Effect of heat exposure on prostaglandin production and expression of COX-2, PGES, PGFS, ITGAV and LGALS15 mRNAs in endometrial epithelial cells of buffalo (Bubalus bubalis).

BACKGROUND: Early embryonic mortality is one of the major intriguing factors of reproductive failure that causes considerable challenge to the mammalian cell biologists. Heat stress is the major factor responsible for reduced fertility in farm animals. The present study aimed to investigate the influence of heat stress on prostaglandin production and the expression of key genes, including COX-2, PGES, PGFS, ITGAV and LGALS15, in buffalo endometrial epithelial cells. METHODS AND RESULTS: Buffalo genitalia containing ovaries with corpus luteum (CL) were collected immediately post-slaughter. The stages of the estrous cycle were determined based on macroscopic observations of the ovaries. Uterine lumens of the mid-luteal phase (days 6-10 of the estrous cycle) were washed and treated with trypsin to isolate epithelial cells, which were then cultured at control temperature (38.5 °C for 24 h) or exposed to elevated temperatures [38.5 °C for 6 h, 40.5 °C for 18 h; Heat Stressed (HS)]. The supernatant and endometrial epithelial cells were collected at various time points (0, 3, 6, 12, and 24 h) from both the control and treatment groups. Although heat stress (40.5 °C) significantly (P < 0.05) increased COX-2, PGES, and PGFS transcripts in epithelial cells but it did not affect the in vitro production of PGF2α and PGE2. The expression of ITGAV and LGALS15 mRNAs in endometrial epithelial cells remained unaltered under elevated temperature conditions. CONCLUSION: It can be concluded that elevated temperature did not directly modulate prostaglandin production but, it promoted the expression of COX-2, PGES and PGFS mRNA in buffalo endometrial epithelial cells.

Expression analysis of pro-apoptotic BAX and anti-apoptotic BCL-2 genes in relation to lactation performance in Deoni and Holstein Friesian crossbred cows.

The study was designed with the objective of expression analysis of pro-apoptotic BAX and anti-apoptotic BCL-2 genes on lactation performance in Bos indicus and HF crossbred cows during early lactation. BAX/BCL-2 mRNA expression ratio in HF crossbreds showed a steady increase from 30th day to 90th day, but in Deoni cows the ratio exhibited a different pattern, which increased from day 30 to day 60, decreased on day 75, and then increased on day 90. BAX/BCL-2 expression ratio in Deoni and HF crossbreds were lowest on day 30 and highest on day 90. On contrary, the milk yield was highest on day 30 and lowest on day 90 suggesting BCL-2 gene favors milk production and BAX gene oppose milk production. In comparison to HF crossbreds, Deoni cows exhibited highest BAX/BCL-2 ratio at the end of early lactation, indicating Bos indicus cows were more sensitive to apoptosis than HF crossbreds. Comparison of daily milk yield with BAX/BCL-2 mRNA expression ratio revealed significant negative correlation with a correlation coefficient of -0.98 (P < 0.01) and -0.95 (P < 0.05) in Deoni and HF crossbred cows, respectively. Our study provides new insights into understanding the genetic control of mammary apoptosis between Bos indicus and HF crossbreds.

Effect of α-tocopherol in the vitrification medium on the viability, lipid peroxidation, expression of key developmental, apoptotic and stress-related genes in ovine secondary follicles.

The present study aimed to evaluate the effect of α-tocopherol on viability, lipid peroxidation and the expression of apoptosis, stress and development-related genes in the vitrified sheep secondary follicles. Ovarian secondary follicles (200-300 µm) were isolated and distributed separately to the vitrification treatment and supplemented with 5 mM, 10 mM, 20 mM and 30 mM of α-tocopherol (while the control fresh group was without vitrification and supplementation of α-tocopherol). After a week, the follicles were thawed and evaluated for follicular viability by trypan blue dye exclusion method, lipid peroxidation and gene expression studies. The results showed that the vitrification with 10 and 20 mM of α-tocopherol positively affected (p < .05) the viability of vitrified follicles in comparison with vitrified ones without α-tocopherol but the higher concentration of α-tocopherol, i.e., 30 mM negatively affected the viability (p < .05) in comparison with the 10 and 20 mM of α-tocopherol groups. The malondialdehyde (MDA) levels were significantly (p < .05) higher in the vitrified without α-tocopherol group in comparison to the vitrified with 20 mM of α-tocopherol group. The expression of apoptotic-related gene, BCL2L1 was significantly higher in 10 mM α-tocopherol group compared to the control fresh and CASPASE 3, 9 expressions were significantly higher in the vitrified group when compared to the vitrified with 10 mM α-tocopherol group. Expressions of BAX, BAD, BAK, BMP-15 and GDF-9 showed no significant difference among the groups. The mRNA expression of SOD1 was significantly higher in the vitrified without α-tocopherol group when compared to other groups. We conclude that the supplementation of 10 and 20 mM α-tocopherol in vitrification solution was the efficient vitrification procedure for the vitrification of ovine secondary follicles.

