Uterine electromyographic activity in horse mares as measured by radiotelemetry

The objective of this study was to determine if radiotelemetry could be used to measure myometrial electromyographic (EMG) activity. A radio transmitter with one pair of biopotential leads was implanted in the flank ipsilateral to the pregnant uterine horn at least five weeks prior to the expected date of parturition in two mares. The biopotential leads were implanted in the base of the pregnant uterine horn. Telemetered data were received by a pair of antennae placed at right angles in a 3.3 by 6.6-m stall. Data were recorded on VHS format videocassette tapes continuously for the 24h prior to and following parturition. Simultaneous physiograph recordings were made as a hard copy reference. In addition, 10 mg of prostaglandin F(2alpha) was administered to two mares in the luteal phase of the estrous cycle. Myometrial EMG during parturition was increased similarly to that of previously published reports that used myometrial electrodes wired directly to a physiograph. Prostaglandin F(2alpha) also caused an increase in myometrial EMG activity within 8 min of administration. This study demonstrated that radiotelemetry can be used for measuring myometrial EMG activity.     

Do parasites express receptors for host lipoproteins?

Although cholesterol is the predominant sterol in parasite tissue, many parasites are unable to synthesize cholesterol or longchain fatty acids, de novo, and must therefore obtain these components from the host. Of particular interest are the plasma lipoproteins, a rich and abundant source of cholesterol, and other lipids that could be used by parasites inhabiting the vascular system of their host or with access to plasma proteins at extravascular sites. It is not inconceivable that parasites may have evolved a variety of receptors for lipoproteins by convergent evolution. Here, Mark Rogers discusses evidence for the presence of lipoprotein receptors in protozoan and metazoan parasites of mammals.     

Calretinin and calbindin in the retina of the developing chick

Summary Calretinin and calbindin-D28k are two calcium-binding proteins that are present in largely different sets of nerve cells in the central nervous system. Their appearance during development of the chick retina was studied by immunohistochemistry and Western blots. The patterns are mature one day before hatching. Each cell type acquires its characteristic calcium-binding protein several days after its differentiation has started, but in most cases before morphological maturation is complete. There is also an early phase of calbindin immunoreactivity in many immature amacrine cells, and of calretinin immunoreactivity in the presumptive photoreceptor layer, suggesting that these proteins may have distinct functions in differentiating cells.Abbreviations CR+ Immunoreactive for calretinin only - CB+ immunoreactive for calbindin only - CR+CB+ immunoreactive for both antisera - IPL inner plexiform layer - OPL outer plexiform layer     

Statistical analysis of the migration component of genetic drift

Statistical methods are introduced for analysis of the migration component of genetic drift, i.e., of the stochastic changes that affect allele frequencies during migration between local groups. Attention focuses on alpha M, a parameter that measures the extent to which this component of drift departs from the ideal of independent random sampling, and which can be interpreted as a measure of the extent to which migration is kin-structured. It is shown that alpha M can be estimated from genetic data, even in the absence of information about the genealogical relationships of migrants, and Monte-Carlo simulations are used to approximate the sampling distribution of the estimator under the null hypothesis of independent random sampling. Application of these methods to data from the Aland Islands, Finland, shows that the migration pattern there is consistent with the hypothesis of independent random sampling.     

Purification and properties of the inducible coenzyme A-linked butyraldehyde dehydrogenase from Clostridium acetobutylicum.

The coenzyme A (CoA)-linked butyraldehyde dehydrogenase (BAD) from Clostridium acetobutylicum was characterized and purified to homogeneity. The enzyme was induced over 200-fold, coincident with a shift from an acidogenic to a solventogenic fermentation, during batch culture growth. The increase in enzyme activity was found to require new protein synthesis since induction was blocked by the addition of rifampin and antibody against the purified enzyme showed the appearance of enzyme antigen beginning at the shift of the fermentation and increasing coordinately with the increase in enzyme specific activity. The CoA-linked acetaldehyde dehydrogenase was copurified with BAD during an 89-fold purification, indicating that one enzyme accounts for the synthesis of the two aldehyde intermediates for both butanol and ethanol production. Butanol dehydrogenase activity was clearly separate from the BAD enzyme activity on TEAE cellulose. A molecular weight of 115,000 was determined for the native enzyme, and the enzyme subunit had a molecular weight of 56,000 indicating that the active form is a homodimer. Kinetic constants were determined in both the forward and reverse directions. In the reverse direction both the Vmax and the apparent affinity for butyraldehyde and caproaldehyde were significantly greater than they were for acetaldehyde, while in the forward direction, the Vmax for butyryl-CoA was fivefold that for acetyl-CoA. These and other properties of BAD indicate that this enzyme is distinctly different from other reported CoA-dependent aldehyde dehydrogenases.     

Effect of the structure of phospholipid on the kinetics of intravesicle scooting of phospholipase A2

Action of pig pancreatic phospholipase A2 on vesicles of over 50 synthetic 1,2-diacylglycerol-3-phosphate derivatives and analogs is examined in the absence of any additives. In general, shorter acyl chains and small substituents on the phosphate make a better substrate, while phospholipids with large apolar substituents are not hydrolyzed. The interfacial turnover rate constant for scooting kinetics, ki, for the various phospholipids were from less than 0.1 to 1 per min. Intervesicle exchange of the bound enzyme is faster in vesicles of phospholipids with larger polar substituents, and it is promoted in the presence of anions like chloride, sulfate and thiocyanate. These factors lower the residence time of the enzyme on the bilayer and therefore effectively decrease the rate of hydrolysis. The apparent Km for the enzyme in the interface of anionic phospholipids in the presence of salts is in the 40 to 100 microM range which is 3- to 7-times larger than the dissociation constants for the bound enzyme measured by fluorescence enhancement of Trp-3. The quantum yield of the bound enzyme in vesicles of the various lipids is found to be up to 4-fold different. It is suggested that this difference is due to the E* + S to E*S equilibrium, where E*S has higher fluorescence intensity. The role of calcium in generating the enzyme binding site at the anionic interface, the role of anion anchoring site on the enzyme, and the relationship between the catalytic efficiency and the fluorescence quantum yields are discussed.     

