Effect of antioxidants on post thaw microscopic, oxidative stress parameter and fertility of Boer goat spermatozoa in Tris egg yolk glycerol extender.

This study was conducted to determine the effect of antioxidants on standard semen parameters, lipid peroxidation and fertility of Boer goat semen after cryopreservation. Ejaculates from four bucks were collected, evaluated and pooled at 37. °C. The pooled semen was diluted with Tris citric acid fructose for washing. Semen samples, which were diluted with a Tris-based extender containing the antioxidant ascorbic acid (8.5. mg/ml), butylated hydroxytoluene (2. mM), cysteine (5. mM) and hypotaurine (10. mM) and an extender without antioxidant supplementation were cooled to 4. °C and frozen in 0.25 straws with programmable freezer and finally stored in liquid nitrogen. Data (10 replicates) were analyzed by one-way analysis of variance. Mean (±SEM) progressive motility was significantly higher in ascorbic acid than other supplement groups and control samples (P> 0.05). Best values were observed in ascorbic acid followed by BHT, cysteine, and hypotaurine. Antioxidant supplementation in extender showed significant (P< 0.05) better values than the control group for sperm membrane integrity, acrosome integrity and viability. The ability of antioxidants to reduce the lipid peroxidation (LPO) after freeze thawing was measured by the formation of malondialdehyde (MDA) using the thiobarbituric acid method. Results showed that addition of antioxidants significantly reduced the rate of LPO in comparison to control (P< 0.05). Ascorbic acid exhibited better values (1.27 ± 0.28), than butylated hydroxytoluene, cysteine and hypotaurine 1.32 ± 0.42, 2.27 ± 0.16 and 2.38 ± 0.17 respectively, which are significantly better than control (3.52 ± 0.54). Higher pregnancy rate was observed with ascorbic acid followed by butylated hydroxtolune, hypotaurine and cysteine. However, differences in the fertility rate were non-significant with hypotaurine, cysteine and control group

Alpha-linolenic acid supplementation in Tris extender can improve frozen–thawed bull semen quality

The study was conducted to evaluate the effects of α-linolenic acid (ALA) on frozen–thawed quality and fatty acid composition of bull sperm. For that, twenty-four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25-ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer-assisted semen analysis), membrane functional integrity (hypo-osmotic swelling test), viability (eosin-nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post-thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen–thawed bull spermatozoa

Effect of hypotaurine and cysteine on sperm cytological parameters of cooled and post thaw boer goat semen.

The purpose of this study was to determine the influence of antioxidant additives (hypotaurine and cysteine) in different concentrations to the cryopreserving media on the semen cytological parameters pre freezing and post thawing (motility, membrane integrity, morphology, acrosome integrity and viability).The experiment was done on 30 ejaculates collected by artificial vagina method from 5 boer goat bucks during April to May 2011. After collection, ejaculates qualifying standard criteria were pooled. Pooled ejaculates were washed for seminal plasma removal and then diluted in medium based on Tris in which antioxidants were added in various concentrations (hypotaurine 5, 10 and 20mM; cysteine 5, 10 and 20mM) or without antioxidants (control). The diluted semen was cooled at 4ºC, filled in 0.25ml French straws and then stored in liquid nitrogen. The results showed that semen quality did not differ (P < 0.05) in terms of morphology and acrosome integrity with antioxidants supplementation after cooling. Hypotaurine and cysteine significantly improved the characteristics of boer goat semen motility, membrane integrity, morphology, acrosome integrity and viability after cryopreservation. Addition of hypotaurine at 10mM and cysteine at 5mM concentration leads maximum improvement in liquid and frozen boer goat sperm cytological characteristics

Biocontrol of Bacillus cereus by Lactobacillus plantarum in Kareish cheese and yogurt

This study aims to biocontrol of Bacillus cereus by Lactobacillus plantarum in Kareish cheese and yogurts. The minimum inhibitory concentrations (MICs), antioxidant potentials, total flavonoids content (TFC) and total phenolic content (TPC) of L. plantarum were also estimated. Results showed that incidence of B. cereus in Kareish cheese and yogurt was 16 and 4%, respectively. Four virulence genes were investigated by PCR in B. cereus isolates (n = 10). Two toxin producing genes, cytotoxin K (cytK), and phosphatidylcholine-hydrolyzing phospholipase C (Pc-plc), were detected in all B. cereus, whereas enterotoxigenic (nhe) and hemolysin BL (hbl) genes were detected in 90 and 50%, respectively. All isolates were vulnerable to erythromycin and gentamicin (100%) with intermediate sensitivity to ciprofloxacin and complete resistance to tetracycline (100%). L. plantarum showed antibacterial power against B. cereus EMCC1006 reference strain with MIC at 3.1 mg/mL. From the different concentrations (1.5, 3.1, 6.25, 12.5, 25.0, 50.0, 100.0 mg/mL) of L. plantarum which mixed with Kareish cheese and yogurt samples, the minimum concentrations displayed the excellent sensory parameters. TPC and TFC of L. plantarum CFS were 18.5 (μg GAE/g) and 2.67 (μg QE/g), respectively. Regarding antioxidant activity, IC50 of L. plantarum was 53.84 μg/mL, while IC50 of ascorbic acid was 26.36 μg/mL. In sum, L. plantarum could be used as a promising antibacterial and antioxidant agent for biocontrol of B. cereus to ensure dairy safe without negative impact on sensorial attributes.Universidade de Vigo/CISU

