Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

Seroprevalence of psittacine beak and feather disease in wild psittacine birds in New South Wales

SUMMARY A haemagglutination inhibition assay was used to detect antibody to psittacine beak and feather disease virus in sera from wild sulphur crested cockatoos (Cacatua galerita), galahs (Eolophus roseicapillus), short-billed corellas (Cacatua sanguinea), eastern long-billed corellas (Cacatua tenuirostris) and other psittacine birds in New South Wales. The seroprevalence of psittacine beak and feather disease ranged from 41% to 94% in different flocks, indicating infection with the virus is widespread in wild populations.     

Plasma indicators of muscle damage in a model of nutritional myopathy in weaner sheep

SUMMARY Subclinical nutritional myopathy was induced in 5-month-old sheep by feeding them a diet low in vitamin E and selenium. Subsequently clinical myopathy was induced by dosing with protected polyunsaturated fatty acids. Plasma activities of creatine kinase (CK), pyruvate kinase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase and aldolase, enzymes of muscle origin, all remained above their reference ranges in clinically affected sheep, but fluctuated widely. Similar fluctuations occurred in subclinically affected animals, resulting in some activities being within the reference ranges and some above, at different times. Plasma malondialdehyde, an indicator of lipid peroxidation, proved of no diagnostic value. Terminal plasma CK activities were significantly correlated with microscopic damage in the vastus lateralis (VL), but not the vastus intermedius (VI) or the tensor fascia lata (TFL) muscles. AST was the most highly correlated with damage in VI and VL. In two clinically affected sheep successfully treated with an oral dose of α-tocopherol actetate all enzymes decreased steadily to within their reference ranges, at rates probably related to their plasma half-lives. These results suggest that measurement of plasma CK activity would be useful in monitoring recovery of treated sheep.     

The confidential enquiry into perioperative equine fatalities (CEPEF): mortality results of Phases 1 and 2

Objectives To document the equine perioperative mortality rate and to highlight any factor associated with an increased risk of death up to 7 days after anaesthesia. Study design A prospective observational epidemiological multicentre study. Methods Data were recorded from all equidae undergoing general anaesthesia in 62 clinics. Power calculations indicated that 45 000 cases were required to detect the significance of important variables. Details of each horse, operation, anaesthetic agents and clinic personnel were recorded. Outcome at 7 days was recorded as: alive, put to sleep (PTS) or dead. Data were analysed by a standard multilevel logistic regression approach, considering the effects of clustering at the level of clinic. Results Data were collected from 41 824 cases over 6 years. A total of 39 025 (93.3%) were alive on day 7 and 785 were dead giving an overall death rate of 1.9% (95% CI: 1.8–2.0) and 2014 (4.8%) were PTS. About 5846 horses undergoing emergency abdominal surgeries (‘colics’) were excluded from subsequent analyses. A total of 35 107 ‘noncolic’ horses were alive at 7 days and 328 dead giving a death rate for noncolics of 0.9% (95% CI: 0.8–1.0). Five hundred and forty‐three (1.5%) noncolic horses classified PTS were excluded from further analyses. There were 109 (33%) deaths from cardiac arrest or post‐operative cardiovascular collapse, with 107 (32%) from fractures and myopathies. Fracture repair, out of hours surgery, and age below 1 month was associated with increased risk of dying whereas the use of acepromazine and intravenous anaesthetic agent maintenance of anaesthesia was associated with reduced risk. Conclusions A number of potential contributors to the high risk of anaesthetic‐related mortality have been identified. Further investigation of the underlying mechanism for their apparent harmful effects and development of alternative techniques is merited.     

Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality.     

Antituberculosis activity of alkylated mulinane diterpenoids

Natural azorellane and mulinane diterpenoids show antituberculosis activity, which is increased by methylation of their free carboxyl group. We have systematically investigated the effect of alkylation in this class of diterpenoids and found that the profile of bioactivity is relatively unaffected by the introduction of short alkyl groups, both linear and branched. In this investigation, three semisynthetic diterpenoids, 13 hydroxy-mulin-11-en-20-oic acid n-propyl ester (3) and the n-propyl (19) and n-butyl (20) esters of isomulinic acid, showed the strongest antituberculosis activity (MIC = 6.25 μg/mL) against a drug-resistant strain of Mycobacterium tuberculosis.     

Continental drainage in north america during the phanerozoic from Nd isotopes

Neodymium (Nd) isotopic data show consistent patterns in the sources of sedimentary rocks in North America at a continental scale. Between 600 and 450 million years ago (Ma), ancient continental shield sources dominated. Around 450 Ma, detritus from the Caledonian-Appalachian mountains overwhelmed sediment from all older sources, and is documented over large areas of the southern, western, and northern margins of North America. This material continued to dominate the sediment supply until about 150 Ma, probably due to cannibalistic recycling of sedimentary rocks formed earlier. Around 150 Ma, the rising western Cordillera delivered new and different detritus to the sedimentary system.     

The structure of the upper atmosphere of mars: In situ accelerometer measurements from mars global surveyor

The Mars Global Surveyor (MGS) z-axis accelerometer has obtained over 200 vertical structures of thermospheric density, temperature, and pressure, ranging from 110 to 170 kilometers, compared to only three previous such vertical structures. In November 1997, a regional dust storm in the Southern Hemisphere triggered an unexpectedly large thermospheric response at mid-northern latitudes, increasing the altitude of thermospheric pressure surfaces there by as much as 8 kilometers and indicating a strong global thermospheric response to a regional dust storm. Throughout the MGS mission, thermospheric density bulges have been detected on opposite sides of the planet near 90 degreesE and 90 degreesW, in the vicinity of maximum terrain heights. This wave 2 pattern may be caused by topographically-forced planetary waves propagating up from the lower atmosphere.     

Left-side displacement of the abomasum in dairy cows at pasture

The presentation of approximately 40 dairy cows affected with left-side displacement of the abomasum (LDA) per annum in a cattle practice in East Gippsland, Victoria provided an opportunity to conduct a survey and case-control study of the disease in a grazing environment. The study, involving 37 dairy cows at pasture, revealed significant differences from the pattern of the disease occurring in the northern hemisphere where cows in older age groups, of larger frame size, higher production and fed high grain rations are at increased risk. Affected cows were diagnosed over a 10-month period and represented approximately 0.06% of the dairy cow population. Most cases were diagnosed in the early lactation period. Evidence for a genetic predisposition was suggested by the discovery that one sire generated a disproportionately large number (9) of the cows with LDA. Although affected cows were average producers in their herds, being a member of a high-producing herd was a significant risk factor.     

水稻丝氨酸蛋白酶S8基因家族的分布及分析

水稻丝氨酸蛋白酶S8基因家族在水稻的生长发育过程中起着重要的调控作用。本研究利用公共数据库资源,分析水稻中丝氨酸蛋白酶S8基因家族,在水稻12条染色体上找到46个该类基因。通过其结构分析发现,每个基因的内含子数目从0到10各不相同,但氨基酸序列是非常保守的,都有催化活性位点和底物结合位点。系统进化树分析显示,这46个基因分为3个亚家族,S8-1亚家族最大。该家族基因的进化主要是通过基因重复复制的方式进行,其表达模式发生了变化,并且多个基因在穗部具有表达。