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Biomedical subjects

D James

Publications and source records attributed to D James.

At least 73 records · Page 4Linked to original sources
PubMed1999

A simple and reliable protocol for the detection of apple stem grooving virus by RT-PCR and in a multiplex PCR assay.

Primers were identified which amplify specifically a 499 bp fragment in the coat protein coding region of apple stem grooving virus (ASGV) genome. These primers were used in various RT-polymerase chain reaction (PCR) analyses for the detection of ASGV in Chenopodium quinoa, Nicotiana occidentalis, and in species of Malus and Pyrus. Isolates of ASGV in Malus and Pyrus from locations in Canada, China, Israel, Japan, Nepal, Pakistan, South Africa, and the U.S.A. were reliably detected in leaf and bark (budwood) tissue. Storage of the tissues at -80 degrees C for more than 4 months did not affect the reliability of detection by immunocapture (IC) RT-PCR. Triton-X was not necessary for the detection of ASGV by IC/RT-PCR, and it was also possible to combine the antibody incubation and virus sap incubation into a single step without any obvious loss in sensitivity. A Tube Capture (TC) RT-PCR procedure was developed that eliminated the need for antibody binding of the virus. Phosphate buffered saline with 2% PVP was identified as the most effective sample-grinding buffer for the detection of ASGV by IC/RT-PCR and TC/RT-PCR. TC/RT-PCR facilitated the simultaneous detection (multiplex PCR) of ASGV and cherry mottle leaf virus.

Base Sequence↗
PubMed1999

Single primer pair designs that facilitate simultaneous detection and differentiation of peach mosaic virus and cherry mottle leaf virus.

Peach mosaic virus (PMV) and cherry mottle leaf virus (CMLV) are viruses which are related serologically and share common Prunus hosts, but cause distinct diseases. An RT-PCR procedure using a single oligonucleotide primer pair that allows simultaneous detection and differentiation of the two viruses was developed. A sense primer with 100% complementarity to PMV and 83% complementarity to the corresponding site of the CMLV genome was combined with either of two antisense primers (one of PMV origin and the other of CMLV origin) with 3' end complementarity at variable sites. This allowed the differential amplification of PMV and CMLV specific fragments, 419 and 705 bp, respectively. When oligo (dT) was used to generate the cDNA template, differential amplification was not observed, only amplification of the homologous virus associated with the antisense primer. This indicates polyadenylation of both viruses. Incorporation of the antisense primer into cDNA at the reverse transcription step was shown to be essential for this approach. The PMV primer pair reliably detected all isolates of PMV tested by RT-PCR analysis, both in peach leaf and budwood tissue.

Base Sequence↗
PubMed1999

Computerised analysis of unstimulated and stimulated behaviour in fetuses with intrauterine growth restriction.

Unstimulated (passive) and stimulated behaviour, in the form of fetal heart rate (FHR) and activity (FA)) was studied in 32 normal fetuses and 14 fetuses with intrauterine growth restriction (IUGR). FHR and FA were recorded using a single 1.5 MHz ultrasound transducer and analysed by computer. A 5 s vibroacoustic stimulus (VAS) (electronic artificial larynx) was used for the stimulation studies. The IUGR fetuses had significantly different patterns of both unstimulated and stimulated behaviour compared to normally grown fetuses. When unstimulated they had lower FA rates than the normally grown fetuses but this was only statistically significant at 28-31 weeks. They also spent a significantly lower proportion of time exhibiting high FHR variation at 28-31 weeks. Following VAS the IUGR fetuses had lower FA responses at all gestations and lower FHR responses from 32 weeks, though only the differences in FA response at 28-31 weeks were statistically significant. Of the 14 fetuses with IUGR, all but one exhibited passive behaviour (ERR and/or FA) that was outside the 10-90th range, for normally grown fetuses, whilst 6 of the 14 had responses to VAS that were within the normal range. We do not feel that computerised assessment of stimulated behaviour in preterm IUGR fetuses confers any advantage over passive observations.

Acoustic Stimulation↗
PubMed1999

Expression of c-fos, c-jun, and c-jun N-terminal kinase (JNK) in a developmental model of induced apoptotic death in neurons of the substantia nigra.

