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Biomedical subjects

J Stevens

Publications and source records attributed to J Stevens.

At least 163 records · Page 9Linked to original sources
PubMed1996

Anterior pericardial tracheoplasty for congenital tracheal stenosis: intermediate to long-term outcomes.

BACKGROUND: Although several techniques for the treatment of long-segment stenosis of the trachea have been reported, including slide tracheoplasty, rib grafting, and use of a pericardial patch, the optimal repair remains controversial because of a lack of midterm to long-term follow-up data. METHODS: To assess the intermediate and long-term outcomes of patients having repair with anterior pericardial tracheoplasty, we reviewed case histories of 12 patients (1984 to present). The median age was 6.7 months (range, 1 to 98 months), and the median weight was 6.0 kg (range, 0.97 to 42 kg). All patients underwent anterior pericardial tracheoplasty through a median sternotomy during partial normothermic cardiopulmonary bypass. An average of 13 tracheal rings (range, five to 23) were divided anteriorly, and a patch of fresh autologous pericardium was used to enlarge the trachea by 1.5 times the predicted diameter for patient age and weight. RESULTS: There was one hospital death, and all but 2 patients are long-term survivors. All but 1 current survivor remain asymptomatic, with no bronchoscopic evidence of airway obstruction or granulation on the pericardial patch. All survivors examined have normal tracheal growth and development, with a median follow-up of 5.5 years (range, 1 to 11 years). CONCLUSIONS: Anterior pericardial tracheoplasty for congenital tracheal stenosis provides excellent results at intermediate to long-term follow-up.

Child↗
PubMed1996

A competitive quantitative polymerase chain reaction assay for bovine transforming growth factor-B1 mRNA.

We have developed a flexible reverse transcription (RT) coupled quantitative polymerase chain reaction (PCR) assay for transforming growth factor beta-1 (TGF-b) mRNA. A deletion mutant cDNA internal standard was prepared from the wild type cDNA and used to normalize intersample PCR efficiency differences. The assay is compatible with samples from cow and other species. Using RT-PCR, we determined that TGF-b mRNA in bovine pulmonary artery endothelial cells is increased by TGF-b 7.5-fold within 6h and remains 4-fold above baseline after 12h. In addition, unlike TGF-b bioactivity, mRNA levels in endothelial cells are not decreased upon exposure of the cells to either glutathione (reduced or oxidized), cysteine, or N-acetylcysteine for 24h.

Animals↗
PubMed1996

Inhibition of neointimal proliferation after coronary angioplasty by low-molecular-weight heparin (clivarine) and polyethyleneglycol-hirudin.

Proliferation of medial smooth muscle cells (SMC) plays a major role in restenosis after coronary angioplasty and can be inhibited by heparin. Platelets stimulate SMC proliferation, and their aggregation after angioplasty can be reduced by the direct thrombin inhibitor hirudin. In a porcine coronary stent-angioplasty model (16 animals, 4 per group), we studied the effect of 3- and 14-day treatment with a novel long-lasting polyethyleneglycol-conjugated (PEG-hirudin, LU 57291), at a dose of 1 mg/kg intravenously (i.v.) and then subcutaneously once daily as compared with chronic low-molecular-weight heparin (LMWH) clivarine at a dose of 150 IU/kg as an intravenous bolus, followed by 10 IU/kg/h i.v. for 24 h, followed by 75 IU/kg twice daily, as compared with acute unfractionated heparin (100 IU/kg) as a control. Sixteen animals were randomly assigned to the four treatment groups. Four weeks after angioplasty, hearts were perfusion-fixed and six slices per angioplasty segment were analyzed for neointimal thickness and neointimal area (% of total vessel cross-sectional area). Maximal neointimal thickness was 1.10 +/- 0.2 mm in the control group (mean +/- SD, n = 9 arteries) and was significantly lower in both PEG-hirudin (0.62 +/- 0.22 and 0.86 +/- 0.18 mm, 11 and 10 arteries) and in clivarine-treated animals (0.75 +/- 0.33 mm, 11 arteries, p < 0.01). Similarly, neointimal area was smaller in PEG-hirudin groups (20 +/- 12 and 21 +/- 12%) and in the clivarine group (24.8 +/- 13.2%) as compared with the control group (41 +/- 17%, p < 0.02). We conclude that PEG-hirudin and clivarine reduce neointimal proliferation in a coronary stent-angioplasty model. Prolonged PEG-hirudin has no better effect than therapy limited to 3 days.

