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Biomedical subjects

T E Staley

Publications and source records attributed to T E Staley.

At least 19 recordsLinked to original sources

Receptor mechanisms of the neonatal intestine and their relationship to immunoglobulin absorption and disease.

Immunoglobulin absorption by the calf has been the subject of considerable research. Despite these efforts little is known about the cytological events that occur at the level of the intestinal epithelial cell. These events have been studied extensively and characterized in the laboratory rodent; however, there have been few attempts to make corollaries between the two species. All neonatal animals display certain similarities in their intestinal morphology that may be correlated, with immunoglobulin absorption. Selectivity in absorption appears to be variable among neonatal animal species; however, all demonstrate some selectivity. Selectivity in absorption implies that receptors are a necessary component in the transport of immunoglobulins. Selectivity further requires binding of immunoglobulins to an endocytic vesicle membrane to ensure transport through the cell, circumvention of intracellular digestion, and release at the basolateral cell membrane. A decrease of immunoglobulin absorption may be accomplished in a variety of ways such as competition between intestinal microbes and immunoglobulins for a common receptor on the intestinal epithelial cell. An additional consideration is aberrant synthesis or recycling of the cell membrane receptor, as induced by metabolic decelerators such as cortisol. Failure to recycle immunoglobulin receptors also would decrease efficiency of absorption.

Aging↗

Two methods for administering colostrum to newborn calves.

Comparison was of immunoglobulin G concentrations in blood serum of calves to which colostrum was administered by nipple bottle or by esophageal tube. Pooled batches of colostrum were given by the designated method soon after birth and at 12 and 24 h thereafter. Blood samples to measure immunoglobulin G concentrations were taken before initial feeding and at 4-h intervals thereafter through 32 h. The rate of increase of immunoglobulin G concentration following feeding was essentially the same for calves given colostrum by the two methods, and the concentration attained by 20 h after initial feeding was similar for the two groups and adequate for calf survival.

Animal Feed↗

Recovery of intestinal membrane binding sites for K88 E. coli from pig mucosal organ cultures.

Putative receptors for K88 + E. coli from piglet intestinal epithelium were released into the organ culture medium and were demonstrated by direct binding with K88 + E. coli through the utilization of an in vitro binding procedure or by immunoprecipitation with K88 antigen. Incorporation of 14C-glucosamine by newborn to day old and 3-week to 6-week old piglet jejunal and ileal mucosa, in organ culture, occurred throughout the 24 hr culture period. Uptake in both age groups and both areas of the intestine was similar with a somewhat greater incorporation by the older age group. Secretion of 14C-glucosamine-labeled components into the culture medium was demonstrated by gel filtration of the concentrated medium. Some large molecular weight components eluted in the void volume in excess of 2 X 10(6) daltons. A second peak of activity was spread from approximately 690K to 25K daltons. All eluted fractions demonstrated binding to K88 + E. coli. Antibodies to purified brush borders from susceptible pigs produced prominent precipitation bands following double diffusion with concentrated organ culture media which confirmed that the organ culture media contained labeled proteins of brush border origin. Immunoprecipitation of the intestinal mucosal organ culture media with K88 + pili and pilus antisera, followed by electrophoresis with SDS and reduced conditions, demonstrated a subunit of approximately 35K daltons.

Animals↗

Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct.

Cultures of lactobacilli identified as Lactobacillus acidophilus from the intestinal contents of young calves varied in their ability to grow in broth containing .3% oxgall compared with control broth. Frozen concentrated cultures were prepared from a strain exhibiting low tolerance to bile and from a strain exhibiting high tolerance to bile. Plate counts were comparable from the concentrated cultures before and after frozen storage on lactobacilli MRS agar with and without .15% oxgall. In a feeding trial involving newborn dairy calves supplementation of the diet with the more bile resistant strain of Lactobacillus acidophilus caused greater increases of numbers of facultative lactobabilli in the upper small intestines than did the strain exhibiting lower resistance to bile. It was not possible to determine whether the lactobacilli would prevent intestinal infections in the calves challenged with enteropathogenic Escherichia coli. This portion of the study failed as the challenge with Escherichia coli did not cause infections even in control animals.

