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Biomedical subjects

D L Clarke

Publications and source records attributed to D L Clarke.

At least 19 recordsLinked to original sources
PubMed2003

Octreotide lowers gastric mucosal blood flow in normal and portal hypertensive stomachs.

BACKGROUND: The vasoactive peptide octreotide has an established role in controlling variceal hemorrhage. The mechanism of action is believed to be a reduction in splanchnic blood flow. A decrease in splanchnic blood flow should be mirrored by a decrease in gastric mucosal blood flow (GMBF). Laser Doppler flowmetry (LDF) should detect changes in GMBF. METHODS: In seven normal volunteers and four patients with portal hypertension, 100 micro g of octreotide was administered as an intravenous bolus. Continuous LDF measurements were then made at a single point on the midantrum for at least 10 min and plotted against time for each subject. RESULTS: After a variable period of stabilization, GMBF decreased in all subjects except one. This was statistically significant in both the controls and the patients with portal hypertension. CONCLUSIONS: Octreotide decreases GMBF in normal and portal hypertensive stomachs. Laser Doppler is a useful and minimally invasive tool to assess the effect of drugs on GMBF.

Adult↗
PubMed2003

Neural stem cells.

The adult vertebrate central nervous system (CNS) consists of four major differentiated cell types: neurons, astrocytes, oligodendrocytes and ependymal cells. Historically, there has been a disagreement on how these differentiated cell types are generated in the CNS. Progress remains hindered by the complexity of cell structure in this system, the lack of specific cell surface markers to identify distinct cell types and the presence of numerous transit amplifying cell populations that rapidly generate early progenitors. At present, it is clear that some cells, termed neural stem cells, can generate neurons as well as astrocytes and oligodendrocytes of the glial lineage both in vitro and in vivo. Additionally, specific neural stem cell populations have also exhibited tissue lineage plasticity.

Animals↗
PubMed2001

The Cryptococcus neoformans STE11alpha gene is similar to other fungal mitogen-activated protein kinase kinase kinase (MAPKKK) genes but is mating type specific.

Partial sequence analysis of the Cryptococcus neoformans MATalpha mating type locus revealed the presence of a gene with substantial sequence similarity to other fungal mitogen-activated protein (MAP) kinase kinase kinase (MAPKKK) genes. The C. neoformans gene, designated STE11alpha, showed the highest degree of similarity to the Neurospora crassa nrc-1, Schizosaccharomyces pombe byr2 and Saccharomyces cerevisiae STE11 genes. A polymerase chain reaction-mediated sib-selection technique was successfully adapted for the purpose of disrupting STE11alpha. C. neoformans ste11alphaDelta mutants were found to be sterile, consistent with the phenotypes of ste11 and byr2 mutants in S. cerevisiae and S. pombe respectively. Haploid ste11alphaDelta mutants were also found to be unable to produce hyphae, suggesting that the C. neoformans gene is functionally conserved when compared with its S. cerevisiae MAPKKK counterpart. Comparison of the wild-type STE11alpha strain with a ste11alphaDelta disruptant for virulence using the mouse model showed that the ste11alphaDelta strain was less virulent, but the difference was only minor. In spite of some of the conserved functions of STE11alpha, linkage analysis showed that STE11alpha is only found in mating type alpha strains. These results demonstrate that, although functionally conserved, the mating pathway in C. neoformans has a unique organization.

Amino Acid Sequence↗
PubMed2001

Origin of Cryptococcus neoformans var. neoformans diploid strains.

The basidiomycetous yeast Cryptococcus neoformans is an important human fungal pathogen. Two varieties, C. neoformans var. neoformans and C. neoformans var. gattii, have been identified. Both are heterothallic with two mating types, MATa and MATalpha. Some rare isolates are self-fertile and are considered occasional diploid or aneuploid strains. In the present study, 133 isolates, mostly from Italian patients, were investigated to detect the presence of diploid strains in the Igiene Università Milano culture collection. All of the diploid isolates were further investigated by different methods to elucidate their origins. Forty-nine diploid strains were identified by flow cytometry. PCR fingerprinting using the (GACA)(4) primer showed that the diploid state was associated with two specific genotypes identified as VN3 and VN4. Determination of mating type on V8 juice medium confirmed that the majority of the strains were sterile. PCR and dot blotting using the two pheromone genes (MFa and MFalpha) as probes identified 36 of the 49 diploid isolates as MATa/alpha. The results of pheromone gene sequencing showed that two allelic MFalpha genes exist and are distinct for serotypes A and D. In contrast, the MFa gene sequence was conserved in both serotype alleles. Amplification of serotype-specific STE20 alleles demonstrated that the diploid strains contained one mating locus inherited from a serotype A parent and one inherited from a serotype D parent. The present results suggest that diploid isolates may be common among the C. neoformans population and that in Italy and other European countries serotype A and D populations are not genetically isolated but are able to recombine by sexual reproduction.

