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Biomedical subjects

K Barnes

Publications and source records attributed to K Barnes.

At least 37 records · Page 2Linked to original sources
PubMed2001

Therapeutic efficacy of sulfadoxine-pyrimethamine in uncomplicated Plasmodium falciparum malaria 3 years after introduction in Mpumalanga.

OBJECTIVES: To assess therapeutic efficacy of sulfadoxine-pyrimethamine (SP) in treatment of uncomplicated Plasmodium falciparum malaria 3 years after introduction in Mpumalanga, South Africa. SETTING: Tonga district with a population of 116,418 and subject to seasonal malaria, with an average annual incidence of 3,200 cases. SUBJECTS: One hundred and nineteen malaria patients presenting to a sentinel surveillance clinic and recruited according to World Health Organisation (WHO) criteria. METHODOLOGY: Patients satisfying WHO inclusion criteria were treated with a single oral dose of SP and the response of infection to treatment in each patient was routinely monitored clinically and parasitologically on days 1, 2, 3, 7, 14, 21, 28 and 42 post-treatment. One hundred and ten patients completed follow-up to day 42 or evidence of clinical or parasitological failure. RESULTS: The cure rate at day 42 was 93.6% (103/110). Two patients (1.8%, RII) were early treatment failures on day 3, while recrudescence (4.5%, RI) occurred in 5 patients on day 28 (N = 3) and on day 42 (N = 2). CONCLUSION: In Mpumalanga P. falciparum remains sensitive to SP, with no significant difference between the baseline cure rate (94.5%) and the cure rate in the present study (93.6%).

Adolescent↗
PubMed2000

The vesicle-to-micelle transition of phosphatidylcholine vesicles induced by nonionic detergents: effects of sodium chloride, sucrose and urea.

The vesicle-to-micelle transition of egg phosphatidylcholine LUVs induced by octylglucoside was studied in buffers with 0-4 M sodium chloride, sucrose or urea. We used both light scattering and fluorescent probes to follow the lipid-detergent complexes in these buffers. The vesicle-to-micelle transition process was fundamentally the same in each solute. However, the detergent-to-lipid ratio required for micelle formation shifted in ways that depended on the aqueous solute. The partitioning of octylglucoside between the vesicles and the aqueous phase was primarily determined by the change in its critical micelle concentration (cmc) induced by each solute. Specifically, the cmc decreased in high salt and sucrose buffers but increased in high concentrations of urea. Cmc for two additional nonionic detergents, decyl- and dodecyl-maltoside, and three zwittergents (3-12, 3-14 and 3-16) were determined as a function of concentration for each of the solutes. In all cases NaCl and sucrose decreased the solubility of the detergents, whereas urea increased their solubilities. The effects clearly depended on acyl chain length in urea-containing solutions, but this dependence was less clear with increasing NaCl and sucrose concentrations. The contributions of these solutes to solubility and to interfacial interactions in the bilayers, pure and mixed micelles are considered.

Buffers↗
PubMed2000

A first trial of retrospective collaboration for positional cloning in complex inheritance: assay of the cytokine region on chromosome 5 by the consortium on asthma genetics (COAG).

The central problem of complex inheritance is to map oligogenes for disease susceptibility, integrating linkage and association over samples that differ in several ways. Combination of evidence over multiple samples with 1,037 families supports loci contributing to asthma susceptibility in the cytokine region on 5q [maximum logarithm of odds (lod) = 2.61 near IL-4], but no evidence for atopy. The principal problems with retrospective collaboration on linkage appear to have been solved, providing far more information than a single study. A multipoint lod table evaluated at commonly agreed reference loci is required for both collaboration and metaanalysis, but variations in ascertainment, pedigree structure, phenotype definition, and marker selection are tolerated. These methods are invariant with statistical methods that increase the power of lods and are applicable to all diseases, motivating collaboration rather than competition. In contrast to linkage, positional cloning by allelic association has yet to be extended to multiple samples, a prerequisite for efficient combination with linkage and the greatest current challenge to genetic epidemiology.

