Reproductive performance of lactating dairy cows treated with cloprostenol at the time of insemination

The effect of intravenous cloprostenol treatment at the time of insemination on reproductive performance was consecutively evaluated in three different subpopulations of high producing lactating dairy cows: Study (1) early postpartum synchronized and fixed-time inseminated (about 50 days in milk) cows (n = 379: 187 control and 192 treated cows); Study (2) presumed high fertility cows first inseminated between 90 and 120 days postpartum (n = 248: 124 control and 124 treated cows); and Study (3) heat stressed repeat breeder cows (n = 183: 93 control and 90 treated cows). Data were analyzed using multiple regression methods. Study 1: Parity (primiparous versus multiparous), milk production, body condition score at AI, insemination season (cool versus warm period) and treatment were included in the analysis as potential factors affecting ovulation, double ovulation, return to estrus, and pregnancy to first AI and to second AI (first AI plus return AI) rates. Logistic regression analysis indicated that the final model for ovulation rate only included the interaction (P = 0.002) between insemination season and treatment. Cloprostenol treatment at insemination led to a 4.2-fold increase in the ovulation rate in cows inseminated during the warm period. There were no significant effects of treatment, parity, milk production, body score or the insemination season on the return to estrus rate. The only variables included in the final logistic model for double ovulation and pregnancy to first AI rates were treatment and season, respectively. Treatment led to a 2.6-fold increase (P = 0.001) in the double ovulation rate, whereas cows inseminated in the warm period were 2.1 times less likely (P = 0.007) to become pregnant at first AI compared to those inseminated in the cool season. The variables included in the final logistic model for the pregnancy rate to second AI were treatment and season. Cloprostenol given at AI increased the risk of pregnancy 1.9 times (P = 0.002), and cows inseminated during the warm season were two times less likely to become pregnant (P = 0.003). No significant interactions were found among these three dependent variables (double ovulation and pregnancy to first and to second AI rates). Study 2: Logistic regression analysis of all the dependent variables: return to estrus, and pregnancy to first and to second AI (first AI plus return to AI) rates indicated no significant effects of treatment, parity, days in milk, milk production or body score at AI. No significant interactions were found. Study 3: The final model for the pregnancy rate only included the interaction between parity (primiparous versus multiparous) and treatment. Days in milk, milk production and insemination number showed no significant effect on pregnancy rate. Cloprostenol treatment at insemination increased the pregnancy rate in primiparous repeat breeder cows (odds ratio: 3.6). The treatment group and parity showed significant (P < 0.0001) interaction. This interaction suggests that cloprostenol treatment of primiparous cows at insemination might enhance pregnancy yet have no effect in multiparous cows. Our findings indicate that cloprostenol administered at insemination promotes ovulation and double ovulation in lactating dairy cows. Cloprostenol treatment showed no benefit in cows with acceptable reproductive performance, suggesting that cloprostenol treatment at AI may only be useful in cows in which stress factors affect ovulation and in repeat breeder cows.     

The effect of addition of equine chorionic gonadotropin to a progesterone-based estrous synchronization protocol in buffaloes (Bubalus bubalis) under tropical conditions

Poor estrus expression and anestrus decrease the reproductive efficiency of buffaloes. The objective of this study was to determine whether the addition of equine chorionic gonadotropin (eCG) to an estrous synchronization protocol and timed insemination could improve ovulation and pregnancy rates of anestrous buffalo cows under tropical conditions. The study population comprised 65 lactating Murrah buffalo cows which were assigned to CIDR (n = 33) or CIDR + eCG (n = 32) treatment groups. Cows in the CIDR group were fitted for 8 d with a controlled intravaginal drug release (CIDR) device containing 1.38 g progesterone, received GnRH (10 μg i.m.) on D 0, PGF_2α (750 μg i.m.) on D 7, and GnRH (10 μg i.m.) on D 9; whereas cows in the CIDR + eCG group received the same treatment plus eCG (500 IU, i.m.) at the time of PGF_2α treatment. All cows were inseminated 16-20 h after the second GnRH treatment. Blood samples were obtained 10 d before the start of synchronization treatment (Day -10) and at the onset of treatment (Day 0). Cows with plasma progesterone concentrations <1 ng/mL recorded in both samples (Low-Low levels of P4) were classified as non-cyclic cows. Similarly, when either one or both of the sample pair contained concentrations of serum progesterone ≥1 ng/mL (High-High, Low-High, or High-Low levels of P4), the buffaloes were classified as cyclic cows. Ovulation rate, defined as the number of buffaloes with at least one corpus luteum 10 days after insemination, was significantly higher (P = 0.018) in the CIDR + eCG (84.4%) cows than in the CIDR cows (57.6%). Pregnancy rate was numerically lower in CIDR (27.3%) than CIDR + eCG (40.6%) cows, though differences were not significant (P = 0.25). Pregnancy rates for CIDR + eCG cows were similar to that of cows inseminated after natural estrus (40.9%; 29/71). In the non-cyclic animals, higher ovulation rates (P = 0.026) were recorded for the CIDR + eCG (81%) than for the CIDR cows (47.4%). Our results indicate that the addition of eCG to a progesterone-based estrous synchronization regimen substantially improves the ovulation rate in non-cyclic buffaloes. When this treatment is followed by timed AI, pregnancy rates achieved in anestrous buffaloes, whether cyclic and non-cyclic, may approach the rates observed in cows inseminated at natural estrus.     