Infantile hypertrophic pyloric stenosis: a single institution’s experience

Background/purpose Infantile hypertrophic pyloric stenosis (IHPS) is said to be relatively common in the western world, but its incidence in the Kingdom of Saudi Arabia is unknown. We set out to study the incidence of IHPS in children presenting at our hospital and review the clinical presentation and treatment outcomes of such patients.Patients and methods A retrospective chart review of all cases with IHPS admitted to a tertiary hospital, Riyadh Military Hospital, over 18 years (1990–2008) was conducted. Diagnosis was confirmed by ultrasonography according to the length of the pyloric channel and thickness of the pyloric muscles. An analysis of demographic data, clinical presentation, diagnostic modality, and associated congenital anomalies was carried out. Operative technique, postoperative course, and complication rates were identified.Results The review of the medical records showed 22 patients with a diagnosis of IHPS. The incidence rate of IHPS was 1.4/10 000 live births. Demographically, most cases presented in the first month of life. It is more common in boys. In all, 55% of infants were first borns. Clinically, nonbilious projectile vomiting was the main symptom. Mild dehydration was found in half of the cases with alkalotic changes. Ultrasonography was the main diagnostic modality in 82% of cases. Others were diagnosed by contrast study. Open Ramstedt pyloromyotomy was the surgical approach in 20 (91%) cases and the laparoscopic approach in 2 cases (9%). Fourteen (14%) patients had an associated congenital inguinal hernia. The mean length of hospital stay was 5.5 days. The most common postoperative complication was wound infection in 4.5% of patients. Incidence of IHPS had obvious seasonal association.Conclusion Incidence of IHPS at our hospital (the biggest hospital in the Kingdom of Saudi Arabia with an annual birth record of 14 000) is far less common than international incidences reported in North America and Europe. More reports from Saudi Arabia, the Middle East, and Asia are recommended to support the rarity of IHPS in this region of the world. Seasonal variation suggests a possible etiological role for environmental factors. It is of practical use for both epidemiologists and clinicians for future comparability.Keywords: incidence, infantile pyloric stenosis, risk factor

Effect of ascorbic acid concentrations, methods of cooling and freezing on boer goat semen cryopreservation.

To improve the Boer goat semen quality during cryopreservation process, three experiments were carried out to investigate the effect of (i) different concentration of ascorbic acid supplementation (ii) rate of cooling with chilled semen characteristics and (iii) method of freezing on post-thaw Boer goat sperm using Tris-based extender. Ascorbic acid at 8.5mg/ml improved the sperm parameters (motility, integrity of membrane and acrosome, morphology and viability), compared to control in cooled samples (p<0.05). With regard to other concentrations and post-thawed parameters, ascorbic acid at 2.5-8.5mg/ml led to higher percentages of sperm motility and integrities of membrane and acrosome when compared to control (p<0.05). Slow cooling rises to higher percentages of sperm motility, acrosome integrity and viability, in comparison with fast cooling, in terms of cooled and frozen samples (p<0.05). Programmable freezing method produced the higher percentages of sperm motility, integrities of membrane and acrosome and viability when compared to the freezing method of polystyrene box during goat sperm freezing (p<0.05). In conclusion, chilled and post-thawed sperm quality of Boer goat was improved when a Tris-based extender supplemented with ascorbic acid was used at stages of different cooling rates and freezing methods. © 2012 Blackwell Verlag GmbH

Inequalities in Health Status from EQ-5D Findings: A Cross-Sectional Study in Low-Income Communities of Bangladesh

Background: Measuring health status by using standardized and validated instrument has become a growing concern over the past few decades throughout the developed and developing countries. The aim of the study was to investigate the overall self-reported health status along with potential inequalities by using EuroQol 5 dimensions (EQ-5D) instrument among low-income people of Bangladesh. Methods: A cross-sectional household survey was conducted in Chandpur district of Bangladesh. Bangla version of the EQ-5D questionnaire was employed along with socio-demographic information. EQ-5D questionnaire composed of 2-part measurements: EQ-5D descriptive system and the visual analogue scale (VAS). For measuring health status, UK-based preference weights were applied while higher score indicated better health status. For facilitating the consistency with EQ-5D score, VASs were converted to a scale with scores ranging from 0 to 1. Multiple logistic regression models were also employed to examine differences among EQ-5D dimensions. Results: A total of 1433 respondents participated in the study. The mean EQ-5D and VAS score was 0.76 and 0.77, respectively. The females were more likely to report any problem than the males (P<0.001). Compared to the younger, elderly were more than 2-3 times likely to report any health problem in all EQ-5D dimensions (OR [odds ratio]=3.17 for mobility, OR=3.24 for self-care). However, the respondents of the poorest income group were significantly suffered more from every EQ-5D dimension than the richest income quintile. Conclusion: Socio-economic and demographic inequalities in health status was observed in the study. Study suggests to do further investigation with country representative sample to measure the inequalities of overall health status. It would be helpful for policy-maker to find a new way aiming to reduce such inequalities

