Catalytic antibodies with lipase activity and R or S substrate selectivity

The specific hydrolysis of unactivated esters bearing an R or S enantiomeric alcohol has been achieved by two separate classes of catalytic antibodies induced to bind either the R or S substrates. The antibodies exhibit rate accelerations (10(3) to 10(5] above background hydrolysis that, coupled with their antipodal specificity, provide a novel set of reagents for use in synthesis.     

Small-colon rupture attributable to granulosa cell tumor in a mare

A large granulosa cell tumor was believed to be responsible for causing obstruction and subsequent rupture of the small colon in a 10-year-old Quarter Horse mare. Two months earlier, a mass, tentatively diagnosed as granulosa cell tumor of the left ovary, had been identified by means of rectal palpation and ultrasonography. The mare was evaluated for clinical signs of acute, severe, abdominal pain, increased heart rate, cyanotic mucous membranes, clinical dehydration, with high PCV, leukopenia, and extreme abdominal distension. A large soft tissue mass and taut band that constricted the lumen of the small colon were palpable per rectum. Septic peritonitis was diagnosed on the basis of results of abdominocentesis. Exploratory surgery revealed extensive fecal contamination of the abdominal viscera, and the mare was euthanatized because of the resultant poor prognosis. At necropsy, the small colon was occluded by a taut, left broad ligament and the ovarian mass that was proved to be a granulosa cell tumor. The occlusion had caused impaction of the small colon, with subsequent perforation at the level of the broad ligament.     

Animal medical genetics: a perspective on the epidemiology and control of inherited disorders

This perspective considers genetic disorders of domestic animal populations, in particular their epidemiology and control. Inherited disorders of animals share the same basic molecular biology as those of human beings, but they differ in their epidemiology due largely to the breed structure of the various species, human control of breeding and a greater influence of the founder effect, particularly due to extensive use of a limited number of sires, and inbreeding. Control of genetic disorders in animals is also more practical through extensive screening for disease, or heterozygous animals within defined breed populations, followed by exclusion of affected or carrier animals from breeding. This is assisted by the fact that, within a breed, many inherited monogenic disorders are associated with a single mutation. However some of the more important disorders may be inherited in a non-Mendelian manner, being influenced by multiple genes as well as environmental factors. These aspects are discussed and contrasted with similar aspects in human medical genetics.     

Conversion of Cortisone to Cortisol and Prostaglandin F2α Production by the Reproductive Tract of Cows at the Late Luteal Stage In Vivo

Previous in vitro studies demonstrated that bovine endometrium has the capacity to convert inactive cortisone to biologically active cortisol (Cr) and that Cr inhibits cytokine‐stimulated prostaglandin F_2α (PGF) production. This study was carried out to test the hypothesis that bovine reproductive tract has the capacity to convert cortisone to Cr in vivo and to evaluate the effects of intravaginal application of exogenous cortisone on uterine PGF secretion during the late luteal stage. The temporal relationships between PGF and Cr levels in uterine plasma were also determined. Catheters were inserted into jugular vein (JV), uterine vein (UV), vena cava caudalis (VCC) and aorta abdominalis (AA) of six cows on Day 15 of the oestrous cycle (ovulation = Day 0) for frequent blood collection. On Day 16, the cows were divided randomly into two groups and infused intravaginally with vaseline gel (10 ml; control; n = 3) or cortisone dissolved in vaseline gel (100 mg; n = 3). Blood samples were collected at ?2, ?1, ?0.5, 0, 0.5, 1, 1.5, 2, 3, 4, 5 and 6 h after treatments (0 h). Intravaginal application of cortisone increased plasma concentrations of Cr between 0.5 and 1.5 h in UV, at 0.5 h in VCC, at 1 h in JV and at 1.5 h in AA. The plasma concentrations of PGF in UV and of PGF metabolite in JV were greater at 0.5 and 1 h in the cortisone‐treated animals than in control animals. The levels of PGF in UV blood plasma decreased after Cr reached its highest levels. The overall findings suggest that the female reproductive tract has the capacity to convert cortisone to Cr in vivo. Based on the temporal changes of PGF and Cr levels in the uterine plasma, a biphasic response in PGF secretion was found to be associated to the Cr increase induced by the cortisone treatment at the late luteal stage in non‐pregnant cows.     

Pregnancy termination in dogs with novel nonhormonal compounds.

Resorption of the products of fertilization was induced in bitches given (subcutaneously) single or multiple doses of the nonhormonal compounds L-10492 and L-10503 during the first half of gestation; also resorption or expulsion of the conceptus was induced when these compounds were given during the latter part of pregnancy. The smallest doses were required at a time immediately after implantation of the fertilized ova in the uterus. Effectiveness and appearance of side effects were dose-dependent. These consisted of decreased appetite, loss of body weight, and diarrhea (feces sometimes containing streaks of blood). Bitches which had aborted returned to estrus within normal intervals of time, exhibited normal mating behavior, and were fertile. They had normal deliveries and lactation, and the pups were normal. The mechanism of action does not involve effects on nidation and the compounds were not luteolytic, but probably involved the uteroplacental complex.     

Evidence for antibody-catalyzed ozone formation in bacterial killing and inflammation

Recently, we showed that antibodies catalyze the generation of hydrogen peroxide (H2O2) from singlet molecular oxygen (1O2*) and water. Here, we show that this process can lead to efficient killing of bacteria, regardless of the antigen specificity of the antibody. H2O2 production by antibodies alone was found to be not sufficient for bacterial killing. Our studies suggested that the antibody-catalyzed water-oxidation pathway produced an additional molecular species with a chemical signature similar to that of ozone. This species is also generated during the oxidative burst of activated human neutrophils and during inflammation. These observations suggest that alternative pathways may exist for biological killing of bacteria that are mediated by potent oxidants previously unknown to biology.     

The enzymic nature of antibody catalysis: development of multistep kinetic processing

Detailed kinetic investigations of a catalytic antibody that promotes the hydrolyses of an anilide and phenyl ester show that this catalyst uses a multistep kinetic sequence resembling that found in serine proteases to hydrolyze its substrates, although antibody was elicited to a single transition-state analog. Like the serine proteases the antibody catalyzes the hydrolysis reactions through a putative covalent intermediate, but unlike the enzymes it may use hydroxide ion to cleave the intermediates. Nevertheless, the antibody is a potent catalyst with turnover at higher pH values rivaling that of chymotrypsin. This analysis also reveals that turnover by the antibody is ultimately limited by product desorption, suggesting that improvements in catalytic efficiency may be achieved by judicious changes in the structure of the substrate, so that it is not superimposable on that of the eliciting hapten.