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Biomedical subjects

D B Williams

Publications and source records attributed to D B Williams.

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

Effects of HRT and exercise training on insulin action, glucose tolerance, and body composition in older women.

The independent and combined effects of exercise training and hormone replacement therapy (HRT) on body composition, fat distribution, glucose tolerance, and insulin action were studied in postmenopausal women, aged 68 +/- 5 yr, assigned to control (n = 19), exercise (n = 18), HRT (n = 15), and exercise + HRT (n = 16) groups. The exercise consisted of 2 mo of flexibility exercises followed by 9 mo of endurance exercise. HRT was conjugated estrogens 0.625 mg/day and trimonthly medroxyprogesterone acetate 5 mg/day for 13 days. Total and regional body composition were measured by dual-energy X-ray absorptiometry. Serum glucose and insulin responses were measured during a 2-h oral glucose tolerance test. There were significant main effects of exercise on reductions in total and regional (trunk, arms, legs) fat mass, increase in leg fat-free mass, and improvements in glucose tolerance and insulin action. There were significant main effects of HRT on the reduction of total fat mass (HRT, -3.0 +/- 4.0 kg; no HRT, -1.3 +/- 2.6 kg), with a strong trend for reductions in trunk and leg fat mass (both P = 0.07). There was also a significant improvement in insulin action in response to HRT. These results suggest that there are independent and additive effects of exercise training and HRT on the reduction in fat mass and improvement in insulin action in postmenopausal women; the effect of HRT on insulin action may be mediated, in part, through changes in central adiposity.

Adipose Tissue↗
PubMed2000

Prevalence of Huntington disease in New South Wales in 1996.

OBJECTIVE: To estimate the prevalence of Huntington disease (HD) in New South Wales on Australian Census Day (6 August) 1996. DESIGN: Survey of records of the Huntington Disease Service and major hospitals, and of neurologists, psychiatrists, clinical geneticists and genetic counsellors. SUBJECTS AND SETTING: All patients in NSW who, on Census Day 1996, either had a definite diagnosis of HD (motor signs of chorea or ataxia and family history of HD or positive DNA test result) or would have had signs and later received a definite diagnosis (assessed 1 April 1997 to 1 July 1999). MAIN OUTCOME MEASURES: Prevalence (HD patients per 100,000 population); patient characteristics; year and basis of diagnosis. RESULTS: 380 patients with definite HD were identified, giving a prevalence of HD in NSW in 1996 of 6.29 per 100,000 population (95% CI, 5.68-6.96). A third of HD patients were aged 60 years or older. Diagnosis was confirmed by DNA testing for 171 patients (45%), including 30 (8%) with no recorded family history. Average numbers of new diagnoses per year were 11.8 (1984-1988), 21.8 (1989-1993) and 28.6 (1994-1998). Estimated number of people with a 50% risk of inheriting the HD mutation was 25.2 per 100,000 population. Estimated incidence of HD in 1996 was 0.65 per 100,000 population. CONCLUSIONS: Prevalence of HD in NSW is similar to estimated prevalence in other Australian and Western populations. Increasing numbers of cases are being diagnosed, and the 18 chronic care beds currently designated for HD patients in NSW are unlikely to be sufficient.

Adult↗
PubMed2000

Functional relationship between calreticulin, calnexin, and the endoplasmic reticulum luminal domain of calnexin.

Calnexin is a membrane protein of the endoplasmic reticulum (ER) that functions as a molecular chaperone and as a component of the ER quality control machinery. Calreticulin, a soluble analog of calnexin, is thought to possess similar functions, but these have not been directly demonstrated in vivo. Both proteins contain a lectin site that directs their association with newly synthesized glycoproteins. Although many glycoproteins bind to both calnexin and calreticulin, there are differences in the spectrum of glycoproteins that each binds. Using a Drosophila expression system and the mouse class I histocompatibility molecule as a model glycoprotein, we found that calreticulin does possess apparent chaperone and quality control functions, enhancing class I folding and subunit assembly, stabilizing subunits, and impeding export of assembly intermediates from the ER. Indeed, the functions of calnexin and calreticulin were largely interchangeable. We also determined that a soluble form of calnexin (residues 1-387) can functionally replace its membrane-bound counterpart. However, when calnexin was expressed as a soluble protein in L cells, the pattern of associated glycoproteins changed to resemble that of calreticulin. Conversely, membrane-anchored calreticulin bound to a similar set of glycoproteins as calnexin. Therefore, the different topological environments of calnexin and calreticulin are important in determining their distinct substrate specificities.

Animals↗
PubMed2000

Selection of an immunogenic peptide mimic of the capsular polysaccharide of Neisseria meningitidis serogroup A using a peptide display library.

