Membrane Functional Integrity Research Articles

Effect of superoxide dismutase, catalase, and glutathione reductase supplementation on cryopreservation of Black Bengal buck semen.

The present experiment was carried out with the objectives to study the effects of antioxidants superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GSH) on cryopreservation of Black Bengal buck semen. Semen ejaculates (n = 60) were collected from eight bucks by artificial vagina method and diluted with Tris citrate egg yolk glycerol extender. To study the effect of antioxidants, SOD was added @ 0, 100, and 150IU/ml; CAT was added @ 0, 200, and 400IU/ml while GSH was added @ 0, 1, and 2mM of diluted semen. Semen samples were equilibrated and vapor frozen in liquid nitrogen. Semen samples were evaluated after 48h of storage for post thaw in vitro characters such as motility, viability, functional membrane integrity, and acrosome integrity. Semen extenders supplemented with SOD @ 100 and 150IU/ml and GSH @ 1 and 2mM had a higher (p < 0.01) number of motile cells, viable cells, HOST reacted cells, and acrosome intact cells than their respective controls. Further, semen extenders added with catalase @ 200 and 400IU/ml had more (p < 0.05) number of viable, HOST reacted cells and significantly higher (p < 0.01) acrosome intact sperm cells than its control group. It can be concluded that supplementation of antioxidants SOD, GSH, and CAT had a beneficial effect on cryopreservation of Black Bengal buck semen.

The hypo-osmotic swelling test: Is it a sperm vitality or a viability assay?

The first two editions of the World Health Organization laboratory manual described the determination of live spermatozoa by a dye exclusion method as a sperm "viability" test, whereas subsequent editions classified it as a "vitality" test, without providing an explanation for the reclassification. Additionally, the hypo-osmotic swelling (HOS) test, which assesses the functional integrity of the human sperm membrane, was placed in the same category as the dye exclusion test. Although the two terms might seem synonymous, the term "vitality" merely means "alive," whereas "viability" assesses qualities or physiological functions of a living entity. After comparing the morphological, physiological, and clinical findings obtained from dye exclusion testing vs. the HOS test, we conclude that the HOS test should be classified as a viability test, not merely as a vitality test.

Positive effect of autologous platelet rich plasma on Saanen buck semen cryopreservation in non-breeding season

The objective of the current study was to evaluate the effect of autologous platelet-rich plasma (PRP) addition into soybean lecithin based extender on buck semen at post-thaw. Semen samples were collected from eight Saanen buck, and each semen sample was split into four equal aliquots and diluted with different concentrations of PRP supplemented extenders [no PRP (control), 0.5 × 107/ml PRP, 1 × 107/ml PRP, or 2 × 107/ml PRP]. Motility, plasma membrane functional integrity, acrosome integrity, mitochondrial membrane potential, DNA integrity and malondialdehyde concentrations (MDA) were measured and analyzed at post-thaw. The results showed that 2 × 107/ml PRP group had a positive effect on motility (62.41 ± 4.24), membrane functional integrity (71.11 ± 2.90), mitochondrial membrane potential (69.70 ± 1.99), DNA integrity (7.22 ± 0.93) and MDA levels (2.56 ± 0.73) at post-thaw (P < 0.05). The results of the study demonstrated that autologous PRP has a protective effect on cryopreservation of buck spermatozoa and the fertility effects are worthy of further study.

Effect of L-cysteine Supplementation on Cryopreservation of Black Bengal Buck Semen

Background: Black Bengal goat is one among the valuable goat breeds of India. Semen preservation and artificial insemination are practical technologies for conservation and improving livestock germplasm. Various biochemical changes that occur during cryopreservation of semen, results in loss of sperm cell structure and their functions. Inclusion of antioxidants to the semen extender has been reported to have positive effects on sperm recovery in various livestock species. The present experiment was carried out to evaluate the effect of antioxidant L-cysteine on post freeze thaw sperm characters, lipid peroxide level and superoxide dismutase (SOD) activity during cryo­preservation of Black Bengal buck semen. Methods: Twenty numbers of semen ejaculates were collected from Black Bengal bucks, antioxidant L cysteine was added to the semen extender @ 0, 1 and 2 mM in control group (CC), treatment group 1 (CT1) and treatment group 2 (CT2), respectively. Semen samples were frozen in liquid nitrogen and measured post freeze-thaw in vitro sperm characters, malondialdehyde (MDA) concentration and SOD activity. Result: Though there was no significant improvement in post thaw sperm motility, functional membrane integrity and viability count, supplementation of L-cysteine significantly improved the percentage of sperm cells with intact acrosome and significantly (P less than 0.05) reduced the level of malondialdehyde. Post thaw SOD activity was found significantly lower in the antioxidant treated groups than the control. It is concluded that supplementation of L cysteine is effective in controlling lipid peroxidation and conserving in vitro sperm characters during freezing of Black Bengal buck semen.

