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N Wood

Publications and source records attributed to N Wood.

At least 55 records · Page 3Linked to original sources
PubMed1999

Rapid determination of apolipoprotein E genotype using a heteroduplex generator.

The apoE gene exhibits two common polymorphisms that have been associated with both coronary artery disease and Alzheimer's disease. The polymorphisms create the three allelic isoforms E2, E3, and E4 which are encoded by Cys;-Cys, Cys;-Arg, and Arg;-Arg at amino acid positions 112 and 158, respectively. Numerous methods have been described to identify these three apoE alleles although there are disadvantages and ambiguities associated with all of them. Here we describe a method by which the two common apoE polymorphisms can be identified simultaneously. The method involves PCR of the region containing the two polymorphic sites, followed by hybridization of this PCR product to a synthetic molecule called a universal heteroduplex generator (UHG). The UHG is used to induce heteroduplex formation which is visualized on a non-denaturing mini-gel using ethidium bromide staining. This technique which can also identify other rare mutations in the amplified region of DNA under investigation, is an unequivocal method of genotyping and is simpler and faster than many methods, including using restriction enzyme digestion.

Apolipoproteins E↗
PubMed1998

Expression of alpha2-macroglobulin receptor-associated protein in normal human epidermal melanocytes and human melanoma cell lines.

Alpha2-Macroglobulin receptor/low-density lipoprotein receptor-related protein is a multifunctional cell surface receptor known to bind and internalize a large number of ligands. alpha2-Macroglobulin receptor-associated protein acts as an intracellular "chaperone" for this receptor, and it has been shown to inhibit binding of all its known ligands. In this paper, we characterize the expression of the receptor-associated protein in both normal human epidermal melanocytes and in six different human melanoma cell lines, by the use of flow cytometry and Western blotting analysis. We show that all the melanoma cell lines and the normal melanocytes express the receptor-associated protein at similar levels, with most located intracellularly. No receptor-associated protein was detected at the cell surface in the melanocytes or in three of the cell lines. However, in two of the melanoma cell lines, large amounts of receptor-associated protein were found on the cell surface, these having the largest amounts of it reported to date; in a further melanoma cell line, there was a small amount at the cell surface. We have also shown that the melanocytes and all the melanoma cell lines express the receptor itself at a wide range of levels, the highest levels of both the cell surface receptor and the cell surface receptor-associated protein being found in one particular melanoma cell line. By growing the cell lines under controlled conditions, we have demonstrated that, although the total cellular content of the receptor is markedly increased at high cell culture density, this treatment has no effect on the level of expression of the receptor-associated protein.

Cell Line↗
PubMed1998

Clinical pharmacology of moclobemide during chronic administration of high doses to healthy subjects.

The objectives of this study were to assess the tolerability, safety, pharmacodynamics and pharmacokinetics of high-dose moclobemide in healthy subjects. Two sequential groups of six male and six female subjects (eight on active treatment, four on placebo) received for 8 days moclobemide 450 mg b.i.d. and 600 mg b.i.d., respectively. Intravenous tyramine pressor tests were conducted at baseline, at the beginning of treatment and at steady state. Oral tyramine pressor tests with 50, 100 and 150 mg tyramine were conducted under steady-state conditions. Pharmacokinetic parameters of moclobemide and two of its metabolites in plasma and urine were determined after the first and last dose of moclobemide. The incidence and intensity of adverse events was dose-dependent. The most frequently reported adverse events were insomnia, headache, dizziness and dry mouth. The i.v. tyramine pressor sensitivity during both moclobemide dosing regimens was enhanced 3 to 4-fold. Intake of tyramine 50 mg did not result in systolic blood pressure increases greater than 30 mmHg. With regard to blood pressure increases, tyramine 100 mg is still compatible with moclobemide 450 mg b.i.d. but not with 600 mg b.i.d. The clearance of moclobemide decreased by about 60% on multiple dosing, but no differences were found between both dosing regimens. The urinary excretion of the N-oxide metabolite doubled during multiple dosing. In conclusion, the maximum tolerated dose of moclobemide in healthy subjects is 600 mg b.i.d. provided the tyramine content in a meal is not higher than 50 mg.

Adult↗
PubMed1998

Fine mapping of de novo CMT1A and HNPP rearrangements within CMT1A-REPs evidences two distinct sex-dependent mechanisms and candidate sequences involved in recombination.

