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Biomedical subjects

J R Allen

Publications and source records attributed to J R Allen.

At least 19 recordsLinked to original sources

Coding assignments of the genome of adult diarrhea rotavirus.

Adult diarrhea rotavirus (ADRV) has caused epidemics of diarrhea in China since 1982 and remains the only group B rotavirus associated with widespread disease in humans. We recently characterized the proteins of ADRV and have now proceeded to identify the gene segments encoding each protein. Viral RNA transcripts were synthesized in vitro with the endogenous viral RNA polymerase and separated by electrophoresis in agarose. The individual transcripts were translated in a cell-free system using nuclease-treated rabbit reticulocyte lysates. The translation products were compared with polypeptides found in purified virus and were characterized by SDS-PAGE, immunoprecipitation, and Western blot analysis using antisera to double- and single-shelled virions, virus cores, and monoclonal antibodies. Furthermore, individual RNA transcripts were hybridized to total dsRNA to determine their genomic origin. Based on this analysis, the core polypeptides VP1, VP2 and VP3 are encoded by segments 1, 2, and 3, respectively. The main polypeptides in the inner capsid, VP6, and the outer capsid, VP4 and VP7, are encoded by segments 6, 4, and 8 respectively. Segments 5, 7, and 9 code for 60, 45, and 30 kDa nonstructural polypeptides. Two other nonstructural polypeptides (24 and 25 kDa) are derived from gene segment 11. Gene segment 10 codes for a 26 kDa polypeptide that is precipitated with serum to ADRV and may be a structural protein VP9. With this exception, gene coding assignments of ADRV are comparable to those of the group A rotaviruses. Our results have clear implications for further work in cloning, sequencing, and expression genes of ADRV and can provide direction towards understanding the origin and the evolution of this virus.

Animals

Outbreaks of summer rotavirus linked to laboratory practices. The National Rotavirus Surveillance System.

In temperate regions rotavirus diarrhea is a disease of the cooler months of the year, but little is known about its patterns in the summer. We report on the first year of national surveillance of rotavirus, during which we actively investigated patterns of summer activity. We obtained data on rotavirus testing from 85 laboratories in 48 states, conducted a survey of their testing practices and retested for confirmation positive specimens from laboratories reporting high rates of positivity during the summer. During 1989 participating laboratories reported 4011 specimens tested for rotavirus during July and August, of which 436 (11%) were said to be positive. Most laboratories reported low rates of positivity during these months (median percent positive, 3), but five had very high rates of summer positivity (> 30%). These five laboratories were geographically separated, and neighboring laboratories showed little rotavirus activity. Positive specimens submitted by four of these centers with high rates of summer rotavirus could not be confirmed. A survey of laboratory methods found one commercial assay (TestPack) and two laboratory practices (failure to use controls and involvement of more than six technicians in the testing process) to be associated with high rates of summer positivity. Moderate rates of positivity (11 to 30%) were fond frequently in the southwest during July and August; reference testing of specimens from these laboratories confirmed positivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical Laboratory Techniques

Identification of cognate genes among heterologous strains of group B rotavirus.

The genetic relatedness of group B rotavirus (GBR) strains has previously been documented by hybridization with probes derived from whole genomic sequences, but the relationship of individual genes of heterologous GBR strains has not been evaluated. Definition of cognate GBR genes would facilitate investigation of the determinants of group specificity, serotype identity, and neutralization epitopes. Therefore, we investigated the genetic relatedness of three GBR strains by means of Northern (RNA) blot hybridization with isotopically labeled probes prepared from each of the 11 genes of the IDIR strain of GBR. Under low-stringency conditions, hybridization between each of the IDIR gene probes and genomic RNA from the ADRV strain of GBR was observed. Genomic RNA obtained from a bovine strain of GBR hybridized with 9 of the 11 IDIR gene probes. In most cases, cognate genes of each of the GBR strains appeared to migrate to similar positions following polyacrylamide gel electrophoresis. However, the electropherotype positions of GBR genes 5, 6, and 7 were different for each of the three GBR strains. Identification of these genomic segments among GBR strains should prove helpful in future evaluations of GBR structure and function.

Animals

Responses of rats exposed to polychlorinated biphenyls for fifty-two weeks. II. Compositional and enzymatic changes in the liver.

Male Sprague-Dawley rats consumed a diet to which 100 ppm of various polychlorinated biphenyl (PCB) mixtures (Aroclors 1248, 1254, and 1262) were added for one year. Rats were hepatectomized at 13, 26, and 52 weeks during feeding and at 13 weeks following the discontinuation of the PCB diets. The liver homogenates of these rats had an increase in protein and RNA on a DNA basis and an increase in lipid and a decrease in DNA on the liver weight basis. The hepatic microsomes from these livers also had an increase in protein and cytochrome P-450. The RNA/microsomal protein levels were decreased, and no marked alterations were recorded for the phospholipids and cholesterol on a microsomal protein basis. Increased enzymatic activity was recorded for N-demethylase and nitroreductase. However, the specific activity of glucose-6-phosphatase was decreased throughout the treatment period.