Cryptococcosis↗
PubMed2000

Regulation of repulsion versus adhesion by different splice forms of an Eph receptor.

Eph tyrosine kinase receptors and their membrane-bound ephrin ligands mediate cell interactions and participate in several developmental processes. Ligand binding to an Eph receptor results in tyrosine phosphorylation of the kinase domain, and repulsion of axonal growth cones and migrating cells. Here we report that a subpopulation of ephrin-A5 null mice display neural tube defects resembling anencephaly in man. This is caused by the failure of the neural folds to fuse in the dorsal midline, suggesting that ephrin-A5, in addition to its involvement in cell repulsion, can participate in cell adhesion. During neurulation, ephrin-A5 is co-expressed with its cognate receptor EphA7 in cells at the edges of the dorsal neural folds. Three different EphA7 splice variants, a full-length form and two truncated versions lacking kinase domains, are expressed in the neural folds. Co-expression of an endogenously expressed truncated form of EphA7 suppresses tyrosine phosphorylation of the full-length EphA7 receptor and shifts the cellular response from repulsion to adhesion in vitro. We conclude that alternative usage of different splice forms of a tyrosine kinase receptor can mediate cellular adhesion or repulsion during embryonic development.

Alternative Splicing↗
PubMed2000

Generalized potential of adult neural stem cells.

The differentiation potential of stem cells in tissues of the adult has been thought to be limited to cell lineages present in the organ from which they were derived, but there is evidence that some stem cells may have a broader differentiation repertoire. We show here that neural stem cells from the adult mouse brain can contribute to the formation of chimeric chick and mouse embryos and give rise to cells of all germ layers. This demonstrates that an adult neural stem cell has a very broad developmental capacity and may potentially be used to generate a variety of cell types for transplantation in different diseases.

Animals↗
PubMed2000

Mapping of the Cryptococcus neoformans MATalpha locus: presence of mating type-specific mitogen-activated protein kinase cascade homologs.

In this study we investigated the relationship between the MATalpha locus of Cryptococcus neoformans and several MATalpha-specific mitogen-activated protein (MAP) kinase signal transduction cascade genes, including STE12alpha, STE11alpha, and STE20alpha. To resolve the location of the genes, we screened a cosmid library of the MATalpha strain B-4500 (JEC21), which was chosen for the C. neoformans genome project. We isolated several overlapping cosmids spanning a region of about 71 kb covering the entire MATalpha locus. It was found that STE12alpha, STE11alpha, and STE20alpha are imbedded within the locus rather than closely linked to the locus. Furthermore, three copies of MFalpha, the mating type alpha-pheromone gene, a MATalpha-specific myosin gene, and a pheromone receptor (CPRalpha) were identified within the locus. We created a physical map, based on the restriction enzyme BamHI, and identified both borders of the MATalpha locus. The MATalpha locus of C. neoformans is approximately 50 kb in size and is one of the largest mating type loci reported among fungi with a one-locus, two-allele mating system.

Chemoreceptor Cells↗
PubMed1999

Identification of a neural stem cell in the adult mammalian central nervous system.

New neurons are continuously added in specific regions of the adult mammalian central nervous system. These neurons are derived from multipotent stem cells whose identity has been enigmatic. In this work, we present evidence that ependymal cells are neural stem cells. Ependymal cells give rise to a rapidly proliferating cell type that generates neurons that migrate to the olfactory bulb. In response to spinal cord injury, ependymal cell proliferation increases dramatically to generate migratory cells that differentiate to astrocytes and participate in scar formation. These data demonstrate that ependymal cells are neural stem cells and identify a novel process in the response to central nervous system injury.

Animals↗
PubMed1999

Universal primary colonic repair in the firearm era.