Supraphysiological concentration of urea affects the functional competence of Holstein-Friesian (Bos taurus) sperm

To understand the effects of urea on sperm functional attributes, fresh bull semen (n = 12) was subjected to four different concentrations (mg/mL) of urea to mimic the physiological (0.04 and 0.13), supraphysiological (0.43) concentrations and control (0 mg/mL). Sperm membrane integrity, kinematics, chromatin integrity, and mitochondrial membrane potential were assessed at different time points (before incubation, 0, 1, 2, and 4 h) of incubation. The concentration of urea in serum and seminal plasma was estimated and correlated with the ejaculate rejection rate and sperm functional attributes. The relative expression of urea transporter gene transcripts (UT-A and UT-B) was assessed in sperm and testis (control) using real-time PCR. The supraphysiological concentration of urea affected sperm kinematics, viability, functional membrane integrity, and acrosome integrity within 1 h of incubation (p < 0.05). Sperm head area decreased (p < 0.05) at 0 h and subsequently increased at 1 h of incubation in all media except supraphysiological (0.43 mg/dL) concentration of urea. Seminal plasma urea concentration showed a significant negative correlation with sperm motility, membrane integrity, and mitochondrial membrane potential (p < 0.05), but had a positive correlation with the ejaculate rejection rate (r = 0.69). Relative expression of the urea transporter genes revealed that UT-A was expressed only in the testis. In contrast, UT-B was expressed in both the testis and sperm, suggesting UT-B's role in regulating urea transport in sperm. At a supraphysiological level, urea adversely affected sperm functional attributes, osmoadaptation and may affect fertility.

Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen.

Simple SummaryCryopreserving dog semen allows the long-term availability of male gametes for future artificial insemination and other assisted reproductive techniques. However, freezing causes irreversible damage to sperm that can affect its ability to fertilize and generate a viable pregnancy. Sperm alterations are partly attributed to oxidation produced by reactive oxygen species (ROS); therefore, antioxidants have been included as extenders for seminal cryopreservation. The unconventional natural antioxidants might reduce deleterious changes in cryopreserved dog sperm; therefore, we evaluated the effects of cryopreservation with the antioxidants ergothioneine and isoespintanol on thawed canine sperm. Various concentrations of both antioxidants improved the movement capacity and structure of thawed spermatozoa, possibly by reducing ROS production. The unconventional antioxidants isoespintanol and ergothioneine improved the quality of cryopreserved canine semen and hence improved assisted canine reproduction.Sperm undergo oxidative stress due to excessive production of reactive oxygen species (ROS) during cryopreservation. Some unconventional natural antioxidants can reduce ROS-induced changes in cryopreserved canine sperm. This study aimed to identify the cryoprotective effects of ergothioneine and isoespintanol on the quality of thawed canine semen. Twelve ejaculates from six dogs were cryopreserved in a tris-yolk extender without (control) or with 50 (E50), 100 (E100), or 150 (E150) µM ergothioneine or 20 (I20), 40 (I40), or 60 (I60) µM isoespintanol. We evaluated the motility and kinetics of thawed sperm using computerized analysis; determined morphology by eosin-nigrosin staining; functional membrane integrity using hypoosmotic tests, and structural membrane and acrosome integrity; mitochondrial membrane potential by fluorescence microscopy; and ROS production by spectrophotometry. Data were statistically analyzed using mixed models and Tukey tests. E100 increased total (60.6% vs. 49.6%) and progressive (26.4% vs. 20.1%) motility, straight line velocity (41.3 vs. 35.9 µm/s), and rapid sperm (17.6% vs. 12.3%) compared with controls. However, E150 reduced the numbers of hyperactive sperm. E100, I40, and I60 reduced the abnormal morphology and ROS production, and all concentrations of both antioxidants increased acrosomal integrity. We concluded that ergothioneine and isoespintanol reduce deleterious sperm alterations and oxidative stress in thawed canine semen.

