Phosphofructokinase and Malate Dehydrogenase Participate in the In Vitro Maturation of Porcine Oocytes

Oocyte maturation depends on the metabolic activity of cumulus–oocyte complex (COC) that performs nutritive and regulatory functions during this process. In this work, the enzymes [phosphofructokinase (PFK) and malate dehydrogenase (MDH)] were tested to elucidate the metabolic profile of porcine COCs during the in vitro maturation (IVM). Enzymatic activity was expressed in U/COC and U/mg protein (specific activity) as mean ± SEM. In vitro maturation was performed with 2‐oxoglutarate (5, 10 and 20 mm ) or hydroxymalonate (30, 60 and 100 mm ) inhibitors of PFK and MDH, respectively. The PFK and MDH activities (U) remained constant during maturation. For PFK, the U were (2.48 ± 0.23) 10^?5 and (2.54 ± 0.32) 10^?5, and for MDH, the U were (4.72 ± 0.42) 10^?5 and (4.38 ± 0.25) 10^?5 for immature and in vitro matured COCs, respectively. The specific activities were significantly lower after IVM, for PFK (4.29 ± 0.48) 10^?3 and (0.94 ± 0.12) 10^?3, and for MDH (9.08 ± 0.93) 10^?3 and (1.89 ± 0.10) 10^?3 for immature and in vitro matured COCs, respectively. In vitro maturation percentages and enzymatic activity diminished with 20 mm 2‐oxoglutarate or 60 mm hydroxymalonate (p < 0.05). Viability was not affected by any concentration of the inhibitors evaluated. The U remained unchanged during IVM; however, the increase in the total protein content per COC provoked a decrease in the specific activity of both enzymes. Phosphofructokinase and MDH necessary for oocyte IVM would be already present in the immature oocyte. The presence of inhibitors of these enzymes impairs the meiotic maturation. Therefore, the participation of these enzymes in the energy metabolism of the porcine oocyte during IVM is confirmed in this study.     

ENDOSCOPIC ULTRASOUND INSTRUMENTATION, APPLICATIONS IN HUMANS, AND POTENTIAL VETERINARY APPLICATIONS

Endoluminal scanning under endoscopic guidance, or endoscopic ultrasonography (EUS), has become the most significant advance for imaging the gastrointestinal (GI) tract wall and contiguous organs in the past 20 years. It was originally designed to overcome the limitations in humans to imaging the abdominal organs transabdominally, such as large penetration depths and GI air. This imaging modality provides detailed images of pathological processes both within and outside of the GI wall since a high-frequency transducer can be brought into close proximity with the target regions. It has found most success in humans for the staging of lung, gastric, and esophageal cancer, the detection of both lymphatic and hepatic metastases, and diagnosis of pancreatitis and pancreatic cancer, as well as achieving an important role in interventional and therapeutic procedures. The EUS examination can be performed to examine both the thorax and abdomen in animals when both conventional transthoracic or transabdominal ultrasound are inadequate due to intervening air, bone, large penetration depths, or obesity. The echoendoscope is similar to a conventional endoscope but has an ultrasound transducer at its tip. Both radial and linear multifrequency scanners are available. Linear scanners allow fine-needle aspiration (FNA) of the bowel wall or extraluminal structures. Transducer coupling is either by direct mucosal contact or by inflation of a water-filled balloon surrounding the transducer. Current thoracic applications for EUS in veterinary medicine include examination of the mediastinum, bronchial lymph nodes, esophagus, and pulmonary lesions as well as FNA of pulmonary masses. Abdominal applications include examination of both pancreatic limbs and the liver, including portosystemic shunts, detection of lymphadenomegaly, and examination of the gastric wall, duodenum, and jejunum. Other potential applications in dogs and cats include tumor staging and intrapelvic ultrasound.     

DOPPLER MEASUREMENT OF SPLANCHNIC BLOOD FLOW DURING DIGESTION IN UNSEDATED NORMAL DOGS

The purpose of this study was to measure splanchnic blood flow during digestion in unsedated dogs by using duplex Doppler sonography. The study population consisted of 12 healthy dogs. Blood flow in the cranial mesenteric artery, the celiac artery, and the aorta was measured before a test meal and at 20, 60, and 90 minutes after eating. The following measurements were made or calculated: vessel diameter, peak systolic velocity, end diastolic velocity, mean velocity, resistive index, pulsatility index, and flow volume. There was a significant postprandial decrease in the resistive and pulsatility indices in both the cranial mesenteric (preprandial RI = 0.867, postprandial RI = 0.796, preprandial PI = 3.033, postprandial PI = 2.173) and the celiac (preprandial RI = 0.854, postprandial RI = 0.769, preprandial PI = 2.639, postprandial PI = 1.930) arteries. In both vessels the end diastolic velocity, the mean velocity, and the flow volume increased significantly postprandially. These changes occurred significantly earlier in the celiac artery than in the cranial mesenteric artery. The findings most likely correspond to postprandial splanchnic vasodilation. Doppler ultrasound provide a good methode of detecting changes in postprandial splanchnic blood flow in the dog.     