Cadmium induced mitochondrial injury and apoptosis in vero cells: protective effect of diallyl tetrasufide from garlic

Oxidative stress and mitochondrial injury has been implicated in cadmium-induced apoptosis. In this study, we examined the protective effect of diallyl tetrasulfide from garlic on cadmium induced oxidative stress and apoptosis in vero cells. Exposure of vero cells to cadmium (10 microM) for 18 h showed the apoptotic events such as loss of cell viability, alterations in nuclear morphology and decreased mitochondrial membrane potential with significantly increased levels of reactive oxygen species (super oxide anion and hydrogen peroxide). Treatment of vero cells with cadmium (10 microM) and diallyl tetrasulfide (5-50 microg/ml) showed that diallyl tetrasulfide attenuated the cadmium-induced suppression of cell viability in a dose dependent manner and highly significant effect was observed at 40 microg/ml. The nuclei morphological analysis with 4',6-diamidino-2-phenylindole staining confirmed that diallyl tetrasulfide at 40 microg/ml prevented the Cd (10 microM) induced apoptosis. Flow cytometric analysis with 2',7'-dichlorofluorencein diacetate showed that the inhibitory effect of diallyl tetrasulfide (10-40 microg/ml) on reactive oxygen species generation parallel with its effect on cell viability. In addition, diallyl tetrasulfide (40 microg/ml) remarkably reduced the cadmium-induced accumulation of superoxide radical and hydrogen peroxide with in cells. Further, diallyl tetrasulfide significantly protected the cadmium-induced decrease in mitochondrial membrane potential, an indicator of mitochondrial function. Our study suggest that diallyl tetrasulfide affect the reactive oxygen species generation induced by cadmium, and possesses a novel protective effect on the cytolethality associated with mitochondrial injury, which contributes to the antiapoptotic effect of diallyl tetrasulfide against cadmium.     

A hexanuclear iron(III) complex [Fe6O2(OH)2(PhCOO)10(hedmp)2]·3CH3CN assembled from 2-hydroxyethyl-3,5-dimethyl pyrazole

In order to assemble polynuclear iron(III) complexes, the coordination chemistry of the 2-hydroxyethyl-3,5-dimethylpyrazole (hedmp-H) ligand has been investigated. Reaction of hedmp-H with trinuclear iron carboxylate precursor [Fe₃O(PhCOO)₆(H₂O)₃]Cl in acetonitrile yielded the hexanuclear Fe(III) complex [Fe₆O₂(OH)₂(PhCOO)₁₀(hedmp)₂]·3CH₃CN (1). This aggregate has been characterized by employing various analytical techniques, spectroscopic studies and single crystal X-ray diffraction. Detailed magnetic susceptibility measurements revealed that 1 displays an S_T = 5 ground state.     

Obtaining valid laboratory data in clinical trials conducted in resource diverse settings: lessons learned from a microbicide phase III clinical trial

Background

Over the last decade several phase III microbicides trials have been conducted in developing countries. However, laboratories in resource constrained settings do not always have the experience, infrastructure, and the capacity to deliver laboratory data meeting the high standards of clinical trials. This paper describes the design and outcomes of a laboratory quality assurance program which was implemented during a phase III clinical trial evaluating the efficacy of the candidate microbicide Cellulose Sulfate 6% (CS) [1].