All patients with colonic trauma treated at King Edward VIII Hospital, Durban, from August 1993 to May 1994 underwent primary repair of the colonic wound. They were evaluated prospectively to assess the mode of injury and outcome variables. Colonic injuries were sustained by 102 patients. These were inflicted by gunshots (62), stabs (22), shotguns (14), and blunt trauma (4). The transverse colon was injured most frequently (53). All shotgun injuries were multiple. Average time from admission to theatre was similar for shocked and non-shocked patients. Eighty-seven patients had simple closure (18 deaths) and 15 required resection and anastomosis (eight deaths). Ten patients died in the first 48 h, and 16 died subsequently owing to multiple-organ systems dysfunction. The mortality rates were stabs 9% (2), gunshots 27% (17), shotguns 50% (7), and 0% for blunt trauma. Septic morbidity was seen in 16 but was not related to breakdown of the colonic repair. Implementation of strategies to reduce preoperative time delays and use damage control principles for the management of massive trauma should be evaluated as methods of reducing mortality.

Adolescent↗
PubMed1999

Inferior vena caval injury in the firearm era.

BACKGROUND: This study compared the outcome of intra-abdominal caval injuries in the current era of firearm injuries with the outcome during the previous era of stab wounds. METHODS: Patients with intra-abdominal vena caval injuries treated at King Edward VIII Hospital, Durban, from December 1990 to December 1995 were reviewed. This group was compared with a similar cohort reviewed a decade earlier. RESULTS: The historical group consisted of 28 patients and the current group of 26 patients. Modes of injury in the historical group were: stabs (15, 53%), firearm injuries (7, 33%), blunt trauma (4, 14%) and iatrogenic injuries (2, 7%). Modes of injury in the current group were: stabs (5, 19%), firearm injuries (17, 65%) and blunt trauma (4, 16%). Mortality rose from 35.7% in the historical group to 88% in the current series. This mortality figure included 5 patients who died later from complications of the associated injuries. CONCLUSIONS: Firearm injuries are more destructive than stab wounds. The increase in firearm injuries partly explains this higher mortality. However, the failure to apply current concepts of abbreviated laparotomy and damage control combined with excessive delays in transferring patients to theatre have contributed to this high mortality.

Adolescent↗
PubMed1997

Cloning and characterisation of the adenosyl phosphosulphate kinase gene from Aspergillus nidulans.

The sD gene of Aspergillus nidulans has been cloned by heterologous screening of rationally selected cosmids. Co-transformation of the sD50 mutant JMP1 confirmed the presence of a functional gene. Sequence analysis determined this gene to be 680 bp in length, containing a 59-bp intron and encoding a protein of 206 amino acids. A protein-sequence comparison revealed a similarity to the C-terminal region of ATP sulphurylase, the sC gene product. Further sequence comparison revealed differences in a consensus sequence ATP-binding motif, indicating non-functionality of the APS kinase-like domain of ATP sulphurylase, and confirms sD as the gene encoding APS kinase in A. nidulans.

Amino Acid Sequence↗
PubMed1997

The different forms of the prolactin receptor in the rat corpus luteum: developmental expression and hormonal regulation in pregnancy.

The corpora lutea of pregnancy in the rat are highly dependent on the action of PRL and PRL-like hormones to hypertrophy and to produce progesterone needed for the maintenance of gestation. Two forms of the PRL receptor (PRL-R), designated as long (PRL-RL) and short (PRL-RS), have been described in rat tissues. To determine whether both forms are present in the corpus luteum during pregnancy and to examine the developmental and hormonal regulation of their expression, total RNA isolated from corpora lutea at different stages of pregnancy and from highly luteinized granulosa cells subjected to different hormonal treatments were analyzed by semiquantitative RT-PCR. Immunoblotting of luteal proteins from early and late pregnancy was also performed to determine if the pattern of PRL-R proteins follows that of PRL-R messenger RNA (mRNA) expression. In addition, the correlation between the well characterized PRL-regulated gene, 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD), and PRL-R gene expression was investigated during the time of luteolysis. Both PRL-RL and PRL-RS mRNA and protein were expressed in corpora lutea of pregnancy, with the long form being the most dominant at all stages. Whereas no changes in mRNA level of either PRL-RL or PRL-RS were found until day 20 of gestation, a profound decline in PRL-R mRNA and protein for both receptor types occurred at the end of pregnancy. This drop in PRL-R expression was accompanied by a sharp and abrupt expression of 20alpha-HSD mRNA. Studies performed in vivo and in luteinized cells in culture indicate that PRL can up-regulate the expression of the PRL-RL mRNA, an effect prevented by the tyrosine kinase inhibitor, genistein. PRL-RL mRNA was also selectively increased by cAMP. In summary, the results of this investigation have established that: 1) the corpus luteum of pregnancy expresses both the short and long forms of the PRL-R with the long form being more abundant; 2) the mRNA for both forms of the PRL-R remains at constant levels throughout pregnancy but drops before parturition; 3) the decline in PRL-R mRNA at the end of pregnancy is accompanied by a dramatic rise in 20alpha-HSD; 4) PRL is able to increase the expression of PRL-R mRNA; and that 5) both A kinase and tyrosine kinase mediated pathways appear to participate in the up-regulatory mechanism involved in PRL-R mRNA expression.

