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Biomedical subjects

G Sherman

Publications and source records attributed to G Sherman.

At least 19 recordsLinked to original sources

Effects of embryo transfer and cortical ectopias upon the behavior of BXSB-Yaa and BXSB-Yaa + mice.

The BXSB-Yaa and BXSB-Yaa + inbred strains of mice differ primarily with respect to the Y chromosome, although there is evidence that they differ on several autosomal genes as well. Each strain has ectopic collections of neurons in neocortical layer I (ectopias), with a higher occurrence in males (58%) than females (42%). Conventionally reared mice from these strains were compared to mice that were transferred, as 8-cell embryos, into the uteri of non-autoimmune recipients, who gave birth to and reared the offspring. The transfer procedure did not change the incidence of ectopias in either sex. There were, however, major differences in behavior. Compared to conventionally reared controls, embryo transfer mice had greater behavioral asymmetry, poorer performance in a black-white discrimination, poorer Morris maze learning, better Lashley maze learning, and better performance in a two-way shuttlebox. Within the transfer groups, females differed as much as males, confirming our prior findings and supporting our thesis that the two strains differ on several autosomal genes in addition to the Y chromosome. These findings show that the intra-uterine environment can powerfully and selectively affect later behavior. When ectopic and non-ectopic mice were compared, BXSB-Yaa mice with neocortical ectopias were better able to learn the Morris spatial maze than non-ectopic controls; this was true whether the mice were conventionally reared or embryo transferred. In contrast, BXSB-Yaa + ectopic mice did not differ from their controls if conventionally reared, but were much worse than controls if embryo transferred.

Animals

The covalently closed duplex form of the hepadnavirus genome exists in situ as a heterogeneous population of viral minichromosomes.

Replication of hepadnaviruses requires a persistent population of covalently closed circular (CCC) DNA molecules in the nucleus of the infected cell. It is widely accepted that the vital role of this molecule is to be the sole DNA template for the synthesis by RNA polymerase II of all viral transcripts throughout the infection process. Since the transcriptional activity of eukaryotic nuclear DNA is considered to be determined in part by its specific organization as chromatin, the nucleoprotein disposition of the hepadnavirus CCC DNA was investigated. These studies were undertaken on the duck hepatitis B virus (DHBV) CCC DNA present in the liver cell nuclei of DHBV-infected ducks. The organization and protein associations of the DHBV CCC DNA in situ were inferred from sedimentation, micrococcal nuclease digestion, and DNA superhelicity analyses. These three lines of investigation demonstrate that the DHBV CCC DNA is stably associated with proteins in the nuclei of infected liver cells. Moreover, they provide compelling evidence that the viral nucleoprotein complex is indeed a minichromosome composed of classical nucleosomes but in arrays that are atypical for chromatin. When the DHBV chromatin is digested with micrococcal nuclease, a ladder of viral DNA fragments that exhibits a 150-bp repeat is produced. This profile for the viral chromatin is obtained from the same nuclei in which the duck chromatin shows the standard 200-bp ladder. The superhelicity of the DHBV CCC DNA ranges from 0 to 20 negative supertwists per molecule, with all possible 21 topoisomers present in each DNA preparation. The 21 topoisomers of DHBV CCC DNA are inferred to derive from an identically diverse array of viral minichromosomes. In the DHBV minichromosomes composed of 20 nucleosomes, 96.7% of the viral DNA is calculated to be compacted into these chromatin subunits spaced on average by 5 bp of linker DNA; other minichromosomes contain fewer nucleosomes and proportionately more linker DNA. Two major subpopulations of DHBV minichromosomes are detected with comparable prevalence. The two groups correspond to minichromosomes which contain essentially a full or half complement of nucleosomes. The functional significance of this minichromosome diversity is unknown but is suggestive of transcriptional regulation of the viral DNA template.

Animals

Case-control study of congenital anomalies in children of cancer patients.

OBJECTIVES: To determine whether the offspring of cancer survivors are at an increased risk of congenital anomalies and whether cancer therapy before conception is associated with such an increase. DESIGN: Case-control study using computerised record linkage. SETTING: Ontario, Canada. SUBJECTS: Parents of children born during April 1979 to December 1986 who had a congenital anomaly diagnosed within the first year of life (45,200 mothers and 41,158 fathers) and a matched sample of parents whose children did not have a congenital anomaly (45,200 mothers and 41,158 fathers). MAIN OUTCOME MEASURES: Cancer diagnosed in either parent before conception and radiotherapy to the pelvis or abdomen or chemotherapy with an alkylating agent. RESULTS: Among the mothers, 54 cases and 52 controls were identified as having had cancer diagnosed in Ontario (relative risk = 1.04, 95% confidence interval 0.7 to 1.5) and among the fathers, 61 cases and 65 controls were identified (0.9, 0.7 to 1.4). No significant associations were found between congenital anomalies in the offspring and any type of cancer treatment in either the mothers or the fathers. CONCLUSIONS: The risk of congenital anomalies among liveborn offspring whose parents have had cancer or been treated for cancer is not higher than that in the general population.

