Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue‐type plasminogen activator (t‐PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus‐oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t‐PA and/or its inhibitor epsilon‐aminocaproic acid (control, t‐PA, t‐PA+ε‐ACA, ε‐ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t‐PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε‐ACA. PAA in the treated group was significantly reduced by ε‐ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t‐PA‐modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t‐PA. In conclusion, it appears that excessive t‐PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.     

Treatment of rupture of suspensory ligament and superficial flexor tendon in a bull

Rupture of the suspensory ligament at the insertions on the proximal sesamoid bones, and of the superficial flexor tendon of the left fore limb, occurred in an adult Angus bull as a result of fighting. There was severe hyperextension of the metacarpophalangeal (MCP) joint with the dewclaws almost touching the ground. Radiographs revealed severe hyper-extension of the MCP joint with the sesamoid bones aligned directly distal to the metacarpus. Initially, a full length fiberglass cast was applied with the limb partially flexed within the cast and the heels elevated. The cast was replaced twice. The cast was removed after 136 days and the bull was bearing full weight on the limb. Prolonged immobilisation of the limb produced new bone in the area (a normal response in cattle) to cause ankylosis of the traumatized MCP joint and partial ankylosis of the carpus. The bull was being used for pasture breeding one year after the injury.     

Mechanical comparison of materials used for extra-capsular stabilisation of the stifle joint in dogs

Objective The mechanical properties of three materials (No. 2 polypropylene, No. 5 polybutilate-coated multifilament polyester and 18, 27 and 36 kg test monofilament nylon leader material) commonly used for extra-capsular stabilisation of the stifle in dogs with cranial cruciate ligament insufficiency were determined. The ability of No. 5 polybutilate-coated multifilament polyester and 36 kg test monofilament nylon leader material, when placed as extra-capsular sutures, to mitigate cranial drawer was evaluated in hindlimbs of cadavers. Design An in vitro mechanical study. Animals Seven pairs of hindlimbs harvested from adult greyhound dogs recently euthanased for other reasons. Procedure Samples of each material, including samples of 27 kg test leader material that had been sterilised by one of three methods (ethylene oxide, one or five cycles in an autoclave), were loaded to determine tensile and stress relaxation properties. The effect of cyclic loading on a No. 5 polybutilate-coated multifilament polyester and 36 kg test leader material was also determined. Using the harvested hindlimbs, cranial drawer was measured before and after transection of the cranial cruciate ligament and on the first and twelfth cycle following extra-capsular stabilisation with either No. 5 polybu-tilate-coated multifilament suture or 36 kg test leader material. Results Leader material was found to have the most suitable mechanical characteristics for use as extracapsular stabilisation of the cranial cruciate ligament deficient stifle. Of the sterilisation methods, ethylene oxide was found to have the least detrimental effects on the handling and material characteristics of the leader material. Stifles stabilised with 36 kg test leader material had significantly less drawer than those stabilised with No. 5 polybutilate-coated multifilament polyester suture. Clinical implications Monofilament nylon leader material would appear to have suitable mechanical properties for extra-capsular stabilisation of the cranial cruciate ligament deficient stifle. If possible the material should be sterilised using ethylene oxide.     

Permeability and structure of junctional membranes at an electrotonic synapse

The dye Procion Yellow M4RS crosses junctional membranes from cytoplasm to cytoplasm at electrotonic synapses between segments of the crayfish septate axon. The dye does not enter the cells from extracellular space. Thus permeability of junctional membranes is qualitatively different from that of nonjunctional membranes. Electron microscopy after fixation in the presence of lanthanum hydroxide indicates that these synapses are "gap junctions" and that there is a network of channels continuous with extracellular space between apposed junctional membranes. These channels must be interlaced with intercytoplasmic channels that are not open to extracellular space.     

Temperature-dependence of resistance at an electrotonic synapse

The junctional resistance at septa of the crayfish lateral giant axon is inversely related to temperature with a Q(1l), of about 3 over the range from 5 degrees to 20 degrees C. Nonjunctional axonal membrane is much less affected. Resistance changes occur rapidly with temperature changes. No correlates in ultrastructure of the synapses have been found.     

Atom counting at surfaces

Multiphoton resonance ionization has been combined with energetic ion bombardment to examine dopant concentrations ofindium on the surface of silicon. The results yield a linear relation between the indium concentration and the known bulk values and a detection limit of 9 parts per trillion, at a mass resolution exceeding 160. This measurement, which surpasses the limits of any previous surface analysis by a factor of 100, has been made possible with an experimental configuration that optimizes sampling and detection efficiency while reducing background noise to virtually zero. During the analysis, submonolayer quantities of the surface are removed, so that as few as 180 surface atoms may be counted.     

Fluorogenic substrate detection of viable intracellular and extracellular pathogenic protozoa

Viable Leishmania promastigotes and amastigotes were detected by epifluorescence microscopy with fluorescein diacetate being used to mark living parasites and the nucleic acid-binding compound ethidium bromide to stain dead cells. This procedure is superior to other assays because it is faster and detects viable intracellular as well as extracellular Leishmania. Furthermore, destruction of intracellular pathogens by macrophages is more accurately determined with fluorescein diacetate than with other stains. The procedure may have applications in programs to develop drugs and vaccines against protozoa responsible for human and animal disease.     

Agricultural practices alter sex ratios in a reptile with environmental sex determination

Anthropogenic changes to the environment have the opportunity to impact natural systems, particularly in organisms that exhibit phenotypic plasticity. Species with environmental sex determination (ESD) are uniquely susceptible to changes in the stimuli that affect sexual development, potentially leading to maladaptive sex ratios. We studied the factors affecting sex determination and sex ratios in the common snapping turtle, Chelydra serpentina, in an area heavily impacted by agricultural practices. We investigated the effects of soybean, corn, and sunflower planting on incubation temperatures, sex ratios, and depredation in naturally laid nests. We also identified and analyzed a novel mitochondrial microsatellite in order to examine the presence of natal homing and determine the likelihood that nest sites impacted by agricultural practices could be transmitted across generations. Females frequently chose to nest in agricultural fields over sand prairie sites, and offspring sex ratios and depredation rates were significantly influenced by crop planting. Despite detecting considerable genetic variation in our population, we found no relationship between relatedness and nesting location, suggesting that females are not transmitting nesting sites across generations. Our results suggest that agricultural practices can directly impact populations of animals with ESD, and will need to be considered in management decisions.     

Decay of methidathion on Greek soultanina grapes during storage and on the vines

Decomposition of the pesticide methidathion on and in soultanina (Thomson seedless) grapes was studied. Vines, uncovered and covered were sprayed with the pesticide. Samples were collected and analyzed in regular time intervals. Two lots were collected, the first of them, 2 h after spraying and the second 20 days later, and were stored in a commercial refrigerated room. Degradation of the pesticide was studied for about 50 days for the grapes on the vines and 130 days for the grapes stored in the refrigerated room. The half-life of the pesticide methidathion was found to be 5 days for the uncovered vines, 7 days for the covered vines and 64 days for the grapes stored in the refrigerator.