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Biomedical subjects

D York

Publications and source records attributed to D York.

At least 19 recordsLinked to original sources

Prenatal detection and evaluation of an extralobar pulmonary sequestration in the posterior mediastinum.

Extralobar pulmonary sequestration (EPS) is a rare developmental anomaly with aberrant nonfunctioning parenchymal tissue, associated with an increased risk of perinatal morbidity and, rarely, mortality owing to possible neonatal respiratory distress. In most cases supernumerary lobes are detected as isolated intra- or extrapleural lesions with independent systemic arterial blood supply. We report an atypical case of prenatal detection and perinatal outcome of a mediastinal EPS.

Adult↗

Floor pen study to evaluate the serological response of broiler breeders after vaccination with ts-11 strain Mycoplasma gallisepticum vaccine.

To determine the Mycoplasma gallisepticum (MG) rapid serum plate agglutination (RSPA) test response of broiler breeders after ts-11 strain vaccination, 55 Cobb pullets derived from a nonvaccinated, MG-negative, commercial, broiler breeder grandparent flock were monitored from 8 to 20 wk of age (over a 12-wk trial period). To evaluate the effect of lateral spread of the ts-11 vaccine strain on RSPA test results from commingled and adjacently penned birds, treatment groups included (A) birds vaccinated with ts-11strain MG at 8 wk of age, (B) commingled nonvaccinates in the same pen as the vaccinated birds, (C) nonvaccinates in a second pen separated from the first pen by a distance of 2 m, and (D) birds vaccinated with ts-11 strain MG at 8 wk of age and kept in a separate room. Rapid serum plate agglutination tests were performed once a week for 6 wk and then every 2 wk for 6 more wk, postvaccination. A polymerase chain reaction (PCR) assay specific fbr ts-11 strain MG was used to confirm vaccination, and a second PCR specific for non-ts-11 strain MG was used to confirm the absence of field infection. Seroconversion was first detected by the RSPA test 2 wk postvaccination and attained maximum positive rates of 58% at 12 wk postvaccination in treatment A and 60% at 8 wk postvaccination in treatment D. Seroconversion rates in nonvaccinated, commingled pullets was 10% at 5 wk and 30% at 12 wk after the vaccination of pen mates. The ts-11-specific PCR detected the vaccine strain in 80%-100% of the vaccinated birds 2 wk after vaccination. One of 15 nonvaccinated birds penned 2 m from vaccinated birds yielded ts-11 by PCR assay 12 wk after vaccination, which indicates that the spread of ts-11 over short distances may be possible in situations in which there is a common caretaker. PCR on tracheal swabs taken 12 wk postvaccination detected ts-l1 in 50% and 60% of the vaccinated birds in treatments A and D, respectively; in 30% of the commingled nonvaccinates; and in 6.6% of the separately penned nonvaccinates. In contrast, choanal swabs collected from vaccinated birds at 12 wk were 21% and 40% PCR positive for ts-11 strain MG, while those from nonvaccinates were negative. All samples were PCR negative for field strain MG. The pattern of seroconversion as measured by RSPA test in small groups of broiler breeders was different from that previously reported for leghorns. Lateral spread of the ts-11 strain to commingled pen mates occurred rapidly, causing RSPA seroconversion patterns that mimicked those of the vaccinated pen mates.

Agglutination Tests↗

Variation in breastmilk HIV-1 viral load in left and right breasts during the first 3 months of lactation.

The mechanism and risk factors associated with mother-to-child transmission of HIV-1 through breastfeeding remain unclear; breastmilk viral load may be an important determinant of transmission. Analysis of breastmilk cell-free viral load in samples taken from each breast at 1, 6 and 14 weeks postpartum showed that HIV-1 is shed intermittently and load may differ considerably between breasts of an individual woman at any given time. Breastmilk HIV-1 load was undetectable in approximately one-third of samples.

Breast↗

Spectrum of myelopathies in HIV seropositive South African patients.