Methodology

In order to assess the effectiveness of CS for HIV and STI prevention, a phase III clinical trial was conducted in 5 sites: 3 in Africa and 2 in India. The trial sponsor identified an International Central Reference Laboratory (ICRL), responsible for the design and management of a quality assurance program, which would guarantee the reliability of laboratory data. The ICRL provided advice on the tests, assessed local laboratories, organized trainings, conducted supervision visits, performed re-tests, and prepared control panels. Local laboratories were provided with control panels for HIV rapid tests and Chlamydia trachomatis/Neisseria gonorrhoeae (CT/NG) amplification technique. Aliquots from respective control panels were tested by local laboratories and were compared with results obtained at the ICRL.

Results

Overall, good results were observed. However, discordances between the ICRL and site laboratories were identified for HIV and CT/NG results. One particular site experienced difficulties with HIV rapid testing shortly after study initiation. At all sites, DNA contamination was identified as a cause of invalid CT/NG results. Both problems were timely detected and solved. Through immediate feedback, guidance and repeated training of laboratory staff, additional inaccuracies were prevented.

Conclusions

Quality control guidelines when applied in field laboratories ensured the reliability and validity of final study data. It is essential that sponsors provide adequate resources for implementation of such comprehensive technical assessment and monitoring systems.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00153777","term_id":"NCT00153777"}}NCT00153777 and Current Controlled Trials ISRCTN95638385     

Cytoprotective and antioxidant role of diallyl tetrasulfide on cadmium induced renal injury: an in vivo and in vitro study

Cadmium (Cd) is an environmental and industrial pollutant that affects various organs in humans and animals. A body of evidence has accumulated implicating the free radical generation with subsequent oxidative stress in the biochemical and molecular mechanisms of Cd toxicity. Since kidney is the critical target of Cd toxicity, we carried out this study to investigate the effects of diallyl tetrasulfide (DTS), an organosulfur compound derived from garlic on Cd induced toxicity in the kidney of rats and also in the kidney cell line (vero cells). In experimental rats, subcutaneous administration of Cd (3 mg/kg bw/day) for 3 weeks induced renal damage, which was evident from significantly increased levels of serum urea and creatinine with significant decrease in creatinine clearance. A markedly increased levels of lipid peroxidation markers (thiobarbituric acid reactive substances and lipid hydroperoxides) and protein carbonyl contents with significant decrease in nonenzymic antioxidants (total sulphydryl groups, reduced glutathione, vitamin C and vitamin E) and enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase) as well as glutathione metabolizing enzymes (glutathione reductase, and glucose-6-phosphate dehydrogenase) were also observed in Cd intoxicated rats. Coadministration of DTS (40 mg/kg bw/day) and Cd resulted in the reversal of the kidney function accompanied by a significant decrease in lipid peroxidation and increase in the antioxidant defense system. In vitro studies with vero cells showed that incubation of DTS (5-50 microg/ml) with Cd (10 microM) significantly reduced the cell death induced by Cd. DTS at 40 microg/ml effectively blocked the cell death and lipid peroxidation induced by Cd (10 microM) indicating its cytoprotective property. Further, the flow cytometric assessment on the level of intracellular reactive oxygen species using a fluorescent probe 2', 7'-dichlorofluorescein diacetate (DCF-DA) confirmed the Cd induced intracellular oxidative stress in vero cells, which was significantly suppressed by DTS (40 microg/ml). The histopathological studies in the kidney of rats also showed that DTS (40 mg/kg bw/day) markedly reduced the toxicity of Cd and preserved the architecture of renal tissue. The present study suggests that the cytoprotective potential of DTS in Cd toxicity might be due to its antioxidant and metal chelating properties, which could be useful for achieving optimum effects in Cd induced renal damage.     

The role of K63‐linked polyubiquitination in cardiac hypertrophy

Ubiquitination, also known as ubiquitylation, is a vital post‐translational modification of proteins that play a crucial role in the multiple biological processes including cell growth, proliferation and apoptosis. K63‐linked ubiquitination is one of the vital post‐translational modifications of proteins that are involved in the activation of protein kinases and protein trafficking during cell survival and proliferation. It also contributes to the development of various disorders including cancer, neurodegeneration and cardiac hypertrophy. In this review, we summarize the role of K63‐linked ubiquitination signalling in protein kinase activation and its implications in cardiac hypertrophy. We have also provided our perspectives on therapeutically targeting K63‐linked ubiquitination in downstream effector molecules of growth factor receptors for the treatment of cardiac hypertrophy.