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Biomedical subjects

D J Henderson

Publications and source records attributed to D J Henderson.

At least 37 records · Page 2Linked to original sources
PubMed1997

Development of a lethal congenital heart defect in the splotch (Pax3) mutant mouse.

OBJECTIVE: The splotch (Sp2h) mutation disrupts the Pax3 gene and is lethal in homozygotes. The aim of the present study was to investigate the cause of lethality. METHODS AND RESULTS: Using the splotch (Sp2H) mouse mutant, we demonstrated that approximately 60% of Sp2H homozygotes die in utero at 13.5-14.5 days of gestation. All these embryos have cardiac malformations involving partial or complete failure of septation of the outflow tract. Although the cause of death in utero is unknown, the dying embryos are edematous, their superior caval veins are over-expanded, and the fetal liver is enlarged and engorged with blood, all signs of cardiac failure. The remaining Sp2H homozygotes die around the time of birth, and these embryos have grossly normal hearts. All Sp2H homozygotes have neural tube defects, either spina bifida, exencephaly, or both. Although these defects clearly do not cause death in utero, they are very likely responsible for the perinatal death of homozygotes that survive to late gestation. There is no correlation between the presence or absence of a cardiac defect and the type of neural tube defect. On the other hand, there is a striking correlation between presence of a cardiac defect and reduction or absence of dorsal root ganglia, which are derivatives of the neural crest. CONCLUSIONS: In this paper, we show that the lethality has a biphasic pattern, and the data strongly suggests that mid-gestation lethality is due to cardiac defects and not the associated neural tube defects. This finding supports the idea that 'conotruncal' cardiac defects involving the ventricular outflow tracts develop as a result of failure of the 'cardiac' neural crest to colonise the developing heart in the mid-gestation embryo, and that the resulting heart defects are solely responsible for the observed mortality.

Animals↗
PubMed1997

Over-expression of the chondroitin sulphate proteoglycan versican is associated with defective neural crest migration in the Pax3 mutant mouse (splotch).

Splotch mice, which harbour mutations in the Pax3 gene, exhibit neural crest-related abnormalities including pigmentation defects, reduced or absent dorsal root ganglia and failure of cardiac outflow tract septation in homozygotes. Although splotch neural crest cells fail to colonise target tissues, they initiate migration in vivo and appear to migrate as well as wild type neural crest cells in vitro, suggesting that the neural crest abnormality in splotch may reside not in the neural crest cells themselves, but rather in the extracellular environment through which they migrate. We have examined the expression of genes encoding extracellular matrix molecules in Sp2H homozygous embryos and find a marked over-expression of transcripts for the chondroitin sulphate proteoglycan versican in the pathways of neural crest cell migration. Use of cadherin-6 expression as a marker for neural crest demonstrates a striking correlation between up-regulation of versican expression and absence of migrating neural crest cells, both in the mesenchyme lateral to the neural tube and in the lower branchial arches of Sp2H homozygotes. Pax3 and versican have mutually exclusive expression patterns in normal embryos whereas, in Sp2H homozygotes, versican is generally over-expressed with 'infilling' in regions that would normally express functional Pax3. Versican, like other chondroitin sulphate proteoglycans, is non-permissive for migration of neural crest cells in vitro, and we suggest that over-expression of this molecule leads to the arrest of neural crest cell migration in splotch embryos. Pax3 may serve to negatively regulate versican expression during normal development, thereby guiding neural crest cells into their pathways of migration.

Animals↗
PubMed1997

Role of the extracellular matrix in neural crest cell migration.

Development of the neural crest involves a remarkable feat of coordinated cell migration in which cells detach from the neural tube, take varying routes of migration through the embryonic tissues and then differentiate at the end of their journey to participate in the formation of a number of organ systems. In general, neural crest cells appear to migrate without the guidance of long-range physical or chemical cues, but rather they respond to heterogeneity in the extracellular matrix that forms their migration substrate. Molecules such as fibronectin and laminin act as permissive substrate components, encouraging neural crest cell attachment and spreading, whereas chondroitin sulphate proteoglycans are nonpermissive for migration. A balance between permissive and nonpermissive substrate components seems to be necessary to ensure successful migration, as indicated by a number of studies in mouse mutant systems where nonpermissive molecules are over-expressed, leading to inhibition of neural crest migration. The neural crest expresses cell surface receptors that permit interaction with the extracellular matrix and may also modify the matrix by secretion of proteases. Thus the principles that govern the complex migration of neural crest cells are beginning to emerge.