The authors determined the cause of myelopathies in 33 HIV seropositive individuals in KwaZulu/Natal, South Africa. The main associations were with human T-cell lymphotrophic virus-I, tuberculosis, herpes zoster, and syphilis. A novel association with probable bilharziasis was noted. Only one case of vacuolar myelopathy was identified. Opportunistic infections will probably persist until routine antiretroviral therapy becomes widely available in South Africa.

Adult↗

Estimation and management of genetic diversity in small populations of plains zebra (Equus quagga) in KwaZulu-Natal, South Africa.

Plains zebras (Equus quagga antiquorum) occur in few large, but many small, isolated populations in KwaZulu-Natal. Problems identified in small populations include reduced striping patterns on hind quarters, smaller size, elevated mortality rates and high number of still-births. Inbreeding may be implicated. Population viability analysis (PVA) was conducted with a computer model (VORTEX), and DNA and allozyme analyses were conducted to test the findings of the model. Using standard methods, DNA (PCR-RAPD) and allozyme diversity was assessed in blood samples from 72 plains zebra from four KwaZulu-Natal Nature Conservation Services (KZN-NCS) protected areas: Umfolozi Game Reserve (UGR), Albert Falls (AFNR), Vernon Crookes (VCNR) and Harold Johnson (HJNR) Nature Reserves. Populations of the latter three, small-sized (9-110 individuals) populations were seeded from the same source population (UGR: current population of 2000) during the past 25 years. Information from PCR-RAPD and allozyme analyses were compared with each other as well as to that predicted by population genetic modelling (using VORTEX). Allozyme heterozygosities were consistently high in all populations (12.1-12.9%), with no observable losses associated with reduced population size. On the other hand, percentage loss of polymorphism (20-39%) calculated from the PCR-RAPD study appeared to be positively correlated with the loss of heterozygosity predicted by population viability analysis (PVA), and negatively correlated with population size. On the basis of the above results, a policy of translocation was advocated for small, intensely managed populations of zebras, whereby a harem should be translocated every five years for a population size of nine (HJNR), while for a population size of 110 (VCNR) translocations should take place every 15 years if heterozygosity is to be maintained at more than 90% within each population over 100 years.

Journal Article↗

Peripheral administration of human corticotropin-releasing hormone: a novel method to increase energy expenditure and fat oxidation in man.

CRH increases energy expenditure and decreases food intake in experimental animals. We proposed the hypothesis that peripheral administration of CRH might increase energy expenditure in human subjects. Four men and four women (age, 19-39 yr) were randomized to a double blind, cross-over trial to test the effect of human CRH (hCRH), ovine CRH (oCRH), and placebo on resting energy expenditure measured by indirect calorimetry. CRH was administered by primed continuous infusion at progressively increasing doses of 0.5, 1.0, and 2.0 microg/kg.h at 2-h intervals. hCRH increased resting energy expenditure by 13.9% at the end of the infusion. Respiratory quotient fell from 0.828 to 0.768 during the hCRH infusion compared with a fall from 0.836 to 0.807 during placebo infusion (P < 0.05). Fat oxidation increased by 55% compared with placebo at the highest dose of hCRH. Heart rate increased during hCRH to 10.7 bpm higher than placebo (P < 0.05). oCRH did not increase heart rate. oCRH also had no significant effect on respiratory quotient, and only a small effect on energy expenditure. During hCRH infusion, venous plasma epinephrine, norepinephrine, glycerol, and nonesterified fatty acid levels were not significantly different from those during placebo treatment. Peripheral CRH administration offers a novel strategy to increase energy expenditure.

Adult↗

N-Acetylaspartate and DARPP-32 levels decrease in the corpus striatum of Huntington's disease mice.