Effects of alpha-lipoic acid on ram semen cryopreservation and post-thaw life span.

The aim of the present study was to evaluate the effects of an alpha-lipoic acid-supplemented extender on ram semen at a post-thaw stage and after incubation (6hr). Semen samples were collected from five Kivircik Rams. Pooled semen was diluted with an egg yolk-based extender with different concentrations of alpha-lipoic acid (0.25mmol L-1 , 0.5mmol L-1 and 1mmol L-1 ) and without alpha-lipoic acid. Motility, plasma membrane functional integrity (HOST), acrosome integrity (FITC-Pisum sativum agglutinin) and DNA integrity (TUNEL) were assessed at post-thaw and 6hr after incubation of the frozen-thawed semen. At the post-thaw stage, 0.25mmol L-1 alpha-lipoic acid had a positive effect on sperm motility and plasma membrane functional integrity compared to the control group (p<.05). At the post-incubation stage (6hr), it was determined that the motility and plasma membrane functional integrity of the antioxidant groups were adversely affected compared to the control group (p<.05) and 1mmol L-1 dose of alpha-lipoic acid had a harmful effect on DNA integrity compared to the control group (p<.05). The results of the study demonstrated that alpha-lipoic acid has positive effects at post-thaw but have harmful effects on long term to ram semen.

Exogenous Nitric Oxide Reinforces Photosynthetic Efficiency, Osmolyte, Mineral Uptake, Antioxidant, Expression of Stress-Responsive Genes and Ameliorates the Effects of Salinity Stress in Wheat.

Salinity stress is one of the major environmental constraints responsible for a reduction in agricultural productivity. This study investigated the effect of exogenously applied nitric oxide (NO) (50 μM and 100 μM) in protecting wheat plants from NaCl-induced oxidative damage by modulating protective mechanisms, including osmolyte accumulation and the antioxidant system. Exogenously sourced NO proved effective in ameliorating the deleterious effects of salinity on the growth parameters studied. NO was beneficial in improving the photosynthetic efficiency, stomatal conductance, and chlorophyll content in normal and NaCl-treated wheat plants. Moreover, NO-treated plants maintained a greater accumulation of proline and soluble sugars, leading to higher relative water content maintenance. Exogenous-sourced NO at both concentrations up-regulated the antioxidant system for averting the NaCl-mediated oxidative damage on membranes. The activity of antioxidant enzymes increased the protection of membrane structural and functional integrity and photosynthetic efficiency. NO application imparted a marked effect on uptake of key mineral elements such as nitrogen (N), potassium (K), and calcium (Ca) with a concomitant reduction in the deleterious ions such as Na+. Greater K and reduced Na uptake in NO-treated plants lead to a considerable decline in the Na/K ratio. Enhancing of salt tolerance by NO was concomitant with an obvious down-regulation in the relative expression of SOS1, NHX1, AQP, and OSM-34, while D2-protein was up-regulated.

Do aging, drinking, and having unhealthy weight have a synergistic impact on semen quality?

To evaluate if age, alcohol consumption, and body mass index (BMI) have synergistic effects on seminal quality, and to rank these factors based on their impact on semen. Retrospective study of 9464 patients attending an andrology laboratory. Data on patients' age and daily alcohol intake were provided by the patients. BMI was recorded in the laboratory. Seminal parameters evaluated were volume, sperm concentration and total count, motility, morphology, viability, nuclear maturity, and membrane functional integrity. All the seminal parameters evaluated were affected by the synergistic interaction Age x BMI, suggesting that this combination is more potent in affecting semen quality. The variables sperm morphology and nuclear maturity seemed to be especially susceptible since they were affected by the three synergistic interactions. In the logistic regression analysis, age was the most powerful factor since it impacted first on five of the nine parameters, impacting mainly on sperm motility, viability, and morphology, with no effects on sperm count. On the contrary, BMI impacted first in sperm concentration and total sperm count; which was confirmed also by the logistic predictions analysis. Alcohol consumption impacted first on membrane functional integrity and nuclear maturity. A J-shaped association between BMI or alcohol consumption with semen quality was found in the multivariate analysis. The factors considered in this study showed a synergistic negative impact on semen quality, being age and unhealthy weight the most important ones. Reducing the exposure to lifestyle risk factors may be promising for improving sperm quality in infertile patients.