Regulatory role of Wnt signal in the oestradiol synthesis of different size categories of ovarian follicles in buffalo (Bubalus bubalis).

The aim of the present study was to understand the role of Wnt signal in ovarian oestradiol synthesis in various size categories of ovarian follicles. A six-day cell culture system was adopted to test the effect of a Wnt inhibitor i.e. Inhibitor of Wnt response (IWR) on the ovarian granulosa cell oestradiol synthesis and associated genes related to oestradiol synthesis and Wnt signalling (CYP19A1, CCND2, WNT2, FZD6, DVL1, APC, AXIN2, CTNNB1) in buffalo. It was conducted with four groups: Group 1: control, Group 2: control + FSH, Group 3: IWR, Group 4: IWR + FSH. No significant effect of IWR was observed on the ovarian granulosa cell proliferation. No significant difference in the oestradiol levels was found in the spent media harvested after six days of in vitro culture among different groups in small and large-sized ovarian follicles. However, the oestradiol level varied significantly (p < .05) among different treatment groups in medium-sized follicles. The oestradiol level was significantly lower (p < .05) in IWR group compared with the control group and was also significantly lower in IWR + FSH group compared with the FSH group. The Wnt inhibitor had significantly (p < .05) reduced the gene expression of CYP19A1 in large ovarian follicles. Varied effects of IWR-1 and FSH on the expression of other genes were observed. The results indicated that there is a positive role of Wnt signal in oestradiol synthesis in buffalo, but the positive role was more discernible in medium- and large-sized follicles.

Effect of retinol in the vitrification medium on viability of vitrified ovine preantral follicles and expression of key developmental and apoptosis related genes.

BACKGROUND: Vitrification increases the production of reactive oxygen species (ROS) and the antioxidants in the vitrification solution may be beneficial by reducing excessive ROS production. OBJECTIVE: To evaluate the effect of retinol supplementation in vitrification solution on viability, apoptosis and development-related gene expression in vitrified sheep preantral follicles. MATERIALS AND METHODS: Preantral follicles were isolated and randomly assigned into one of five groups: Group1, control fresh preantral follicles; Group 2, vitrification treatment; Group 3, vitrification + 2 µM retinol; Group 4, vitrification + 5 µM retinol; Group 5, vitrification + 10 µM retinol. Preantral follicles were placed in vitrification solutions and then plunged into liquid nitrogen (-196°C). After a week, the follicles were thawed and analyzed for follicular viability by trypan blue exclusion method and for gene expression. RESULTS: Vitrification with 5 µM retinol positively affected viability in comparison with vitrification without retinol (P < 0.05). There was no significant difference in viability among the Group 1, Group 2, Group 3 and Group 5. Expression of apoptotic genes BAX and Casp 3 were higher in the vitrified group, and vitrification with 5 µM retinol (Group 4) is comparable to the control fresh. Expressions of other apoptosis-related genes (i.e., BCL2L1, BAD and BAK) showed significant difference between the control fresh group and the vitrification group with 5 µM retinol. Expression of Annexin5 was also significantly different among various groups. The expression of development competence genes GDF-9 and BMP-15 were higher (P < 0.05) in the Group vitrified with 5 µM retinol. CONCLUSION: The supplementation of 5 µM retinol in vitrification solution was beneficial for the vitrification of ovine preantral follicles.

Copper and Selenium stimulates CYP19A1 expression in caprine ovarian granulosa cells: possible involvement of AKT and WNT signalling pathways.

