Supplementation of cauda epididymal plasma improves sperm characteristics following liquid preservation of ram semen at 3-5°C.

Mammalian spermatozoa remain immotile and metabolically inactive in the cauda epididymidis, thus maintaining fertility for several weeks. The aim of this study was to functionally characterise and evaluate the effect of cauda epididymal plasma (CEP) on liquid preservation of ram spermatozoa. Four experiments were conducted to investigate the effects of: (1) CEP and its fractions on sperm motility; (2) CEP (10%, 15%, 20% v/v) on liquid preservation of ram spermatozoa; (3) seminal plasma (SP; 20%, 30%, 50% v/v) on liquid-preserved spermatozoa; and (4) both CEP and post-storage SP treatment on sperm characteristics. Biochemical characterisation of ram CEP revealed high protein (30.9mgmL-1), catalase (68.9IUmL-1), alkaline phosphatase (17.5IUmL-1) activities and total antioxidant capacity (1112µM Trolox equivalent). Progressive motility of prewashed cauda spermatozoa was reduced (P<0.05) by CEP or its protein-rich fraction compared with protein-free plasma or phosphate-buffered saline. After 48h storage, total motility, rapid motility (average path velocity >75µms-1; 53.9%, 73.5% and 71.8% with 0, 15% and 20% CEP respectively) and straight line velocity (86.3, 102.1 and 102.4µms-1 with 0, 15% and 20% CEP respectively) were significantly (P<0.05) higher in the CEP-treated groups than the control. Viability and acrosomal integrity were similar between groups; however, functional membrane integrity was higher (P<0.05) in the 15% CEP-treated group. Treatment of liquid-preserved spermatozoa with either 20%, 30% or 50% SP improved (P<0.05) rapid motility and kinematics at each time point of storage compared with control. In conclusion, liquid preservation of ram spermatozoa in the presence of 15% or 20% CEP and post-storage treatment with SP significantly improve sperm characteristics.

Antioxidants protect proteins' anchorage to the bilayer by improving plasma membrane integrity of ram spermatozoa during liquid preservation in a soya lecithin-based diluent.

Antioxidants are known to prevent the reactive oxygen species (ROS)-mediated peroxidative damage to the membrane lipids during hypothermic storage of mammalian spermatozoa. We hypothesized here that ROS also affect the lipid-protein interactions, thereby diminishing the membrane's integrity and proteins' anchorage to the bilayer. Antioxidants prevent these damages by scavenging the ROS. Ejaculates from Patanwadi rams were pooled after subjective evaluation and centrifuged using Percoll® . Sperm pellet was resuspended in soya lecithin-Tris-fructose diluent (400 × 106  cells/ml) containing either antioxidants (100 IU/ml catalase + 10 mM reduced glutathione) or no antioxidant. Aliquots were chilled to 5°C in a cabinet and stored in a refrigerator at 3-5°C for 72 hr. Sperm motility, viability, lipid peroxidation (LPO) and hypo-osmotic swelling test (HOST) were performed at 0, 24, 48 and 72 hr. Sperm proteins extracted with 0.5% Triton X-100 were resolved by SDS-PAGE and quantified using Quantity One software (Bio-Rad, USA). The rapid motility, linearity and straight-line velocity (VSL) were found significantly (p < .05) higher in the antioxidant-treated group compared to the control at 48 hr of storage. Sperm viability was found comparable between the groups. Higher HOST response and lower LPO were found in the antioxidant-treated sample compared to the control both at 48 and at 72 hr. Overall, the proteins P1 (106.09 kDa), P2 (87.00 kDa) and P4 (51.14 kDa) were lower (p < .05) in the sperm extract of antioxidant-treated group compared to the control. The content of P4 (51.14 kDa) in sperm extract was found to increase (p < .05) earlier (48 vs. 72 hr) in the control group compared to the antioxidant-treated group. Altogether, the results suggested that antioxidants reduced LPO in spermatozoa, resulting in higher sperm motility, plasma membrane integrity and protection of proteins' anchorage to the plasma membrane at 48 and 72 hr of storage.

Impact of Adrenocortical Insufficiency on Biochemical Parameters in Haemodynamically Stable Cirrhotic Patients with Ascites.

Cirrhosis has many complications regardless of the aetiology. Among them, adrenal insufficiency is recently identified entity. A prospective cohort study was done to evaluate the biochemical impact of adrenocortical insufficiency in haemodynamically stable, non-septic, cirrhotic patients with ascites and had been performed at the inpatient of GHPD department, BIRDEM, Dhaka from April 2011 to March 2012. A total of fifty three (53) patients fulfilling inclusion criteria were included in the study. Patients were divided into two groups: Group A (patients of normal adrenal function) and Group B (patients of insufficient adrenal function) and those were followed up for the next 6 months. In Group A, the total number of patients was 25(47%) and in Group B it was 28(53%). Between two groups, mean age difference and gender difference were not statistically significant. (p value was 0.278 and 0.933, respectively). At enrollment, there was significant lower mean Hb concentration in Group B (p=0.008). There was no significant difference of means of WBC count and platelet count between two groups (p value was 0.829 and 0.333, respectively). There were significant abnormalities in serum bilirubin, serum albumin, INR, SBP, HRS, Serum Na concentration, TCO2 concentration in Group B patients at follow up after 6 months. Adrenal insufficient decompensated cirrhotic patients have higher biochemical abnormalities, thus higher morbidities.