Investigation of phytochemical profile and in vivo anti-proliferative effect of Laetiporus versisporus (Lloyd) Imazeki mushroom against diethylnitrosamine-induced hepatocellular carcinoma

The purpose of the study is to explore the bioactive compounds present in Laetiporus versisporus (LVEE) Lloyd Imazeki ethanolic extract and its anticancer activity against hepatocellular carcinoma (HCC) induced by DEN in rats. The bioactive compounds present in the LVEE was analyzed by GC–MS. The anticancer activity of LVEE was analyzed with the DEN induced rats were orally treated with LVEE (250 mg/kg), cyclophosphamide (50 mg/kg) to the respective treatment groups for 45 days. The levels of tissue markers, phase-I metabolizing enzymes was measured calorimetrically. ELISA was used to determine the level of AFP and CEA. The expression of apoptotic genes Bcl-2, p53, caspase 3 and caspase 9 was assessed by real time RT-PCR. Results of the phytochemical screening analysis showed that 12 bioactive molecules were existing in LVEE. Results of pharmacodynamics analysis showed that the 45 days of treatment with LVEE and cyclophosphamide (CPA) therapy significantly (p < 0.05) decreased the levels of MDA, AFP, CEA, cytochrome P450 and cytochrome b5, and increased levels of tissue markers and bilirubin in serum. The findings of RT-PCR analysis showed that the LVEE and CPA were significantly (p < 0.05) downregulated the pro-apoptotic genes Bcl-2 and upregulated the p53, caspase 3 and caspase 9 mRNA expression through mechanisms involved in promoting the cell apoptosis, and preserving the functional integrity of the liver membrane. L. versisporus (LV) can be considered as new anticancer agent that offers new opportunities for anticancer therapy.

Freezability of Goat Epididymal Sperm using Aloe vera Extract and Trehalose in Diluents

Background: Cryopreservation process causes oxidative stress on sperm membranes, which in turn damages sperm organs and enzymatic activities which thereby decrease motility, functional membrane integrity and sperm fertility. Therefore, current study carried out to evaluate the effect of Aloe vera Ethanolic Extract (AEE), alone and with trehalose in diluents on cryopreserved epididymal goat sperm. Methodology: Epididymal sperm isolated from testes with motility &gt;70% and total morphological abnormalities &lt;10%. The experimental treatments consist of control (no additives) and basic diluents plus 5, 10, 20 or 50 μg/ml of AEE (AEE1, AEE2, AEE3 and AEE4, respectively), tr (150 mM trehalose), tr+AEE1, tr+AEE2, tr+AEE3 and tr+AEE4. Results: Obtained data show that the extender containing AEE3, AEE1+tr, AEE2+tr and AEE3+tr improved significantly the cryopreserved sperm. The combined treatments indicate also a decrease in MDA than control. In addition, AEE2+tr and AEE3+tr showed the lowest (P&lt;0.05) DNA fragmentation compared to the other treatments. Extender containing AEE3+tr resulted in higher total motility and viability than the extender containing tr alone, as well as AEE1, AEE2 and AEE4 treatments. Conclusion: The present study indicates that ethanolic extract of Aloe vera could be used for goat sperm cryopreservation. Also, it can be concluded that trehalose in combination with 20 μg/ml of Aloe vera extract can be promised cryoprotectant in goat epididymal sperm freezing.