20-Hydroxysteroid Dehydrogenases↗
PubMed1996

The decidual prolactin receptor and its regulation by decidua-derived factors.

Decidualization of the endometrial stroma in the rat gives rise to two different cell populations, located either mesometrially or antimesometrially in the uterus. We have previously shown that the rat decidua is the site of production and action of a PRL-like hormone. In this investigation we examined, using reverse transcription-PCR, whether and which type of PRL receptor (PRL-R) messenger RNA (mRNA) is expressed in the decidua, whether the receptor is confined only to one cell population, and whether the PRL-R expression is regulated by decidua-derived factors. The results indicate that the uterus of pseudopregnant rats does not express the PRL-R and that decidualization does not trigger a rapid appearance of PRL-R mRNA. It is only 3 days after the induction of decidualization that the long form of the PRL-R was first expressed. Thereafter, mRNAs for both the short (PRL-RS) and the long (PRL-RL) form became detectable in both antimesometrial and mesometrial decidua, although PRL-RL mRNA was much more abundant than PRL-RS. As development proceeded, PRL-R mRNA decreased and disappeared specifically from the antimesometrial decidua, whereas the mesometrial decidua continued to express this receptor mRNA. Concomitant with down-regulation of the PRL-R in the antimesometrial tissue was a rather abrupt expression of activin A. In contrast, the mesometrial tissue that maintained high levels of PRL-R mRNA expressed little activin A, but produced an activin-binding protein, alpha(2)-macroglobulin (alpha(2)MG). To determine whether activin A and alpha(2)MG regulate PRL-R expression, antimesometrial and mesometrial cells were separated by elutriation and maintained in culture in the presence or absence of activin A, alpha(2)MG, or follistatin. Just after cell separation, both cell populations expressed PRL-R, but not activin A. Within 6 h, activin A mRNA and protein became highly expressed in the mesometrial cells, whereas PRL-RL mRNA became undetectable. In contrast, activin A mRNA was at very low levels in the antimesometrial cells, and no activin A protein could be detected in the medium for at least 12 h. In these cells PRL-RL mRNA remained elevated. Addition of activin A to antimesometrial cells caused a marked down-regulation of PRL-RL mRNA expression, whereas addition of alpha(2)MG and follistatin to mesometrial cells prevented the disappearance of PRL-R. In summary, the results of this investigation 1) indicate that decidualization of the endometrial stroma induces the appearance of both forms of the PRL-R mRNAs; 2) show differential expression of the PRL-R mRNA in the two-cell population forming the decidua; 3) establish that this differential expression is due to two key decidual molecules, activin A and alpha(2) macroglobulin; and 4) demonstrate that activin A can cause the decidual cells to lose the PRL-R and that the disappearance of the decidual PRL-R can be prevented by addition to the culture of two activin binding proteins, follistatin and alpha(2) MG.

Activins↗
PubMed1993

Changes in prolactin receptor expression during pregnancy in the mouse ovary.

We have isolated the cDNA encoding the cytoplasmic domain of a long form of the mouse PRL receptor (PRL-R). The mRNA for this long form PRL-R and the three mRNAs encoding short mouse PRL-R that have been previously characterized are all expressed in both the liver and ovary. The relative amounts of these receptor forms differ between tissues, however. In addition, the structure of one of the short receptor forms may not be identical in the liver and ovary. Within the ovary, the abundance and sites of synthesis of the four PRL-R mRNAs vary during pregnancy. Expression of the two most abundant PRL-R mRNAs increases significantly at midgestation. Expression of PRL-R mRNA is detected in the corpus luteum throughout pregnancy, while increased receptor mRNA levels are evident in the granulosa cells of a subset of Graafian follicles toward the end of pregnancy and during lactation. Some differences are also observed in the expression patterns of the individual receptor forms. Most notably, one of the short form PRL-R mRNAs is uniquely detected in atretic follicles in early to midgestation.

Amino Acid Sequence↗