Animals↗
PubMed1997

Pax3 is required for cardiac neural crest migration in the mouse: evidence from the splotch (Sp2H) mutant.

Neural crest cells originating in the occipital region of the avian embryo are known to play a vital role in formation of the septum of the cardiac outflow tract and to contribute cells to the aortic arches, thymus, thyroid and parathyroids. This 'cardiac' neural crest sub-population is assumed to exist in mammals, but without direct evidence. In this paper we demonstrate, using RT-PCR and in situ hybridisation, that Pax3 expression can serve as a marker of cardiac neural crest cells in the mouse embryo. Cells of this lineage were traced from the occipital neural tube, via branchial arches 3, 4 and 6, into the aortic sac and aorto-pulmonary outflow tract. Confirmation that these Pax3-positive cells are indeed cardiac neural crest is provided by experiments in which hearts were deprived of a source of colonising neural crest, by organ culture in vitro, with consequent lack of up-regulation of Pax3. Occipital neural crest cell outgrowths in vitro were also shown to express Pax3. Mutation of Pax3, as occurs in the splotch (Sp2H) mouse, results in development of conotruncal heart defects including persistent truncus arteriosus. Homozygotes also exhibit defects of the aortic arches, thymus, thyroid and parathyroids. Pax3-positive neural crest cells were found to emigrate from the occipital neural tube of Sp2H/Sp2H embryos in a relatively normal fashion, but there was a marked deficiency or absence of neural crest cells traversing branchial arches 3, 4 and 6, and entering the cardiac outflow tract. This decreased expression of Pax3 in Sp2H/Sp2H embryos was not due to down-regulation of Pax3 in neural crest cells, as use of independent neural crest markers, Hoxa-3, CrabpI, Prx1, Prx2 and c-met also revealed a deficiency of migrating cardiac neural crest cells in homozygous embryos. This work demonstrates the essential role of the cardiac neural crest in formation of the heart and great vessels in the mouse and, furthermore, shows that Pax3 function is required for the cardiac neural crest to complete its migration to the developing heart.

Animals↗
PubMed1996

Transferrin and its receptor in the development of genetically determined neural tube defects in the mouse embryo.

The iron-binding growth factor transferrin is taken up and localised in the hindgut of midgestation mouse embryos. We investigated whether the distribution of transferrin may be disturbed in mutant curly tail embryos, a proportion of which exhibit a cell proliferation defect affecting the hindgut endoderm, as part of the pathogenetic sequence leading to development of neural tube defects. Immunostaining revealed a reduction in the binding and/or uptake of transferrin by hindgut epithelial cells in affected curly tail embryos compared with their unaffected littermates. There was no apparent difference between the two embryo types, however, in the distribution or level of expression of the transferrin receptor. The receptor is expressed specifically in the hindgut endoderm of the 10.5-day embryo, although its mRNA is present in all tissues of the posterior neuropore region, suggesting posttranscriptional control of gene expression. These findings may indicate a role for transferrin binding and/or uptake in the regulation of cell proliferation in the hindgut endoderm, with a defect in this process in the curly tail mutant. However, an alternative explanation is suggested by our finding that transferrin immunostaining is more intense in the hindgut of unaffected curly tail embryos than in nonmutant CBA/Ca and CD-1 embryos. Thus, mutant embryos may increase their uptake of transferrin in an attempt to compensate for defective cell proliferation in the hindgut resulting from a defect in another pathway. Only a proportion of embryos are able to mount this compensatory response leading to the observed partial penetrance of developmental defects in the curly tail mutant mouse.

Animals↗
PubMed1996

Does pre-incisional thoracic extradural block combined with diclofenac reduce postoperative pain after abdominal hysterectomy?

In a double-blind, randomized study, we investigated 40 patients undergoing abdominal hysterectomy; patients received 0.5% plain bupivacaine 20 ml via a low thoracic extradural catheter and a diclofenac suppository (100 mg), either 30 min before incision (group 1) or 30 min after incision (group 2). All patients received a standard general anaesthetic and no opioid was used before or during operation. Postoperative analgesic requirements were measured using a patient-controlled analgesia (PCA) system. Pain was assessed using a visual analogue scale (VAS) and a verbal pain score (VPS) on movement up to 48 h after operation. There was no significant difference in the time to first request for morphine but consumption of morphine was significantly greater in group 1 at all times except 24 h. There were no significant differences in VAS and VPS pain scores, although both scores were consistently higher in group 1. Patient satisfaction with the quality of analgesia, at 24 h, demonstrated no significant difference between the two groups. The combination of extradural block and diclofenac suppository given before operation did not appear to produce a clinically effective pre-emptive analgesic effect.