The presently available meningococcal vaccine is poorly immunogenic in infants and fails to induce long-lasting immunity in adults. Efforts to convert this TI-2 type vaccine into a T dependent vaccine are being actively pursued and include conjugate vaccine development. Alternatively, the meningococcal polysaccharide can be rendered into a T dependent antigen through the use of peptides which mimic the capsular polysaccharide complexed or conjugated to potent protein carrier molecules. We have previously developed an anti-idiotypic monoclonal antibody (mAb) based peptide mimic of meningococcal group C polysaccharide (MCPS). A direct approach to identification of peptide mimics of antigen is through the use of peptide display libraries. We have utilized a phage library and a mAb with specificity for meningococcal group A polysaccharide (MAPS) to screen for a peptide mimic of MAPS. Six different peptide motifs were selected with the use of the mAb. Thirty-eight of the 60 sequenced phage clones were represented by motif 1 and 2 which differed only in three amino acids at the carboxy terminus. Immunological assays were performed. Phage clones with motif 1 and 2 were capable of binding human hyperimmune sera and inhibiting the binding of human hyperimmune sera to nominal antigen. Immunization with motif 1 peptide complexed to proteosomes resulted in an anti-MAPS antibody response. Priming with the peptide proteosome complex induced an anamnestic response indicating the formation of immunological memory.

Amino Acid Sequence↗
PubMed2000

Electrical impedance imaging at multiple frequencies in phantoms.

We have recently built and tested a 32 channel, multi-frequency (1 kHz to 1 MHz) voltage mode system to investigate electrical impedance spectroscopy (EIS) imaging. We completed a series of phantom experiments to define the baseline imaging performance of our system. Our phantom consisted of a plastic circular tank (20 cm diameter) filled with 0.9% aqueous NaCl solution. Conductors and nonconductors of decreasing width (W5: 3.4 cm, W4: 2.54 cm, W3: 0.95 cm, W2: 0.64 cm and WI: 0.32 cm) were positioned at various distances from the tank edge (1 cm, 2 cm, 4 cm and 8 cm). The results suggest that the detection of objects less than 1 cm in width is limited to the first 1 to 2 cm from the tank edge for absolute images, but this depth can extend to 8 cm in difference images. Larger 3.4 cm wide objects can be detected in absolute images at depths up to 8 cm from the tank edge. Generally, conductor images were clearer than their nonconductor counterparts. Not only did electrode artefacts lessen as the frequency increased, but the system's maximum resolution was attained at the highest operating frequencies. Although the system recovered the value of the electrical conductivity at the correct order of magnitude, it tended to smooth out large property discontinuities. The calculated electrical permittivity in these phantom studies was inconclusive due to the presence of electrode artefacts.

Electric Impedance↗
PubMed2000

Multifrequency electrical impedance imaging: preliminary in vivo experience in breast.

We have deployed a recently completed spectroscopic electrical impedance tomography (EITS) imaging system in a small series of women (13 participants accrued to date) in order to investigate the feasibility of delivering EITS breast examinations on a routine basis. Hardware is driven with sinusoidally varying spatial patterns of applied voltage delivered to 16 electrodes over the 10 kHz to 1 MHz spectral range using a radially translating interface which couples the electrodes to the breast through direct contact. Imaging examinations have consisted of the acquisition of multi-channel measurements at ten frequencies on both breasts. Participants lie prone on an examination table with the breast to be imaged pendant in the electrode array that is located below the table. Examinations were comfortable and easy to deliver (about 10 minutes per breast including electrode-positioning time). Although localized near-surface electrode artefacts are evident in the acquired images, several findings have emerged. Permittivity images have generally been more informative than their conductivity counterparts, except in the case of fluid-filled cysts. Specifically, the mammographically normal breast appears to have characteristic absolute EITS permittivity and conductivity images that emerge across subjects. Structural features in the EITS images have correlated with limited clinical information available on participants with benign and malignant abnormality, cysts and scarring from previous lumpectomy and follow-up radiation therapy. Several cases from this preliminary experience are described.

Breast↗
PubMed2000

A multichannel continuously selectable multifrequency electrical impedance spectroscopy measurement system.

There is increasing evidence that alterations in the electrical property spectrum of tissues below 10 MHz is diagnostic for tissue pathology and/or pathophysiology. Yet, the complexity associated with constructing a high-fidelity multichannel, multifrequency data acquisition instrument has limited widespread development of spectroscopic electrical impedance imaging concepts. To contribute to the relatively sparse experience with multichannel spectroscopy systems this paper reports on the design, realization and evaluation of a prototype 32-channel instrument. The salient features of the system include a continuously selectable driving frequency up to 1 MHz, either voltage or current source modes of operation and simultaneous measurement of both voltage and current on each channel in either of these driving configurations. Comparisons of performance with recently reported fixed-frequency systems is favorable. Volts dc (VDC) signal-to-noise ratios of 75-80 dB are achieved and the noise floor for ac signals is near 100 dB below the signal strength of interest at 10 kHz and 60 dB down at 1 MHz. The added benefit of being able to record multispectral information on source and sense signal amplitudes and phases has also been realized. Phase-sensitive detection schemes and multiperiod undersampling techniques have been deployed to ensure measurement fidelity over the full bandwidth of system operation.

