoparticles causes functionality and DNA damage in buffalo (Bubalus bubalis) sperm in vitro.

The present study has examined the effect of different concentrations (1 µg/ml, 10 µg/ml and 100 µg/ml) of titanium oxide (TiO2) nanoparticles (NPs) (<100 nm) on viability, membrane integrity, capacitation status and DNA integrity of buffalo spermatozoa. Characterization of NPs was done by the transmission electron microscopy (TEM) and dynamic light scattering (DLS). Sperm chromatin dispersion (SCD) test and acridine orange test (AOT) were employed to detect DNA fragmentation in sperm treated with NPs. There was significant (p < 0.05) decrease in cell viability and membrane integrity (assessed by enzyme leakage) at 6 h of incubation with NPs. However, significant (p < 0.05) increase in sperm capacitation was observed for TiO2 NP albeit at lower concentrations. In DNA fragmentation assay, there was dose-dependent increase in the DNA fragmentation (r = 0.96). Ultrathin cross-sections revealed TiO2 NPs inside head and plasma membrane of the buffalo spermatozoa as assessed by TEM. These studies suggest that TiO2 NPs may have cytotoxic effect on buffalo spermatozoa by affecting sperm functionality and causing high amount of DNA fragmentations.

MsbA ATP-binding cassette (ABC) transporter of E. coli: structure and possible flippase mechanism.

ATP-binding cassette (ABC) transporters utilize the energy present in cellular ATP to drive the translocation of structurally diverse set of solutes across the membrane barriers of eubacteria, archaebacteria and eukaryotes. In bacteria, these transporters are considered to be important virulence factors because they play role in nutrient uptake and in the secretion of toxins. The advances in structural determination and functional analysis of bacterial transporters have greatly increased our understanding of the mechanism of transport of these ABC transporters. Although progress in the field of structural biology has been made with the prokaryotic family members, it is likely that eukaryotic transporters will utilize the same mechanisms for translocation process. In this review, we summarize the function of the known MsbA ABC transporters in E. coli and mechanistic insights from structural and possible flippase mechanism studies.

Eukaryotic elongation factor-2 (eEF2): its regulation and peptide chain elongation.

Regulation at the level of translation in eukaryotes is feasible because of the longer lifetime of eukaryotic mRNAs in the cell. The elongation stage of mRNA translation requires a substantial amount of energy and also eukaryotic elongation factors (eEFs). The important component of eEFs, i.e. eEF2 promotes the GTP-dependent translocation of the nascent protein chain from the A-site to the P-site of the ribosome. Mostly the eEF2 is regulated by phosphorylation and dephosphorylation by a specific kinase known as eEF2 kinase, which itself is up-regulated by various mechanisms in the eukaryotic cell. The activity of this kinase is dependent on calcium ions and calmodulin. Recently it has been shown that the activity of eEF2 kinase is regulated by MAP kinase signalling and mTOR signalling pathway. There are also various stimuli that control the peptide chain elongation in eukaryotic cell; some stimuli inhibit and some activate eEF2. These reports provide the mechanisms by which cells likely serve to slow down protein synthesis and conserve energy under nutrient deprived conditions via regulation of eEF2. The regulation via eEF2 has also been seen in mammary tissue of lactating cows, suggesting that eEF2 may be a limiting factor in milk protein synthesis. Regulation at this level provides the molecular understanding about the control of protein translocation reactions in eukaryotes, which is critical for numerous biological phenomenons. Further the elongation factors could be potential targets for regulation of protein synthesis like milk protein synthesis and hence probably its foreseeable application to synthetic biology.

Role of heat shock proteins in diseases and their therapeutic potential.

Heat shock proteins are ubiquitously expressed intracellular proteins and act as molecular chaperones in processes like protein folding and protein trafficking between different intracellular compartments. They are induced during stress conditions like oxidative stress, nutritional deficiencies and radiation. They are released into extracellular compartment during necrosis. However, recent research findings highlights that, they are not solely present in cytoplasm, but also released into extracellular compartment during normal conditions and even in the absence of necrosis. When present in extracellular compartment, they have been shown to perform various functions like antigen presentation, intercellular signaling and induction of pro-inflammatory cytokines. Heat shock proteins represents as dominant microbial antigens during infection. The phylogenetic similarity between prokaryotic and eukaryotic heat shock proteins has led to proposition that, microbial heat shock proteins can induce self reactivity to host heat shock proteins and result in autoimmune diseases. The self-reactivity of heat shock proteins protects host against disease by controlling induction and release of pro-inflammatory cytokines. However, antibodies to self heat shock proteins haven been implicated in pathogenesis of autoimmune diseases like arthritis and atherosclerosis. Some heat shock proteins are potent inducers of innate and adaptive immunity. They activate dendritic cells and natural killer cells through toll-like receptors, CD14 and CD91. They play an important role in MHC-antigen processing and presentation. These immune effector functions of heat shock proteins are being exploited them as therapeutic agents as well as therapeutic targets for various infectious diseases and cancers.

Carbon nanotubes as a scaffold for spermatogonial cell maintenance.

Carbon nanotubes (CNTs) are viewed as a class of nanomaterials with high potential for many biomedical applications due to their unique properties. One of the prospective bio-application of CNTs is in the area of tissue engineering as a scaffold for cell culture. Supporting the use of CNTs as scaffold in tissue engineering a few cell types have been successfully grown on it but no report for any germline cell. In the present study, the in vitro maintenance of spermatogonial cells on carbon nanotube scaffold has been evaluated for the first time. Protein adsorption on the CNT scaffolds along with other substrates has also been evaluated because it is very likely that adsorbed proteins in serum influence the properties for the cell growth. Microscopic analysis (Light and SEM) showed that cells maintain their proper shape and adhered on all CNT scaffolds during in vitro culture. It was observed that the cells were maintained for at least 21 days on all CNT scaffolds compared to the positive control (Sertoli feeder layer). The results express a degree of biocompatibility between spermatogonial cells and CNTs and the possibility for CNTs to be used as substratum for in vitro growth of these cells.

A cost-utility analysis of amputation versus salvage for Gustilo type IIIB and IIIC open tibial fractures.

BACKGROUND: Lower extremity trauma is common. Despite an abundance of literature on severe injuries that can be treated with salvage or amputation, the appropriate management of these injuries remains uncertain. In this situation, a cost-utility analysis is an important tool in providing an evidence-based practice approach to guide treatment decisions. METHODS: Costs following amputation and salvage were derived from data presented in a study that emerged from the Lower Extremity Assessment Project. The authors extracted relevant data on projected lifetime costs and analyzed them to include discounting and sensitivity analysis by considering patient age. The utilities for the various health states (amputation or salvage, including possible complications) were measured previously using the standard gamble method and a decision tree simulation to determine quality-adjusted life-years. RESULTS: Amputation is more expensive than salvage, independently of varied ongoing prosthesis needs, discount rate, and patient age at presentation. Moreover, amputation yields fewer quality-adjusted life-years than salvage. Salvage is deemed the dominant, cost-saving strategy. CONCLUSION: Unless the injury is so severe that salvage is not a possibility, based on this economic model, surgeons should consider limb salvage, which will yield lower costs and higher utility when compared with amputation.