Huntington's disease (HD) is an autosomal dominant condition involving progressive neurodegeneration, primarily the corpus striatum and cerebral cortex. We have used in vivo magnetic resonance spectroscopy (MRS) to assess specific neuronal markers in transgenic mice (R6/1 line) expressing exon I of the human huntingtin gene with an expanded CAG repeat. Levels of N-acetylaspartate (NAA), an indicator of healthy neuronal function, were significantly reduced (26%) in the corpus striatum of HD mice relative to wild-type littermates at 5 months of age. However, levels of cholines and creatine-phosphocreatine were not altered in the HD mice. Expression of dopamine- and cAMP-regulated phosphoprotein, 32 kDa (DARPP-32), was assessed by immunohistochemistry in the striatum of HD mice and found to be downregulated by 5 months and, even more dramatically, at 11 months of age. In contrast, expression of calbindin was not significantly decreased in HD mice. Our results suggest that the observed decreases in DARPP-32 and NAA may contribute to aberrant receptor signalling and neuronal dysfunction in HD.

Animals↗

Characterization of a Tn5 pre-cleavage synaptic complex.

Protein catalyzed DNA rearrangements typically require assembly of complex nucleoprotein structures. In transposition and integration reactions, these structures, termed synaptic complexes, are mandatory for catalysis. We characterize the Tn5 pre-cleavage synaptic complex, the simplest transposition complex described to date. We identified this complex by gel retardation assay using short, linear fragments and have shown that it contains a dimer of transposase, two DNA molecules, and is competent for DNA cleavage in the presence of Mg(2+). We also used hydroxyl radical footprinting and interference techniques to delineate the protein-DNA contacts made in the Tn5 synaptic and monomer complexes. All positions (except position 1) of the end sequence are contacted by transposase in the synaptic complex. We have determined that positions 2-5 of the end sequence are specifically required for synaptic complex formation as they are not required for monomer complex formation. In addition, in the synaptic complex, there is a strong, local distortion centered around position 1 which likely facilitates cleavage.

Bacteria↗

Cell-free virus in breast milk of HIV-1-seropositive women.

To examine the prevalence, quantification, and factors that influence HIV in the cell-free compartment of breast milk, we performed reverse transcription polymerase chain reaction (RT-PCR) on samples obtained from HIV-1-infected study subjects. Virus was detected in 86 of 136 samples (63.2%) from 79 study subjects. HIV RNA quantity ranged from undetectable to 227,586 copies/ml. Prevalence and mean viral load were not affected by postnatal ages or maternal vitamin A supplementation. Among study subjects with multiple samples, breast milk viral load did not change at different postnatal ages. Breast milk viral load correlated positively with plasma viral load (r = 0.47; p =.005) and negatively with maternal CD4 count at entry to the study (r = -0.26; p =.02). Mothers of HIV-infected children had a higher proportion of detectable HIV RNA in their breast milk than mothers of uninfected children (p =.03) and higher mean log10 HIV RNA quantities (p =.04). In a multivariate logistic regression model, log10 HIV RNA quantity in breast milk was significantly associated with the risk of mother-child transmission (odds ratio [OR], 2.82; 95% confidence interval [CI], 1.22-6.51). Thus, prevention and treatment of opportunistic infections and of mastitis and early weaning may be important elements of a public health policy that is relevant to women in developing countries with HIV infection. Where available, antiretrovirals may also have an impact on opportunistic infections and mastitis.

Adult↗

GB virus C/hepatitis G virus (GBV-C/HGV): still looking for a disease.

GB Virus C and Hepatitis G Virus (GBV-C/HGV) are positive, single-stranded flaviviruses. GBV-C and HGV are independent isolates of the same virus. Transmission via the blood-borne route is the commonest mode, although vertical and sexual transmission is well documented. GBV-C/HGV is distributed globally; its prevalence in the general population is 10 fold higher in African countries than in non-African countries. High prevalences of GBV-C/HGV have been found in subjects with frequent parenteral exposure and in groups at high risk of exposure to blood and blood products. The clinical significance of human infection with GBV-C/HGV is currently unclear. The virus can establish both acute and chronic infection and appears to be sensitive to interferon. Only some 12-15% of chronic Non-A, B, C hepatitis cases are infected with GBV-C/HGV. A direct association with liver pathology is still lacking and it is not yet clear as to whether GBV-C/HGV is indeed a hepatotropic virus. Current evidence suggests that the spectrum of association of GBV-C/HGV infection with extrahepatic diseases ranges from haematalogical diseases, aplastic anaemia, human immunodeficiency virus (HIV)-positive idiopathic thrombocytopenia and thalassemia, through to common variable immune deficiency and cryoglobunemia.