The role of copper and selenium on activation of estradiol synthesis pathways viz. PKA/AKT/WNT is not clearly elucidated. On this background we attempt to elcuiated the role of copper and selenium on mRNA expression of genes associated with estradiol synthesis in caprine ovarian granulose cell models. Ovarian granulosa cells from medium (3-5 mm) sized follicles were aspirated and distributed separately to different groups. Group I: control, Group II: cupric chloride (Cu: 0.5 mM), Group III: sodium selenite (Se: 100 ng/ml), Group IV: Cu + Se. The cells (105/well) were cultured in 96 well plate in the base culture medium of MEMα comprising of nonessential amino acids (1.1 mM), FSH (10 ng/mL), transferrin (5 µg/mL), IGF-I (2 ng/mL), androstenedione (10-6 M), penicillin (100 IU/mL), streptomycin (0.1 mg/mL) and fungizone (0.625 µl/mL) and insulin (1 ng/mL). The cells were incubated in a carbondioxide incubator (38 °C, 5% CO2, 95% RH). The medium was changed on alternate days and cells were harvested on day 6. Day 6 media was used for estimation of estradiol. The RNA isolated form harvested cells was used for qPCR assay. There was no significant (p > 0.05) difference in estradiol concentration between groups. The mRNA expression of AKT1, CYP19A1, WNT2 & 4, FZD6 and APC2 were significantly (p < 0.05) higher in Cu and Cu + Se groups compared to control. Whereas, the mRNA transcript of DVL1 and CSNK1 was significantly (p < 0.05) higher in Cu + Se group compared to control. Incontrast, no significant difference in mRNA expression of PRKAR1A and CTNNB1 was noticed. Our study support a key role of copper and selenium in activation of AKT and WNT signalling pathway that further lead to increase in the mRNA expression of CYP19A1.

Effect of retinol as antioxidant on the post-thaw viability and the expression of apoptosis and developmental competence-related genes of vitrified preantral follicles in buffalo (Bubalus bubalis).

The present study evaluated the effect of supplementation of retinol in the vitrification solution on the viability, apoptosis and development-related gene expression in vitrified buffalo preantral follicles. Preantral follicles isolated from cortical slices of ovaries were randomly assigned into three groups: Group1-Control fresh preantral follicles; Group 2-Vitrification treatment (Vitrification solution 1 (VS1) -TCM-199 + 25 mM HEPES + Foetal bovine serum (FBS) 10%, Ethylene glycol (EG): 10%, Dimethyl sulphoxide (DMSO): 10%, Sucrose-0.3 M for 4 min; VS2- TCM-199 + 25 mM HEPES + FBS10%, EG:25%, DMSO: 25%, Sucrose:0.3 M for 45 s); Group3-vitrification treatment +5 µM of Retinol. Preantral follicles were placed in corresponding vitrification medium and plunged into liquid nitrogen (-196°C). After a week, the follicles were thawed and analysed for follicular viability and gene expression. There was no significant difference in the viability rates among the Group 1(Fresh preantral follicles) (91.46 ± 2.39%), Group 2 (89.59 ± 2.46%) and Group 3 (87.19 ± 4.05%). There was a significantly (p < .05) higher mRNA expression of BCL2L1, GDF-9 and BMP-15 in the vitrification + retinol group compared with the control group. There was a significantly (p < .05) higher expression of Caspase-3 and Annexin-5 in the vitrification group and Vitrification + retinol group compared with control group of follicles. It is concluded that the supplementation of 5 µM of Retinol in Vitrification solution was an efficient vitrification procedure for the vitrification of buffalo preantral follicles.

Influence of tillage based crop establishment and residue management practices on soil quality indices and yield sustainability in rice-wheat cropping system of Eastern Indo-Gangetic Plains.

Rice-wheat cropping system (RWCS) is the most important system occupying around 26 M ha spread over the Indo Gangetic Plains in South Asia and China. Many long-term trials were led to assess the agronomic productivity and economic profitability of various combinations of conservation agricultural (CA) practices (zero tillage, residue management and crop establishment) in RWCS of Eastern Indo-Gangetic Plains (EIGP) of India. The purpose of this study was to investigate the best management practices involving different tillage-based crop establishment and residue retention techniques and their contribution to agricultural system sustainability through improvement in soil health by developing soil quality index (SQI). We have used SQI as an instrument based on physical [macro aggregate stability (MAS), available water capacity (AWC) and soil penetration resistance (SPR)], chemical [soil organic carbon (OC), available N, available P and available K] and biological [microbial biomass carbon (MBC), fluorescein diacetate (FDA) and dehydrogenase activity (DHA)] properties of soil, because these are very useful indicators of soil's functions for agronomic productivity and soil fertility. Soil properties like MAS, OC, MBC, FDA and DHA were higher by 47, 18, 56, 48 and 53%, respectively, under ZTDSR-ZTW (T7: Zero-till direct seeded rice - Zero-till wheat) than RPTR-CTW (T1: Random puddled transplanted rice - Conventional till broadcasted wheat), at 0-10 cm. CA based treatment T7 also recorded lower SPR (126 N cm-1). SQI for different treatments were calculated by performing principal component analysis based on the total data set method. The higher system rice equivalent yield of 12.41 t ha-1 was observed at SQI value of 0.90 at 0-10 cm and 0.86 at 10-20 cm in T7. It can be concluded that crop residue retention on the surface with zero tillage is beneficial for the sustainability and productivity of the RWCS in EIGP of India.

