Cryoprotectant with a mitochondrial derived peptide, humanin, improves post-thaw quality of buffalo spermatozoa.

BACKGROUND: Semen cryopreservation results in deleterious effects on spermatozoa, including lipid peroxidation and a reduction in the total antioxidant components of seminal plasma. The ultimate outcome of these changes is a reduction in post-thaw semen quality. A mitochondrial derived peptide, humanin, a potent cytoprotective and antioxidant agent was used in the present study. OBJECTIVE: To evaluate the efficacy of a mitochondrial-derived peptide, humanin to improve the post-thaw quality of buffalo spermatozoa. MATERIALS AND METHODS: A total of 18 ejaculates from three Murrah buffalo bulls (n=6 each) were collected. Each ejaculate was divided into four aliquots. The first aliquot was diluted with standard EYTG dilutor (Group I, control), whereas the other three aliquots were diluted with EYTG supplemented with 2 µM (Group II), 5 µM (Group III) and 10 µM humanin (Group IV), respectively. Semen was evaluated for physico-morphological and functional attributes such as progressive motility, viability, abnormality, acrosome integrity, plasmamembrane integrity of fresh samples, pre-freeze and post-thaw stages. Oxidative stress parameters [lipid peroxidation (LPO) and total antioxidant capacity (TAC)] were also measured at the pre-freeze and post-thaw stages. RESULTS: Humanin supplementation resulted in significantly higher (p < 0.05) post-thaw motility in all treatment groups and, higher (p < 0.05) viability in Groups III and IV in comparison to the control at the post-thaw stage. Spermatozoa with intact acrosome and plasma membrane were higher (p < 0.05) in Groups III and IV as compared to Groups I and II. The LPO levels at the post-thaw stage were found to be lower (p < 0.05) in all treatment groups versus the control group, whereas, higher (p≤0.05) TAC values were recorded in Groups III and IV in comparison to the control and Group II. CONCLUSION: Humanin supplementation in the extender improved the freezabilty of buffalo spermatozoa.

Effect of endogenous hormones, antisperm antibody and oxidative stress on semen quality of crossbred bulls.

The present study was designed to evaluate the effect of factors like hormones, antisperm antibody (ASA), and oxidative stress and its relation with semen quality in crossbred bulls. Ejaculates from two bulls were categorized into good (n = 12) and poor (n = 12) based on initial progressive motility, that is, ≥70% and ≤50%, respectively. The level of hormones like Testosterone (p < 0.05) and PGE2 (p < 0.01) was significantly higher in good-quality ejaculates compared to poor-quality ejaculates; however, estradiol (p < 0.05), progesterone, oxidative stress, and ASAs were significantly higher (p < 0.01) in poor-quality ejaculates compared to good-quality ejaculates. Therefore, it could be concluded that oxidative stress and hormonal imbalance might have resulted in high number of dead and defective spermatozoa which was ultimately responsible for poor quality semen ejaculates.

Addition of oral fexofenadine to topical therapy leads to a significantly greater reduction in the serum interleukin-31 levels in mild to moderate paediatric atopic dermatitis.

BACKGROUND: Recent evidence has suggested that oral antihistamines could have a beneficial role in atopic dermatitis (AD) because of their anti-inflammatory action. AIM: To evaluate the effectiveness of adding an oral second-generation, nonsedating, H1-receptor antihistamine (fexofenadine) to topical treatment in AD. METHODS: In this prospective randomized study, 50 patients with a diagnosis of mild to moderate AD were recruited and randomized into two groups: Group A was given appropriate topical treatment (topical tacrolimus 0.03-0.1% ointment once daily along with topical fluticasone propionate 0.05% cream once daily, as well as paraffin-based emollients) combined with oral fexofenadine, while Group B was given appropriate topical treatment only. Both groups received the respective treatments for 8 weeks. RESULTS: There was no significant difference between the two groups in terms of the SCORing Atopic Dermatitis and the 5-dimensions Itch Scale at any of the time points (Weeks 2, 4 and 8). However, in the fexofenadine group, the level of serum interleukin (IL)-31 decreased significantly from baseline to Week 8 of treatment. CONCLUSIONS: Although we could not conclusively confirm the clinical efficacy of adding oral fexofenadine to topical treatment in AD, serological evaluation indicates that fexofenadine treatment can lead to significant lowering of serum IL-31 levels in patients with AD.

