Effects of testosterone implants in pregnant ewes on their female offspring

Optimal conditions for using high-performance liquid chromatography (HPLC) in the size exclusion mode have been determined for measuring the molecular-weight (MW) distribution of chitosan samples. Physical separation according to molecular size was accomplished on the stationary phase of glass supports having controlled pore sizes ranging from 2500 to 40 A. Selection of column combinations was based on the requirements to resolve the higher MW fraction of chitosan and to give a linear calibration curve within the required MW range. The best combination of glass pore sizes and column lengths in two foot sections joined sequentially was: 2500 A (2 ft.), 1500 A (4 ft.), 550 A (6 ft.), 250 A (2 ft), 100 A (2 ft.), and 40 A (2 ft.). A loading study showed that an injection load of 500 mug, i.e. 100 mul at 5 g/l or 50 mul at 10 g/l (w/v), was the optimal load to give reproducible elution volumes, precision in quantitation, and minimum viscosity effects. The best calibration curve using defined dextran standards was obtained from the geometric mean of Mw (weight average MW) and Mn (number average MW) values and peak elution volumes. Precision in determining MW distribution of chitosan as well as dextran standards was better than 5% relative standard deviation, and the differences between these results and the manufacturer's data on the dextran standards were 6 to -17%. The MW distribution of a selected chitosan samples in 2% acetic acid thus determined was Mw = 2,055,000, Mn = 936,000, dispersity = 2.16, and the most abundant species was around 1,103,000. Analysis time for the HPLC separation was less than 20 min per sample. Chitosan is an effective coagulating agent for the treatment of food processing wastes and activated sludge from biological treatment systems. It is manufactured from chitin in shrimp and crab wastes. The rapid methods developed here for determining the MW distribution of chitosan preparations will be used to optimize the manufacturing process and guide the selection of more effective chitosan products.     

An evaluation of transovarian uptake of metabolites using arterio-venous difference methods in dairy cattle

Arterio-venous (A-V) difference techniques were used in cattle to examine ovarian energy metabolism, cholesterol uptake and steroid hormone outputs. Catheters were inserted into the ovarian vein and facial artery, and Transonic flow transducers were placed around the ovarian A-V plexus. Further, in some cows, the effects of a challenge with GnRH were examined. Glucose uptake and lactate output were significant in most individual cows. Nonesterified fatty acids (NEFA) uptake were not significant in any cow in dioestrus. Ovarian uptake of beta-Hydroxy-butyrate (3-OHB) was significant in 4 cows in dioestrus. Cholesterol uptake was significant in only 1 cow. Oxygen uptake was significant in all cows at all stages of the oestrous cycle. All cows had significant output of progesterone and oestradiol-17 beta. These data show that the bovine ovary utilises significant amounts of glucose, and Respiratory quotient (RQ) estimates demonstrated that glucose was the primary fuel used by the ovary. The significant output of lactate suggested that anaerobic pathways were mainly used for glucose oxidation. The observed uptakes of 3-OHB indicated that the ovary utilises 3-OHB as a source of energy. Cholesterol uptake was not a rate-limiting factor for steroid hormone production in the ovary. Despite the high metabolic rate in the luteal ovary, the small difference in PO2 between arterial and ovarian venous blood indicated that the ovary consumes only a small proportion of available oxygen. GnRH had no significant effect on the uptake of metabolites and energy metabolism, but it increased OBF and the output of progesterone and oestradiol-17 beta. The use of A-V methods to determine the metabolic needs of the ovary is useful in understanding the means by which nutrition can influence fertility.     

A biochemical basis for obligate methylotrophy: properties of a mutant of Pseudomonas AM1 lacking 2-oxoglutarate dehydrogenase

The effects of PGE1 on the dog heart were studied using the blood-perfused sinus node and papillary muscle preparations isolated separately from the same animal. PGE1 administered into the papillary muscle artery as bolus injections in doses of 1-1000 ng caused a dose-dependent increase of the developed tension and dT/dt of the papillary muscle. The effect was not inhibited by the beta-adrenoceptor blocking agent pindolol. PGE1 injected into the sinus node artery in doses of 3-300 ng did not change the rate of contraction of the sinus node preparation. PGE1 in the blood concentrations of 4.4 X 10(-9) to 1.7 X 10(-7) M enhanced the positive inotropic responses to noradrenalin and field stimulation as well as to calcium. The influence of PGE1 on the positive inotropic effect of perivascular nerve stimulation was not consistent: the action of perivascular nerve stimulation was enhanced by PGE1 in the majority of preparations but was reduced in one third of preparations. PGE1 in the same blood concentrations as used in the papillary muscle significantly depressed the positive chronotropic responses to noradrenaline and dopamine. The present results indicate that PGE1 induces multiple actions on the dog heart. Its predominant effect on the ventricular myocardium appears to be enhancement of the adrenergic stimuli probably via the facilitation of calcium movement through the myocardial cell membrane. In addition, PGE1 may decrease the sensitivity of beta-adrenoceptors to adrenergic stimuli in the sinus node.     

A comparison of the effect of two regimens of infestation on the development of resistance by cattle to the cattle tick, Boophilus microplus (Can.)