Carcinoma, Hepatocellular↗

Molecular cloning and functional analysis of three type D endogenous retroviruses of sheep reveal a different cell tropism from that of the highly related exogenous jaagsiekte sheep retrovirus.

Integrated into the sheep genome are 15 to 20 copies of type D endogenous loci that are highly related to two exogenous oncogenic viruses, jaagsiekte sheep retrovirus (JSRV) and enzootic nasal tumor virus (ENTV). The exogenous viruses cause infectious neoplasms of the respiratory tract in small ruminants. In this study, we molecularly cloned three intact type D endogenous retroviruses of sheep (enJS56A1, enJS5F16, and enJS59A1; collectively called enJRSVs) and analyzed their genomic structures, their phylogenies with respect to their exogenous counterparts, their capacity to form viral particles, and the expression specificities of their long terminal repeats (LTRs). In addition, the pattern of expression of enJSRVs in vivo was studied by in situ hybridization. All of the three enJSRV proviruses had open reading frames for at least one of the structural genes. In particular, enJS56A1 had open reading frames for all structural genes, but it could not assemble viral particles when highly expressed in human 293T cells. We localized the defect for viral assembly in the first two-thirds of the gag gene by making a series of chimeras between enJS56A1 and the exogenous infectious molecular clone JSRV(21). Phylogenetic analysis distinguished five ovine type D retroviruses: enJSRV groups A and B, ENTV, and two exogenous JSRV groups (African versus United Kingdom/North America isolates). Transient transfection assays indicated that the LTRs of the three enJSRVs were not preferentially active in differentiated lung epithelial cells. This suggests that the pulmonary tropic JSRV developed from a type D retrovirus that did not have lung specificity. Consistent with this, in situ hybridization of a panel of normal ovine tissues revealed high expression of enJSRV mRNA in the luminal epithelium and glandular epithelium of the uterus; lower expression was localized in the lamina propria of the gut and in the bronchiolar epithelium of the lungs.

Amino Acid Sequence↗

How obesity develops: insights from the new biology.

Molecular and genetic studies of animal models have identified numerous genes that may cause or contribute to the development of obesity. They have also provided significant insight into the peripheral and central pathways that control energy intake and expenditure. Genetic studies of families and populations have generated useful information on genes and mutations associated with or linked to obesity, body fat distribution, and other relevant phenotypes. This information, combined with knowledge of the chromosomal location of genes identified from animal studies, has made it possible to identify specific mutations that contribute to the development of obesity in humans.

Adolescent↗

Comparison of a maedi-visna virus CA-TM fusion protein ELISA with other assays for detecting sheep infected with North American ovine lentivirus strains.