Asthma↗
PubMed2000

The neprilysin family in health and disease.

The mammalian neprilysin (NEP) family comprises at least seven members: NEP itself, Kell blood group antigen (KELL), the endothelin-converting enzymes (ECE-1 and ECE-2), the enzyme PEX, associated with X-linked hypophosphataemia, "X-converting enzyme" (XCE) a CNS-expressed orphan peptidase and a soluble, secreted endopeptidase (SEP). These zinc metallopeptidases are all type II integral membrane proteins. Where identified, these enzymes have roles in the processing or metabolism of regulatory peptides and therefore represent potential therapeutic targets. A distinct feature of ECE-1 species is their existence as distinct isoforms differing in their N-terminal cytoplasmic tails. These tails play a role in enzyme targeting and turnover with di-leucine and tyrosine-based motifs affecting localization. Additional anchorage of these enzymes can also occur through palmitoylation. Bacterial homologues of the neprilysin family exist, for example the products of the pepO genes from L. lactis and S. parasanguis, and a recently described gene product of P. gingivalis which is an ECE-1 homologue that can catalyse the conversion of big endothelin to endothelin. A genomics based approach to understanding the functions of this proteinase family is aided by the completion of the C. elegans and Drosophila genomes, both of which encode multiple copies of NEP-like enzymes.

Acylation↗
PubMed2000

Functional significance of the isoforms of endothelin-converting enzyme-1.

The subcellular localization of endothelin-converting enzyme-1 (ECE-1) is a matter of some controversy, further complicated by the discovery of its multiple isoforms. ECE-1 is a critical enzyme in the biosynthesis of the potent vasoconstrictor peptide endothelin (ET), and, as such, represents a potential target for drug therapy in the control of disease states involving the ET system. Knowledge of the precise locations of the isoforms and their regulation would aid in the design of drugs to target specifically ECE-1. In this study, the subcellular localization and potential targeting pathways of the ECE-1 isoforms were investigated. Antipeptide antibodies were raised to the unique N-terminal sequence of ECE-1b and were then used in the investigation of its subcellular distribution. Mutagenesis of proposed targeting sequences within the cytoplasmic tails of the isoforms was carried out to determine their significance in subcellular localization.

Animals↗
PubMed1999

Impact of community volunteers on immunization rates of children younger than 2 years.

OBJECTIVE: To assess the effectiveness of a volunteer-driven outreach program on immunization rates in children younger than 2 years. DESIGN: Randomized controlled trial. SETTING: Pediatric ambulatory clinics in New York, NY. PATIENTS: A total of 163 children were randomly assigned to receive services from the volunteer-driven outreach program or to serve as control subjects. All children were (1) younger than 2 years, (2) no-shows for a scheduled appointment in the pediatric clinic, and (3) due or overdue for a vaccine. INTERVENTION: Immunization outreach, tracking, and follow-up were provided by community volunteers throughout follow-up (mean, 6.5 months). Control children were notified of immunization status at enrollment but received no further contact until the conclusion of follow-up (mean, 6.4 months). MAIN OUTCOME MEASURE: Immunization status 6 months after enrollment. RESULTS: Significantly more intervention children were up-to-date with their vaccination series than controls (75% vs. 54%; P = .03). Children in the control group were 2.8 times more likely to be late for a vaccine than intervention children (odds ratio = 2.8; P = .02). In addition, an immunization delay of longer than 30 days at enrollment was a significant predictor of final immunization delay (odds ratio = 2.6; P = .02). CONCLUSIONS: This volunteer-driven program significantly improved immunization rates among intervention children compared with controls. Results confirm previous findings that indicate an increased risk of an incomplete immunization series by 2 years of age among children who fall behind early in their primary vaccination series. However, control children were almost 3 times more likely to be late (for > or =1 vaccines) than intervention children, regardless of whether an earlier immunization delay was present.

Ambulatory Care↗
PubMed1999

Endothelin converting enzyme is located on alpha-actin filaments in smooth muscle cells.

