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W King

Publications and source records attributed to W King.

At least 73 records · Page 4Linked to original sources

Human vascular endothelial cells process and present autoantigen to human T cell lines.

The effectiveness of cultured human umbilical vein endothelial cells as accessory cells for T cell activation has been investigated using T cell clones and lines derived from patients with myasthenia gravis which were specific for different epitopes on the alpha subunit of the human acetylcholine receptor. The endothelial cells were induced with IFN-gamma to express HLA-DR and -DQ at high and low levels respectively. They could then efficiently present specific peptides of the alpha subunit to an HLA-DR- and an HLA-DQw5-restricted T cell line. They could also process epitopes for both T cell lines from the full-length recombinant alpha subunit (r1-437) of the human acetylcholine receptor, where the known epitopes are 80 amino acid residues apart. The endothelial presentation of r1-437, but not of the peptides, was sensitive to chloroquine inhibition. Presentation appeared slightly less efficient (by 1.5- to 3.0-fold) with endothelial cells than with presenting cells from peripheral blood. This may reflect differences in accessory signalling since mAb blocking studies suggested that ligands for CD28 provided important accessory signalling by peripheral blood presenting cells while LFA-3 was used by endothelial cells.

Antigen Presentation↗

Comparison of characteristics of Q beta replicase-amplified assay with competitive PCR assay for Chlamydia trachomatis.

In order to study infections due to Chlamydia trachomatis, we have compared semiquantitative PCR and Q beta replicase-amplified assays for detection of this organism. The PCR assay was directed against the C. trachomatis 16S rRNA gene. Quantitation was accomplished by adding known amounts of a plasmid containing a truncated segment of the 16S rRNA gene target to chlamydia-containing samples and then amplifying with a common primer set. The Q beta replicase assay consisted of reversible target capture of C. trachomatis 16S rRNA, which was followed by amplification of an RNA detector probe in the presence of the enzyme Q beta replicase. In a clinical matrix, the lower limit of detection of both the PCR and Q beta replicase assays was five elementary bodies. The Q beta replicase and PCR assays were quantitative over 10,000- and 1,000-fold ranges of organisms, respectively. Analysis of the effects of endocervical matrix on amplification was accomplished by examining 94 endocervical specimens by each technique. Both assays detected five of six culture-confirmed specimens as well as three culture-negative specimens. PCR inhibitors were detected in 13 specimens. The Q beta replicase assay, in contrast, showed no evidence of sample inhibition. The Q beta replicase and PCR assays should allow quantitative investigation of infections due to C. trachomatis. In addition, because it targets highly labile RNA, the Q beta replicase assay may facilitate investigations into the role of active persisting infection in culture-negative inflammatory conditions.

Base Sequence↗

Detection of Mycobacterium tuberculosis directly from spiked human sputum by Q-beta replicase-amplified assay.

We report on a rapid, sensitive, Q-Beta replicase-amplified nucleic acid hybridization assay for the detection of Mycobacterium tuberculosis directly from spiked human sputum. Specimens were processed by either an N-acetyl-L-cysteine-NaOH or a 2% NaOH digestion-decontamination method and then washed to neutralize the pH of the cell pellet. The washed sputum pellets were heated at 100 degrees C to inactivate the M. tuberculosis organisms. The heat-inactivated samples were mechanically lysed at 5,000 rpm for 6 min in the GENE-TRAK Sample Processing Instrument in the presence of zirconium oxide beads and a buffer containing guanidine thiocyanate. The released nucleic acid was subjected to the GENE-TRAK Q-Beta replicase-amplified, dual-capture assay. The assay sensitivity was 10(3) purified rRNA targets or 1 CFU of M. tuberculosis spiked into M. tuberculosis-negative human sputum. There was a low level of noise because of the limitations of performing a signal amplification assay in an open system. High levels of other mycobacterial rRNA (approximately 10(7) organisms), including rRNAs of Mycobacterium avium and Mycobacterium gordonae, did not interfere with the sensitivity of the assay.

Bacteriological Techniques↗

Comparison of amplified Q beta replicase and PCR assays for detection of Mycobacterium tuberculosis.