Adult

The UL8 subunit of the herpes simplex virus helicase-primase complex is required for efficient primer utilization.

The herpes simplex virus (HSV) type 1 helicase-primase is a three-protein complex, consisting of a 1:1:1 association of UL5, UL8, and UL52 gene products (J.J. Crute, T. Tsurumi, L. Zhu, S. K. Weller, P. D. Olivo, M. D. Challberg, E. S. Mocarski, and I. R. Lehman, Proc. Natl. Acad. Sci. USA 86:2186-2189, 1989). We have purified this complex, as well as a subcomplex consisting of UL5 and UL52 proteins, from insect cells infected with baculovirus recombinants expressing the appropriate gene products. In confirmation of previous reports, we find that whereas UL5 alone has greatly reduced DNA-dependent ATPase activity, the UL5/UL52 subcomplex retains the activities characteristic of the heterotrimer: DNA-dependent ATPase activity, DNA helicase activity, and the ability to prime DNA synthesis on a poly(dT) template. We also found that the primers made by the subcomplex are equal in length to those synthesized by the UL5/UL8/UL52 complex. In an effort to uncover a role for UL8 in HSV DNA replication, we have developed a model system for lagging-strand synthesis in which the primase activity of the helicase-primase complex is coupled to the activity of the HSV DNA polymerase on ICP8-coated single-stranded M13 DNA. Using this assay, we found that the UL8 subunit of the helicase-primase is critical for the efficient utilization of primers; in the absence of UL8, we detected essentially no elongation of primers despite the fact that the rate of primer synthesis on the same template is undiminished. Reconstitution of lagging-strand synthesis in the presence of UL5/UL52 was achieved by the addition of partially purified UL8. Essentially identical results were obtained when Escherichia coli DNA polymerase I was substituted for the HSV polymerase/UL42 complex. On the basis of these findings, we propose that UL8 acts to increase the efficiency of primer utilization by stabilizing the association between nascent oligoribonucleotide primers and template DNA.

Adenosine Triphosphatases

Body awareness and medical care utilization among older adults in an HMO.

This study investigated the association between the disposition of body awareness and medical care utilization among older adult members of a health maintenance organization (HMO). Results indicated that higher levels of body awareness are associated significantly with longitudinal increases in the volume of patient-initiated illness visits to the HMO, and with a greater likelihood of patient-initiated contact with the hospital emergency room, controlling for prior utilization, self-reported health status, and other factors. In contrast, body awareness was not associated significantly with longitudinal changes in physician-initiated follow-up visits, internal referrals, external referrals, or hospital inpatient days. Other findings indicated that higher levels of patient-initiated utilization were associated with greater physician-initiated utilization, controlling for prior utilization. These results illustrate how patient-initiated utilization may influence subsequent physician-initiated utilization.

Aged

Running limb varus.

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Foot Deformities

Chemicals, birth defects and stillbirths in New Brunswick: associations with agricultural activity.

We describe a series of investigations that were conducted in support of the Task Force on Chemicals in the Environment and Human Reproductive Problems in New Brunswick. Geographic and temporal analyses and case-control studies, with the use of vital statistics, hospital records, the Canadian Congenital Anomalies Surveillance System and chemical databases, revealed no association between pesticides used in forestry and reproductive problems. Evidence of an association between the potential exposure to agricultural chemicals and three major anomalies combined as well as spina bifida without hydrocephalus was found. More plausible was an association between stillbirths and such exposure during the second trimester of pregnancy. This finding, along with the cyclic patterns of stillbirth in the agricultural Saint John River basin and the somewhat higher stillbirth rates in New Brunswick than in adjacent provinces or in Canada as a whole, suggests that further attention should focus on possible associations between agricultural activity and stillbirths.

Abnormalities, Drug-Induced

Characterization of intranuclear capsids made by ts morphogenic mutants of HSV-1.

We have characterized capsids made by seven temperature-sensitive (ts) mutants of HSV-1 previously shown to be defective in viral DNA processing and packaging at the nonpermissive temperature (NPT). The empty capsids isolated from mutant-infected cells at the NPT were devoid of DNA, cosedimented in sucrose with wt B capsids, and contained the same structural proteins found in wt B capsids (W. Gibson and B. Roizman (1972). J. Virol. 10, 1044-1052). The presence of VP22a in empty capsids suggests that the processing of this protein from higher-molecular-weight precursors and its association with capsids is required, but not sufficient, for DNA encapsidation. Mutants made no detectable A capsids at the NPT, but did so at the permissive temperature (PT), suggesting that A particles are generated during or subsequent to, rather than prior to, encapsidation. In temperature-shift experiments, it was demonstrated that capsids of one of the mutants, F18, made at the NPT did not participate in DNA encapsidation when cells were subsequently shifted to the PT. Only those capsids made after temperature shift to the PT acquired viral DNA, implying that the ts mutation in F18 may lie in a gene coding for a structural protein, or in a protein involved in the processing of viral DNA.