OBJECTIVE: Endothelin converting enzyme is the key enzyme in the generation of endothelin-1 from big-endothelin-1. The mature endothelin-1 is a potent vasoconstrictor which also promotes mitogenesis and proliferation of smooth muscle cells. The objectives were to demonstrate in smooth muscle cells the presence of a phosphoramidon-sensitive endothelin converting enzyme activity, reveal the subcellular localization of the enzyme protein and determine the effects of the metalloproteinase inhibitor, phosphoramidon, the lysosomotrophic drug, chloroquine, and colchicine on the cycling pathway of the enzyme. METHODS: Subcellular localization of endothelin converting enzyme on human smooth muscle cells and the rat cell line, A7r5, was by immunofluorescence and confocal microscopy or by biotinylation of cell cultures and immunoblotting, after treatment of cell cultures with cytochalasin D, colchicine, chloroquine and phosphoramidon. Converting enzyme activity was determined by high performance liquid chromatographic assay. RESULTS: We detected phosphoramidon-sensitive endothelin converting enzyme activity in smooth muscle cells. In addition to its plasma membrane location, for the first time we revealed a striking co-localization of endothelin converting enzyme and alpha-actin filaments in smooth muscle cells. Colchicine treatment results in a perinuclear accumulation of endothelin converting enzyme. An increased level of endothelin converting enzyme protein was shown to be present in smooth muscle cells which had been grown in the presence of phosphoramidon or chloroquine. CONCLUSION: The 120 kDa endothelin converting enzyme co-localizes with alpha-actin in smooth muscle cells and resembles that found in endothelial cells in that it is present on both the plasma membrane and intracellularly.

Actins↗
PubMed1999

Ochratoxin A in dried vine fruit: method development and survey.

A method is described for the determination of concentrations of the mycotoxin ochratoxin A in dried vine fruits (currants, raisins and sultanas) using acidic methanolic extraction, immunoaffinity chromatography clean-up and HPLC determination. The limit of detection was estimated as 0.2 microgram/kg, and recoveries of 63-77% were achieved at 5 micrograms/kg. HPLC-mass spectrometric confirmation of the identity of ochratoxin was obtained. Ochratoxin A and aflatoxins were determined in 60 samples of retail dried vine fruits purchased in the United Kingdom. Ochratoxin A was found in excess of 0.2 microgram/kg in 19 of 20 currant, 17 of 20 sultana and 17 of 20 raisin samples examined, an overall incidence of 88%. The maximum level found was 53.6 micrograms/kg. No aflatoxin was found in any sample analysed, using a method with a detection limit of 0.2 microgram/kg for each of aflatoxin B1, B2, G1 and G2.

Aflatoxins↗
PubMed1999

Reproductive axis after discontinuation of gonadotropin-releasing hormone analog treatment of girls with precocious puberty: long term follow-up comparing girls with hypothalamic hamartoma to those with idiopathic precocious puberty.