Cryoprotection of humanin-like peptides in seminal plasma for ejaculated spermatozoa of crossbred bulls.

BACKGROUND: Cryopreservation process negatively affects spermatozoa functions. Humanin, a small polypeptide encoded in the mitochondrial genome, is well known for its role in cell survival. OBJECTIVE: To quantify the endogenous levels of humanin in seminal plasma of crossbred Frieswal bulls and to study its role in cryoprotection. The presence of humanin in bull spermatozoa was also investigated. MATERIALS AND METHODS: A total of 40 semen samples were separated into two groups based on the initial progressive motility (IPM): Good (IPM >70%) and Poor (IPM <50%) groups; and/or based on the post-thaw motility (PTM): Freezable (PTM>50%) and Non-freezable (PTM < 50%) groups. Humanin concentration in seminal plasma (SP-HN) was quantified using ELISA. RESULTS: SP-HN concentration ranged from undetectable to 67.6 pg/mL with a median level of 35.2 pg/mL. SP-HN level was significantly higher in the good quality semen group than in the poor quality semen group (p<0.001), and also significantly higher in the freezable group than in the non-freezable group (p<0.001). SP-HN level was positively correlated with initial progressive motility, post-thaw semen motility, viability, acrosome intactness and plasma membrane integrity, but negatively correlated the level of reactive oxygen species and malondialdehyde content. Immunochemical localization showed the presence of humanin in the proximal region of the middle piece of spermatozoa. CONCLUSION: Endogenous humanin level had significant correlation with semen quality and might protect sperm cells against freeze-induced oxidative stress. doi.org/10.54680/fr22510110712.

Effect of Mito-TEMPO Incorporated Semen Extender on Physico-morphological Attributes and Functional Membrane Integrity of Frozen Thawed Buffalo Spermatozoa.

BACKGROUND: Sperm mitochondria are the major site of reactive oxygen species (ROS) production and excess production during freezing-thawing process inflicts oxidative damages to spermatozoa. Buffalo spermatozoa are more prone to oxidative damage due to inherently more polyunsaturated fatty acids and low cholesterol to phospholipids ratio in the plasma membrane. A mitochondrial targeted antioxidant, Mito-TEMPO was used in this study. OBJECTIVE: To study the effect of Mito-TEMPO incorporated semen extender on the post-thaw semen quality in buffalo. MATERIALS AND METHODS: A total of 18 ejaculates from three murrah buffalo bulls with ≥70% individual progressive motility were utilized for the study. Each semen sample was equally divided and extended with five groups: Group I (Control, without Mito-TEMPO addition); Group II (10 µM Mito-TEMPO); Group III (50 µM Mito-TEMPO); Group IV (100 µM Mito-TEMPO); Group V (500 µM Mito-TEMPO) to have 80×106 progressive motile sperm/mL of extender, filled and sealed in French mini straws (0.25 mL) and frozen following equilibration. The effect of Mito-TEMPO was assessed at fresh/post-dilution and post-thaw stages by evaluating physico-morphological attributes and functional membrane integrity such as hypo-osmotic swelling test (HOST). RESULTS: Initial progressive motility, viability, acrosomal integrity and HOS response was significantly (p<0.05) improved and sperm abnormality was significantly (p<0.05) reduced in extended semen with Mito-TEMPO (50 µM) compared to control at post-thaw stage, although improvement was also observed at 10 and 100 µM in post-thaw samples. CONCLUSION: Mito-TEMPO incorporated semen extender at 50 µM concentration, could be part of a rationale for improving post-thaw semen quality in buffalo.