The development of resistance to Boophilus microplus by cattle was studied using sets of cattle twins, in stalls. One twin from each set received 3 infestations of 40,000 larvae and the other a continuous infestation of 1,000 larvae a day over the same period. Sets of twins were then challenged with 1,000 larvae a day for 40 days and 2 field infestations of 20,000 larvae. Correlations of ranking for resistance made at 40-day periods during daily infestations of 1,000 larval ticks or for corresponding intermittent infestations of 40,000 larvae were low until animals had received 120,000 larvae. Thereafter irrespective of method of infestation correlations of ranking were relatively high (r = 0.61-0.96) between periods of infestations or method of infestation including 2 field infestations. Fewer adult female ticks matured on the daily infested animals than on the intermittently infested animals during the treatment period, but animals developed a similar resistance level whether infested with either technique.     

Operative correction of pectus excavatum: an evolving perspective

Volatile free fatty acids (VFA) in blood increased approximately twofold in dogs subjected to total hepatectomy. The average total plasma VFA preoperatively was 1,585 mug percent and shortly before death, postoperatively, was 2,798 mug percent. The corresponding red cell concentrations were essentially the same. Acetic acid was 81 percent of the total VFA, propionic acid 7 percent, isobutyric acid 4 percent, butyric acid one percent, and isovaleric acid 5 percent. There was little or no isovalerate in red cells. The increments in the individual fatty acids after hepatectomy were highly variable, but the average increase with time was almost linear. The increase in VFA probably reflects an increased utilization of the branched-chain aminoacids by extrahepatic tissues.     

T cells of patients with the Wiskott-Aldrich syndrome have a restricted defect in proliferative responses

The Wiskott-Aldrich syndrome (WAS) is a disease of profound thrombocytopenia and severe immune defects caused by an unidentified defective X chromosome gene. In this study, T lymphocyte function is examined using a panel of allospecific WAS patient T cell lines, previously found to express the abnormal disease gene and the cytoarchitectural defect characteristic of the disease. Although T cell lines from normal individuals proliferate vigorously in response to immobilized anti-CD3 mAb OKT3 and SPV-T3b, five of seven WAS patient T cell lines failed to proliferate and two lines showed significantly decreased proliferation when challenged with the immobilized anti-CD3 mAb. The deficient responsiveness of the WAS T cell lines to immobilized anti-CD3 mAb is a restricted defect, because the cells proliferate normally when challenged with allospecific Ag, PHA, or PMA plus ionomycin. Addition of anti-CD28 mAb did not correct the deficient proliferation of the WAS cells challenged with immobilized anti-CD3. Deficient response of the WAS T cell lines to immobilized anti-CD3 was detected also when earlier events of the proliferation process, IL-2 production and up-regulation of activation Ag CD69 and CD28, were measured. On the other hand, WAS cell lines did not differ from normal cell lines in binding of anti-CD3 mAb, mobilization of Ca2+ in response to soluble OKT3, and tyrosine phosphorylation and GTP binding of the CD3 zeta-chain in response to OKT3. Cumulatively, these findings demonstrate a striking restricted defect in the proliferative response of WAS T cells, which because it is found in cell lines free of secondary changes that occur in the patient circulation must be a reflection of the inherited defective disease gene product.     

Abnormal activation of lipoprotein lipase by non-equilibrating apoC-II: further evidence for the presence of non-equilibrating pools of apolipoproteins C-II and C-III in plasma lipoproteins

Using artificial triglyceride emulsions, we have demonstrated the presence of non-equilibrating pools of apolipoproteins C-II and C-III in human plasma lipoproteins. As the concentrations of acceptor triglycerides were increased, a greater fraction of both apoC-II and apoC-III shifted away from the native plasma lipoproteins to the artificial lipid emulsions. All of the apoC-II and apoC-III in very low density and high density lipoproteins (VLDL and HDL), however, could not be removed from native plasma lipoproteins. The percent of total plasma apoC-II and apoC-III that could be recovered in the VLDL and HDL density fractions varied when plasma from different individuals was used. When plasma samples from normotriglyceridemic subjects were used, HDL was the primary donor of apoCs. The percent of total plasma apoCs associated with HDL decreased from 60% to 25% for apoC-II and from 65% to 15% for apoC-III. When plasma samples from hypertriglyceridemic subjects were incubated with artificial lipid emulsions, VLDL was the primary donor of apoCs. HDL from hypertriglyceridemic subjects only accounted for 5-10% of total fasting plasma apoCs and did not contribute significantly to the final apoC contents of the artificial triglyceride emulsions. To evaluate the significance of the depletion of exchangeable apoCs from plasma HDL, we also examined the ability of control and apoC-depleted HDL to serve as activator for bovine milk lipoprotein lipase (LPL) in vitro. When HDL depleted of exchangeable apoCs were used as the source of plasma apolipoproteins for the activation of LPL in vitro, only 5-10% of the maximal activity obtained with native HDL was demonstrated. In fact, in the presence of comparable concentrations of HDL apoC-II, activation of LPL was the least with HDL which lacked exchangeable apoCs. Our data thus indicated that the presence of exchangeable apoC-II on HDL is necessary for the activation of LPL in vitro. This finding is consistent with our data that suggest that HDL from hypertriglyceridemic subjects do not stimulate LPL as well as HDL from normolipidemic subjects.