Although the GnRH agonist analogs have become an established treatment for precocious puberty, there have been few long term studies of reproductive function and general health after discontinuation of therapy. To this end, we compared peak LH and FSH after 100 microg sc GnRH, estradiol, mean ovarian volume (MOV), age of onset and frequency of menses, body mass (BMI), and incidence of neurological and psychiatric problems in 2 groups of girls: those with precocious puberty due to hypothalamic hamartoma (HH; n 18) and those with idiopathic precocious puberty (IPP; n = 32) who had been treated with deslorelin (4-8 microg/kg x day, s.c.) or histrelin (10 microg/kg x day, s.c.) for 3.1-10.3 yr and were observed at 1, 2, 3, and 4-5 yr after discontinuation of treatment. The endocrine findings were also compared to those in 14 normal perimenarcheal girls. There were no differences between the HH and IPP groups in age or bone age at the start of treatment, at the end of treatment, or during GnRH analog therapy. We found that whereas the peak LH level was higher in HH than in IPP girls before (165.5 +/- 129 vs. 97.5 +/- 55.7; P < 0.02) and at the end (6.8 +/- 6.0 vs. 3.9 +/- 1.8 mIU/mL; P < 0.05) of therapy, this difference did not persist at any of the posttherapy time points. LH, FSH, and estradiol rose into the pubertal range by 1 yr posttherapy in both HH and IPP. However, the mean posttherapy peak LH levels in both HH and IPP groups tended to be lower than normal, whereas the peak FSH levels were not different from normal, so that the overall posttherapy LH/FSH ratio was decreased compared to that in the normal girls (HH, 2.7 +/- 0.3; IPP, 2.6 +/- 0.1; normal, 5.2 +/- 4.8; P < 0.05). The MOV was larger in HH than IPP at the end of treatment (3.7 +/- 3.5 vs. 2.0 +/- 1.2 mL; P < 0.05) and tended to increase in both groups over time to become larger than that in normal girls by 4-5 yr posttherapy (HH, 14.9 +/- 12.9; IPP, 7.6 +/- 2.2; normal, 5.4 +/- 2.5 mL; P < 0.05). Whereas the onset of spontaneous menses varied widely in both groups, once menses had started, the HH group had a higher incidence of oligomenorrhea. Pelvic ultrasonography revealed more than 10-mm hypoechoic regions in 4 HH patients, 15 IPP patients, and 3 normal girls, all of whom were reporting regular menses. Live births of normal infants were reported by 2 HH and 2 IPP patients, and elective terminations of pregnancy were reported by 1 HH and 2 IPP patients. BMI was greater than normal in HH and IPP both before treatment and at all posttherapy time points and tended to be higher in the HH patients. Marked obesity (BMI, +2 to +5.2 SD score) was observed in 5 HH and 6 IPP patients, 1 of whom had a BMI of +2.5 SD score and developed acanthosis nigricans, insulin resistance, and hyperglycemia. Seizure disorders developed during GnRH analog therapy in 5 HH and 1 IPP patient, and 2 additional HH girls developed severe depression and emotional lability posttherapy. Although the mean anterior-posterior dimension of the hamartoma was larger in the HH patients with seizure than in those who were seizure free (1.7 +/- 1.2 vs. 0.9 +/- 0.4 cm; P < 0.05), no change in hamartoma size was observed either during or after therapy, and no patient has reported the onset of a seizure disorder posttherapy. Other than a tendency toward a larger MOV, a higher incidence of oligomenorrhea, obesity, and frequency of neurological disorders, recovery of the reproductive axis after GnRH analog therapy was not markedly different in HH compared to IPP. Continued follow-up of these patients may determine whether the decreased LH responses and increased BMI in both groups compared to those in normal girls remain clinically significant problems.

Adolescent↗
PubMed1998

Junctioning of lateral and anterior fields in head and neck cancer: a dosimetric assessment of the monoisocentric technique (including reproducibility).

PURPOSE: The matching or junctioning of two lateral fields with an anterior field is commonly performed in the treatment of head and neck cancer. A monoisocentric technique utilising asymmetric collimation is potentially associated with improved dosimetry in the plane of the junction due to decreased reliance on operator skill and the avoidance of couch movement. The aim of this study was not only to assess the average dose delivered in the plane of the junction, but also the reproducibility of this dose for the monoisocentric technique and two other commonly used techniques. METHODS AND MATERIALS: An anthropomorphic head and neck wax phantom was fashioned to allow the placement of 22 TLD chips in a 2-mm thick transverse plane positioned superior to the potential site of the larynx. Three different treatment techniques were used with the phantom being treated by four different operators a minimum of 20 times for each technique: (1) "straight fields"--using isocentric laterals with an anterior field junctioned in the midline. This technique makes no allowance for divergence; (2) "angled fields"--couch and gantry rotation are used to account for divergence; (3) "monoisocentric"--using asymmetric collimators to create a single isocenter. RESULTS: For an applied dose of 1 Gy the monoisocentric technique produced a mean dose measured of 1.01 Gy compared with 1.23 and 0.92 Gy for techniques 1 and 2. The reproducibility of the mean dose measured was better for the monoisocentric technique by a factor of 2. The superior reproducibility of the monoisocentric technique was not found to be operator dependent. CONCLUSIONS: A monoisocentric technique for the treatment of two laterals and an anterior field in head and neck cancer is likely to be associated with more accurate and reproducible dosimetry in the plane of the junction. Our center has subsequently adopted this technique for matching such fields.

