Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Histofarcin test for the diagnosis of epizootic lymphangitis in Ethiopia: development, optimisation, and validation in the field

Histofarcin, a skin test antigen for the diagnosis of epizootic lymphangitis, was locally produced from the mycelial form of Histoplasma capsulatum var. farciminosum (HCF) in disease-endemic districts of Ethiopia and tested for its application in the field between April 2002 and May 2003. The test was evaluated using 108 mules, 84 in endemic and 24 in disease-free districts. Microscopic and mycological examinations of clinical lesions were used as the "gold standard" for the validation of the test. The concentration of histofarcin that caused an optimum reaction was 0.2-0.4 mg/mL in a 0.1 mL dose and this was attained 24-48 h post-injection. The sensitivity and specificity of the histofarcin test were 90.3% (CI = 73.1, 97.5) and 69% (95%, CI = 48.1, 84.9%) in disease-endemic districts. On the other hand, specificity was 100% (CI=94.8, 100) in disease-free districts. Positive and negative predictive values of the histofarcin test were 77.78% (95% CI = 60.4, 89.3) and 85.71% (95% CI = 62.6, 96.2), respectively. A marginal substantial agreement (kappa = 0.61, P = 0.0000) was observed between the clinical status and the result of the histofarcin test. A large proportion (31%) of 'false positives' was recorded in endemic districts, which could be due to the pre-clinical stage of the disease. The latter ended in lower specificity and positive predictive value of the test since the true infection status of a 'false positive' could not be known on the basis of clinical features. Therefore, standard test validation procedures including slaughtering and isolation of HCF is required. After proper validation, we conclude that the histofarcin test could play a significant role in detecting early infection, and differentiating of EL from glanders, ulcerative lymphangitis, and sporotrichosis.     

Progression of chocolate spot (Botrytis fabae) and grain yield of faba bean as influenced by integration of fungicide rate and host resistance in Southern Ethiopia

Journal of Crop Science and Biotechnology - Chocolate spot is economically the most important and destructive disease of faba bean. Field studies were conducted to develop and determine an...     

Antagonism of detomidine with tolazoline in isoflurane-anaesthetised ponies

The effects of tolazoline (4.0 mg/kg iv) antagonism of detomidine (0.02 mg/kg iv) were evaluated in isoflurane-anaesthetised, ventilated ponies. Each of 6 ponies received both tolazoline and saline treatment during separate anaesthetic episodes only (no surgery was performed). Detomidine administration produced an increase in blood pressure, decrease in heart rate and decrease in P_aO₂ Tolazoline treatment transiently increased heart rate while blood pressure returned to baseline after both treatments. Arterial oxygenation decreased further after tolazoline treatment while oxgenation recovered towards baseline with saline treatment. No other cardiopulmonary effects were detected. Recovery from anaesthesia tended to be more rapid when detomidine was antagonized. The potential benefit of antagonizing detomidine-induced bradycardia with tolazoline, during isoflurane anaesthesia should be weighed against the potential to produce a decrease in arterial oxygenation. The mechanism for this effect is not clear.     

Distribution and association of faba bean gall (Olpidium viciae) disease with agro-ecological factors in northwestern Ethiopia

Journal of Plant Diseases and Protection - Faba bean (Vicia faba L.) is an important food legume crop used for various dishes due to its high protein and essential nutrients content. In recent...     

Effect of pH on rheotaxis of bull sperm using microfluidics

The aim of the present research is to study the effect of pH values on the sperm rheotaxis properties. Semen collected from bulls was diluted with SOF medium (1:10). pH of the medium was adjusted using a digital pH meter to the following pH values: 6.0, 6.2, 6.4, 6.4, 6.8, 7.0. All kinetic parameters of sperm (n = 3,385) were determined through a computer‐assisted sperm analysis (CASA) system using microfluidic devices with controlled flow velocity. The following parameters were determined: total motility (TM%), positive rheotaxis (PR%), straightline velocity (VSL, μm/s), average path velocity (VAP, μm/s), linearity (LIN, as VSL/VCL, %), beat cross‐frequency (BCF, Hz) and curvilinear velocity (VCL, μm/s). Nitric oxide, calcium and potassium were estimated in semen at different pH values. To confirm the effect of nitric oxide and K⁺, we used sodium nitroprusside (an NO donor) and KCL as (a K⁺ donor) to see their effect on sperm PR%. The results showed no difference in TM% at pH (6–7). The PR% was the lowest at pH 6 and 7. The best parameters for the PR% were at pH 6.4–6.6. The concentration of Ca⁺² did not change at different pH values. The mean NO values decreased with the increase of pH; however, the mean values of K⁺ increased with the increase of pH. Addition of high concentration of NO and K⁺ to the semen media at fixed pH level had a negative effect on TM% and PR%. In conclusion, the bull sperm had the best rheotaxis properties at pH 6.4–6.6 and sensitive to the change of seminal NO and K⁺.     

Simrank: Rapid and sensitive general-purpose k-mer search tool

Background  

Terabyte-scale collections of string-encoded data are expected from consortia efforts such as the Human Microbiome Project . Intra- and inter-project data similarity searches are enabled by rapid k-mer matching strategies. Software applications for sequence database partitioning, guide tree estimation, molecular classification and alignment acceleration have benefited from embedded k-mer searches as sub-routines. However, a rapid, general-purpose, open-source, flexible, stand-alone k-mer tool has not been available.