Adult↗
PubMed1995

Cloning and characterisation of a bovine P2Y receptor.

Using a chick P2Y1 receptor cDNA probe we have isolated a mammalian P2Y receptor clone from a bovine aortic endothelial cell library. The sequence has a high degree of similarity to the chick P2Y1 clone. When transfected into the Jurkat cell line, the cDNA conferred sensitivity to purinoceptor agonists. Using fura-2 loaded cells the potency order at the receptor was found to be 2-methylthioadenosine 5' triphosphate = adenosine 5' diphosphate > adenosine 5' triphosphate >> alpha,beta-methyleneadenosine 5' triphosphate and uridine 5' triphosphate. This corresponds to the agonist potency order expected for the bovine aortic endothelial cell P2Y receptor.

Adenosine Diphosphate↗
PubMed1995

Effect of i.v. diamorphine on the regression of spinal block.

Twenty patients undergoing transurethral prostatectomy under spinal anaesthesia were allocated randomly in one of two groups. After operation dermatomal levels to cold were measured every 30 min until they had receded to T10. Patients in group 1 were then given diamorphine 5 mg in 0.9% saline 5 ml i.v. and in group 2 0.9% saline 5 ml i.v. Block level to cold and degree of motor block were assessed at 15-min intervals for 1 h after injection. Block regression continued in the control group while there was no decrease in the diamorphine group for 30 min (P < 0.01) after which it then receded at a similar rate as the control group. There was no significant difference in motor block between the two groups.

Aged↗
PubMed1995

Consciousness raising in participatory research: method and methodology for emancipatory nursing inquiry.

Nursing practice and research that are grounded in emancipatory inquiry challenge the inequalities and injustices that affect health in this society. Increasing numbers of nurse scholars have explored the use of emancipatory inquiries (particularly feminist theory and critical theory) as philosophic bases for nursing practice and as theoretical or methodologic perspectives for nursing research. This article presents participatory research as a methodology and consciousness raising as a method for nursing research that addresses the emancipatory goals of feminist and critical theories. An example of consciousness raising in a participatory nursing research project is offered. It is argued that participatory research can produce a revolutionary emancipatory knowledge for the future of nursing and health care.

Data Collection↗
PubMed1994

Gender, relationships, and crack cocaine: a content analysis.

Past research has indicated gender differences among narcotic users that have implications for the prevention and treatment of substance abuse; however, little is known about these differences among crack cocaine users. The purpose of this secondary analysis was to compare the relational context of male and female crack cocaine use. Data from open-ended, structured interviews with 46 predominantly African-American women (n = 23) and men (n = 23) were compared using manifest content analysis. Women were more likely to begin, use and/or maintain their use of crack in the context of more intimate opposite sex relationships, while men were more likely to begin their use with male friends and associates and to maintain drug use with income from jobs and selling drugs. Overall, relationships (both sexual and familial) were a more prominent aspect of crack use for women while enterpreneurship was more salient for men. These findings suggest the need for gender-sensitive prevention and treatment strategies.

Adult↗
PubMed1994

Early embryonic failure associated with uniparental disomy for human chromosome 21.

As many as 16% of all recognized pregnancies may be anembryonic, with failure of the embryo at a very early stage of development leaving only the extraembryonic components of the conceptus to proliferate. Studies in the mouse have shown that the maternal and paternal contributions to the genome of the zygote are not functionally equivalent, due to parental genomic imprinting. Uniparental disomy can reveal imprinting effects, as in this phenomenon both members of a chromosome pair are inherited from the same parent. We have carried out a systematic search for uniparental disomy in tissues from 23 cases of early embryonic failure, using variable number tandem repeat (VNTR) analysis and PCR amplification of polymorphic short sequence repeats. Two cases of maternal uniparental heterodisomy for chromosome 21 were identified. One case occurred in conjunction with trisomy for chromosomes 7 and 9, but in the other case maternal uniparental heterodisomy for chromosome 21 was the only chromosomal abnormality found. We therefore postulate that there may be developmentally important genes on human chromosome 21 which are imprinted such that both parental copies are essential for normal embryogenesis.