Carboxymethyl cellulose and glycerol act synergistically as cryoprotectant during cryopreservation of ram semen

Wider implementation of AI in sheep in the field condition has not been possible till date due to very poor conception rate after cervical insemination with cryopreserved semen. Poor cervical penetrability in ewe and diminished sperm functions in cryopreserved semen are considered responsible for it. In the present study, effect of carboxymethyl cellulose (CMC) on post-thaw qualities of ram semen was investigated. Ejaculates from eight adult Malpura rams were pooled and diluted (800 × 106 sperm mL−1) with TES-Tris-fructose-egg yolk extender having either 5 or 6% glycerol and supplemented with 0, 0.25, 0.5, 0.75 and 1.0% (w/v) CMC and packaged into 0.25 mL French mini straws. The straws were progressively cooled to 5 °C inside a cold cabinet (5 °C) and then equilibrated for 22 h inside a refrigerator (2–5 °C). Straws were frozen at −25 °C min−1 up to −125 °C using a programmable cell freezer (Planer Biomed R-204, UK) and finally plunged into liquid nitrogen. The post-thaw progressive motility was higher (P < 0.05) in 0.75% CMC-treated group compared to control. Overall, both pre-freeze and post-thaw sperm kinetics was comparable between CMC-treated and control groups. The post-thaw sperm viability, acrosomal integrity and sperm with high mitochondrial membrane potential (hMMP) were relatively higher while sperm with high membrane cholesterol was significantly (P < 0.05) higher in presence of 0.25% CMC compared to the control. Both sperm having hMMP and non-capacitated sperm were significantly (P < 0.05) higher in presence of 5% glycerol than 6% glycerol. Similarly, functional membrane integrity (FMI) was higher in presence of 5% glycerol than 6% glycerol when CMC was added at 0.5% to extender. In conclusion, both 0.25% CMC and 5% glycerol resulted in improvement in several post-thaw sperm functions in cryopreserved ram semen. Thus CMC demonstrated cryoprotective effect on ram sperm in a synergistic manner with glycerol.

Boron decreases cadmium influx into root cells of Capsicum annuum by altering cell wall components and plasmalemma permeability.

Large areas of soil are boron (B) deficient and contaminated with cadmium (Cd) in southern China. The aim of this study was to select the optimal B supply level and elucidate the underlying physiological and biochemical mechanisms to understand how B reduces Cd influx into root cells of hot pepper (Capsicum annuum). An experiment was conducted to investigate the changes in Cd accumulation with B supply. Hot pepper seedlings were grown in two nutrient solutions containing 0.05- and 0.2-mg Cd L-1 and supplied with six different B concentrations for 2 weeks. The other experiment was conducted to determine the Cd2+ flux into cells, cell wall components, antioxidative ability, and plasmalemma permeability of root tips of hot pepper exposed to 0.1-mg Cd L-1 in the presence and absence of B. The results showed that the optimal B concentration to promote plant growth and reduce Cd accumulation was 0.25 mg L-1. Moreover, B application significantly decreased Cd2+ influx into cells, increased the contents of lignin and pectin, enhanced the activities of antioxidant enzymes, reduced the production of reactive oxygen species, and decreased membrane peroxidation and permeability. Overall, boron in moderation can promote plant growth, maintain the normal structures and functions of the cell wall and membrane, and thus decrease Cd2+ influx into root cells and subsequently Cd translocation to shoots. Consequently, B is a reliable inhibitor of Cd uptake, and the functional and structural integrity of cell walls and membranes may have some relevance to reduced Cd uptake after B application.