Kinetic Concepts for Measuring Microbial Rate Constants: Effects of Nutrients on Rate Constants

We investigated the effect of preincubation of environmental waters amended with inorganic nutrients (nitrogen, phosphorus, and traces of iron and magnesium) on the kinetics of the microbial transformation of phenol, propanil, propyl ester of (2,4-dichlorophenoxy)acetic acid, methyl parathion, Ronnel, and methoxychlor in pond and river waters. No effect on the second-order rate constants for these compounds was observed, although there was an increase in the bacterial populations and the pseudo-first-order rate constants. The use of nutrient-amended waters could be a useful tool for estimating second-order rate constants for an expanded number of compounds. This technique would provide a larger data base for predicting the behavior of xenobiotic compounds in the environment by using currently available mathematical models.     

Three distinct H-2Ks molecules differing at the carboxy terminus are expressed on a tumor from SJL/J mice

A dimethylbenzanthracene-induced leukemia of H-2s origin expressed at least two class I molecules on the cell surface that were precipitated by anti-H-2.19, an alloantiserum prepared against the private H-2Ks specificity. Mapping studies in recombinant inbred strains along with comparisons of tryptic peptide maps and N-terminal sequences indicated that the proteins were virtually identical and probably encoded by the same class I gene. When cells were labeled in the presence of tunicamycin, the proteins precipitated by anti-H-2.19 were further resolved into three distinct peptides. Experiments were performed to determine which of these various proteins were phosphorylated and which were recognized by an anti-synthetic peptide serum directed against the ultimate C-terminus of H-2K class I molecules. The results indicate that a single class I gene from the H-2Ks region may encode three class I molecules that differ only at the C-terminus due to alternative splicing of pre-mRNA.     

Acyl-CoA reductase and acyl-CoA: fatty alcohol acyl transferase in the microsomal preparation from the bovine meibomian gland

Biosynthesis of wax esters, one of the two major products of the meibomian gland, was found to be catalyzed mainly by the microsomes of the bovine meibomian gland. The microsomal preparation catalyzed hexadecanoyl-CoA reduction to hexadecanol without any accumulation of the aldehyde intermediate. Maximal rates of reduction occurred at pH 6.5 and required both NADH and NADPH; the latter alone gave considerable rates whereas NADH alone was ineffective. Exogenous hexadecanal reduction catalyzed by the same preparation showed a preference for NADH. The hexadecanoyl-CoA saturation pattern was slightly sigmoidal and concentrations higher than 125 microM inhibited reduction. The fatty alcohol generated from hexadecanoyl-CoA was found as free alcohol and as wax esters. Esterification of hexadecanol to wax esters catalyzed by the meibomian gland microsomal preparation required exogenous acyl-CoA or ATP and CoA and was not affected by exogenous cholesterol. Maximal rates of esterification were observed at neutral pH. Hexadecanoyl-CoA concentrations higher than 125 microM inhibited esterification. Hexadecanol showed a typical substrate saturation pattern with an apparent Km of 125 microM. Radio gas-liquid chromatography showed that, in the presence of exogenous hexadecanoyl-CoA, hexadecanol gave hexadecyl hexadecanoate whereas in the presence of ATP and CoA both C16 and C18 endogenous acids were used to esterify the alcohol. Consistent with the composition of the meibomian gland secretion, exogenous acyl-CoA longer than C14 and shorter than C20 gave maximal rates of esterification of hexadecanol.     

Relationship between mitogenic activity of influenza viruses and the receptor-binding specificity of their hemagglutinin molecules

The relationship between the mitogenic activity of influenza type A viruses for murine B lymphocytes and the receptor-binding specificity of their hemagglutinin was examined. Receptor-binding specificity was determined by the ability of the virus to agglutinate erythrocytes that had been sialidase treated and then enzymatically resialylated to contain sialyloligosaccharides with defined sequences. Distinct differences in receptor-binding specificity were observed between strongly and weakly mitogenic viruses of the H3 subtype, with strong mitogenic activity correlating with the ability of the virus to recognize the sequence N-glycolylneuraminic acid alpha 2,6 galactose (NeuGc alpha 2,6Gal). Viruses isolated early in the evolution of the H3 subtype (from 1968 to 1971) are relatively weak mitogens and recognize the sequence N-acetylneuraminic acid alpha 2,6 galactose (NeuAc alpha 2,6Gal) but not NeuGc alpha 2,6Gal. H3 viruses isolated since 1972 are strongly mitogenic, and these viruses recognize both NeuGc alpha 2,6Gal and NeuAc alpha 2,6Gal. The amino acid substitution of Tyr for Thr at residue 155 of HA1 may be critical to this change in receptor-binding specificity and mitogenic activity of the later H3 viruses. Horse serum-resistant variants of H3 viruses, which bind preferentially to the sequence NeuAc alpha 2,3Gal, are poorly mitogenic. Differences were also observed between the receptor-binding specificity of the strongly mitogenic H3 viruses and viruses of the H2 and H6 subtypes, the mitogenic activity of which is limited to strains of mice that express the class II major histocompatibility complex glycoprotein I-E. The results indicate that the receptor-binding specificity of the hemagglutinin plays a critical role in determining the mitogenic activity of influenza viruses.