Impact of Adrenocortical Insufficiency on Clinical Parameters in Haemodynamically Stable Cirrhotic Patients with Ascites.

Cirrhosis has many complications regardless of the aetiology. Complications include splenomegaly, ascites, hepatic encephalopathy, spontaneous bacterial peritonitis, hepatorenal syndrome and hepatocellular carcinoma and also linked to abnormalities in the endocrine system, including abnormal sex hormone metabolism, thyroid disease, osteoporosis, and, most recently identified, adrenal insufficiency. This prospective cohort study was done to evaluate the impact of adrenocortical insufficiency on clinical parameters in haemodynamically stable cirrhotic patients with ascites and had been performed at the inpatient of GHPD Department, BIRDEM, Dhaka, Bangladesh from April 2011 to March 2012. A total of fifty three (53) patients fulfilling inclusion criteria were included in the study. Patients were divided into two groups: Group A (patients of normal adrenal function) and Group B (patients of insufficient adrenal function) and those were followed up for the next 6 months. In Group A, the total number of patients was 25(47%) and in Group B it was 28(53%). Between two groups, mean age difference and gender difference were not statistically significant (p value was 0.278 and 0.933, respectively). Group B patients had significant higher CLD duration (p=0.004). Haematemesis and/or maelena was significantly lower in Group B at follow up (p=0.0001) due to significant higher number of band ligation in this group (p=0.009). Hepatic encephalopathy was significantly higher in Group B at enrollment (p=0.028) and at follow up (p<0.001). During the period of follow up, significant higher number of patients had developed hepatic encephalopathy in Group B compared to Group A (p<0.05). There was statistically significant higher number of patients had SBP (p=0.031) in Group B at follow up. During the period of follow up, only 1(4%) patient in Group A and 5(18%) patients in Group B died. There was no significant difference of number of death between two groups (p=0.196). Adrenal insufficient decompensated cirrhotic patients have higher morbidities.

Comparative Study between Pentoxifylline and Pioglitazone in the Treatment of Non-Alcoholic Fatty Liver Disease among Newly Detected Glucose Intolerant Patients.

This quasi experimental study was carried out to compare the efficacy of Pentoxifylline versus Pioglitazone in non-alcoholic fatty liver disease (NAFLD) among newly detected glucose intolerant patients attended at GHPD, BIRDEM, Dhaka, Bangladesh from March 2011 to May 2012. Sixty patients with newly detected abnormal glucose tolerance, naive to any antidiabetic drugs were randomly selected, with the findings of USG changes of fatty liver and raised ALT. Patients were divided into Group A (51.7%; mean age - 44.45±7.34 years, BMI - 26.76±3.65kg/m(2)) receiving Pioglitazone 30mg/day and Group B (48.3%; mean age - 43.97±10.13 years, BMI - 27.52±4.44kg/m(2)) receiving Pentoxifylline 1200mg/day along with dietary and lifestyle modification. Baseline, follow up 1 & follow up 2: Serum AST in Group A (66.58±40.78U/L, 45.00±19.43U/L and 33.25±9.92U/L) respectively and in Group B (54.13±20.11U/L, 38.31±12.90U/L and 30.62±9.63U/L) respectively. Serum ALT in Group A (113.48±61.38U/L, 61.16±19.45U/L and 42.45±13.84U/L) respectively & in Group B (99.13±37.95U/L, 50.27±15.95U/L and 37.24±9.51U/L) respectively. No patient with normal USG finding was included in the study. Baseline USG finding: Grade I - Group A (35.5%), Group B (27.6%); Grade II - Group A (54.8%), Group B (48.3%); Grade III - Group A (9.7%), Group B (24.1%). Final Follow up: Normal - Group A (16.1%), Group B (10.3%); Grade I - Group A (38.7%), Group B (34.5%); Grade II - Group A (41.9%), Group B (44.8%); Grade III - Group A (3.2%), Group B (10.3%). Within groups findings were statistically significant (P<0.001) but between groups not significant (P>0.001). Biochemical and USG grading were improved in follow ups in both the groups. Such findings were observed due to synergistic effect of both therapeutic intervention along with supplementary dietary and lifestyle modification. Both Pioglitazone and Pentoxifylline have similar therapeutic outcome combined with supplementary dietary and lifestyle modification.

Investigation on CMB monopole and dipole using blackbody radiation inversion.

The COBE/FIRAS dataset is used to calculate the Cosmic Microwave Background temperature and the uncertainty using the Blackbody Radiation Inversion (BRI) method. In this research work, the procedure is somewhat comparable to the mixing of weighted blackbodies in the case of the dipole. The temperature and its spreading for the monopole and dipole, respectively, are 2.741 ± 0.018 K and 2.748 ± 0.270 K. This dipole spreading exceeds the spreading predicted by taking relative motion into account (i.e., 3.3 × 10-3 K). The comparison of the probability distributions for the monopole spectrum, dipole spectrum, and their resultant is also displayed. It is shown that the distribution is symmetrically orientated. We estimated the µ and y-distortions by interpreting the spreading as the distortion and found that they are of the order of 10-4 and 10-5, respectively, for the monopole spectrum and 10-2 for the dipole spectrum. The paper also highlights the effectiveness of the BRI method and hints at future applications in the thermal nature of the early universe.

