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The kinetic patterns of CpG methylation of the cis-regulatory region of heat stress–related genes on exposed to heat stress (at 42 °C) between the Sahiwal and Frieswal cattle was compared in the present study. Using an in vitro whole blood culture model, cells were continuously exposed to heat stress (at 42 °C) for 6 h. Methylation levels of five genes, viz., GPX1, HSP70, HSP90, c-FOS, and JUN were estimated by SyberGreen-based quantitative methylation-specific PCR (qMSP) assay. CpG methylation kinetics at different time points of heat stress (0.5, 1, 2, 4, 6 h) were analyzed using mixed ANOVA. The initial methylation level, estimated at 37 °C, of HSP70 was significantly high in the Sahiwal breed. A significant (p<0.001) time-dependent hypomethylation of an antioxidant gene (GPX1) CpG islands was detected at the acute phase of the stress. Heat shock protein gene (HSP70) showed a similar CpG methylation kinetics where the hypomethylation was prominent from 1 h and persisted up to 4 h. The heat stress responses of both Sahiwal and Frieswal cattle were identical as there was no distinctiveness in the methylation kinetics of CpG islands of studied genes. The acclimatization of Frieswal cattle—a breed developed in India over the years to the tropical climatic conditions, maybe one of the reasons for this similarity. Thus, the present study results could pave a path to understand the molecular mechanism of heat stress and adaptation of indigenous and crossbred cattle populations to the changing scenario in tropical climate conditions. [ABSTRACT FROM AUTHOR]

In cattle production systems, an intense selection pressure for production traits has resulted in the decline of fertility traits. To optimize an efficient reproduction system, the inclusion of both male and female fertility traits in the selection process is very much essential. RAPD (Random Amplified Polymorphic DNA) was developed as a molecular biology tool and has been extensively used, to study intra‐ and interspecific genetic diversity. The present study was undertaken to utilize RAPD primers to investigate the association between DNA markers and semen quality traits viz. Sperm concentration, total sperm count ejaculate and initial sperm motility and thereby to identify good/poor semen producers. DNA isolated from the blood samples of healthy bulls was subjected to RAPD‐PCR. The multiple regression analysis followed by independent t test was carried out to identify suitable markers. Based on the results, only 12 bands were identified as marker suitable for any of the quality trait. This includes, OPA2 ~ 760, OPA2 ~ 700, OPA6 ~ 1,200, OPA9 ~ 400, OPA9 ~ 380, OPA12 ~ 970, OPA14 ~ 715, OPA14 ~ 605, OPA16 ~ 485, OPA17 ~ 860 and OPA18 ~ 480. Multiple regression analysis selected, OPA2 ~ 760 and OPA2 ~ 1,750 for sperm concentration and OPA2 ~ 760, OPA2 ~ 700, OPA9 ~ 620, OPA4 ~ 670 and OPA18 ~ 1,015 for total sperm count/ejaculate. But the t test revealed a significant association between OPA2 ~ 760 and total sperm count. Further, discriminant function analysis also identified this marker in the first step itself. The results of the present study can be exploited as a low‐cost alternative strategy for identification of good /poor semen producers in crossbred bulls at an early age. [ABSTRACT FROM AUTHOR]

Faced with the increasing need for correctly designed hybrid and cyber-physical systems today, the problem of including provision for continuously varying behaviour as well as the usual discrete changes of state is considered in the context of Event-B. An extension of Event-B called Hybrid Event-B is presented, that accommodates continuous behaviours (called pliant events) in between familiar discrete transitions (called mode events in this context). The continuous state change can be specified by a combination of indirect specification via ordinary differential equations, or direct specification via assignment of variables to values that depend on time, or indirect specification by demanding that behaviour obeys a time dependent predicate. The syntactic elements of the extension are discussed, and the semantics is described in terms of the properties of time dependent valuations of variables. Refinement is examined in detail, with reference to the notion of refinement inherited from discrete Event-B. A full suite of proof obligations is presented, covering all aspects of the new framework. A selection of examples and case studies is presented. A particular challenge — bearing in mind the desirability of conforming to existing intuitions about discrete Event-B, and the impact on tool support (as embodied in tools for discrete Event-B like Rodin) — is to design the whole framework so as to disturb as little as possible the existing structures for handling discrete Event-B. [ABSTRACT FROM AUTHOR]