T11TS immunotherapy potentiates the repressed calcineurin-NFAT signalling pathway of T cells in Cryptococcus neoformans infected rats: a cue towards T-cell activation for antifungal immunity.

AIMS: To examine the modulation of the interacting partners of the calcineurin (CaN)-NFAT pathway in T cells during Cryptococcus neoformans fungal infection and post-T11TS immunotherapy. METHODS AND RESULTS: Wistar rats were infected with C. neoformans and followed by immunotherapy with immune-potentiator T11TS. T cells were analysed by flow cytometry, immunoblotting and nuclear translocation study. The signalling proteins LCK, FYN, LAT, PLCγ1 and CaN in T cells were regulated by C. neoformans infection resulting in reduced nuclear translocation of NFAT and IL-2 expression. Following T11TS immunotherapy, the expressions of the above-mentioned proteins were boosted and thus resulting in the clearance of C. neoformans from lung and spleen. CONCLUSIONS: The precise mechanism of suppression of the T-cell function by C. neoformans is still unknown. Previously, we have shown that T11TS positively regulates the function of T cells to abrogate glioma and other immunosuppressive conditions. T11TS immunotherapy increased the expression of the above signalling partners of the CaN-NFAT pathway in T cells and improved nuclear retention of NFAT. As a result, an increased IL-2 expression leads to activation and proliferation of T cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results demonstrate the role of T11TS in restoring the CaN-NFAT signalling pathway in T cells. It identifies T11TS as an immunotherapeutic agent with potential clinical outcomes to counteract C. neoformans infection.

A singular value decomposition Bayesian multiple-trait and multiple-environment genomic model.

Today, breeders perform genomic-assisted breeding to improve more than one trait. However, frequently there are several traits under study at one time, and the implementation of current genomic multiple-trait and multiple-environment models is challenging. Consequently, we propose a four-stage analysis for multiple-trait data in this paper. In the first stage, we perform singular value decomposition (SVD) on the resulting matrix of trait responses; in the second stage, we perform multiple trait analysis on transformed responses. In stages three and four, we collect and transform the traits back to their original state and obtain the parameter estimates and the predictions on these scale variables prior to transformation. The results of the proposed method are compared, in terms of parameter estimation and prediction accuracy, with the results of the Bayesian multiple-trait and multiple-environment model (BMTME) previously described in the literature. We found that the proposed method based on SVD produced similar results, in terms of parameter estimation and prediction accuracy, to those obtained with the BMTME model. Moreover, the proposed multiple-trait method is atractive because it can be implemented using current single-trait genomic prediction software, which yields a more efficient algorithm in terms of computation.

Effect of Tris-egg Yolk, Soya Milk, and Liposome-based Extenders on Sahiwal (Bos indicus) Sperm Quality during Pre- and Post-Cryopreservation Stages.

BACKGROUND: Even though there are plenty of semen cryopreservation extenders available, their adoption is limited. Although normal tris-based egg yolk (EYC) extender is widely used, it leads to compromised post-thaw sperm quality. OBJECTIVE: To find a standard semen extender, six different semen extenders were validated. METHODS: In a split study, six aliquots of zebu cattle fresh semen ejaculate were cryopreserved in extenders containing egg yolk obtained from hen which was reared either in 1) normal, 2) omega-3 enriched, and 3) herbal enriched diet supplementation, and egg yolk free extenders such as 4) soya lecithin, 5) Bioxcell and 6) Optixcell. RESULT: Significantly poor sperm quality and kinematics were observed in extender containing herbal egg yolk. However, omega-3 enriched egg yolk extender was on par with EYC. Among all extenders, soya lecithin and bioxcell have shown better sperm quality. Sperm motility was significantly higher in semen extended in liposome-based extender Optixcell. CONCULSION: Optixcell can be considered as a standard extender for cattle semen cryopreservation to maintain adequate sperm quality required for artificial insemination.