Head and Neck Neoplasms↗
PubMed1998

Anti-peptide antibodies specific to rat endothelin-converting enzyme-1 isoforms reveal isoform localisation and expression.

Endothelin-converting enzyme-1 (ECE-1) is a critical enzyme in the biosynthesis of the potent vasoconstrictor peptide endothelin and exists in several isoforms. Anti-peptide antibodies raised against epitopes in the distinct N-terminal cytoplasmic tails of the rat ECE-1 isoforms have been obtained. By using these antibodies in Western blot analysis and immunofluorescence studies, we have shown that cultures of transformed rat lung vascular endothelial cells treated with the metalloprotease inhibitor phosphoramidon and untreated cells express ECE-1alpha only, whereas human umbilical vein endothelial cells express ECE-1alpha and ECE-1beta. The ECE-1 isoforms expressed in CHO-K1 cells transfected with rat cDNA to ECE-1alpha and ECE-1beta could be immunoprecipitated by using the appropriate isospecific antibody.

Amino Acid Sequence↗
PubMed1998

Endothelin-converting enzyme: ultrastructural localization and its recycling from the cell surface.

The potent vasoconstrictor endothelin-1 (ET-1) is secreted constitutively by endothelial cells and has been implicated in the pathophysiology of several cardiovascular diseases. It is generated from its inactive intermediate, big ET-1, through the action of endothelin-converting enzyme (ECE). Using several complementary techniques, we have demonstrated that ECE is present at the cell surface and on intracellular vesicles and that it recycles from the cell surface in endothelial cells. This is the first ultrastructural localization of ECE in lung and the first time big ET-1 and ECE have been colocalized by immunogold in a vesicular population, 50 to 100 nm in diameter. In addition, by double immunogold staining of ultrathin cryosections, we have localized ECE together with angiotensin-converting enzyme on the luminal membrane of endothelial cells. With cell surface biotinylation of a transformed rat endothelial cell line and of human umbilical vein endothelial cells, we have confirmed the presence of ECE on the plasma membrane. After treatment of endothelial cells with chloroquine, ECE and trans-Golgi network 38 protein were shown by immunofluorescence staining to localize to the same intracellular compartment.

Animals↗
PubMed1998

Novel CYP11B1 mutations in congenital adrenal hyperplasia due to steroid 11 beta-hydroxylase deficiency.

The second most common cause of congenital adrenal hyperplasia is 11 beta-hydroxylase deficiency, an autosomal recessive disorder. We performed genetic analysis of CYP11B1, the gene encoding steroid 11 beta-hydroxylase, in three patients with classic 11 beta-hydroxylase deficiency. Herein we describe the first splice donor site mutation, a new nonsense mutation, and a new missense mutation in this disorder. An African-American patient was found to be a compound heterozygote for a codon 318 + 1G --> A substitution at the 5'-splice donor site of intron 5, in combination with Q356X, a nonsense mutation previously reported in an African-American patient. A Caucasian patient was found to be a compound heterozygote with a novel missense mutation, T318R, in combination with a previously reported 28-bp deletion in exon 2. A different mutation at codon 318 (T318M) has been described previously. A Caucasian patient was heterozygous for a novel nonsense mutation (Q19X) in exon 2. The second mutation was not identified in this patient. Multiple apparent polymorphisms were also observed. Two of these polymorphisms in CYP11B1 represent sequences from CYP11B2, suggesting that gene conversion may have occurred. In summary, we have identified three novel mutations and two previously reported mutations in CYP11B1 patients with 11 beta-hydroxylase deficiency. Our data suggest the presence of a mutational hot spot at codon 318 of CYP11B1, and the possibility of a founder effect in frequently identified mutations.

Adrenal Hyperplasia, Congenital↗