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J Walker

Publications and source records attributed to J Walker.

At least 19 recordsLinked to original sources

Chemical modification of a cloned glutathione S-transferase from Schistosoma japonicum: evidence for an essential histidine residue.

Diethylpyrocarbonate (DEP) inhibits the catalytic activity of a cloned glutathione S-transferase from Schistosoma japonicum (Sj26GST) with a second-order rate constant of 474 M-1 min-1 at pH 7.0 and 25 degrees C. There is an accompanying increase in absorbance at 242 nm due to the formation of N-carbethoxyhistidyl derivatives. There was no evidence that tyrosine or cysteine residues were modified by DEP treatment nor did the enzyme undergo any major conformational change. Activity can be restored by treating the DEP-modified enzyme with hydroxylamine and the pH curve for inactivation indicates involvement of a residue with a pKa of 7.3. Complete inactivation of Sj26GST requires the modification of six histidine residues per subunit. Statistical analysis of residual enzyme activity versus number of groups modified showed that of the six modifiable groups, only one is critical for activity. Substrate protection suggests that this essential histidine residue is at or near the active site.

Animals

New closure technique for lateral operative laparoscopic trocar sites. A report of 80 closures.

The reported frequency of incisional hernias, after operative gynecological laparoscopy, at extraumbilical trocar sites is one per 32 puncture sites created by a 12-mm trocar. A new closure technique of suturing with the Grice Needle (Ideas for Medicine, Inc., Clearwater, FL) before removing the trocars was utilized to close 80 lateral trocar sites (42 consecutive laparoscopic myomectomies). The trocar sizes in this study were 12 mm and 18 mm. This is the largest reported series of lateral trocar-site closures. No hematomas or bleeding or incisional hernias have resulted from use of this technique. This closure allows the surgeon to completely close both peritoneum and fascia, of the lateral trocar sites, under direct laparoscopic visualization without the loss of pneumoperitoneum or risk of inadvertent injury to the small bowel.

Hernia

Issues of recruitment, retention, and compliance in community-based clinical trials with traditionally underserved populations.

Site-specific sociodemographic recruitment, retention, and compliance (RRC) data were solicited at two points in time from the 18 National Institutes of Health-funded Community Programs for Clinical Research on AIDS (CPCRA). Based on their experiences delivering primary care to human immunodeficiency virus-infected individuals, nurses at each site identified organizational and client-centered factors functioning as barriers to protocol participation. In addition, the clinicians described the nature, frequency, and relative success of strategies used to enhance recruitment, retention, and protocol compliance. CPCRA units where nurses had clearly identified RRC barriers related to protocol design also were the sites that had accrued the most research participants. This study suggests that as the CPCRA units evolve, the most successful programs will be those in which the clinical and research staff can identify and develop innovative strategies that will successfully overcome RRC barriers.

Acquired Immunodeficiency Syndrome

Neuroendoscopic third ventriculostomy: a nursing perspective.

Endoscopic third ventriculostomy is an old technique that has gained greater acceptance in recent years with the advent of new endoscopic technology. This procedure, which involves fenestrating the floor of the third ventricle to communicate the basilar cisterns with the third ventricle offers new hope to patients with non-communicating hydrocephalus. Until recently, the only accepted treatment for these patients was the placement of a ventriculoperitoneal shunt to drain the accumulating cerebrospinal fluid. It is well known that shunting carries the risks of infection, blockage and overdrainage. This technique provides an alternative to a less than perfect treatment for reestablishing CSF pathways. Patients best suited for the procedure have delayed onset aqueductal stenosis caused by a tumor, cyst or hemorrhagic event. Patients with aqueductal stenosis as a consequence of myelomeningeocele fare less well, but show better success with the technique if it is performed as a follow-up to shunting rather than as an initial procedure. Several techniques may be used to perform third ventriculostomy. An understanding of the special needs of the patient undergoing the procedure is essential to optimum care.

Critical Care

A collapsing form of glomerulopathy.

An unusual presentation of glomerular disease occurring in a 30 year old black female is described. The case was characterized by a rapid deterioration of renal function. Associated glomerular manifestations were diffuse collapse of the glomerular capillary lumina, extracapillary hypercellularity without obliteration of the urinary space and without proliferation of parietal epithelial cells, electron-dense deposits, and deposition of IgG and C3. There was little loss of tubules and minor interstitial fibrosis. This case demonstrated a collapsing form of glomerulopathy with immunoglobulin and electron-dense deposits.

Adult

The effect of radiofrequency catheter ablation on myocardial creatine kinase activity.