The transcription factors c-fos and c-jun have been proposed to play a role in the initiation of programmed cell death in neurons. We have shown that programmed cell death, with the morphology of apoptosis, occurs in dopamine neurons of the substantia nigra (SN) during normal postnatal development and that this death event can be induced by early striatal target injury. We have investigated the relationship between c-fos and c-jun protein expression and induced death in neurons of the SN. Although c-fos is induced, it is unlikely to play a role in cell death, because its expression is not well correlated with apoptotic death either temporally or at a cellular level. Expression of c-jun, however, is both temporally and regionally correlated with induction of death, and, at a cellular level, it colocalizes with apoptotic morphology. The increased expression of c-jun is likely to be functionally significant, because it is associated with increased c-jun N-terminal kinase (JNK) and phosphorylated c-jun expression. JNK expression also colocalizes with apoptotic morphology. We conclude that c-jun is likely to play a role in the initiation of apoptotic cell death in these neurons.

Animals↗
PubMed1999

Efficient human IFN-gamma expression in the mammary gland of transgenic mice.

Two hybrid genes (BLG-HuIFN-gamma2 and BLG-HuIFN-gamma3) were constructed on the basis of sheep beta-lactoglobulin (BLG) and human interferon-gamma (HuIFN-gamma) gene sequences. They were used to direct HuIFN-gamma synthesis in the mammary gland of transgenic mice. HuIFN-gamma was efficiently produced in the mammary gland of transgenic mice. BLG-HuIFN-gamma2 transgenic females expressed HuIFN-gamma in the milk at concentrations up to 570 mg/ml, and BLG-HuIFN-gamma3 transgenic females expressed up to 350 mg/ml. All females carrying the BLG-HuIFN-gamma3 gene expressed HuIFN-gamma in their milk. No significant changes were observed in the HuIFN-gamma expression level during the lactation period. Using RT-PCR analysis, ectopic expression for both hybrid genes was found in transgenic mice. Despite ectopic expression of HuIFN-gamma in transgenic mice, their development and pregnancy were normal. The heritability of the HuIFN-gamma expression level in milk was demonstrated up to the F2 generation. This work demonstrates that hybrid genes have the potential to develop in transgenic domestic animals producing HuIFN-gamma in milk.

Animals↗
PubMed1999

Larsen scoring of digitized X-ray images.

OBJECTIVE: To determine how Larsen scores from digitized X-rays compare to those from film originals. METHODS: A hundred sets of radiographs of patients recruited with early rheumatoid arthritis (RA) were assessed using the Larsen scoring system. Digitized copies of these sets were then viewed on a computer screen and scored according to Larsen in a random order. The quality of the digitized image was also recorded. For each set of X-rays, the signed difference between the score from film and the score from the digitized images was calculated. RESULTS: A total of 95% of the digitized X-ray sets were scored successfully; 5% were not scored due to the images being unreadable. The mean difference between the two sets of scores was -1.2 (95% CI [-2.06, -0.37]). There was no trend in the difference with respect to the mean of the two scores (P>0.1). CONCLUSION: The Larsen scoring of digitized X-ray images has been validated.

Analog-Digital Conversion↗
PubMed1999

Population forecasting with endogenous migration: an application to trans-Tasman migration.

"This article focuses on forecasting migration between Australia and New Zealand (trans-Tasman migration), which is largely visa-free and therefore resembles internal migration. Net trans-Tasman migration is a major component of New Zealand population change and is embedded in this article in a Bayesian or unrestricted vector autoregression (VAR) model, which includes foreign and domestic economic variables. When time series of net migration are available, this approach provides a useful input into forecasting population growth in the short run in the absence of major policy changes. This conclusion applies equally to interregional migration and to unrestricted international migration between economically integrated nations."

Australia↗
PubMed1998

Fetal medicine.

Explore the source record for details and available documents.

Amnion↗
PubMed1998

A GR-motif functions in nuclear accumulation of the large FGF-2 isoforms and interferes with mitogenic signalling.