Hepatoprotective activity of Phyllanthus niruri Linn. endophytes

BackgroundThe liver is the vital organ which plays a major role in metabolism with numerous functions in the human beings such as protein synthesis, hormone production, and detoxification. Present research work is focused on hepatoprotective potential of chloroform (PNFC) and ethyl acetate (PNFEA) endophytic fractions from Phyllanthus niruri Linn. against CCl4-induced hepatotoxicity in albino Wistar rats. To test our hypothesis, both endophytic fungal fractions were tested for vitro antioxidant and in vivo hepatoprotective activity. Serum biochemical parameters like SGOT, SGOT, SALP, cholesterol, bilirubin, and protein were estimated to assess hepatoprotective activity.ResultsGroup of rats treated with CCl4 possess marked hepatic damage and oxidative stress which indicates that cellular leakage and loss of functional integrity of cell membrane in liver. PNFC and PNFEA fractions of endophyte from Phyllanthus niruri Linn. stem have significantly reduced the elevated levels of biomarkers like SGPT, SGOT, SALP, bilirubin, cholesterol, and total protein in CCl4-induced hepatotoxicity in rats. The results obtained confirm hepatoprotective activity of endophytic fractions (PNFC and PNFEA) mediated through the stabilization of plasma membrane, repair of hepatic tissue damage, return of biochemical marker levels to normal, and regeneration of hepatocytes. Histopathological observations revealed improvement in the liver architecture after the treatment of secondary metabolites of endophytic fractions against CCl4-induced liver damage. Both fungal endophytes PNFC and PNFEA showed DPPH scavenging activity with IC50 of 97.79 μg/ml and 108.40 μg/ml, respectively, and possess antioxidant potential. Presence of flavonoids in the both fractions of endophytes may be a possible reason for its antioxidant potential and identified as Eurotium amstelodami strain.ConclusionBoth fungal endophytes PNFC and PNFEA possess hepatoprotective potential due to the presence of secondary metabolites of fungi, i.e., Eurotiumam stelodami strain which support the claim endophytes and act as a potent biomedicine for treatment of various chronic diseases.

Deciphering the complexity of sperm transcriptome reveals genes governing functional membrane and acrosome integrities potentially influence fertility

Deciphering sperm transcriptome is the key to understanding the molecular mechanisms governing peri-fertilization, embryonic development, and pregnancy establishment. This study aimed to profile sperm transcriptome to identify signature transcripts regulating male fertility. Semen samples were collected from 47 bulls with varied fertility rates. The sperm total RNA was isolated (n = 8) and subjected to transcriptome sequencing. Based on the expression pattern obtained from RNA profiling, the bulls were grouped (p = 0.03) into high-fertile and sub-fertile, and signature transcripts controlling sperm functions and fertility were identified. The results were validated using the OMIM database, qPCR, and sperm function tests. The sperm contains 1100 to 1700 intact transcripts, of which BCL2L11 and CAPZA3 were abundant and associated (p < 0.05) with spermatogenesis and post-embryonic organ morphogenesis. The upregulated genes in the acrosome integrity and functional membrane integrity groupshad a close association with the fertility rate. The biological functions of these upregulated genes (p < 0.05) in the high-fertile bulls were associated with spermatogenesis (AFF4 and BRIP1), sperm motility (AK6 and ATP6V1G3), capacitation and zona binding (AGFG1), embryo development (TCF7 and AKIRIN2), and placental development (KRT19). The transcripts involved in pathways regulating embryonic development such as translation (EEF1B2 and MTIF3, p = 8.87E-05) and nonsense-mediated decay (RPL23 and RPL7A, p = 5.01E-27) were upregulated in high-fertile bulls. The identified transcripts may significantly impact oocyte function, embryogenesis, trophectoderm development, and pregnancy establishment. In addition, the study also reveals that the genes governing sperm functional membrane integrity and acrosome integrity have a prospective effect on male fertility.

Effect of Butylated Hydroxytoluene and Tocopherol Supplementation on In vitro Sperm Characters during Cryopreservation of Black Bengal Buck Semen

Background: Black Bengal goat is one of the important goat breeds of India. Cryopreservation of semen and artificial insemination are effective techniques for improving goat breeding programs. Various biochemical and functional changes that occur during freezing-thawing process results in poor post thaw sperm recovery. Supplementation of antioxidants to the extender has been reported to have positive effects on semen cryopreservation in various species. The present study was undertaken to assess the effect of antioxidants butylated hydroxyl toluene (BHT), α tocopherol on post thaw in vitro sperm characters, lipid peroxide level and superoxide dismutase (SOD) activity during cryo­preservation of Black Bengal buck semen. Methods: Semen was collected from bucks by artificial vagina, antioxidant BHT was added to the Tris egg yolk extender @ 0, 1 and 2 mM/ml in control group (BHTC), treatment group 1 (BHTT1) and treatment group 2 (BHTT2), respectively. Similarly, tocopherol was added @ 0, 1 and 2 mg/ml in control group (TFC), in treatment group 1 (TFT1) and treatment group 2 (TFT2), respectively. Each antioxidant was tested with 20 semen ejaculates. Semen samples were frozen in liquid nitrogen and post freeze-thaw in vitro sperm characters, malondialdehyde (MDA) concentration and SOD activity were measured. Result: Post thaw sperm motility, functional membrane integrity and viable count were significantly (P less than 0.05) higher in the BHT and α tocopherol supplemented groups than control groups. Significantly (P less than 0.05) higher acrosome intact cells were recovered in BHTT1, BHTT2 and TFT2 groups than the control and TFT1 groups. Further, BHT supplemented groups had significantly lower level of MDA than the control, while the supplementation of α tocopherol could not control the generation of MDA. Post thaw SOD activity was found significantly lower in the antioxidant treated groups than their respective controls. It is concluded that supplementation of BHT and α tocopherol were found to be more promising in conserving in vitro sperm characters from cryo-damages during freezing of Black Bengal bucks semen.