INTRODUCTION: The primary mechanism of myocardial injury during radiofrequency (RF) catheter ablation in the heart is presumed to be thermal. Creatine kinase has been measured in serum to assess the volume of myocardial injury after ablation. However, its thermal inactivation by RF ablation could lead to underestimation of the true volume of injury. METHODS AND RESULTS: Serial RF lesions were created in 10 canine left ventricles in vivo, and serial serum creatine kinase activities were measured and compared to lesion volume. To assess the stability of myocardial creatine kinase during RF catheter ablation, 29 RF ablations were made on the epicardial surface of porcine left ventricle in vivo and a 2-mm core biopsy was rapidly removed. The cores were rapidly frozen, sectioned longitudinally in 1-mm slices, and homogenized in 0.3 M Tris buffer solution containing EDTA and dithiothreitol for subsequent analysis of creatine kinase activity. An additional 19 tissue cores from RF lesions were stained and used to determine mean lesion depth. Normal tissue biopsies were exposed to 60 seconds of hyperthermia (37 degrees to 85 degrees C, n = 190), or high-density RF current at 50 degrees C (0 to 100 mA/mm2, n = 50), and tissue creatine kinase activity was measured. There was no evidence of creatine kinase washout within the first 2 hours, and peak values were measured 5 to 7 hours postablation. Tissue creatine kinase activity in the first mm depth of RF lesions averaged 10% of control values and increased over the first 5 mm of lesion depth. The mean creatine kinase activity within the hemisphere of ablated myocardium was calculated to be 31% of control. Creatine kinase activity declined significantly at temperatures above 65 degrees C, but no difference in tissue creatine kinase activity was observed among differing levels of RF current exposure in the absence of significant heating. CONCLUSIONS: Creatine kinase activity in myocardial tissue is significantly diminished within the RF lesion. Creatine kinase activity is not stable at temperatures above 65 degrees C, which are routinely achieved within the central zone of RF ablation, and is unaffected by RF current in the absence of hyperthermia. Measurements of serum creatine kinase activity after RF catheter ablation may significantly underestimate the volume of myocardial injury.

Animals

Molecular and biochemical characterization of a protective 40-kilodalton antigen from Corynebacterium pseudotuberculosis.

A 40-kDa protein from Corynebacterium pseudotuberculosis has been previously identified as a protective antigen against ovine caseous lymphadenitis. From genomic DNA libraries of C. pseudotuberculosis, we have cloned and sequenced the 40-kDa protein gene, which was found to contain an open reading frame of 1,137 bp encoding a protein of 379 amino acids. No significant homology with previously published DNA or amino acid sequence data was found in databases, suggesting that this is a novel protein. Recombinant 40-kDa protein was overexpressed as a fusion protein to 15% of total cell proteins in Escherichia coli. Biochemical analysis of native and recombinant 40-kDa proteins has revealed associated proteolytic activity, which was shown to be of the serine protease type through the use of specific inhibitors. We suggest that this novel protective antigen be termed corynebacterial protease 40 (CP40).

Amino Acid Sequence

Cloning and sequence analysis of a protective M-like protein gene from Streptococcus equi subsp. zooepidemicus.

Streptococcus equi subsp. zooepidemicus, a Lancefield group C streptococcus, is a frequently isolated opportunist pathogen from a variety of animal hosts, including the horse. Previous studies have indicated that equine strains carry antigens with characteristics of the antiphagocytic M proteins on the Lancefield groups A and G streptococci. We have cloned a protective M-like protein gene (SzPW60) of an equine strain of S. equi subsp. zooepidemicus W60 and determined its sequence. This gene encodes a protein with a molecular weight of 40,123 which protects mice against subsp. zooepidemicus but not subsp. equi, stimulates antibodies which opsonize subsp. zooepidemicus but not equi, and reacts with antiserum to the protein of the parent strain. The predicted amino acid structure shows significant homology with the carboxy termini of groups A and G M proteins but no other homology. The M-like protein, although showing an extensive region of alpha helix, lacks the A, B, and C repeats found in group A M proteins and has a shorter signal sequence. A proline-rich region upstream from the LPSTGE motif contains 20 repeats of the tetrapeptide PEPK. The presence of this repeat region may account for the slow migration of the M-like protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Amino Acid Sequence

Portal and systemic haemodynamic response to acute and chronic administration of low and high dose isosorbide-5-mononitrate in patients with cirrhosis.