Because of the long time required to isolate Mycobacterium tuberculosis in culture, there is an acute need for simple rapid methods for direct detection of M. tuberculosis from human sputum specimens. We have developed and characterized quantitative manual Q beta replicase and PCR assays for M. tuberculosis. The Q beta replicase assay was based on reversible target capture of M. tuberculosis 23S rRNA followed by amplification of a replicatable detector probe with Q beta replicase. For PCR assays, primers generating a 370-bp amplification product from the IS6110 insertion element were used in combination with a control plasmid containing an internal deletion in the IS6110 amplicon. Serial dilutions of M. tuberculosis were spiked into sputum and subjected to digestion and decontamination with N-acetyl-L-cysteine and NaOH. Assay conditions were optimized for hybridization and sample processing chemistries in order to maximize sample utilization. Following assay optimization, the sensitivities of the Q beta replicase and PCR assays of spiked sputum samples were 0.5 and 5.0 CFU per assay reaction, respectively. The effects of sputum matrix on each assay were examined by testing 20 patient sputum samples which had been cultured for M. tuberculosis. The culture-positive samples included smear-positive and smear-negative samples. The results of the Q beta replicase assay were not inhibited by sputum and were in 100% agreement with those of culture, including detection of 10 culture-positive specimens. However, using an internal control plasmid coamplified with each PCR as an indicator, we detected PCR inhibition in 9 of 20 samples tested. Decreasing the amount of sample assayed in the PCR 24-fold alleviated the inhibitory effects in all but two specimens, one of which was culture positive. The decreased sample utilization also resulted in a false-negative result with a third specimen which was culture positive for M. tuberculosis. Quantitative smear results and QB replicase assay estimates of the number of organisms present in these specimens were in close agreement. The QB replicase assay performed well in comparison with both culture and PCR and should offer a rapid means for detecting and controlling infection due to M. tuberculosis.

Bacteriological Techniques↗

Q-beta replicase-amplified assay for detection of Mycobacterium tuberculosis directly from clinical specimens.

We report the results of a study conducted to evaluate the performance of manual Q-Beta replicase-amplified Mycobacterium tuberculosis complex assay compared with that of culture for detecting M. tuberculosis directly from digested sputum pellets. A total of 261 specimens submitted to three tuberculosis testing laboratories were analyzed. Culture and acid-fast bacillus smear results were provided by the tuberculosis testing laboratories. Of these 261 specimens, 34 (13% prevalence rate) were positive for M. tuberculosis by culture. The samples were digested and decontaminated by the testing laboratories by using their standard digestion and decontamination procedures. An aliquot of the digested and decontaminated pellet was sent to GENE-TRAK. The digested and decontaminated pellet was neutralized by washing it with 0.067 M phosphate buffer (pH 6.8), and the bacteria present in the washed pellet were heat inactivated at 100 degrees C for 15 min. The samples were combined with sample processing buffer containing GuSCN and were treated for 6 min in the GENE-TRAK Sample Processing Instrument to release the nucleic acids. The release rRNA was analyzed in a manual Q-Beta replicase assay format which incorporates elements of sandwich hybridization, reversible target capture, and Q-beta replicase signal amplification technologies. In comparison with culture, the overall assay sensitivity and specificity were 97.1 and 96.5%, respectively. The positive predictive value was 80.5%, and the negative predictive value was 99.5%. After analysis of discrepant results, the assay sensitivity and specificity were 97.3 and 97.8, respectively, and the prevalence rate was 14%. The positive predictive value and the negative predictive value were 87.8 and 99.5%, respectively. The Q-Beta replicase assay is rapid sensitive, semiquantitative, and specific for the direct detection of M. tuberculosis from clinical specimens.

Bacteriological Techniques↗

Ultrasonographic demonstration of normal axillary lymph nodes: a learning curve.

High frequency transducers with near field resolution allow visualization of superficial structures in the axilla, such as the lymph nodes, which could not be visualized with older equipment. We have been able to observe normal axillary nodes in 61.5% of 26 women who had histologic correlation of normal lymph nodes at axillary dissection. We studied 663 women with a clinically palpable breast lump using breast and axillary ultrasonography. They were divided into four groups. The first group consisted of all women who had undergone surgery and had histologic correlation of axillary nodes. The second, third, and fourth groups were made up of three consecutive series of 221 women examined. Women from group one were included in groups two, three, and four. A steep learning curve of 7.1 to 41.9 to 64.7% was observed in the detection of normal axillary nodes, suggesting that these appearances can be recognized easily.

Axilla↗

Information. Counting what counts.

The NHS changes are highlighting the need for the collection of meaningful data in the community health services, writes Wendy King. She describes how health visitors can help supercede the simplifications of Korner statistics by helping the NHSE's information management group to devise systems which count what really counts.

Community Health Nursing↗

Plasma levels of gonadal steroids during final oocyte maturation of striped bass, Morone saxatilis L.

