Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Cardiopulmonary effects of a halothane/oxygen combination in hypovolemic cats.

The cardiopulmonary effects of a halothane/oxygen combination were studied in eight cats subjected to a 25% whole blood volume loss. Test parameters included cardiac output measured via thermodilution, heart rate, respiratory rate, arterial blood pressure (systolic, diastolic and mean) and blood gas analysis. Values for cardiac index, stroke volume and systemic vascular resistance were calculated from these data. Posthemorrhage cardiac output, cardiac index, stroke volume and measurements of arterial blood pressure were significantly decreased (p less than 0.05). Heart rate remained unchanged. Following induction of halothane anesthesia the above parameters experienced a further significant decline (p less than 0.05) from their immediate preanesthetic (i.e. posthemorrhage) values. Heart rate also significantly decreased (p less than 0.05). Thirty minutes following the cessation of halothane anesthesia these values returned to near-hemorrhage levels, being above their respective preanesthetic values. Systemic vascular resistance initially rose, peaking ten minutes into halothane anesthesia, before gradually falling to prehemorrhage values at the end of halothane anesthesia. Following hemorrhage, respiratory rate demonstrated a transient increase, associated with an arterial CO2 tension fall, before returning to initial values at the preanesthetic time. During halothane anesthesia respiratory rate remained unchanged whereas arterial CO2 tension rose significantly (p less than 0.05) and pH declined slightly from preanesthetic readings. These returned to prehemorrhage values 30 minutes following the cessation of halothane anesthesia.     

Infertility in female rabbits immunized with lactate dehydrogenase X

Immunization of female rabbits with the sperm-specific lactate dehydrogenase (LDH-X) resulted in a highly significant reduction of pregnancies compared to nonimmunized controls. This is the first demonstration of immunosuppression of fertility by a crystalline protein shown to be strictly homogeneous by ultracentrifugation, polyacrylamide gel electrophoresis, immunodiffusion, and micro complement fixation.     

Repeated drying–rewetting cycles and their effects on the emission of CO2, N2O,NO, and CH4 in a forest soil

Prolonged summer droughts due to climate change are expected for this century, but little is known about the effects of drying and wetting on biogenic trace‐gas fluxes of forest soils. Here, the response of CO₂, N₂O, NO, and CH₄ fluxes from temperate forest soils towards drying–wetting events has been investigated, using undisturbed soil columns from a Norway spruce forest in the “Fichtelgebirge”, Germany. Two different types of soil columns have been used for this study to quantify the contribution of organic and mineral horizons to the total fluxes: (1) organic horizons (O) and (2) organic and mineral soil horizons (O+M). Three drying–wetting treatments with different rewetting intensities (8, 20, and 50 mm of irrigation d^–1) have been compared to a constantly moist control to estimate the influence of rainfall intensity under identical drying conditions and constant temperature (+15°C). Drought significantly reduced CO₂, N₂O, and NO fluxes in most cycles. Following rewetting, CO₂ fluxes quickly recovered back to control level in the O columns but remained significantly reduced in the O+M columns with total CO₂ fluxes from the drying–wetting treatment ranging approx. 80% of control fluxes. Fluxes of N₂O and NO remained significantly reduced in both O and O+M columns even after rewetting, with cumulative fluxes from drying–wetting treatments ranging between 20% and 90% of the control fluxes, depending on gas and cycle. Fluxes of CH₄ were small in all treatments and seem to play no significant role in this soil. No evidence for the release of additional gas fluxes due to drying–wetting was found. The intensity of rewetting had no significant effect on the CO₂, N₂O, NO, and CH₄ fluxes, suggesting that the length of the drought period is more important for the emission of these gases. We can therefore not confirm earlier findings that fluxes of CO₂, N₂O, and NO during wetting of dry soil exceed the fluxes of constantly moist soil.