Angioplasty, Balloon, Coronary↗
PubMed1996

Real time quantitative PCR.

We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude). Real-time quantitative PCR is extremely accurate and less labor-intensive than current quantitative PCR methods.

Cell Line↗
PubMed1996

Ketamine and its use in the pig. Recommendations of the Consensus meeting on Ketamine Anaesthesia in Pigs, Bergen 1994. Ketamine Consensus Working Group.

Ketamine, when used as a mono-anaesthetic, does not appear to induce surgical anaesthesia in the pig. The addition of other drugs such as opioids, benzodiazepines and alpha 2-adrenergic agonists deepens anaesthesia and enables major surgery to be performed. A decision-tree for the rational use of ketamine for both short and long-term anaesthesia in the pig under three different levels of procedure severity is presented.

Anesthesia↗
PubMed1996

Pictorial review: the radiological spectrum of vertically-acquired HIV infection.

Anonymous antenatal and newborn screening demonstrates a positive HIV seroprevalence of one in 400 in areas of inner city London. European Collaborative Study data show a vertical transmission rate of 15% and suggests that 23% of infected children will develop clinical manifestations of AIDS before reaching 1 year of age [1], meaning that recognition of the radiological findings in vertically-acquired HIV is important. 75 serial cases of vertically-acquired HIV (age range 6 weeks to 12 years, mean age 35 months) presenting to a national referral centre over 48 months were reviewed; the radiographic findings we describe constitute a core of typical abnormalities together with a selection of the rarer manifestations of vertically-acquired HIV.

AIDS-Related Complex↗
PubMed1996

Predictors of outcome in the quadriparetic nonambulatory myelopathic patient with rheumatoid arthritis: a prospective study of 55 surgically treated Ranawat class IIIb patients.

The functional results of surgery in patients with myelopathic nonambulatory rheumatoid arthritis (Ranawat Class IIIb) are often disappointing, with high rates of postoperative morbidity and mortality. The authors therefore undertook a detailed investigation of a cohort of 55 Ranawat Class IIIb patients (11 men and 44 women) with a mean age of 64.7 years who were recruited prospectively over a 10-year period (1983-1993), to determine what factors may accurately predict a good surgical outcome. Only 14 patients (25.5%) were judged to have had a favorable outcome as determined by an improvement to Ranawat Class I or II or an improvement of at least 0.5 points in the Stanford Health Assessment Questionnaire disability index. The early postoperative mortality rate was high (12.7%) in this group and almost one-quarter of the patients were dead within 6 months. These poor results mirror those already published in the existing literature. Univariate analysis revealed that age (p = 0.02), degree of vertical translocation (p = 0.05), and, more importantly, spinal cord area (p = 0.006) were significant predictors of outcome. Multiple logistic regression analysis showed that spinal cord area (p = 0.026) was, in fact, the major determinant of outcome and, indeed, of long-term survival (p = 0.001). The mean spinal cord area of those patients not achieving a good outcome was 44 mm2. The atlantodens interval (ADI) was not shown to be a significant outcome determinant, which may be explained by the correlation between an increasing vertical translocation and a decreasing ADI (r = 0.4, p = 0.01). Furthermore, as the degree of vertical translocation increased, the space available for the cord was observed to decrease (p = 0.003) commensurate with a reduction in spinal cord area (p = 0.02). Together, these findings strongly argue for earlier surgical intervention, before the development of vertical translocation, permanent neurological damage, and spinal cord atrophy can occur.

Adult↗
PubMed1996

13-year-old female with imperforate hymen.

Imperforate hymen is a rare occurrence in younger women resulting in hydrocolpos and hydrometrocolpos. Though imperforate hymen may be diagnosed in infancy, females ages 9-15 years most often present with the problem. Imperforate hymen usually is a congenital anomaly, but has been reported as a result of sexual abuse. This case report reviews the presentation and treatment of a 13-year-old female with imperforate hymen.