Effect of Cryopreservation on Semen Quality Parameters in Relation to Lipid Peroxidation and Antioxidant Profile in Indian Buffalo.

BACKGROUND: Lipid peroxidation (LPO) due to oxidative stress leads to structural and functional changes in spermatozoa. OBJECTIVE: To evaluate any association of various seminal characteristics at the pre- and post-cryopreservation stages with LPO and total antioxidant capacity (TAC) in Murrah buffalo semen samples. MATERIALS AND METHODS: Sixty-five ejaculates from seven bulls were processed for cryopreservation in liquid nitrogen. RESULTS: Only 31 (47.7%) samples were found satisfactory for inclusion in the further artificial insemination. A strong negative correlation was observed between LPO and individual progressive motility, TAC, viability, plasma membrane integrity as well as acrosome integrity of fresh spermatozoa. At the post-thaw stage, post-thaw motility, viability, plasma membrane integrity and acrosome integrity had strong positive correlation with TAC. CONCLUSION: The effort to minimize LPO and enhance TAC shall play a pivotal role in improving buffalo semen quality upon cryopreservation.

Exploring the role of E.coli derived enzyme, Oxyrase, as an oxygen scavenger to improve the cryotolerance of spermatozoa of Sahiwal bull.

The current study intended to optimize the concentration of Oxyrase in the semen dilutor and to evaluate its effect on freezability of spermatozoa of Sahiwal bulls. Supplementation of Oxyrase at 0.125 IU/mL concentration significantly reduced dissolved oxygen (DO) in the dilutor to 4 ppm in 16-18 min at 35 °C. For supplementation studies, a total of 24 ejaculates were categorized into poor and good ejaculates categories (n = 12 each) based on their initial progressive motility. Each ejaculate was further divided into two aliquotes. The first aliquote was diluted with tris-egg yolk extender without Oxyrase (control group) whereas, in the treatment group, Oxyrase was supplemented at the concentration of 0.125 IU/mL of extender. The parameters evaluated include cholesterol and plasma membrane phospholipids (PMP) at fresh, while IPM, acrosomal and plasma membrane integrity, cholesterol, PMP and oxidative stress parameters like lipid peroxidation (LPO), total antioxidant capacity (TAC) and reactive oxygen species (ROS) were evaluated at pre-freeze and post-thaw stages. The IPM and acrosomal intactness were higher (p < 0.05) in treatment group at post-thaw stage in good ejaculates. Oxyrase supplementation resulted in lower (p < 0.05) cholesterol leakage in both categories and lower (p < 0.05) LPO in good ejaculates at post-thaw stage. No statistical difference in ROS was observed between control and treatment groups at all stages whereas, level of TAC was higher (p < 0.05) in the treatment group compared to control group at post-thaw stage of both categories. Therefore, Oxyrase as an oxygen scavenging agent could preserve the post-thaw quality of Sahiwal bull spermatozoa.

The nucleus acts as a ruler tailoring cell responses to spatial constraints.

The microscopic environment inside a metazoan organism is highly crowded. Whether individual cells can tailor their behavior to the limited space remains unclear. In this study, we found that cells measure the degree of spatial confinement by using their largest and stiffest organelle, the nucleus. Cell confinement below a resting nucleus size deforms the nucleus, which expands and stretches its envelope. This activates signaling to the actomyosin cortex via nuclear envelope stretch-sensitive proteins, up-regulating cell contractility. We established that the tailored contractile response constitutes a nuclear ruler-based signaling pathway involved in migratory cell behaviors. Cells rely on the nuclear ruler to modulate the motive force that enables their passage through restrictive pores in complex three-dimensional environments, a process relevant to cancer cell invasion, immune responses, and embryonic development.