Animals

The effects of salt concentration and H-1 depletion on the digestion of calf thymus chromatin by micrococcal nuclease.

We have removed histone H1 specifically from calf thymus nuclei by low pH treatment, and studied the digestion of such nuclei in comparison with undepleted nuclei. By a number of criteria the nuclei do not appear damaged. The DNA repeat-length in nuclear chromatin is found to be the same (192 +/- 4 bp) in the presence or absence of H1. These experiments demonstrate that the core histone complex of H2A, H2B, H3, and H4 can itself protect DNA sequences as long as 168 bp from nuclease. Our interpretation is that this represents an important structural element in chromatin, carrying two full turns of superhelical DNA. Depending on conditions of digestion this 168 bp fragment may be metastable and is normally rapidly converted by exonucleolytic trimming to the well-known "core-particle" containing 145 bp. Larger stable DNA fragments observed indigestion of H-1 depleted nuclei appear to arise from oligomers assembled from 168 bp cores in close contact exhibiting trimming of 0-20 bp at the ends. Electrophorograms of undepleted nuclear digests reveal oligomer bands in several size classes, each corresponding to one or more combinations of 168 bp particles, H1-protected spacers of about 20 bp length, and particles with ends trimmed to varying degrees.

Animals

Pathophysiology of dehydromonocrotaline-induced pulmonary fibrosis in the beagle.

The purpose of this study was to characterize the sequential hemodynamic alterations and pulmonary vascular lesions produced by a single pulmonary artery injection of the vasotoxic pyrrolic alkaloid dehydromonocrotaline in the young beagle. Normotensive pulmonary pressure was replaced by hypertension 21 days after injection. By 28 days, the pulmonary pressure and total pulmonary vascualr resistance of the experimental animals were significantly greater than the controls (p less than 0.01). Right ventricular work increased from a baseline mean of 0.58 to 1.40 kg . m/min. Morphological and morphometrical analyses revealed alveolar edema, increased numbers of alveolar macrophages, cellular hyperplasia in the alveolar septa, and a progressive interstitial fibrosis. The precise mechansims by which dehydromonocrotaline injection initiates and promotes pulmonary hypertension and pulmonary fibrosis still needs clarification; however, our data indicate that the fraction of air space is reduced relative to the fraction of tissue space, and this change occurs with concurrent fibrosis in the alveolar septa and an increased pulmonary arterial pressure although hypoxia was not clinically detectable.

Animals

Absence of fatty livers in rhesus monkeys fed orotic acid.

Pairs of rhesus monkeys were fed for 10 wk a basal diet containing 1% orotic acid or 10% nonfat milk powders. Amounts of total lipids in the liver and hepatic morphology were normal after 10 wk indicating that orotic acid in the diet did not induce fatty livers in rhesus monkeys.

Animals

Fatal nosocomial Legionnaires' disease: clinical and epidemiologic characteristics.

The occurrence, clinical, and epidemiologic characteristics of fatal nosocomial Legionnaires' disease were ascertained by examining a sample of autopsy specimens submitted by hospitals participating in the National Nosocomial Infections Study. Sections of lung tissue from 263 patients were submitted by 40 hospitals in 24 states. Legionnaires' disease was identified in 10 patients (3.8%) from six hospitals in five states. The clinical and epidemiologic characteristics of sporadic fatal nosocomial Legionnaires' disease were similar to those from cases that occurred during epidemics of Legionnaires' diseases. A case-control study showed that cases of Legionnaires' disease were associated with an increased occurrence of Legionnaires' disease during the period June through November, temperatures greater than 39.2 degrees C, and administration of corticosteroids before onset of pneumonia. Legionnaires' disease can occur as a fatal nosocomial pneumonia and has clinical and epidemiologic features that may be helpful in differentiating it from nosocomial pneumonia of other causes.

Adolescent

DNA single strand breaks caused by 2,2',5,5'-tetrachlorobiphenyl and its metabolites.

Polychlorinated biphenyls (PCBs) are widespread environmental pollutants which have been shown to be carcinogenic. One possible mechanism for the apparent carcinogenicity of PCBs is their metabolism to phenolic metabolites through an arene oxide intermediate, with the arene oxide that is formed being the ultimate mutagenic or carcinogenic species. The ability of a tetrachlorobiphenyl and its phenolic and arene oxide metabolites to cause single strand breaks in DNA was therefore examined. 2,2',5,5'-TCB-3,4-epoxide was more potent in causing DNA single strand breaks than either 2,2',5,5'-TCB or a mixture of 3-hydroxy and 4-hydroxy-2,2',5,5'-TCB. These results support the idea that PCB epoxide may be involved in PCB carcinogenesis.

DNA, Single-Stranded

Langerhans cells trap tick salivary gland antigens in tick-resistant guinea pigs.

In the infested skin of tick-resistant guinea pigs, indirect immunofluorescence techniques have revealed that antigens from the ticks' salivary glands are associated with discrete dendritic cells in the epidermis. Evidence is presented to support the suggestion that these antigen-retaining cells are Langerhans cells.

Adenosine Triphosphatases