Animal Feed↗

Soluble pig intestinal cell membrane components with affinities for E. coli K88+ antigen.

Pig intestinal brush borders (BB) were radiolabeled by iodination using the lactoperoxidase-hydrogen peroxide procedure. The BB were then detergent solubilized, centrifuged to remove particulate material, and chromatographed on Sepharose CL-4B. The fractions were incubated with K88+ E. coli using an in vitro binding assay. Binding of the iodinated membranes to K88+ E. coli occurred throughout a wide range of molecular weight components, in excess of 690K daltons to near 25K daltons. The system utilizing intact K88+ E. coli and solubilized BB was shown to be saturable. Prior contact of K88+ E. coli with nonradiolabeled membranes or specific antibodies to K88+ pili inhibited binding of the radiolabeled BB. Simple sugars were tested for their ability to block binding of the labeled BB; partial inhibition occurred with galactose (17.9%), galactosamine (32%), glucose (10.6%), and N-acetylglucosamine (32%). Calcium enhanced binding with as little as 10 microM. A 10 x increase in binding occurred with 500 microM calcium. Affinity chromatography using K88+ pili coupled on agarose beads avidly bound the labeled BB. The receptor membranes were eluted with high molar concentrations of salt, however considerable degradation occurred. Despite low yields from the affinity system, receptor membranes with higher binding activities were recovered. Protein: glycoprotein ratios were 1:4. Elution with SDS and electrophoresis on 12.5% polyacrylamide gels in the presence of a reducing agent produced two major subunits 35--32K and 23K daltons. These components were recovered from the gels and retained their binding activity. This information suggests that the intestinal receptor responsible for binding of K88+ E. coli is a glycoprotein, that in the native state exists in multimeric forms.

Animals↗

Metastatic basal cell carcinoma of the scrotum.

We report a rare case of basal cell carcinoma of the scrotum presenting with distant metastases. Rationale and treatment with bacillus Calmette-Guerin immunotherapy and subsequent combination chemotherapy are detailed.

Antineoplastic Agents↗

[14C]glucosamine incorporation into intestinal glycoproteins by calf intestinal epithelium in organ culture.

The uptake and incorporation of [14C]glucosamine into secreted intestinal glycoproteins was examined in the calf intestinal mucosa and isolated epithelial cells, using an organ culture system. Incorporation varied with the area of the intestine, ie duodenum greater than jejunum greater than ileum. Secreted glycoproteins were examined after 3 and 24 hours in organ culture. Initially, a glycoprotein approximately 68,000 molecular weight (mol wt) was secreted by all areas of the intestine. This was followed by the appearance of a larger glycoprotein 10(6) to 10(7) mol wt. Enzymatic cleavage of jejunal brush borders released a glycocalyx glycoprotein similar in size to the secreted large mol wt glycoprotein. These similarities in mol wt are only suggestive evidence that the 2 large glycoproteins are the same. This procedure allows for the study of the maturation and development of a poorly understood component of the intestinal epithelial cell which has a wide range of functional considerations.

Animals↗

Reduction of reactivity of Escherichia coli enterotoxins by intestinal mucosal components.

Incubation studies involving rabbit and piglet small intestinal mucosal components and Escherichia coli and Vibrio cholerae enterotoxins were conducted at 37 and 4 degrees C. Mucosal homogenate cytosol from rabbits did not significantly alter the reactivities of either cholera enterotoxin (CT) or E. coli labile enterotoxin (LT). However, mucosal homogenate cytosol from piglets was capable of neutralizing LT, though it had no effect on E. coli stable enterotoxin. LT became bound to piglet and rabbit microvillous membranes at 4 degrees C in the presence of a protective protein. In rabbits, the binding of LT was not dependent upon an intact glycocalyx or free unbound CT-receptors, although some binding was apparently associated with the glycocalyx and CT-receptors. These results indicated the presence of two different LT-receptors in microvillous membranes one being associated with the membrane proper and the other with the glycocalyx. Stable enterotoxin did not bind to in vitro preparations of piglet mucosal components, which is evidence for a different mechanism for inducing intestinal secretion.