Updated results from MONALEESA-2, a phase III trial of first-line ribociclib plus letrozole versus placebo plus letrozole in hormone receptor-positive, HER2-negative advanced breast cancer.

Background: The phase III MONALEESA-2 study demonstrated significantly prolonged progression-free survival (PFS) and a manageable toxicity profile for first-line ribociclib plus letrozole versus placebo plus letrozole in patients with hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) advanced breast cancer. Here, we report updated efficacy and safety data, together with exploratory biomarker analyses, from the MONALEESA-2 study. Patients and methods: A total of 668 postmenopausal women with HR+, HER2- recurrent/metastatic breast cancer were randomized (1 : 1; stratified by presence/absence of liver and/or lung metastases) to ribociclib (600 mg/day; 3-weeks-on/1-week-off; 28-day treatment cycles) plus letrozole (2.5 mg/day; continuous) or placebo plus letrozole. The primary end point was locally assessed PFS. The key secondary end point was overall survival (OS). Other secondary end points included overall response rate (ORR) and safety. Biomarker analysis was an exploratory end point. Results: At the time of the second interim analysis, the median duration of follow-up was 26.4 months. Median PFS was 25.3 months [95% confidence interval (CI) 23.0-30.3] for ribociclib plus letrozole and 16.0 months (95% CI 13.4-18.2) for placebo plus letrozole (hazard ratio 0.568; 95% CI 0.457-0.704; log-rank P = 9.63 × 10-8). Ribociclib treatment benefit was maintained irrespective of PIK3CA or TP53 mutation status, total Rb, Ki67, or p16 protein expression, and CDKN2A, CCND1, or ESR1 mRNA levels. Ribociclib benefit was more pronounced in patients with wild-type versus altered receptor tyrosine kinase genes. OS data remain immature, with 116 deaths observed; 50 in the ribociclib arm and 66 in the placebo arm (hazard ratio 0.746; 95% CI 0.517-1.078). The ORR was 42.5% versus 28.7% for all patients treated with ribociclib plus letrozole versus placebo plus letrozole, respectively, and 54.5% versus 38.8%, respectively, for patients with measurable disease. Safety results, after a further 11.1 months of follow-up, were comparable with those reported at the first analysis, with no new or unexpected toxicities observed, and no evidence of cumulative toxicity. Conclusions: The improved efficacy outcomes and manageable tolerability observed with first-line ribociclib plus letrozole are maintained with longer follow-up, relative to letrozole monotherapy. Clinical trials number: NCT01958021.

Harmonic Generation from Neutral Manganese Atoms in the Vicinity of the Giant Autoionization Resonance.

High harmonics from laser-ablated plumes are mostly generated from ionic species. We demonstrate that with ultrashort infrared (∼1.82 µm) driving lasers, high harmonics from laser-ablated manganese are predominantly generated from neutral atoms, a transition metal atom with an ionization potential of 7.4 eV. Our results open the possibility to advance laser-ablation technique to study the dynamics of neutral atoms of low ionization potential. Moreover, as manganese contains giant autoionizing resonance, intense and broadband high harmonics have been demonstrated from this resonance at energies from 49 to 53 eV. This opens the possibility to generate intense attosecond pulses directly from the giant resonances, as well as to study these resonances using high-harmonic spectroscopy.

Molecular cloning and expression of FGF2 gene in pre-implantation developmental stages of in vitro-produced sheep embryos.