Chromosome Aberrations↗
PubMed1994

Transposition of IS117, the 2.5 kb Streptomyces coelicolor A3(2) 'minicircle': roles of open reading frames and origin of tandem insertions.

IS117 is a 2527 bp transposable element from Streptomyces coelicolor A3(2) with a circular transposition intermediate. Disruption of ORF1 of IS117, presumed to encode a transposase, abolished transposition. Deletion or mutation of ORF2 and ORF3, which overlap each other on opposite strands of IS117, caused a c. 20-fold reduction in integration frequency of the circular form of IS117 into the Streptomyces lividans chromosome or into the preferred chromosomal target site cloned on a plasmid in transformation experiments. In contrast, inactivation of ORF2/3 did not significantly influence transposition of IS117 derivatives from an already integrated state in the chromosome to the preferred target site cloned on a plasmid. ORF2 mutants apparently excised readily from the S. lividans chromosome, whereas excision of integrated wild-type IS117 derivatives to yield the unoccupied site was not detected; presumably, therefore, the circular transposition intermediate normally arises replicatively. Attempts to promote integration of a plasmid carrying the attachment site of IS117 by providing the ORF1 product in trans were unsuccessful. Most transformation of S. lividans with circular IS117 derivatives yielded tandem chromosomal insertions, which arose by co-transformation rather than dimerization of a monomeric insert. Typically, two to three transforming elements gave a transformed strain, suggesting a local concentration of transposase as a limit on integration.

Base Sequence↗
PubMed1994

Linkage analysis of 62 X-chromosomal loci excludes the X chromosome in an Icelandic family showing apparent X-linked recessive inheritance of neural tube defects.

We report here the findings of a linkage analysis, involving numerous markers from the human X chromosome, in an attempt to localise a putative gene causing apparent X-linked spina bifida and anencephaly (SBA) in a large Icelandic pedigree. Two-point linkage analysis was performed using markers from 62 informative loci in this family. Although small positive lod scores were found at a number of these loci, none reached the significance level for linkage. Haplotypes were extensively analysed and found to exclude linkage to the X chromosome.

Adult↗
PubMed1992

Changes in expression of the cyclooxygenase gene in human fetal membranes and placenta with labor.

OBJECTIVE: Our objective was to examine the expression of the gene coding for cyclooxygenase, the central enzyme in prostaglandin synthesis, in human placenta and fetal membranes during pregnancy and before and after labor at term. STUDY DESIGN: Expression of the gene for cyclooxygenase was examined with Northern hybridization to ribonucleic acid from human placenta throughout pregnancy and human amnion and chorion decidua in the late third trimester. RESULTS: Expression was undetectable in trophoblast during the first and second trimesters. Expression in amnion and trophoblast increased 3.5- and 2.5-fold, respectively, in association with labor. CONCLUSIONS: Our results suggest that the increase in prostaglandin synthesis within the uterus that is seen with the onset of labor is associated with an increase in the expression of the gene cyclooxygenase.

Amnion↗
PubMed1992

Expression of human chorionic gonadotrophin alpha and beta subunits is depressed in trophoblast from pregnancies with early embryonic failure.

The expression of the placental proteins human chorionic gonadotrophin alpha (HCG alpha), beta (HCG beta) and human placental lactogen (HPL) was examined in trophoblast from human normal pregnancy and early embryonic failure (EEF) using Northern blot analysis. Trophoblast from EEF expressed significantly depressed levels of HCG alpha (P < 0.05) and HCG beta (P < 0.01) when compared to normal pregnancy. Levels of expression of HPL in EEF were not significantly different to those from normal pregnancy. The gestational ages of the two groups were not significantly different. Immunocytochemistry on paraffin-fixed tissue sections supported these data and showed that mRNA levels reflected protein production of HCG within the tissue. It appears from these data that the depressed level of HCG found in the serum of women with early pregnancy failure is not solely a consequence of diminished placentation in these pregnancies, or placental necrosis, but that the genes of the alpha and beta HCG subunits are down-regulated. This study further suggests that there are two subsets of women with early pregnancy failure; those with apparently normal levels of placental proteins, and those with severely depressed levels. It would seem likely that these two groups have different aetiologies.

Abortion, Spontaneous↗