The molecular mechanism resulting in the duplication or deletion of a 1.5 Mb region of 17p11.2-p12, associated, respectively, with Charcot-Marie-Tooth type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP), has been proposed to be an unequal crossing-over during meiosis between the two chromosome 17 homologues generated by misalignment of the proximal and distal CMT1A-REP repeats, two homologous sequences flanking the 1.5 Mb CMT1A/HNPP monomer unit. In a recent study of a large series of de novo cases of CMT1A and HNPP, two distinct sex-dependent mechanisms were identified. Rearrangements of paternal origin, essentially duplications, were indeed generated by unequal meiotic crossing-over between the two chromosome 17 homologues, but duplications and deletions of maternal origin resulted from an intrachromosomal process, either unequal sister chromatid exchange or, in the case of deletion, excision of an intrachromatidal loop. In order to determine how these recombinations occur, 24 de novo crossover breakpoints were localized within the 1.7 kb rearrangement hot spot by comparing the sequences of the parental CMT1A-REPs with the chimeric copy in affected offspring. Nineteen out of 21 paternal crossovers were found in a 741 bp hot spot. All the breakpoints of maternal origin (n = 3), however, were located outside this interval, but in closely flanking sequences, supporting the hypothesis that two distinct sex-dependent mechanisms are involved. Several putative recombination promoting sequences in the hot spot, which are rare or absent in the surrounding 7.8 kb, were identified.

Charcot-Marie-Tooth Disease↗
PubMed1998

De novo expansion of intermediate alleles in spinocerebellar ataxia 7.

Spinocerebellar ataxia 7 (SCA7) is the eighth neurodegenerative disorder caused by a translated CAG repeat expansion. Normal SCA7 alleles carry from four to 35 CAG repeats, whereas pathological alleles carry from 37 to approximately 200. Intermediate alleles (IAs), with 28-35 repeats in the SCA7 gene are exceedingly rare in the general population and are not associated with the SCA7 phenotype, although they have been found among relatives of four SCA7 families. In two of these families, IAs bearing 35 and 28 CAG repeats gave rise, during paternal transmission, to SCA7 expansions of 57 and 47 repeats, respectively, that were confirmed by haplotype reconstructions in one case and by inference in the other. Furthermore, the four haplotypes segregating with IAs were identical to the expanded alleles in each kindred, but differed among the families, indicating multiple origins of the SCA7 mutation in these families with different geographical origins. Our results provide the first evidence of de novo SCA7 expansions from IAs that are not associated with the phenotype but can expand to the pathological range during some paternal transmissions. IAs that segregate in unaffected branches of the pedigrees might, therefore, constitute a reservoir for future de novo mutations that occur in a recurrent but random manner. This would explain the persistence of the disease in spite of the great anticipation (approximately 20 years/generation) characteristic of SCA7. So far, de novo expansions among the disorders caused by polyglutamine repeats have only been demonstrated in Huntington's disease.

Alleles↗
PubMed1998

Symptomatic unilateral pleural effusion as a presentation of ovarian hyperstimulation syndrome.

Isolated acute unilateral pleural effusion has twice been reported as the only symptom of ovarian hyperstimulation syndrome (Kingsland et al, 1989; Jewelewicz and Vande Wiele, 1975). The pathogenesis of this disorder is not fully understood and the presence of an isolated pleural effusion lends support to the role of systemic factors rather than purely the transudation of fluid from grossly enlarged ovaries in the progression of this disease. This article describes a second case of an isolated pleural effusion following in-vitro fertilization and embryo transfer.

Adult↗
PubMed1998

Response rate in patient satisfaction research: an analysis of 210 published studies.