Strategies to Minimize Various Stress-Related Freeze-Thaw Damages During Conventional Cryopreservation of Mammalian Spermatozoa.

The aim of the article is to report a review on different sperm cryopreservation techniques, various stress-related freeze-thaw damages altering sperm structure and function during conventional cryopreservation, and strategies to minimize these stresses. Sperm cryopreservation has allowed indefinite storage and successful transportation of valuable germplasm from proven sites at distant locations, for genetic upgradation through implementation of reproductive techniques, such as artificial insemination. Different techniques for sperm cryopreservation have been proposed such as conventional freezing techniques, directional freezing, and sperm vitrification. Drawbacks related to conventional freezing methods, such as heterogeneous ice nucleation and repeated freeze-thaw cycles at the ice front that disrupts and kill sperm cells, led to the emergence of the directional freezing technique. Sperm vitrification is advantageous as there is no ice crystal-induced physical damages to sperm. However, sperm vitrification has less applicability as encouraging results are only reported in human, dog, and cat. In spite of several drawbacks, conventional freezing techniques are still most widely used for sperm cryopreservation. Spermatozoa experience stresses in the form of cold shock, osmotic stress, and mainly oxidative stress during conventional cryopreservation ultimately reduces the sperm viability and fertility. Several attempts have been made in the past to minimize all these stresses individually or in combination. Membrane fluidity was increased to prevent the cold shock and cryocapacitation-like changes by the addition of cholesterol to the membrane. Antifreeze proteins were added in semen extender to minimize freeze-thaw damages due to heterogeneous ice nucleation and ice recrystallization. Oxidative stress was reduced either by neutralizing reactive oxygen species (ROS) through enzymatic, nonenzymatic, plant-based antioxidants or reductants; or by minimizing the level of sources like the semen radiation exposure, leucocytes, and dead and defective spermatozoa, which lead to ROS production during the semen cryopreservation process. A novel approach of minimizing oxidative stress was to reduce the oxygen tension in sperm microenvironment that is, extender by partial deoxygenation process, as a number of literatures pointed out direct link of O2 with ROS production. When compared with other strategies, partial deoxygenation of semen extender with N2 gassing is found as a cost-effective, comparatively easy and a potential approach to large-scale frozen semen production.

Interaction of antioxidant gene variants and susceptibility to type 2 diabetes mellitus.

Background: Diabetes is the seventh most common disease leading to death with a global estimate of 425 million diabetics, expected to be 629 million in 2045. The role of reactive metabolites and antioxidants, such as glutathione, glutathione peroxidase, superoxide dismutase and catalase in type 2 diabetes mellitus (T2DM) provides an opportunity for identifying gene variants and risk genotypes. We hypothesised that certain antioxidant gene-gene interactions are linked with T2DM and can model disease risk prediction.Materials and methods: Genotyping of single nucleotide polymorphisms (SNPs) in antioxidant genes for glutathione (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) was performed in 558 T2DMs and 410 age and sex matched healthy controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), routine lab indices by standard techniques.Results: The null/null allele combination of GSTM1del and GSTT1del increased disease risk up to 1.7-fold. The combination of SNPs in GSTM1del, GSTT1del, GSTP1 + 313A/G and in CAT-21A/T, SOD2 + 47C/T, GPx1 + 599C/T increased the risk of diabetes 13.5 and 2.1-fold, respectively. Interaction of SNPs GSTM1del, GSTT1del, GSTP1 + 313A/G (105Ile/Val), CAT-21A/T, SOD2 + 47C/T, GPx1 + 599C/T were significantly linked with disease risk >5 × 103 fold.Conclusion: As the number of gene combinations increase, there is a rise in the odds ratio of disease risk, suggesting that gene-gene interaction plays an important role in T2DM susceptibility. Individuals who possess the GSTM1del, GSTT1del, GSTP1 105I/V(+313A/G), CAT-21A/T, SOD2 + 47C/T and GPx1 + 599C/T are at very high risk of developing T2DM.