Animals↗

Influence of colostrum on transepithelial movement of Escherichia coli 055.

The effect of colostrum on transepithelial migration of live Escherichia coli 055:B5:H7 in the neonatal calf intestine was determined by microbiological and microscopic investigations. Colostrum-deprived calves (2 to 6 h old) were given E. coli suspended in saline, E. coli suspended in colostrum, or E. coli in saline 1 h after colostrum. Twenty-four hours after exposure, tissues were collected for examination. Escherichia coli were numerous in mesenteric lymph nodes of calves given this organism in saline. Fewer were recovered from nodes of calves that received the bacteria in colostrum. Escherichia coli were not recovered from mesenteric lymph nodes of calves given colostrum before dosage with the organism. Electron microscopic studies of small intestines from calves that received E. coli in saline revealed bacterial attachment sites surrounded by exfoliation of microvilli. Bacteria appeared to enter epithelial cells by invagination of apical plasma membrane or dilation of apical tubules. Intracellular E. coli were enclosed in a surrounding membrane. The organisms were not observed adhering to or penetrating intestinal epithelium of calves that received E. coli in colostrum or 1 h after colostrum.

Animals↗

Absorption of horseradish peroxidase by neonatal pig intestinal epithelium: effect of Escherichia coli (055B5) on absorption.

The pathway of macromolecular transport through the neonatal pig small intestinal epithelium was examined, utilizing the cytochemical marker horseradish peroxidase (HRP). The marker was found adsorbed to the apical microvillous of the enterocytes, within apical tubules, and cytoplasmic vacuoles. Extracellular absorbed marker assumed a spherical appearance within the subepithelial spaces but was dispersed within the capillary lumens. Uptake of HRP into the enterocyte occurred in 48-hour old neonatal pigs, but transport into the circulation was not observed. Adherence of Escherichia coli to the surface of the ileal enterocyte did not totally inhibit HRP uptake. The E coli adhered to the surface of the enterocyte or within intercellular vacuoles appeared to be static in as much as they were not involved in the transepithelial migration of envacuolated HRP.

Animals↗

Polynucleotide sequence relationships among Japanese and American strains of Vibrio parahaemolyticus.

Polynucleotide sequence relationships between two reference Vibrio parahaemolyticus strains isolated from Japanese and American gastroenteritis patients were investigated by use of (32)P-DNA/DNA reassociation in free solution. In addition, these strains were similarly compared with 22 other strains of estuarine and marine vibrios, including 11 strains previously identified as V. parahaemolyticus (2 Japanese, 1 of unknown location, and 8 American strains obtained from diverse geographical locations and sources in North America), 3 strains of V. alginolyticus, and 8 of Vibrio spp. Deoxyribonucleic acid (DNA) from the Japanese and American gastroenteritis isolates showed high relative levels of intraspecific duplex formation (92 to 93%) when reassociated, reciprocally, at 60 C. Heterologous DNA duplexes exhibited thermal elution midpoint [Tm(e)] values comparable to those obtained from homologous duplexes (88.0) when thermally eluted from hydroxyapatite, thus indicating high base-pair complementarity. Other V. parahaemolyticus strains showed DNA homologies of 85% or greater, with correspondingly high Tm(e) values (86.0 to 88.0) for the heteroduplexes formed. DNA of two of three V. alginolyticus strains (ATCC 17749 and 166-70) was 55 to 60% homologous to reference V. parahaemolyticus DNA preparations; Vibrio sp. strain 5144 (originally classified as V. parahaemolyticus biotype 2 and subsequently as V. alginolyticus strain 5144) showed only 24 to 26% DNA homology to the same reference DNA. These data provide evidence that Vibrio sp. strain 5144 is genetically distinct from the other V. alginolyticus strains used in this study. Three bioluminescent strains thought to be closely related to V. parahaemolyticus demonstrated only 24 to 31% DNA homology to the reference V. parahaemolyticus DNA. These data firmly establish the existence in some Atlantic and Gulf Coast estuaries of organisms genetically very similar to V. parahaemolyticus, the causative agent of "shirasu" food poisoning in Japan.

Base Sequence↗

E. coli diarrhoea.

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Adenylyl Cyclases↗