Levels of estradiol-17 beta (E2), testosterone (T), 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (DHP), and 17 alpha,20 beta,21-trihydroxy-4-pregnen-3-one (20 beta-S) were measured by radioimmunoassay (RIA) in blood plasma of striped bass undergoing final oocyte maturation (FOM). Females were captured just prior to, or in the early stages of, FOM and induced to complete maturation and ovulation with injected human chorionic gonadotropin, synthetic salmon gonadotropin-releasing hormone analogue (sGnRHa; [D-Arg6-Pro9 NEt]-sGnRH), sGnRHa plus the dopamine receptor antagonist, domperidone (DOM), or OVAPRIM, a commercial preparation of sGnRHa + DOM. Their plasma levels of immunoreactive DHP and 20 beta-S were significantly greater at ovulation relative to the time of hormone injection, whereas the plasma levels of E2 and T were greatest at injection and decreased by ovulation and 24 hr thereafter. Plasma levels of 20 beta-S, but not DHP, were sustained at high levels after ovulation. Fish injected only with DOM did not undergo FOM, its associated changes in plasma steroid levels, or ovulation. In females captured at various natural stages of FOM, plasma levels of 20 beta-S and DHP were low during germinal vesicle migration (GVM), peaked coincident with germinal vesicle breakdown, and then decreased near the time of ovulation. Plasma levels of E2 and T were greatest during GVM and decreased as DHP and 20 beta-S levels increased. Analyses of conjugated versus free plasma steroids showed 64-79% of the various hormones to be in the free fraction. RIA of plasma fractionated by reversed-phase HPLC showed that half of the 20 beta-S immunoreactivity coeluted with 5 beta-pregnan-3 alpha,17,20 beta,21-tetrol, a putative 20 beta-S metabolite with 99.7% cross-reactivity in the 20 beta-S RIA. These results indicate that striped bass follow the typical profile of changing plasma steroid levels seen in other teleosts during FOM, with a clear shift from C18 and C19 steroids to C21 steroids. They suggest that both DHP and 20 beta-S, both potent inducers of striped bass FOM in vitro, may play a role in regulating FOM in this species.

Animals↗

Hormonal regulation of final maturation of striped bass oocytes in vitro.

An in vitro culture system was developed to investigate hormonal control of final oocyte maturation (FOM) in striped bass (Morone saxatilis). Isolated ovarian fragments exposed to human chorionic gonadotropin (hCG), dibutyryl cAMP, or forskolin produced significant amounts of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (DHP) and the oocytes underwent germinal vesicle breakdown (GVBD). Slight increases in 17 alpha,20 beta,21-trihydroxy-4-pregnen-3-one (20 beta-S) production were also observed. Production of testosterone and estradiol-17 beta was relatively high at the beginning of in vitro treatment with hCG but decreased as production of DHP increased and GVBD was initiated. Inhibitors of protein transcription (actinomycin-D), translation (cycloheximide), and steroidogenesis (trilostane) completely blocked hCG-induced DHP and 20 beta-S production and the associated GVBD. FOM, assessed from the progress of GVBD, proceeded in trilostane-treated but not in cycloheximide-treated follicle-enclosed oocytes when DHP or 20 beta-S was added to the cultures. Structure-activity experiments revealed that DHP and 20 beta-S were more potent at inducing GVBD than 14 other structurally similar C21 steroids that were tested. These results demonstrate that FOM in striped bass is induced by gonadotropin-mediated production of a delta 4 steroid through an adenylate-cyclase pathway which requires protein synthesis. DHP and 20 beta-S are implicated as final oocyte maturation-inducing steroid hormones in striped bass.

Animals↗

Role of ultrasonography in thyroid metastases.

The ultrasound features in eight cases of metastases to the thyroid are reported. The primary tumours were bronchogenic carcinoma (4 cases), breast carcinoma (2 cases), oesophageal carcinoma (1 case) and carcinoma of the stomach (1 case). In this series, none of the metastases were from malignant melanoma or a renal neoplasm. Malignant cervical lymphadenopathy was seen in four cases, and five patients had evidence of disseminated disease.

Breast Neoplasms↗

Novel, ultrasensitive, Q-beta replicase-amplified hybridization assay for detection of Chlamydia trachomatis.

A sensitive, nonisotopic hybridization assay termed "dual capture" is described. The assay rapidly and specifically detects very low levels of target nucleic acids and organisms. The assay is based on the principles of sandwich hybridization, reversible target capture, and Q-Beta replicase amplification. The assay can be completed in less than 4 h, and in the described model format, it detects Chlamydia trachomatis rRNA or rDNA. Up to 96 samples can be analyzed simultaneously. The assay employs two types of probes: a test-specific capture probe, which mediates the cycling of the target probe complex on and off derivatized magnetic beads, and a replicatable RNA detector molecule containing a sequence complementary to and adjacent to the capture probe site on the target. Following reversible target capture, detection of the signal is accomplished by replication of the detector molecule by Q-Beta replicase in the presence of propidium iodide. A specific assay signal can be detected from as few as 1,000 molecules above the background. In a limited study of 94 urogenital samples the assay detected five of the six culture-positive samples and did not detect the C. trachomatis target in 85 of the 88 culture-negative samples.