A maedi-visna virus CA-TM fusion protein ELISA (MVV ELISA) was evaluated for the detection of antibody in sheep infected with North American ovine lentivirus (OvLV). The results of the MVV ELISA were compared with other assays for OvLV antibody and with viral infection in an intensively studied group of 38 sheep with a high prevalence of OvLV infection and disease. The sensitivity, specificity, and concordance of assays for OvLV antibody (MVV ELISA, indirect ELISA, Western blot, and AGID), virus (virus isolation, PCR, antigen ELISA), and OvLV-induced disease in each animal were compared with OvLV infection status as defined by a positive result in two or more of the assays. Five sheep met the criteria for absence of OvLV infection. The sensitivity of the MVV ELISA in detecting OvLV infected sheep was 64%, whereas the sensitivity of the other three tests for antibody ranged from 85 to 94%. All the antibody assays were 100% specific in this group of animals. Of the assays for virus, the PCR test had the highest sensitivity and the best concordance with OvLV infection, but it also had the lowest specificity of any of the virus or antibody assays. Among the antibody tests, the concordance of the MVV ELISA compared most favorably with the AGID test for detecting OvLV-infected sheep. Analysis of serum samples from 28 lambs experimentally-infected with one of three North American strains of OvLV suggested that there were no significant strain differences detectable by antibody assay. Twenty virus-inoculated lambs were positive by both the MVV ELISA and the AGID test, five lambs were MVV ELISA negative and AGID test positive, and three lambs were MVV ELISA positive and AGID test negative. No pre-inoculation samples were positive by either assay. In a longitudinal study involving seven lambs, antibodies to OvLV were detected by AGID 3-5 weeks post-inoculation, but were not detected by MVV ELISA until 5-10 weeks post-inoculation. Among 128 naturally and experimentally-infected sheep that were seropositive in the AGID test, the overall sensitivity of the MVV ELISA was higher in the naturally infected sheep (84%) than in the experimentally infected sheep (69%). The data indicated that the MVV ELISA represents a less sensitive, but specific alternative for the detection of OvLV antibodies.

Animals↗

Sequence of the env gene of some KwaZulu-Natal, South African strains of HTLV type I.

Phylogenetic analysis of HTLV-I suggests three main subtypes, namely, cosmopolitan, Central African, and Australo-Melanesian. HTLV-I is endemic in KwaZulu-Natal, South Africa. However, sequence data on the local strains are limited to the LTR region. The env gene of the local strain was amplified and sequenced from the peripheral blood of five seropositive individuals. Four had HTLV-I-associated myelopathy and one had infective eczema. The sequence analysis of the env gene showed a greater then 99% homology of the local strains. They were closely related to the North American strains (99.3%), followed by the Japanese strains (98.3-98.9%). Phylogenetic studies linked the local strains to the cosmopolitan subtype. This study provides new sequence data on the env gene of the local HTLV-I strain.

Amino Acid Sequence↗

Simple and efficient generation in vitro of nested deletions and inversions: Tn5 intramolecular transposition.

We have exploited the intramolecular transposition preference of the Tn 5 in vitro transposition system to test its effectiveness as a tool for generation of nested families of deletions and inversions. A synthetic transposon was constructed containing an ori, an ampicillin resistance (Ampr) gene, a multi-cloning site (MCS) and two hyperactive end sequences. The donor DNA that adjoins the transposon contains a kanamycin resistance (Kanr) gene. Any Amprreplicating plasmid that has undergone a transposition event (Kans) will be targeted primarily to any insert in the MCS. Two different size targets were tested in the in vitro system. Synthetic transposon plasmids containing either target were incubated in the presence of purified transposase (Tnp) protein and transformed. Transposition frequencies (Ampr/Kans) for both targets were found to be 30-50%, of which >95% occur within the target sequence, in an apparently random manner. By a conservative estimate 10(5) or more deletions/inversions within a given segment of DNA can be expected from a single one-step 20 microl transposition reaction. These nested deletions can be used for structure-function analysis of proteins and for sequence analysis. The inversions provide nested sequencing templates of the opposite strand from the deletions.

Chromosome Inversion↗

Single-grain 40Ar-39Ar ages of glauconies: implications for the geologic time scale and global sea level variations

The mineral series glaucony supplies 40% of the absolute-age database for the geologic time scale of the last 250 million years. However, glauconies have long been suspected of giving young potassium-argon ages on bulk samples. Laser-probe argon-argon dating shows that glaucony populations comprise grains with a wide range of ages, suggesting a period of genesis several times longer ( approximately 5 million years) than previously thought. An estimate of the age of their enclosing sediments (and therefore of time scale boundaries) is given by the oldest nonrelict grains in the glaucony populations, whereas the formation times of the younger grains appear to be modulated by global sea level.

Journal Article↗