Health risks from PAHs and potentially toxic elements in street dust of a coal mining area in India.

Polycyclic aromatic hydrocarbons (PAHs) and potentially toxic elements (PTEs) (Ba, Zn, Pb, Cu, Cr, Ni, As, Co) were determined in the road dusts of a coal mining area (Dhanbad, India) to assess their content and potential human health risks. Dust samples were collected from sign boards of the heavy traffic road connecting Dhanbad and Sindri. The total PAHs (∑PAHs, all values in mg/kg) content in the road dust samples varied from 3.98 to 13.1, with carcinogenic PAHs content of 14.8-34.4% of the ∑PAHs. Phenanthrene (2.72), fluorene (0.715) and pyrene (0.575) are the major PAHs. Principal component analysis revealed that these PAHs are probably originated from pyrogenic (coal combustion and traffic emission) and petrogenic (coal dust, tyre and road particles) sources. Among the PTEs, the mean content was higher for Ba (293 mg/kg) followed by Zn (224), Pb (128), Cu (52.6), Cr (45.2), Ni (22.0), As (17.5) and Co (8.11). The overall pollution load index varied from 0.43 to 1.0. Source analysis showed that PTEs in the road dust of the study site were derived from traffic emission (Zn, Fe, Mn, Co and Pb), coal dust (Cr, As and Ni) and soil (K, Mg, Ba, Sr and Ca). In general, the PTEs are lower, but the PAHs contents were elevated in the road dust samples. Although the exposure risks from PTEs are low, the risk to children (expressed as hazardous quotient) for As and Pb is near to the permissible limit of 1.0. Cancer risk from PAHs for adult (4.8 × 10-6) and child (5.3 × 10-6) has exceeded the acceptable limit of 10-6.

Four compartment method as an efficacious and simplified technique for autologous non-cultured epidermal cell suspension preparation in vitiligo surgery: A randomized, active-controlled study.

BACKGROUND: Autologous non-cultured epidermal cell suspension (NCES) is a successful surgical method for repigmentation of stable, refractory vitiligo. The need for laboratory equipment and expertise restricts its use to only a few research centres; hence, there is a requisite to simplify the technique of NCES preparation. OBJECTIVES: To evaluate the efficacy of NCES prepared by four compartment (4C) method compared to the laboratory-based method (lab-NCES). METHODS: Anatomically based matched lesions (41 pairs) in 30 stable vitiligo patients were randomized to receive NCES prepared by 4C method or lab-NCES. Each patient was evaluated at 4, 8, 16 weeks after surgery by a blinded observer with regard to extent of repigmentation, colour match, patient global assessment (PGA) and pattern of repigmentation. RESULTS: Repigmentation outcome in 4C method as compared to lab-NCES was as follows: excellent (≥90%) repigmentation: 34% vs. 37%, P = 1.000, good (≥75%) repigmentation: 68% vs. 71%, P = 1.000; colour match: 59% vs. 54%, P = 0.794, patient satisfaction based on PGA score - 23.02 vs. 23.39 (P = 0.210) and major pattern of repigmentation (diffuse) - 76% vs. 71% (P = 0.618). LIMITATIONS: Short follow-up period of 16 weeks. CONCLUSION: Four compartment method is a simple and effective technique for vitiligo surgery in routine clinical practice.

Reduction of dissolved oxygen in semen extender with nitrogen gassing reduces oxidative stress and improves post-thaw semen quality of bulls.

