Molecular dynamics simulation study on Bacillus subtilis EngA: the presence of Mg2+ at the active-sites promotes the functionally important conformation.

EngA, a GTPase contains two GTP binding domains [GD1, GD2], and the C-terminal KH domain shown to be involved in the later stages of ribosome maturation. Association of EngA to the ribosomal subunit in the intermediate stage of maturation is essential for complete ribosome maturation. However, this association was shown to be dependent on the nucleotide bound combinations. This nucleotide dependent association tendency is attributed to the conformational changes that occur among different nucleotide bound combinations. Therefore, to explore the conformational changes, all-atom molecular dynamics simulations for Bacillus subtilis EngA in different nucleotide bound combinations along with the presence or absence of Mg2+ in the active-sites were carried out. The presence of Mg2+ along with the bound nucleotide at the GD2 active-site dictates the GD2-Sw-II mobility, but the GD1-Sw-II mobility has not shown any nucleotide or Mg2+ dependent movement. However, the GD1-Sw-II secondary conformations are shown to be influenced by the GD2 nucleotide bound state. This allosteric connection between the GD2 active-site and the GD1-Sw-II is also observed through the dynamic network analysis. Further, the exploration of the GD1-KH interface interactions exhibited a more attractive tendency when GD1 is bound to GTP-Mg2+. In addition, the presence of Mg2+ stabilizes active-site water and also increases the distances between the α- and γ- phosphates of the bound GTP. Curiously, three water molecules in the GD1 active-site and only one water molecule in the GD2 active-site are stabilized. This indicates that the probability of GTP hydrolysis is more in GD1 compared to GD2.Communicated by Ramaswamy H. Sarma.

Efficacy of different methods for detecting the onset of ovarian cyclicity in post-partum Murrah buffaloes.

This study was designed to investigate the sensitivity and specificity of three methods for detecting the onset of cyclicity in post-partum Murrah buffaloes. The methods investigated were visual signs, transrectal ultrasonography, and serum progesterone (P4) assay. For this study, 102 post-partum Murrah buffalo cows were grouped for monitoring their ovarian activity. The first group of buffaloes was between 26 and 35 days post-partum. Thereafter, the buffalo cows that calved were grouped after every 10 days for the study sample. Thus, the study animals were adjudged between 26 and 35, 36-45, 46-55, 56-65, 66-75, 76-85, and 86-95 days post-partum with an average of 30, 40, 50, 60, 70, 80, and 90 days post-partum, respectively. Visual estrus signs were monitored twice daily, and simultaneously, ultrasound examination was carried out at 10 days interval for accessing the presence of corpus luteum (CL). Serum P4 was estimated in the animals which were adjudged cyclic by ultrasound examination, and the assay was repeated after 10 days. The buffalo cows in estrus were inseminated artificially, and pregnancy status was assessed after 30 days post-insemination. In this study, the sensitivity and specificity of visual observation were low (39.37 and 70.73%, respectively) when compared to P4 assay (98.80 and 96.47%) and ultrasound examination (single, 97.59 and 97.59%; double, 100 and 100%), respectively. Furthermore, the sensitivity and specificity of single and double ultrasound examination and P4 assay were comparable. In conclusion, this study reports that single and double ultrasound examination and P4 assay are more efficient than visual observation in detecting the onset of ovarian cyclicity in post-partum Murrah buffaloes.

Ovulatory and fertility response using modified Heatsynch and Ovsynch protocols in the anovular Murrah buffalo (Bubalus bubalis).

We studied the effect of modified Heatsynch and Ovsynch protocols on the ovulatory response (OR), estrus induction rate (EIR) and conception rate (CR) in the anovular postpartum Murrah buffalo (n = 35). In the modified Heatsynch protocol (Group I; n = 12), buffaloes were given two GnRH at 2 h interval on treatment day 0, PGF (PGF2α) on day 7 and estradiol (E2) 1 mg on day 8. Two FTAI were done at 20 h intervals after E2 administration. In the modified Ovsynch protocol (Group II; n = 15), GnRH was given on day 0, 7 and 16 with a PGF on day 14. Two FTAI were done; one at last GnRH and the other 20 h later. Group III served as untreated negative control (n = 8). During the treatment, ovarian changes were monitored by transrectal ultrasonography and plasma progesterone (P4) and E2. Administration of two GnRH at 2 h interval neither increased the OR nor strengthened the subsequent P4 priming. Interestingly, in group I, none of the buffalo ovulated to E2 though the EIR was 100% indicating the occurrence of behavioral, but not ovulatory estrus. Administration of GnRH 7 day prior to the commencement of Ovsynch protocol (Group II) did not improve the CR (21.4%), though the OR was 71.4%. No significant difference was found in the diameter of largest follicle between the ovulated and non-ovulated buffalo in response to GnRH suggesting that follicle of ≥9.5 mm is necessary but not sufficient to induce ovulation in the anovular buffalo. In both the protocols, the plasma P4 was higher on day 7 in those buffaloes that ovulated to GnRH. Buffaloes treated with modified Ovsynch regimens were 5.27 times more likely to become pregnant than modified Heatsynch protocol. It is concluded that modified Ovsynch protocol is superior to modified Heatsynch protocol in terms of OR and CR.

Advances in reproductive biotechnologies.

In recent times, reproductive biotechnologies have emerged and started to replace the conventional techniques. It is noteworthy that for sustained livestock productivity, it is imperative to start using these techniques for facing the increasing challenges for productivity, reproduction and health with impending environment conditions. These recent biotechniques, both in male and female, have revolutionized and opened avenues for studying and manipulating the reproductive process both in vitro and in vivo in various livestock species for improving tis efficiency. This review attempts to highlight pros and cons, on the recent developments in reproductive biotechnologies, both in male and female in livestock species.