Revisiting cosmic microwave background radiation using blackbody radiation inversion.

Blackbody radiation inversion is a mathematical process for the determination of probability distribution of temperature from measured radiated power spectrum. In this paper a simple and stable blackbody radiation inversion is achieved by using an analytical function with three determinable parameters for temperature distribution. This inversion technique is used to invert the blackbody radiation field of the cosmic microwave background, the remnant radiation of the hot big bang, to infer the temperature distribution of the generating medium. The salient features of this distribution are investigated and analysis of this distribution predicts the presence of distortion in the cosmic microwave background spectrum.

Combining molecular docking and molecular dynamics studies for modelling Staphylococcus aureus MurD inhibitory activity.

The ATP-dependent bacterial MurD enzyme catalyses the formation of the peptide bond between cytoplasmic intermediate UDP-N-acetylmuramoyl-L-alanine and D-glutamic acid. This is essential for bacterial cell wall peptidoglycan synthesis in both Gram-positive and Gram-negative bacteria. MurD is recognized as an important target for the development of new antibacterial agents. In the present study we prepared the 3D-stucture of the catalytic pocket of the Staphylococcus aureus MurD enzyme by homology modelling. Extra-precision docking, binding free energy calculation by the MM-GBSA approach and a 40 ns molecular dynamics (MD) simulation of 2-thioxothiazolidin-4-one based inhibitor $1 was carried out to elucidate its inhibition potential for the S. aureus MurD enzyme. Molecular docking results showed that Lys19, Gly147, Tyr148, Lys328, Thr330 and Phe431 residues are responsible for the inhibitor-protein complex stabilization. Binding free energy calculation revealed electrostatic solvation and van der Waals energy components as major contributors for the inhibitor binding. The inhibitor-modelled S. aureus protein complex had a stable conformation in response to the atomic flexibility and interaction, when subjected to MD simulation at 40 ns in aqueous solution. We designed some molecules as potent inhibitors of S. aureus MurD, and to validate the stability of the designed molecule D1-modelled protein complex we performed a 20 ns MD simulation. Results obtained from this study can be utilized for the design of potent S. aureus MurD inhibitors.

Novel approach to Abel inversion.

Simple yet versatile, physically valid emissivity functions for peaked and hollow profiles with only two determinable parameters are proposed for performing Abel inversion. The advantages of the proposed functions have been explored. The inversion is very fast, accurate, convenient, and viable, in contrast to the existing methods. The validation of these functions has been confirmed by using simulated data under various conditions. The error in the process has been computed and found to depend on the functional form of the model emissivity. A comprehensive comparison has been drawn with the existing method and it has been found to offer a definite advantage over the existing technique in some respects, especially for real time applications. Limitation of this technique has also been discussed. The soft x-ray and visible light emissivity profile of SINP tokamak has been successfully obtained by using this method.

Mass spectrometry-compatible ICH (International Conference on Harmonization) impurity analysis with a high-pH mobile phase advantages and pitfalls.

Recent advances in bonding chemistry and novel silica synthesis have significantly extended the pH range of silica-based HPLC columns. This extended range now enables the analysis of water-soluble basic drugs at high pH without ion-pairing reagents, thus offering an alternative approach to assay or impurity analyses. This paper describes the many advantages and potential pitfalls of using high-pH mobile phases in the development of MS-friendly LC gradient impurity analytical methods for water-soluble basic drugs under International Conference on Harmonization (ICH) guidelines. Operating at high-pH provides excellent peak shapes and retention, and accentuates selectivity differences of structurally similar impurities and degradants. However, several problems unique to the use of high-pH mobile phases, such as column lifetime, robustness of pH adjustments, peak fronting, and on-column dimerization, were encountered. Each of these problems is discussed with its respective remedy.

Antiulcer activity of Mikania cordata.

The alkaloidal fraction obtained from an ethanolic extract of the leaves of Mikania cordata exhibited significant in vivo antiulcer activity in diclofenac sodium-induced gastric erosions in Long Evans rats.

Gene expression of ecdysteroid-regulated gene E74 of the honeybee in ovary and brain.

To facilitate studies of hormonal control in the honeybee (Apis mellifera L.), a cDNA for a honeybee homologue of the ecdysteroid-regulated gene E74 (AmE74) was isolated and its expression was analysed. Northern blot analysis indicated strong expression in the adult queen abdomen, and no significant expression in the adult drone and worker abdomens. In situ hybridization demonstrated that this gene was expressed selectively in the ovary and gut in the queen abdomen. Furthermore, this gene was also expressed selectively in subsets of mushroom body interneurones in the brain of the adult worker bees. These findings suggest that AmE74 is involved in neural function as well as in reproduction in adult honeybees.