Animals

DNA processing in temperature-sensitive morphogenic mutants of HSV-1.

The anatomy of DNA synthesized by five HSV-1 mutants previously shown to accumulate predominantly empty capsids at the nonpermissive temperature (NPT) was analyzed with Bg/II restriction digestion. At the NPT, all five generated DNA lacking termini, indicating that in the absence of packaging, viral DNA is not processed to unit length. One mutant, F18, was able to process DNA made at the NPT to unit length molecules during a 6-hr period after shift to the permissive temperature. The appearance of unit length molecules correlated with the appearance of staphylococcal nuclease-resistant F18 DNA.

Capsid

Selectivity of cholestyramine treatment on hepatic mixed function oxidase activity.

The possibility that oral cholestyramine treatment might indirectly increase hepatic drug oxidations was investigated in rats using pentobarbital, antipyrine, zoxazolamine, and aminopyrine as probes of the hepatic mixed function oxidase system and in humans using amobarbital and antipyrine as probes. Cholestyramine pretreatment of rats for 5 days (87.5 mg/kg twice daily by stomach tube) shortened pentobarbital sleep times, decreased antipyrine-induced hypothermia, but did not influence either zoxazolamine paralysis times or the in vitro N-demethylation of aminopyrine. Neither pentobarbital nor antipyrine pharmacokinetics in rats were affected by the cholestyramine pretreatment. Similarly, in two-way crossover studies with human subjects, a 5-day oral cholestyramine pretreatment (4 g 3 times daily) had no demonstrable effect on the pharmacokinetics of single doses of amobarbital (200 mg i.v.) or antipyrine (500 mg per os). After cholestyramine pretreatment, a trend toward diminished CNS depression produced by amobarbital was observed, but the effect was not statistically significant. The results suggest that hepatic mixed function oxidases in rats for which aminopyrine, antipyrine, pentobarbital, and zoxazolamine are substrates and that hepatic mixed function oxidases in humans for which amobarbital and antipyrine are substrates are not significantly affected by cholestyramine pretreatment.

Adult

Discriminative response control by naloxone in morphine pretreated rats.

In an operant procedure using a lever press response 12 male, hooded rats were trained to discriminate 1.25 mg/kg naloxone from a saline injection. On certain days, according to a counterbalanced training schedule, naloxone was administered 8 h after 40 mg/kg morphine and 10 min prior to a trail in which food was available on an FR10 schedule from one of two levers in a dual lever operant chamber. On other days saline was administered 10 min prior to a trial in which food was made available by pressing the other lever. After criterion performance for acquisition of the discrimination had been reached, tests were carried out to determine its nature. Discrimination of naloxone was dose-dependent and was significantly diminished when naloxone was administered 36 h after morphine. Partial generalization of cyclazocine with naloxone was observed. Spontaneous withdrawal from morphine, tested during trials preceeded by an injection of saline instead of naloxone at various time intervals after morphine, did not generalize with the naloxone discriminative stimulus.

Animals

Thick filament size changes in contraction of human muscles.

Measurements done on electron micrographs shows that in myofibres with sarcomeres contracted to below 2.1 micron, proportional shortening of the A bands occurs. In muscles from patients with idiopathic scoliosis very short A bands are especially prominent.

Cytoskeleton

Primary malignant lymphoma of the small intestine in Israel. Changing incidence with time.

One hundred and eighty-one cases of primary small intestinal lymphoma were diagnosed in residents of Israel--145 Jews and 36 Arabs--during a 16-year period, 1960--75. The male:female ratio was 1.8:1. Incidence rates were higher in children, the middle-aged and the elderly than in teenagers or young adults. Crude and age-adjusted incidence rates were higher in Arabs than in Jews. The pattern among the subgroups of the Jewish population varied with age. In young adults of both sexes the rates were much higher among Jews born in North Africa or Asia than among those born in Europe or Israel. In elderly males, however, the rates were almost twice as high in European-born Jews than among those born in North Africa or Asia. The mean annual incidence of primary small intestinal lymphoma fell from 4.8 per million during 1960--67 to 3.6 per million during 1968--75. This was due to a marked fall in the rates in children and young adults, whereas the rates in those aged 40 years or more rose with time. The pattern of the change with time suggests that environmental conditions are an important factor in the causation of primary intestinal lymphoma in young adults.

Adolescent