Fluoride induced testicular toxicities in adult Wistar rats

Fluoride is essential for the development of teeth and bone but its excessive exposure causes reprotoxic effects. We have studied the graded effects of different doses of sodium fluoride (NaF) on 24 adult Wistar rats which were randomly divided into four groups (n = 6). All the rats were given normal diet prepared in our laboratory. Control (group I) rats were received vehicle only and treated rats (group II, III, and IV) were administered NaF orally at 10, 15, and 20 mg/kg/d doses, respectively, for 30 consecutive days. Gravimetric index of testis was decreased significantly in group III and IV rats but such changes were not observed in the accessory reproductive organs. Marked alterations in number, motility, viability, and plasma membrane functional integrity of caudal spermatozoa were noted in most of the treated groups. Noticeable alterations in testicular histoarchitecture were observed in group III and IV rats. Graded Changes of testicular redox homeostasis were noted by assessment of enzymatic (superoxide dismutase [SOD], catalase, glutathione s-transferase [GST], and glutathione peroxidase [GPx]) and nonenzymatic (malondialdehyde [MDA] and protein carbonyl [PC]) markers. Changes of testicular functional markers (acid phosphatase [ACP], alkaline phosphatase [ALP], and lactate dehydrogenase (LDH) activity) were noted in group III and IV rats. DNA fragmentation assay proved the possibility of DNA damage caused by oxidative stress, as evidenced by prominent trailing pattern of fragmented testicular genomic DNA from group III and IV rats. The study concludes that fluoride-mediated oxidative stress not only impedes the structural and functional facets of testis but also results in testicular DNA damage.

Docosahexaenoic acid associated vitamin E in the diluent for cryopreservation of goat semen

The objective of this study was to evaluate the effect of and determine the optimum level of inclusion of docosahexaenoic acid (DHA) in the diluent for goat semen cryopreservation. Five Boer males underwent semen collection, totaling 10 viable collections per animal. After evaluation, the ejaculates were pooled and fractionated in Tris-yolk medium with the addition of 0; 30; 45; or 60ng mL-1 of DHA and 0.4 mmol of alpha-tocopherol (Vitamin E). The semen was cryopreserved in a freezing machine (TK 3000TM) and placed in a cryogenic cylinder for subsequent analysis. Data were evaluated by regression analysis at 5% significance. There were no differences (P &gt; 0.05) in sperm kinetic parameters evaluated by computer assisted sperm analysis: total motility (79.17 ± 17.31%), progressive motility (14.04 ± 5.73%), curvilinear speed (58.82 ± 6.35µm/s), progressive linear speed (22.49 ± 3.63µm/s), mean path speed (35.17 ± 4.52µm/s), linearity (38.69 ± 5.79%), rectilinearity (63.99 ± 6.64%), and oscillation index (59.68 ± 2.99%). There were no differences (P &gt; 0.05) found from the membrane functional integrity test for reactive spermatozoa (69.66 ± 9.76%), plasma and acrosomal membrane integrity of intact spermatozoa (29.86 ± 7.57%), mitochondrial potential of Class I cryopreserved goat semen (72.75 ± 9.81%), and chromatin compaction of intact chromatin (96.87 ± 4.37%). Thus, the inclusion of up to 60ng mL-1 of DHA did not promote any improvement in the seminal quality parameters of post-thawed goat semen.