Oral isosorbide-5-mononitrate (Is-5-Mn) was given in doses of 10 and 40 mg acutely and chronically (twice daily for four weeks), allowing a nitrate free interval to 25 patients with cirrhosis. Both 10 mg and 40 mg Is-5-Mn reduced the hepatic venous pressure gradient acutely and chronically, without evidence of tolerance. This was achieved by a reduction in the wedged hepatic venous pressure. The effect on mean azygos blood flow was variable with no significant mean change seen acutely or after chronic use with either dose. The variability was dependent not on the dose used but on the initial azygos flow; the flow in patients with initially low values increased and those with high azygos flows decreased after nitrate challenge. The development of the porto-collateral flow seems an important parameter in predicting haemodynamic response to Is-5-Mn.

Adult

Relation of granulomas to lymphatic vessels in Crohn's disease.

AIMS: To examine the relation between granulomas and lymphatic vessels in Crohn's disease. METHODS: Formalin fixed, paraffin wax embedded sections were selected from surgical resection specimens from 10 cases diagnosed as Crohn's disease. The block that showed the most granulomas was selected from each case. Sections 5 microns thick were immunostained with antibodies directed against the endothelial markers factor VIII related antigen and Ulex europaeus lectin, and against the vascular wall components collagen IV and laminin. Granulomas were counted on each slide in the serosa, muscularis propria, submucosa, and mucosa. In each area granulomas were classified according to their relation to lymphatic or blood vessels. RESULTS: Overall, an average of 46.1% of granulomas (range 15.3-90.4%) was related to lymphatic vessels, with the majority of these being adjacent to the vessel, rather than in the lumen or distorting the wall. A smaller percentage (10.1%, range 2.4-25.8%) was related to blood vessels. CONCLUSIONS: A significant proportion of granulomas in Crohn's disease is associated with lymphatic vessels. Blood vessel involvement may be a secondary phenomenon, rather than the primary event.

Antigens

Production and characterisation of monoclonal and polyclonal antibodies to different regions of the cystic fibrosis transmembrane conductance regulator (CFTR): detection of immunologically related proteins.

We have raised mouse monoclonal antibodies to eight synthetic peptides corresponding to different regions of the human cystic fibrosis transmembrane conductance regulator (CFTR) and rabbit polyclonal antisera to beta-galactosidase fusion proteins which encompass three different regions of CFTR. Immunoblot, immunoprecipitation, immunofluorescence and immunocytochemical experiments demonstrate that, in addition to recognising CFTR, these antibodies recognise one or more immunologically related proteins with a similar molecular mass, calcium responsiveness and tissue distribution to CFTR.

Amino Acid Sequence

Targeting seniors.

Health promotion is not just for the young. There has been far too much emphasis on the so-called burden that Canada's aging population puts on our health care system. It's time we focused on keeping our seniors well.

Aged

Bipartite atlas and hypertrophy of its anterior arch. A case report.

On the lateral cervical spine radiographs of a 60-year-old woman complaining of neck pain, diffuse hyperostosis or calcification was present anterior to the anterior arch of the atlas. CT of atlas revealed it to be bipartite and the hyperostosis to be due to hypertrophy of its anterior arch. We report this as a new sign associated with a bipartite atlas.

Calcinosis

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Humans

Expression of EGFR, HER-2/neu, P53, and PCNA in endometrioid, serous papillary, and clear cell endometrial adenocarcinomas.

Expression of four biologic markers was studied in 69 cases of endometrial cancer to identify their association with cell type, decreased survival, and increased tumor metastasis. Cell types included endometrioid (n = 45), serous papillary (n = 16), and clear cell (n = 8). Immunohistochemical stains were employed to detect the presence of epidermal growth factor receptor (EGFR), HER-2/neu, p53, and proliferating cell nuclear antigen (PCNA). Analysis revealed that EGFR was expressed in 49%, HER-2/neu in 59%, p53 in 9%, and PCNA in 16% of tumor specimens. HER-2/neu overexpression was significantly associated with depth of myometrial invasion. p53 and PCNA immunoreactivity significantly correlated with nonendometrioid histology, although PCNA was less specific in labeling these less favorable cell types. EGFR immunoreactivity also significantly correlated with nonendometrioid cell types and tumor metastases at time of diagnosis. Seventy-seven percent of patients with metastatic disease were EGFR-positive versus 36% positivity in patients with no evidence of metastases (P < 0.002). For patients with endometrioid adenocarcinoma, evidence of EGFR overexpression decreased survival from 89 to 69% (P < 0.04). In the serous papillary and clear cell category, EGFR positivity decreased survival from 86 to 27% (P < 0.03). EGFR strongly correlates with tumor metastasis and patient survival in endometrial cancer. Altered expression of this oncoprotein may serve as a guide to prognosis and treatment in these patients.

Adenocarcinoma