Abdominal Pain↗
PubMed1995

Stimulation of the herpes simplex virus type I protease by antichaeotrophic salts.

The herpes simplex virus type 1 protease is expressed as an 80,000-dalton polypeptide, encoded within the 635-amino acid open reading frame of the UL26 gene. The two known protein substrates for this enzyme are the protease itself and the capsid assembly protein ICP35 (Liu, F., and Roizman, B. (1991) J. Virol. 65, 5149-5156). In this report we describe the use of a rapid and quantitative assay for characterizing the protease. The assay uses a glutathione S-transferase fusion protein containing the COOH-terminal cleavage site of ICP35 as the substrate (GST-56). The protease consists of N0, the NH2-terminal 247 amino acid catalytic domain of the UL26 gene product, also expressed as a GST fusion protein. Upon cleavage with N0, a single 25-mer peptide is released from GST-56, which is soluble in trichloroacetic acid. Using this assay, the protease displayed a pH optimum between 7 and 9 but most importantly had an absolute requirement for high concentrations of an antichaeotrophic agent. Strong salting out salts such as Na2SO4 and KPO4 (> or = 1 M) stimulated activity, whereas NaCl and KCl had no effect. The degree of stimulation by 1.25 M Na2SO4 and KPO4 were 100-150- and 200-300-fold, respectively. Using the fluorescent probe 1-anilino-8-naphthalene sulfonate, the protease was shown to bind the dye in the presence of 1.25 M Na2SO4 or KPO4, but not at low ionic strength or in the presence of 1.25 or 2.2 M NaCl. This binding was most likely at the protease active site because a high affinity cleavage site peptide, but not a control peptide, could displace the dye. In addition to cleaving GST-56, the herpes simplex virus type I protease also cleaved the purified 56-mer peptide. Circular dichroism and NMR spectroscopy showed the peptide to be primarily random coil under physiological conditions, suggesting that antichaeotrophic agents affect the conformation of the substrate as well as the protease.

Acetates↗
PubMed1995

The removal of 2-oxoacyl residues from the N-terminus of peptides and cystatin in non-denaturing conditions.

The N-terminal residue of a protein or peptide may be converted into a 2-oxoacyl group by non-enzymic transamination. This group may then be removed, to obtain the peptide chain shortened by one residue, by treatment with phenylene-1,2-diamine. Hitherto this scission has required a pH of 4-5, but we find that the reaction will proceed well at pH 7 in the presence of concentrated phosphate buffer. We describe a method using reverse-phase HPLC for determining the extent of scission in model peptides; this method also allows products to be isolated and identified. The new scission conditions have been tested by removing the N-terminal residue from cystatin, an inhibitor of cysteine peptidases; electrospray mass spectrometry was used to assess how this protein reacted.

Amino Acid Sequence↗
PubMed1995

Agonist-induced desensitization of dopamine D1 receptor-stimulated adenylyl cyclase activity is temporally and biochemically separated from D1 receptor internalization.

The regulation of the dopamine D1 receptor was investigated by using c-myc epitope-tagged D1 receptors expressed in Sf9 (fall armyworm ovary) cells. Treatment of D1 receptors with 10 microM dopamine for 15 min led to a loss of the dopamine-detected high-affinity state of the receptor accompanying a 40% reduction in the ability of the receptor to mediate maximal dopamine stimulation of adenylyl cyclase activity. After 60 min of agonist exposure, 45 min after the occurrence of desensitization, 28% of the cell surface receptors were internalized into an intracellular light vesicular membrane fraction as determined by radioligand binding and supported by photoaffinity labeling, immunocytochemical staining, and immunoblot analysis. Pretreatment of cells with concanavalin A or sucrose completely blocked agonist-induced D1 receptor internalization without preventing agonist-induced desensitization, indicating a biochemical separation of these processes. Collectively, these findings indicate that the desensitization of D1 receptor-coupled adenylyl cyclase activity and D1 receptor internalization are temporarily and biochemically distinct mechanisms regulating D1 receptor function following agonist activation.

Adenylyl Cyclases↗
PubMed1995

Isolation and mapping of a gene encoding a novel human ADP-ribosylation factor on chromosome 17q12-q21.