Single electrode piezoelectric nanogenerator for intelligent passive daytime radiative cooling

Passive daytime radiative cooling (PDRC) can cool down objects by simultaneously reflecting sunlight and radiating heat to the cold external space, which has no energy consumption. Traditional PDRC design usually needs to rely on micro-nano structures, which are easily damaged and lose function. On the one hand, the mechanical strength of PDRC can be enhanced. On the other hand, by integrating self-monitoring character in the same membrane fault part can be easily distinguished for fixing or replacing. Monitoring the damage by using traditional sensors will cause additional energy loss. Therefore, an intelligent PDRC that can monitor the failure itself is urgently needed to be developed. Here we found that the polyvinylidene fluoride-co-hexafluoropropylene (PVDF-HFP) nanofiber membrane obtained by electrospinning technology can effectively scatter sunlight, while emitting infrared rays matching the atmospheric window to achieve effective heat dissipation. In the preparation process, the electrostatic field can effectively polarize the PVDF-HFP to obtain good piezoelectric performance, which can be used to monitoring the integrity of membrane function. Meanwhile, the membrane can be used as a nanogenerator to collect mechanical energy in the environment, such as raindrop energy when used on the roof, thereby not only reducing cooling energy loss, but also collecting energy additionally, achieving a dual energy-saving design. Its excellent self-cleaning and recyclable characteristic bring more benefits to its green and energy-saving features. Therefore, this intelligent PDRC provides ideas for new energy-saving designs. At the same time, a simple preparation method is more conducive to the promotion of this technology. • Nanogenerator realizes good passive daytime radiation cooling for the first time. • The single-electrode piezoelectric nanogenerator gives PDRC the self-monitoring function to accurately monitor the working status of each area. • Efficiently collect raindrops' mechanical energy to achieve dual energy-saving design. • Self-cleaning and recyclable capabilities bring additional benefits to energy-saving design.

Seminal plasma and serum fertility biomarkers in Ossimi rams and their relationship with functional membrane integrity and morphology of spermatozoa

This study aimed to explore some fertility biomarkers in seminal plasma (SP) and blood of rams with different field fertility, based on their conception and lambing rates. Ejaculates were collected from fertile (n = 20) and subfertile (n = 10) Ossimi rams by electroejaculation for 16 replicates. Each ejaculate was analyzed for volume and sperm characteristics before centrifugation at 12,000 × g for 30 min at 4 °C to collect SP. Blood samples were collected from all rams and centrifuged to collect serum which was frozen stored at – 30 °C until analysis. Seminal and serum levels of superoxide dismutase, nitric oxide, testosterone, thyroxin and malondialdehyde as well as seminal fructose and serum glucose were determined. Ovine SP proteins were extracted and separated by gel electrophoresis. Both ejaculate volume (P < 0.05) and % hypo-osmotic swelling test (HOST)-positive spermatozoa (P < 0.001) were greater in fertile than subfertile rams. On the other hand, stimulation time (P < 0.05) and % abnormal spermatozoa (P < 0.0001) were greater in subfertile than fertile rams. Intriguingly, seminal and serum levels of superoxide dismutase, nitric oxide, testosterone and thyroxin as well as seminal fructose and serum glucose were greater (P < 0.05) in fertile than subfertile rams. On contrary, seminal and serum malondialdehyde were higher (P < 0.01) in subfertile than fertile rams. The proportion of HOST-positive spermatozoa has positive association (P < 0.05) with seminal and serum levels of superoxide dismutase, nitric oxide, testosterone and thyroxin as well as with seminal fructose and serum glucose and negative association (P < 0.001) with malondialdehyde content, while % abnormal spermatozoa was positively correlated (P < 0.05) with only malondialdehyde content either in serum or SP. Noteworthy, protein band of 14 kDa was prominent in fertile rams and faint to absent in subfertile rams. In conclusion, determination of seminal and/or serum superoxide dismutase, nitric oxide, testosterone, thyroxin and malondialdehyde as well as seminal fructose and serum glucose might be beneficial in assessment of male fertility during breeding soundness examination. Moreover, SP protein of 14 kDa might be a candidate of fertility-associated protein in ovine SP at least for the Ossimi breed.