Early embryonic mortality is one of the main sources of reproductive loss in domestic ruminants including sheep. Fibroblast growth factor-2 (FGF-2) is a member of FGFs family that mediates trophoblast activities and regulates embryonic development in various species. In this study, we have cloned, characterized sheep FGF2 cDNA (KU316368) and studied the expression in sheep embryos. Ovaries of non-pregnant sheep were collected from local abattoir and matured in culture medium at 38.5ºC, 5% CO2 , 95% humidity for 22-24 hr. The matured oocytes were inseminated with capacitated spermatozoa in Brackett and Oliphant medium and resulted embryos were cultured in CO2 incubator for 6-7 days to complete the developmental stages from two cells to blastocyst stage. Total RNA was extracted from immature oocytes (n = 100), mature oocytes (n = 100) and different stages of embryos such as 2 cell (n = 50), 4 cell (n = 25), 8 cell (n = 12), 16 cell (n = 6), morula (n = 5) and blastocyst (n = 3). The total RNA isolated from the oocytes and embryos was reverse transcribed and subjected to real-time polymerase chain reaction using sequence-specific primers and SYBR green as the DNA dye. On sequence analysis, the nucleotide sequence of sheep FGF2 exhibited highest sequence similarity with cattle (100%) and least with rat and mouse (69.2%). At the deduced amino acid level, a highest degree of similarity was noticed with cattle, buffalo, goat, pig, camel and horse (100%) and lowest degree of identity with rat, human and mouse (98.2%). The FGF2 mRNA expression was higher in immature and mature oocytes and gradually decreases from 2-cell stage of embryo to the blastocyst stage. More over a significant differences in FGF2 mRNA expression (p < .05) were observed between immature oocytes and all pre-implantation stages of embryo. It can be concluded that FGF-2 plays a significant role in pre-implantation and early development of embryos in sheep.

Time-Resolved Measurement of Interatomic Coulombic Decay Induced by Two-Photon Double Excitation of Ne_{2}.

The hitherto unexplored two-photon doubly excited states [Ne^{*}(2p^{-1}3s)]_{2} were experimentally identified using the seeded, fully coherent, intense extreme ultraviolet free-electron laser FERMI. These states undergo ultrafast interatomic Coulombic decay (ICD), which predominantly produces singly ionized dimers. In order to obtain the rate of ICD, the resulting yield of Ne_{2}^{+} ions was recorded as a function of delay between the extreme ultraviolet pump and UV probe laser pulses. The extracted lifetimes of the long-lived doubly excited states, 390(-130/+450) fs, and of the short-lived ones, less than 150 fs, are in good agreement with ab initio quantum mechanical calculations.

Inhibition of Escherichia coli ribosome subunit dissociation by chloramphenicol and Blasticidin: a new mode of action of the antibiotics.

The ability of the ribosome to assist in folding of proteins both in vitro and in vivo is well documented and is a nontranslational function of the ribosome. The interaction of the unfolded protein with the peptidyl transferase centre (PTC) of the bacterial large ribosomal subunit is followed by release of the protein in the folding competent state and rapid dissociation of ribosomal subunits. Our study demonstrates that the PTC-specific antibiotics, chloramphenicol and blasticidin S inhibit unfolded protein-mediated subunit dissociation. During post-termination stage of translation in bacteria, ribosome recycling factor (RRF) is used together with elongation factor G to recycle the 30S and 50S ribosomal subunits for the next round of translation. Ribosome dissociation mediated by RRF and induced at low magnesium concentration was also inhibited by the antibiotics indicating that the PTC antibiotics exert an associative effect on ribosomal subunits. In vivo, the antibiotics can also reduce the ribosomal degradation during carbon starvation, a process requiring ribosome subunit dissociation. This study reveals a new mode of action of the broad-spectrum antibiotics chloramphenicol and blasticidin. SIGNIFICANCE AND IMPACT OF THE STUDY: Ribosome synthesizes protein in all organisms and is the target for multiple antimicrobial agents. Our study demonstrates that chloramphenicol and blasticidin S that target the peptidyl transferase centre of the bacterial ribosome can then inhibit dissociation of 70S ribosome mediated by (i) unfolded protein, (ii) translation factors or (iii) low Mg+2 concentrations in vitro and thereby suppresses ribosomal degradation during carbon starvation in vivo. The demonstration of this new mode of action furthers the understanding of these broad-spectrum antibiotics that differentially inhibit protein synthesis in prokaryotic and eukaryotic cells.

Synchronous behaviour of two interacting oscillatory systems undergoing quasiperiodic route to chaos.