Nuclear translocation has been documented for members of the fibroblast growth factor (FGF) family in addition to their roles as extra-cellular signalling molecules. Fusing different parts of the chicken FGF-2 open reading frame to pyruvate kinase shows that direct nuclear accumulation is mediated by the amino-termini of the two leucine initiated FGF-2 isoforms (Leu-isoforms; 21.5 and 20.0 kDa). An evolutionarily conserved glycine-arginine (GR)-motif is present in the 21.5 kDa Leu-isoform and a shorter GR-repeat in the 20.0 kDa Leu-isoform, whereas no such repeats are present in the 18.5 kDa FGF-2 isoform (Met-isoform). Expression in NIH3T3 fibroblasts shows that the 21.5 kDa Leu-isoform is predominantly nuclear, whereas the Met-isoform is predominantly cytoplasmic. Most importantly, insertion of the GR-motif into the Met-isoform results in a protein with characteristics similar to the Leu-isoforms, as shown by nuclear accumulation of the chimeric MGR-protein. Furthermore, only NIH3T3 fibroblasts expressing the Met-isoform proliferate under serum starvation conditions, whereas cells expressing either the MGR- or Leu-isoforms stay growth arrested. These studies show that the GR-signal mediates nuclear translocation of endogenous Leu-isoforms and blocks their mitogenic activity.

3T3 Cells↗
PubMed1998

Lymph node genesis is induced by signaling through the lymphotoxin beta receptor.

We investigated lymphotoxin (LT) and TNF function in lymph node genesis and cellular organization by manipulating LTbeta-R and TNF-R signaling. Lymph nodes developed in LTalpha-/- mice treated in utero with agonist anti-LTbeta-R monoclonal antibody. Thus, LTbeta-R signaling mediates lymph node genesis. Surprisingly, mucosal lymph nodes that can develop independently of LTalphabeta/LTbeta-R interaction were generated. Normal mice treated in utero with LTbeta-R-Ig and TNF-R55-Ig or anti-TNF lacked all lymph nodes, indicating that TNF signaling contributes to lymph node genesis. Lymph nodes generated in LTalpha-/- mice had disrupted cellular organization. Therefore, LTbeta-R signaling during gestation is not sufficient to establish normal cellular microarchitecture. We conclude that LT and TNF play critical roles in the genesis and cellular organization of lymph nodes.

Animals↗
PubMed1998

A comparison of two alcohol craving questionnaires.

AIM: To compare two multi-dimensional questionnaires to measure cravings and urges for alcohol: the Alcohol Craving Questionnaire (ACQ: Singleton, Henningfield and Tiffany, 1994a) and the Desires for Alcohol Questionnaire (DAQ: Clark et al., 1996). DESIGN, SETTING, AND PARTICIPANTS: Both questionnaires were administered, in a counterbalanced order, to a total of 380 recreational drinkers. In a further study, a shortened version of the DAQ was administered to a sample of 131 drinkers attending AA or a treatment centre. Exploratory factor analyses were carried out on the data and relationships between questionnaire score and other variables were assessed. FINDINGS: In recreational drinkers both instruments appeared to have a three-factor structure. The DAQ appeared superior to the ACQ in a number of respects: it produced more reliable factors; its structure accounted for a higher proportion of the variance; the factor inter-correlations were somewhat lower; in a combined analysis of both instruments most of the factors retained came from the DAQ; and the DAQ discriminated better between bing and non-binge drinkers and excessive and moderate drinkers. A similar factor structure was found for the DAQ in the alcoholic subjects with addition of a factor of "controllability". CONCLUSIONS: The results support a multifactorial account of alcohol craving, and indicate that the DAQ has some advantages over the ACQ as a research tool.

Adolescent↗
PubMed1998

The glycosylation heterogeneity of recombinant human IFN-gamma.

The cloning of the cDNA for human interferon-gamma (IFN-gamma) has resulted in its expression in Escherichia coli, baculovirus-infected insect cells, Chinese hamster ovary (CHO) cells, and the mammary gland of transgenic mice. Large quantities of highly purified recombinant IFN-gamma have been generated, aided by the use of highly specific neutralizing monoclonal antibodies, with a view to its production as a human therapeutic protein. The primary source of structural heterogeneity for IFN-gamma during its production in mammalian expression systems is glycosylation, which can profoundly affect the three-dimensional structure of a glycoprotein and its biological function. A number of analytical approaches have been developed recently to allow a detailed analysis of the carbohydrate structures associated with IFN-gamma, the principal advances being in the areas of capillary electrophoresis and mass spectrometry. The implementation of these high-resolution analytical tools to determine the glycosylation profile of IFN-gamma makes it one of the best characterized recombinant glycoproteins. Recombinant human IFN-gamma acts as a model secretory glycoprotein, typifying the intrinsic glycosylation processing events associated with production of a potential therapeutic glycoprotein.

Animals↗