Base Sequence↗

Attitudes to body weight, weight gain and eating behavior in pregnancy.

The eating behavior and attitudes to body weight of 100 healthy women were studied 3 days after the birth of their first child. During pregnancy women 'watch their weight' and use a range of methods of weight control which include cigarette smoking and inducing vomiting. During pregnancy 41 women reported weight control problems and 20 women considered their weight and eating problems to be greater than at any previous time. Picking was the most common unwanted behavior. Binge eating was experienced by 44 women, nine of whom reported it to be a 'severe' problem. Although women were ambivalent about being weighed at each antenatal visit, 81 recommended weighing once each month. The women held differing opinions on the effects of breastfeeding on body weight and on the need for nutritional supplements during pregnancy. Women reporting 'disordered eating' were more likely to have antenatal complications and give birth to low birthweight babies. The results suggest good obstetric care should include a history of the woman's eating behavior and body weight.

Adolescent↗

An educational program for neonatal intensive care unit developmental care.

An innovative educational program on developmental care of the neonate was successfully implemented within the high-tech environment of a Level III NICU. Devised by a multidisciplinary team, the program provided appropriate levels of education to all personnel--from neonatologists to housekeeping staff--involved on a day-to-day basis with the infants. To assist those wishing to implement a similar program in their institution, details of program development, content, implementation, evaluation, and future goals are discussed.

Child Development↗

Quantitative comparison of the stress response of striped bass (Morone saxatilis) and hybrid striped bass (Morone saxatilis x Morone chrysops and Morone saxatilis x Morone americana).

Striped bass (Morone saxatilis) exposed to a standardized confinement stress had markedly different clinical and endocrinologic responses, compared with hybrid striped bass exposed to the same stress. Plasma cortisol concentration increased at a faster rate and appeared to reach a higher value in striped bass than in hybrid bass. Mean plasma cortisol concentration was 742 +/- 43 ng/ml in striped bass, compared with 490 +/- 37 and 531 +/- 40 ng/ml in striped bass x white perch (M americana) and striped bass x white bass (M chrysops) hybrids, respectively, after a 45-minute net confinement. Plasma cortisol concentration also remained significantly (P = 0.003) higher in striped bass for at least 48 hours after the net confinement. These hormonal differences were associated with a markedly lower survival and resistance to infection in striped bass, compared with the hybrids.

Analysis of Variance↗

A comparison of the iron-clearing properties of parabactin and desferrioxamine.

A comparative study of the iron-clearing properties of subcutaneously administered desferrioxamine and parabactin is presented. The evaluation was performed in both a non-iron-overloaded bile-duct cannulated rat and an iron-loaded Cebus monkey model. Parabactin was superior to desferrioxamine in both the rodent and the primate. Unlike desferrioxamine, nearly all of the parabactin-induced iron clearance occurred in the bile. This study represents an evaluation of the iron-clearing properties of parabactin, a hexacoordinate catecholamide siderophore.

Animals↗

A comparative study of the iron-clearing properties of desferrithiocin analogues with desferrioxamine B in a Cebus monkey model.

A comparative study of the iron-clearing properties of subcutaneously administered desferrioxamine B (DFO) with those of orally administered desferrithiocin sodium salt (1), desmethyl desferrithiocin (2), desazadesmethyl desferrithiocin sodium salt (3), desazadesmethyl desferrithiocin pivaloyloxymethyl ester (4), and desazadesmethyl-5,5-dimethyl desferrithiocin (5) in an iron-loaded Cebus monkey model and a non-iron overloaded bile duct-cannulated rat model is presented. All six drugs, which performed well in rodent studies, demonstrated increased efficiency in the Cebus monkey model. When administered to rodents at a daily dosage of 384 mumol/kg over a period of 10 days, drug 1 demonstrated severe renal toxicity. whereas drugs 3, 4, and 5 exhibited severe gastrointestinal (GI) toxicity. Under the same experimental protocol, drug 2 did not show significant toxic side effects. In addition, to further evaluate the iron-clearing properties of analogue 2, a dose-response study was performed in the primates that showed that iron excretion increased in a dose-dependent fashion.

Animals↗

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Organizational Innovation↗