A gene encoding a low-molecular-weight GTP-binding protein was isolated from a retinal cDNA library and mapped to human chromosome 17q12-q21. Comparison of the predicted protein with the protein databases revealed striking homology to the family of ADP-ribosylation factors (ARFs), which are thought to be involved in membrane trafficking and protein secretion. The greatest homology observed was with the rat ARF-like 4 protein (ARL4), with which it shared 58% identity, while the more highly conserved human ARF1 and ARF3 proteins each shared 46% identity. Inspection of the predicted new protein showed that it contained each of the six conserved motifs that are required for guanine nucleotide binding and hydrolysis, and thus it is probably a novel ARF isoform. We have designated the new protein and its corresponding gene ARF4L.

ADP-Ribosylation Factor 1↗
PubMed1995

Genomic organization of the neurofibromatosis 1 gene (NF1).

Neurofibromatosis 1 maps to chromosome band 17q11.2, and the NF1 locus has been partially characterized. Even though the full-length NF1 cDNA has been sequenced, the complete genomic structure of the NF1 gene has not been elucidated. The 5' end of NF1 is embedded in a CpG island containing a NotI restriction site, and the remainder of the gene lies in the adjacent 350-kb NotI fragment. In our efforts to develop a comprehensive screen for NF1 mutations, we have isolated genomic DNA clones that together harbor the entire NF1 cDNA sequence. We have identified all intron-exon boundaries of the coding region and established that it is composed of 59 exons. Furthermore, we have defined the 3'-untranslated region (3'-UTR) of the NF1 gene; it spans approximately 3.5 kb of genomic DNA sequence and is continuous with the stop codon. Oligonucleotide primer pairs synthesized from exon-flanking DNA sequences were used in the polymerase chain reaction with cloned, chromosome 17-specific genomic DNA as template to amplify NF1 exons 1 through 27b and the exon containing the 3'-UTR separately. This information should be useful for implementing a comprehensive NF1 mutation screen using genomic DNA as template.

Amino Acid Sequence↗
PubMed1995

Obesity, fat patterning and cardiovascular risk.

Investigators have refined the study of the relationship of adiposity to disease by differentiating body shapes on the basis of the physical location of adipose tissue on the body. Different combinations of anthropometric measurements have been investigated as indicators of fat distribution. Recently combinations of easily obtained anthropometric measurements have been studied as correlates of visceral fat as measured by more complex measures such as computed tomography and magnetic resonance imaging. Despite difficulties in the measurement of the regional distribution of adipose tissue, several studies have shown that an upper body, centralized or abdominal fat pattern is correlated with increased CVD risk factors and CVD risk. The majority of studies have been cross-sectional, although several prospective cohort studies have examined relationships between fat distribution and mortality or incidence of morbidity over rather long time intervals. Results from the Charleston Heart Study indicate these relationships may differ by race and gender. More research is needed to define the role of fat patterning and changes in fat patterning on disease. Also, the relationship between total fatness and fat distribution to morbidity and mortality in specific populations and ethnic groups needs further exploration.

Adipose Tissue↗
PubMed1995

Demonstration that bone mineral density of the lumbar spine, trochanter, and femoral neck is higher in black than in white young men.

The incidence of osteoporosis and fractures of the hip and spine is lower in black than in white subjects. To determine whether bone mass is increased in black men and to assess the influence of body weight and age, bone mineral density (BMD) of the lumbar spine, trochanter, and femoral neck was measured by dual-photon absorptiometry in 59 normal white men and 40 black men between the ages of 20 and 50 years. Body weight and age were not different from each other in the two groups. BMD of the midradius was measured by single-photon absorptiometry. Multivariate regression was used for independent analysis of each group and for analysis of the two groups together. After adjusting for body weight, age was inversely related to BMD of the femoral neck in both blacks and whites and of the trochanter in blacks. When body weight was analyzed independently of age, it was a positive predictor for BMD of the midradius of black men and of the femoral neck in white men. Despite the racial differences in age and weight on BMD, there were no significant interactions between race and age or race and weight when the data from black and white men were combined. Race had a highly significant effect on BMD of the lumbar spine, trochanter, and femoral neck midradius, and BMD was higher in blacks than in whites at these sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorptiometry, Photon↗