The objective of the present study was to investigate the effects of two different concentrations of dissolved oxygen (DO, 4 and 8) ppm in the extender on oxidative stress affecting plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), and deoxyribonucleic acid (DNA) damage of bull spermatozoa following cryopreservation. For the experiment, nitrogen (N2) gassing of the extender for varied time intervals yielded extender with DO concentration of 4 ppm and 8 ppm (Groups II and III, respectively). For the Control (Group I) without N2 gassing, a DO concentration of 11.7 ppm was recorded. Following sample selection, ejaculates were divided into three aliquots and were extended to have 80 × 106 spermatozoa/mL of extender in the three groups. Semen samples were evaluated for reactive oxygen species (ROS), lipid peroxidation (LPO), total antioxidant capacity (TAC) and superoxide dismutase (SOD) at the fresh, pre-freeze, and post-thaw stages. Evaluation of PMI, MMP, and DNA damage were conducted on frozen-thawed samples. There were greater (P < 0.05) increase in ROS and LPO and decrease in TAC concentrations in Group I than Groups II and III. Mean values of SOD at the post-thaw stage was greater (P < 0.05) in Group II than Group I. There was a similar trend in the PMI in Groups II and III; MMP and DNA integrity in Group II was greater compared with Group I. In conclusion, results indicate there was a beneficial effect of maintaining DO concentrations at 4 rather than of 8 or 11.7 ppm in extender for sustaining post-thaw semen quality.

Administration of slow release exogenous melatonin modulates oxidative stress profiles and in vitro fertilizing ability of the cryopreserved mithun (Bos frontalis) spermatozoa.

Mithun (Bos frontalis) is a unique domestic free range bovine species of North Eastern hilly regions of India. The present study was designed to assess the seasonal effect of slow release exogenous melatonin (MT) implant on semen quality parameters (SQP) and in vitro zona binding ability (IVZ) of spermatozoa. The experimental animals were divided into Gr I: Control (n = 5) and Gr II: Treatment (n = 5; melatonin implant @ 18mg/50 kg bwt). A total of 20 semen samples/group in winter, spring, autumn and summer seasons (n = 160), twice per week were collected. Following cryopreservation, samples were evaluated for motility parameters (forward progressive, mobility & velocity by computer assisted sperm analyser (CASA), viability, acrosome integrity, plasma membrane and nuclear abnormality, functional status of mitochondria, enzymatic, antioxidant and oxidative profiles, and IVZ. The study revealed significant (p < 0.05) improvement in total motility, viability, acrosome-, plasma membrane-, and nuclear-integrity, and antioxidant profiles; with highest values in spring and lowest in summer season in the fresh semen in Gr II than the Control. A significant (p < 0.05) improvement in motility parameters, membrane potential of mitochondria, antioxidant profiles and reduction in sperm and nuclear abnormalities, leakage of intracellular enzymes and oxidative stress and IVZ index & binding percentage in post-thaw semen samples in melatonin supplemented than in un-supplemented control group was observed. It can be concluded from the study that slow-release melatonin supplementation can be effectively utilized to improve the antioxidant profiles and reduction of oxidative stress, with cascading beneficial effects on semen quality parameters and fertility status of the mithun bull.

Effect of season and age on scrotal circumference, testicular parameters and endocrinological profiles in mithun bulls.