OBJECTIVES: To examine the quality of response rate reporting and to identify methodological factors influencing response rates in published patient satisfaction studies. DESIGN: Examination and analysis of 210 studies from 200 papers published in 1994 in 141 different health journals. Papers were located in the following databases: British Nursing Index, CINAHL, EMBASE, MedLine, Popline, and PsycLIT. MAIN MEASURES: Reported and calculated response rates, collection and recruitment procedures of published studies, and type of instruments used for data collection. RESULTS: Forty-eight per cent of studies reported a response rate. The mean response rate was 72.1%. There was no association between response rate and the type of instrument used for data collection. Studies which used a face-to-face approach to either subject recruitment (mean response rate, 76.7%) or data collection (mean response rate, 76.9%) were associated with significantly higher response rates than those in which subjects were recruited by mail (mean response rate, 66.5%) or data were collected by mail (mean response rate, 67%). Response rate was not related to questionnaire length. CONCLUSION: Patient satisfaction studies generally show poor awareness of the importance of methodological issues relevant to response rate. Far more attention to this aspect is needed if findings in this field are to be accepted as valid and useful.

Australia↗
PubMed1998

Controlled trial of the subjective patient benefits of accompanied walking to the operating theatre.

A trial was carried out to determine if walking unpremedicated patients to the operating theatre would prove acceptable to the patients. One hundred surgical patients from a short-stay-ward were randomised into experimental (walked to theatre by ward nurse) and control (taken to theatre on a hospital trolley) groups. Seventy-eight patients responded to a questionnaire; a large majority indicated they would like to be given the choice of mode of conveyance and perceived this as an improvement in patient care. The results showed that a more favourable impression of walking to the theatre was given by patients who had actually experienced it, and of those given the opportunity to walk, almost all reported that it made them feel more relaxed. The findings are discussed in relation to patient choice, efficiency gains and a reduction in manual handling.

Adult↗
PubMed1997

Expression of alpha 2 macroglobulin receptor/low density lipoprotein receptor-related protein is cell culture density-dependent in human breast cancer cell line BT-20.

alpha 2Macroglobulin receptor/low density lipoprotein receptor-related protein (alpha 2MR/LRP) is a multifunctional cell plasma membrane receptor that binds and facilitates the endocytosis of a number of ligands involved in protease regulation and lipoprotein metabolism. In the invasive breast cancer cell line BT-20 we show that the expression of alpha 2MR/LRP can be strongly affected by cell culture density. By comparing measurements of mRNA, total cellular alpha 2MR/LRP antigen, and cell surface alpha 2MR/LRP expression we have demonstrated a marked cell density-dependent regulation of this receptor expression. Increasing the cell density results in a 3.4-fold increase in cell surface alpha 2MR/LRP expression. This corresponds to a marked increase in alpha 2MR/LRP mRNA in Northern blots of total RNA from cells cultured at high density and to consistent increases in alpha 2MR/LRP heavy chain in Western blots of cell lysates from high density cultures. These studies together demonstrate the significant up-regulation of alpha 2MR/LRP expression in BT-20 by increased cell density.

Blotting, Northern↗
PubMed1997

Expression of alpha2-macroglobulin receptor/low density lipoprotein receptor-related protein on surfaces of tumour cells: a study using flow cytometry.

alpha2-Macroglobulin receptor/low density lipoprotein receptor-related protein (alpha2 MR/LRP) is a multifunctional cell surface receptor that binds and endocytoses several structurally and functionally distinct ligands. Very little is known about the expression and function of alpha2 MR/LRP in tumour cells. The aim of this study was to quantify the number of alpha2 MR/LRP on surfaces of human tumour cells by flow cytometry. Using human alpha2 MR/LRP monoclonal antibody 8G1, human peripheral blood lymphocytes (negative control cells), monocytes (positive control cells), human neonatal foreskin fibroblast cells (NFF) (positive control cells), three human breast cancer cell lines (BT-20, T-47D, and MCF-7), two human ovarian tumour cell lines (JAM, and CI80-13S), and five human melanomas (MM418c1, MM253c1, A2058, MM138, MM370) were indirectly labelled with goat anti-mouse IgFITC. The fluorescent signals of stained cells were measured by flow cytometry. Using Quantum Simply Cellular bead standards, the number of alpha2 MR/LRP binding sites per cell was assessed. The flow cytometric method to quantify of alpha2 MR/LRP described here is simple and reliable. All the human tumour cell lines so far examined express alpha2 MR/LRP at different levels from approximately 300 to approximately 10000 sites per cell.

Fibroblasts↗
PubMed1997

Simultaneous genotyping for all three known structural mutations in the human mannose-binding lectin gene.