Thermoacoustic instability, caused by a positive feedback between the unsteady heat release and the acoustic field in a combustor, is a major challenge faced in most practical combustors such as those used in rockets and gas turbines. We employ the synchronization theory for understanding the coupling between the unsteady heat release and the acoustic field of a thermoacoustic system. Interactions between coupled subsystems exhibiting different collective dynamics such as periodic, quasiperiodic, and chaotic oscillations are addressed. Even though synchronization studies have focused on different dynamical states separately, synchronous behaviour of two coupled systems exhibiting a quasiperiodic route to chaos has not been studied. In this study, we report the first experimental observation of different synchronous behaviours between two subsystems of a thermoacoustic system exhibiting such a transition as reported in Kabiraj et al. [Chaos 22, 023129 (2012)]. A rich variety of synchronous behaviours such as phase locking, intermittent phase locking, and phase drifting are observed as the dynamics of such subsystem change. The observed synchronization behaviour is further characterized using phase locking value, correlation coefficient, and relative mean frequency. These measures clearly reveal the boundaries between different states of synchronization.

Differential expression of ISG 15 mRNA in peripheral blood mononuclear cells of nulliparous and multiparous pregnant versus non-pregnant Bos indicus cattle.

Embryonic mortality is found to be the main source of reproductive wastage in domestic ruminants. Many genes are involved in the growth and development of the embryo, and the interferon-stimulated gene 15 (ISG 15) is one of the major gene stimulated by interferon tau, the maternal recognition of pregnancy signal in ruminants. In this study, both genomic and cDNA sequences of ISG 15 from Bos indicus (Deoni breed) were amplified and characterized. The genomic sequence of Deoni ISG 15 exhibited 99% identity with Bos taurus and 97% identity with that of Bos mutus and Bubalus bubalis. Moreover qRT-PCR analysis revealed constitutive expression of the ISG 15 mRNA in peripheral blood mononuclear cells of Deoni heifers and multiparous cows during early pregnancy. Fourteen Deoni heifers and fifteen multiparous Deoni cows were synchronized for timed AI by CIDR-Ovsynch protocol, and six animals were kept as cyclic control in each group. Blood samples were collected on days 7, 14, 16, 18, 21, 30 and 45 from the day of AI. Pregnancy was confirmed by plasma progesterone level through ELISA. A significantly higher expression of ISG 15 mRNA was found on day 16 (p < .05) and day 18 (p < .05) of pregnancy in nulliparous heifers. Although in multiparous Deoni cows ISG 15 expression was greater in pregnant cows, difference was statistically non-significant. The result of this study indicates that ISG 15 gene expression is upregulated during 16-18 days of pregnancy and could be used as an early pregnancy marker in dairy cows especially in heifers.

Evaluation of long-term conservation agriculture and crop intensification in rice-wheat rotation of Indo-Gangetic Plains of South Asia: Carbon dynamics and productivity.

In the context of deteriorating soil health, stagnation of yield in rice-wheat cropping system (RWCS) across Indo- Gangetic plains (IGP) and environmental pollution, a long term field experiment was conducted during 2009-2016 taking four crop scenarios with conservation agriculture (CA), crop intensification and diversified cropping as intervening technology aiming to evaluate the sustainability of the systems. Scenario 1 (S1) represented conventional farmers' practice of growing rice and wheat with summer fallow. In scenario 2 (S2) and scenario 3 (S3), legume crop was taken along with rice and wheat with partial CA and full CA, respectively. Conventional RWCS was replaced with rice-potato + maize- cowpea cropping system with partial CA in scenario 4 (S4). The S3 scenario registered highest total organic carbon (TOC) stock of 47.71 Mg C ha-1 and resulted in significant increase of 14.57% over S1 (Farmer's practice) in 0-30 cm soil depth after 7 years of field trial. The S4 scenario having intensified cropping systems recorded lowest TOC of 39.33 Mg C ha-1 and resulted in significant depletion of 17.56% in C stock with respect to S3 in 0-30 cm soil depth. The TOC enrichment was higher in S2, S3 and S4 scenario in the surface soil (0-10 cm) compared to S1. At lower depth (20-30 cm), the TOC enrichment was significantly higher in S2 (12.82 Mg C ha-1) and S3 (13.10 Mg C ha-1 soil) over S1 scenario. The S2 and S3 scenario recorded highest increased allocation of TOC (3.55 and 6.13 Mg C ha-1) to passive pool over S1. The S2 (15.72 t ha-1), S3 (16.08 t ha-1) and S4 (16.39 t ha-1) scenarios recorded significantly higher system rice equivalent yield over S1 (10.30 t ha-1). Among the scenarios, S3 scenario had greater amount of total soil organic carbon, passive pool of carbon and higher system rice equivalent yield, thus, is considered the best cropping management practice to maintain soil health and food security in the middle IGP.