Mithun is a domesticated free-range bovine species primarily used as a meat animal and is a pride of North Eastern Hilly regions of India. The present study was conducted to measure the effect of seasons on scrotal & testicular biometrics and endocrinological profiles for different age groups at different seasons in mithun bulls. A total of 30 mithun males were selected from the mithun breeding farm, ICAR-NRC on Mithun, Medziphema, Nagaland, India and were equally distributed into five groups based on their age. Each group consisted of six animals and the groups were Gr A (0.1-1.0 year), Gr B (1.1-2.0 years), Gr C (2.1-3.0 years), Gr D (3.1-5.0 years) and Gr E (5.1-6.0 years). The seasons were grouped into winter, spring, summer and autumn based on the meteorological data such as temperature humidity index (THI) and sunshine hours. Scrotal circumference (SC) & testicular biometrics and endocrinological profiles such as follicle stimulating hormone (FSH), luteinizing hormone/interstitial cell stimulating hormone (LH/ICSH), testosterone, thyroxine (T4), cortisol and insulin like growth factor 1 (IGF 1) were estimated during different seasons for different age groups. Statistical results revealed that the SC & testicular biometrics and endocrinological profiles differed significantly (p < 0.05) among the different age groups for the same season whereas SC and endocrinological profiles significantly (p < 0.05) differed among the seasons for same age group. Significantly (p < 0.05) greater SC and testicular weight were observed in Gr E and D, lower in Gr A and B and intermediate in Gr C and increased as age advances. Significantly (p < 0.05) greater SC was observed in winter and spring and lowest was in summer season. The hormone profiles such as FSH, LH/ICSH, testosterone & thyroxine were significantly (p < 0.05) greater and IGF-1 & cortisol were significantly (p < 0.05) lower in spring & winter than in summer season. The hormones, FSH, LH/ICSH and thyroxine increased significantly in Gr A followed by reduced in Gr B and then increased to Gr D and E, whereas concentration of testosterone, cortisol and IGF-1 increased according to age advanced. It was concluded that the spring and winter seasons has significantly greater beneficial effects than summer season on reproduction and artificial breeding programme in semi-intensive management of mithun in the present location.

Antisperm antibodies in repeat-breeding cows: Frequency, detection and validation of threshold levels employing sperm immobilization, sperm agglutination and immunoperoxidase assay.

Antisperm antibodies have been found in repeat-breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat-breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut-off value) peroxidase-positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer

Effectiveness of Long Term Supervised and Assisted Physiotherapy in Postsurgery Oral Submucous Fibrosis Patients.

Oral submucous fibrosis is one of the leading potentially malignant disorders prevailing in India. A number of conservative and surgical treatment options have been suggested for this potentially malignant disorder (Arakeri and Brennan, 2013). While the role of physiotherapy has been highlighted in the conservative management, its importance in postsurgical cases to avoid scar contracture and subsequent relapse has not been given due importance in the literature. The following is a case report of a male patient surgically treated for OSMF (oral submucous fibrosis) and meticulously followed up for recalls and physiotherapy. The constant supervision and motivation for physiotherapy along with the constant assistance helped achieve satisfying results.

Effects of low-density lipoproteins as additive on quality parameters and oxidative stress following cryopreservation of mithun (Bos frontalis) spermatozoa.

Artificial breeding of mithun poses several challenges including lack of standard protocol for cryopreservation of spermatozoa. This is further complicated by harmful effects of hen's egg yolk (EY) as additive in extender. Purified low-density lipoproteins (LDL) extracted from EY have been shown as beneficial over EY extender for long-term semen storage in several species. This investigation explored use of LDL versus EY on semen quality and oxidative stress following freezing-thawing of spermatozoa. A total of 25 of 50 ejaculates based on biophysical parameters were selected for the experiment. After diluting with the Tris-citrate-glycerol (TCG) extender, each sample was split into three equal aliquots: Group I, control, EY; Group II and Group III contained 8% and 10% purified LDL, respectively. Frozen-thawed samples were evaluated for motility parameters (progressive, and in the bovine cervical mucus penetration test [BCMPT]), viability, sperm and nuclear abnormality, acrosome integrity, and enzymatic (leakage of intracellular contents) and biochemical (oxidative stress) profiles and in vitro fertility (IVF) assay. Study revealed a significant (p < .05) improvement in viability, sperm and nuclear abnormality, acrosome integrity, motility (progressive and in cervical mucus), cholesterol content, and reduction in the leakage of intracellular enzymes in Group II. Moreover, intactness of acrosome and biochemical membranes was protected significantly (p < .05) in addition to significant (p < .05) improvement in binding per cent and binding index in IVF assay in extender containing 8% LDL. These results demonstrate that although cryopreservation of mithun's spermatozoa in EY was comparable with other species, addition of 8% LDL holds a clear advantage over EY or 10% LDL.