Head and neck cancer predictive risk estimator to determine control and therapeutic outcomes of radiotherapy (HNC-PREDICTOR):development, international multi-institutional validation, and web implementation of clinic-ready model-based risk stratification for head and neck cancer

Background: Personalised radiotherapy can improve treatment outcomes of patients with head and neck cancer (HNC), where currently a ‘one-dose-fits-all’ approach is the standard. The aim was to establish individualised outcome prediction based on multi-institutional international ‘big-data’ to facilitate risk-based stratification of patients with HNC. Methods: The data of 4611 HNC radiotherapy patients from three academic cancer centres were split into four cohorts: a training (n = 2241), independent test (n = 786), and external validation cohorts 1 (n = 1087) and 2 (n = 497). Tumour- and patient-related clinical variables were considered in a machine learning pipeline to predict overall survival (primary end-point) and local and regional tumour control (secondary end-points); serially, imaging features were considered for optional model improvement. Finally, patients were stratified into high-, intermediate-, and low-risk groups. Results: Performance score, AJCC8th stage, pack-years, and Age were identified as predictors for overall survival, demonstrating good performance in both the training cohort (c-index = 0.72 [95% CI, 0.66–0.77]) and in all three validation cohorts (c-indices: 0.76 [0.69–0.83], 0.73 [0.68–0.77], and 0.75 [0.68–0.80]). Excellent stratification of patients with HNC into high, intermediate, and low mortality risk was achieved; with 5-year overall survival rates of 17–46% for the high-risk group compared to 92–98% for the low-risk group. The addition of morphological image feature further improved the performance (c-index = 0.73 [0.64–0.81]). These models are integrated in a clinic-ready interactive web interface: https://uic-evl.github.io/hnc-predictor/ Conclusions: Robust model-based prediction was able to stratify patients with HNC in distinct high, intermediate, and low mortality risk groups. This can effectively be capitalised for personalised radiotherapy, e.g., for tumour radiation dose escalation/de-escalation

The PKR-binding domain of adenovirus VA RNAI exists as a mixture of two functionally non-equivalent structures

VA RNAI is a non-coding adenoviral transcript that counteracts the host cell anti-viral defenses such as immune responses mediated via PKR. We investigated potential alternate secondary structure conformations within the PKR-binding domain of VA RNAI using site-directed mutagenesis, RNA UV-melting analysis and enzymatic RNA secondary structure probing. The latter data clearly indicated that the wild-type VA RNAI apical stem can adopt two different conformations and that it exists as a mixed population of these two structures. In contrast, in two sequence variants we designed to eliminate one of the possible structures, while leaving the other intact, each formed a unique secondary structure. This clarification of the apical stem pairing also suggests a small alteration to the apical stem–loop secondary structure. The relative ability of the two apical stem conformations to bind PKR and inhibit kinase activity was measured by isothermal titration calorimetry and PKR autophosphorylation inhibition assay. We found that the two sequence variants displayed markedly different activities, with one being a significantly poorer binder and inhibitor of PKR. Whether the presence of the VA RNAI conformation with reduced PKR inhibitory activity is directly beneficial to the virus in the cell for some other function requires further investigation

Biophysical, Biochemical, and Molecular Docking Investigations of Anti-Glycating, Antioxidant, and Protein Structural Stability Potential of Garlic.

Garlic has been reported to inhibit protein glycation, a process that underlies several disease processes, including chronic complications of diabetes mellitus. Biophysical, biochemical, and molecular docking investigations were conducted to assess anti-glycating, antioxidant, and protein structural protection activities of garlic. Results from spectral (UV and fluorescence) and circular dichroism (CD) analysis helped ascertain protein conformation and secondary structure protection against glycation to a significant extent. Further, garlic showed heat-induced protein denaturation inhibition activity (52.17%). It also inhibited glycation, advanced glycation end products (AGEs) formation as well as lent human serum albumin (HSA) protein structural stability, as revealed by reduction in browning intensity (65.23%), decrease in protein aggregation index (67.77%), and overall reduction in cross amyloid structure formation (33.26%) compared with positive controls (100%). The significant antioxidant nature of garlic was revealed by FRAP assay (58.23%) and DPPH assay (66.18%). Using molecular docking analysis, some of the important garlic metabolites were investigated for their interactions with the HSA molecule. Molecular docking analysis showed quercetin, a phenolic compound present in garlic, appears to be the most promising inhibitor of glucose interaction with the HSA molecule. Our findings show that garlic can prevent oxidative stress and glycation-induced biomolecular damage and that it can potentially be used in the treatment of several health conditions, including diabetes and other inflammatory diseases