Comparison of some selection procedures and objectives in faba beans (Vicia faba L.). I. direct and index selection for seed yield

Summary Starting with two heterogeneous F₂ populations, each based on 20 cross combinations, different selection procedures were applied for four consecutive selection cycles. Three of the selection procedures involved seed yield per plant (SYP): (i) direct selection for SYP (DSY); (ii) indexselection for SYP (ISY) with plant length, the ratio of the vegetative top to plant length, end of grain filling and duration of the generative stage as auxiliary characters and finally; (iii) independent culling levels selection (ICL) for the index described above and crude protein content. For ICL-selection the attention is focussed on SYP in this paper.For all three selection procedures, predicted and realized responses were calculated and expressed as percentage of a corresponding non-selected population. The intensities of selection differed over selection methods and generations, mostly due to practical constraints.As far as predicted responses are concerned, ISY-selection seemed to be slightly superior to DSY-selection and DSY-selection to ICL-selection. This order may partly be due to differences in selection intensity. However, none of the predictions was very accurate, but the predictions of ICL-selection seemed to be the most accurate. This has been ascribed also to the relative mild selection intensity applied in ICL-selection. It is concluded that a decision in favour of a complicated selection procedure like index selection based on predicted responses only cannot be defended. As far as realized responses are concerned, DSY-selection seems to be slightly superior to the other two selection procedures, as it is the only procedure that resulted in a more or less stable positive response to selection.The auxiliary characters, especially plant length, showed a marked correlated response to all three selection procedures. In case of direct selection for SYP this seemed paradoxical with the inefficiency of index selection, but, it may reflect the differences in the genuine genotypic correlations and the estimated genotypic correlations.     

Nitrous oxide emissions from fertilised grassland: A 2-year study of the effects of N fertiliser form and environmental conditions

The aim was to investigate the effects of different N fertilisers on nitrous oxide (N₂O) flux from agricultural grassland, with a view to suggesting fertiliser practices least likely to cause substantial N₂O emissions, and to assess the influence of soil and environmental factors on the emissions. Replicate plots on a clay loam grassland were fertilised with ammonium sulphate (AS), urea (U), calcium nitrate (CN), ammonium nitrate (AN), or cattle slurry supplemented with AN on three occasions in each of 2 years. Frequent measurements were made of N₂O flux and soil and environmental variables. The loss of N₂O-N as a percentage of N fertiliser applied was highest from the supplemented slurry (SS) treatment and U, and lowest from AS. The temporal pattern of losses was different for the different fertilisers and between years. Losses from U were lower than those from AN and CN in the spring, but higher in the summer. The high summer fluxes were associated with high water-filled pore space (WFPS) values. Fluxes also rose steeply with temperature where WFPS or mineral N values were not limiting. Total annual loss was higher in the 2nd year, probably because of the rainfall pattern: the percentage losses were 2.2, 1.4, 1.2, 1.1 and 0.4 from SS, U, AN, CN and AS, respectively. Application of U in the spring and AN twice in the summer in the 2nd year gave an average emission factor of 0.8% – lower than from application of either individual fertiliser. We suggest that similar varied fertilisation practices, modified according to soil and crop type and climatic conditions, might be employed to minimise N₂O emissions from agricultural land. Received: 30 August 1996     

Wnt3a-mediated formation of phosphatidylinositol 4,5-bisphosphate regulates LRP6 phosphorylation

The canonical Wnt-beta-catenin signaling pathway is initiated by inducing phosphorylation of one of the Wnt receptors, low-density lipoprotein receptor-related protein 6 (LRP6), at threonine residue 1479 (Thr1479) and serine residue 1490 (Ser1490). By screening a human kinase small interfering RNA library, we identified phosphatidylinositol 4-kinase type II alpha and phosphatidylinositol-4-phosphate 5-kinase type I (PIP5KI) as required for Wnt3a-induced LRP6 phosphorylation at Ser1490 in mammalian cells and confirmed that these kinases are important for Wnt signaling in Xenopus embryos. Wnt3a stimulates the formation of phosphatidylinositol 4,5-bisphosphates [PtdIns (4,5)P2] through frizzled and dishevelled, the latter of which directly interacted with and activated PIP5KI. In turn, PtdIns (4,5)P2 regulated phosphorylation of LRP6 at Thr1479 and Ser1490. Therefore, our study reveals a signaling mechanism for Wnt to regulate LRP6 phosphorylation.     

The dispersal of Culicoides brevitarsis in eastern New South Wales and associations with the occurrences of arbovirus infections in cattle

Distributions of the vector Culicoides brevitarsis Kieffer (Diptera: Ceratopogonidae) (determined from light trap data) and 2 arboviruses (determined from seroconversions in sentinel cattle) were studied in eastern New South Wales in 1993–94. C brevitarsis was recorded progressively from endemic areas on the north coast, to Nowra on the south coast, and westward to Scone, in the Hunter Valley. C brevitarsis also survived through winter at Paterson, in the Hunter Valley. Its apparently focal reappearance in this marginal area had no obvious effect on the broad pattern of its progression or the dispersal of Akabane and bluetongue viruses. These viruses were first recorded from foci near Coffs Harbour, on the mid-north coast. Their first occurrences at different locations were associated with those of C brevitarsis, but not with each other. The viruses were found only within the recorded limits of the vector's distribution. Delays between the initial occurrence of C brevitarsis and first evidence of virus transmissions at locations ranged from 2 to 7 months. The delays decreased away from the points of focus and were negatively associated with the time of initial occurrence of the vector. Seroconversions to the viruses were related to the presence of C brevitarsis. However, the densities of C brevitarsis had no apparent effect on the initial numbers of cattle seroconverting to either virus. The results support the conclusion that the progressions of C brevitarsis and Akabane and bluetongue viruses were the result of gradual movements by the vector.