We describe a rapid and simple method for genotyping the three known structural mutations within exon 1 of the mannan-binding lectin (MBL) gene. A PCR-amplifiable synthetic DNA (Universal Heteroduplex Generator) was annealed to genomic PCR product from exon 1 to generate unique DNA heteroduplexes for each mutation. Heteroduplexes were then resolved by non-denaturing polyacrylamide gel electrophoresis. The technique was initially validated with previously typed samples and then applied to previously untyped samples with the results confirmed by DNA sequencing.

Amino Acid Sequence↗
PubMed1997

Patient satisfaction: a review of issues and concepts.

This review presents issues arising from an analysis of over 100 papers published in the field of patient satisfaction. The published output appearing in the medical and nursing literature which incorporated the term "patient satisfaction" rose to a peak of over 1000 papers annually in 1994, reflecting changes in service management especially in the U.K. and U.S.A. over the past decade. An introductory section discusses the setting and measurement of patient satisfaction within this wider context of changes in service delivery. Various models are examined that have attempted to define and interpret the idea of determining individual perceptions of the quality of health care delivered. Determinants of satisfaction are examined in relation to the literature on expectations, and demographic and psychosocial variables. These are distinguished from the multidimensional components of satisfaction as aspects of the delivery of care, identified by many authors. The review highlights the complexity and breadth of the literature in this field, the existence of which is often not acknowledged by researchers presenting the findings of studies.

Humans↗
PubMed1997

Cloning of the SCA7 gene reveals a highly unstable CAG repeat expansion.

The gene for spinocerebellar ataxia 7 (SCA7) has been mapped to chromosome 3p12-13. By positional cloning, we have identified a new gene of unknown function containing a CAG repeat that is expanded in SCA7 patients. On mutated alleles, CAG repeat size is highly variable, ranging from 38 to 130 repeats, whereas on normal alleles it ranges from 7 to 17 repeats. Gonadal instability in SCA7 is greater than that observed in any of the seven known neuro-degenerative diseases caused by translated CAG repeat expansions, and is markedly associated with paternal transmissions. SCA7 is the first such disorder in which the degenerative process also affects the retina.

Adult↗
PubMed1997

Rapid mutation screening in type 2A von Willebrand's disease using universal heteroduplex generators.

Patients with type 2A von Willebrand's disease (VWD) commonly have missense mutations in the A2 domain of the von Willebrand factor (VWF) protein. This domain is encoded by the 3' region of VWF gene exon 2 8 and the large majority of patients have heterozygous mutations clustered in the sequence between codons 742 and 909. We describe a DNA-based diagnostic technique which enables at least 10 previously described mutations to be rapidly identified. The method involves polymerase chain reaction (PCR) amplification of two exon 28 gene segments between codons 717-788 and 803-893, respectively. Each fragment is then hybridized with a synthetic complementary DNA molecule of similar size, termed a Universal Heteroduplex Generator (UHG). The UHG contains base deletions contiguous to the sites of known mutations and, following hybridization, allele-specific heteroduplexes are generated which can be detected by simple polyacrylamide gel electrophoresis and ethidium bromide staining. A small panel of UHG molecules covering the 3' region of exon 28 should enable the large majority of type 2A VWD patients to be rapidly diagnosed by genotype.

Base Sequence↗
PubMed1997

Chorea-acanthocytosis: genetic linkage to chromosome 9q21.

Chorea-acanthocytosis (CHAC) is a rare autosomal recessive disorder characterized by progressive neurodegeneration and unusual red-cell morphology (acanthocytosis), with onset in the third to fifth decade of life. Neurological impairment with acanthocytosis (neuroacanthocytosis) also is seen in abetalipoproteinemia and X-linked McLeod syndrome. Whereas the molecular etiology of McLeod syndrome has been defined (Ho et al. 1994), that of CHAC is still unknown. In the absence of cytogenetic rearrangements, we initiated a genomewide scan for linkage in 11 families, segregating for CHAC, who are of diverse geographical origin. We report here that the disease is linked, in all families, to a 6-cM region of chromosome 9q21 that is flanked by the recombinant markers GATA89a11 and D9S1843. A maximum two-point LOD score of 7.1 (theta = .00) for D9S1867 was achieved, and the linked region has been confirmed by homozygosity-by-descent, in offspring from inbred families. These findings provide strong evidence for the involvement of a single locus for CHAC and are the first step in positional cloning of the disease gene.

Acanthocytes↗