Calibration↗
PubMed2000

Benzodiazepines induce a conformational change in the region of the gamma-aminobutyric acid type A receptor alpha(1)-subunit M3 membrane-spanning segment.

Benzodiazepine binding to gamma-aminobutyric acid type A (GABA(A)) receptors allosterically modulates GABA binding and increases the currents induced by submaximal GABA concentrations. Benzodiazepines induce conformational changes in the GABA-binding site in the extracellular domain, but it is uncertain whether these conformational changes extend into the membrane-spanning domain where the channel gate is located. Alone, benzodiazepines do not open the channel. We used the substituted-cysteine-accessibility method to investigate diazepam-induced conformational changes in the region of the alpha(1)-subunit M3 membrane-spanning segment. In the absence of diazepam or GABA, pCMBS(-) did not react at a measurable rate with cysteine-substitution mutants between alpha(1)Phe296 and alpha(1)Glu303. In the presence of 100 nM diazepam, pCMBS(-) reacted with alpha(1)F296C, alpha(1)F298C, and alpha(1)L301C but not with the other cysteine mutants between alpha(1)Phe296 and alpha(1)Glu303. These three mutants are a subset of the five residues that we previously showed reacted with pCMBS(-) applied in the presence of GABA. The pCMBS(-) reaction rates with these three cysteine mutants were similar in the presence of diazepam and GABA. Thus, diazepam, which binds to the extracellular domain, induces a conformational change in the membrane-spanning domain that is similar to a portion of the change induced by GABA. Because diazepam does not open the channel, these results provide structural evidence that the diazepam-bound state represents an intermediate conformation distinct from the open and resting/closed states of the receptor. The diazepam-induced conformational change in the M3 segment vicinity may be related to the mechanism of allosteric potentiation.

4-Chloromercuribenzenesulfonate↗
PubMed1999

Calreticulin functions in vitro as a molecular chaperone for both glycosylated and non-glycosylated proteins.

Calreticulin (CRT) is thought to be a molecular chaperone that interacts with glycoproteins exclusively through a lectin site specific for monoglucosylated oligosaccharides. However, this chaperone function has never been directly demonstrated nor is it clear how lectin-oligosaccharide interactions facilitate glycoprotein folding. Using purified components, we show that CRT suppresses the aggregation not only of a glycoprotein bearing monoglucosylated oligosaccharides but also that of non-glycosylated proteins. Furthermore, CRT forms stable complexes with unfolded, non-glycosylated substrates but does not associate with native proteins. ATP and Zn(2+) enhance CRT's ability to suppress aggregation of non- glycoproteins, whereas engagement of its lectin site with purified oligosaccharide attenuates this function. CRT also confers protection against thermal inactivation and maintains substrates in a folding-competent state. We conclude that in addition to being a lectin CRT possesses a polypeptide binding capacity capable of discriminating between protein conformational states and that it functions in vitro as a classical molecular chaperone.

Adenosine Triphosphate↗
PubMed1999

Interaction of murine MHC class I molecules with tapasin and TAP enhances peptide loading and involves the heavy chain alpha3 domain.

In human cells the association of MHC class I molecules with TAP is thought to be mediated by a third protein termed tapasin. We now show that tapasin is present in murine TAP-class I complexes as well. Furthermore, we demonstrate that a mutant H-2Dd molecule that does not interact with TAP due to a Glu to Lys mutation at residue 222 of the H chain (Dd(E222K)) also fails to bind to tapasin. This finding supports the view that tapasin bridges the association between class I and TAP and implicates residue 222 as a site of contact with tapasin. The inability of Dd(E222K) to interact with tapasin and TAP results in impaired peptide loading within the endoplasmic reticulum. However, significant acquisition of peptides can still be detected as assessed by the decay kinetics of cell surface Dd(E222K) molecules and by the finding that prolonged viral infection accumulates sufficient target structures to stimulate T cells at 50% the level observed with wild-type Dd. Thus, although interaction with tapasin and TAP enhances peptide loading, it is not essential. Finally, a cohort of Dd(E222K) molecules decays more rapidly on the cell surface compared with wild-type Dd molecules but much more slowly than peptide-deficient molecules. This suggests that some of the peptides obtained in the absence of an interaction with tapasin and TAP are suboptimal, suggesting a peptide-editing function for tapasin/TAP in addition to their role in enhancing peptide loading.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1999

Gamma-aminobutyric acid increases the water accessibility of M3 membrane-spanning segment residues in gamma-aminobutyric acid type A receptors.

Gamma-aminobutyric acid type A (GABA(A)) receptors are members of the ligand-gated ion channel gene superfamily. Using the substituted cysteine accessibility method, we investigated whether residues in the alpha(1)M3 membrane-spanning segment are water-accessible. Cysteine was substituted, one at a time, for each M3 residue from alpha(1)Ala(291) to alpha(1)Val(307). The ability of these mutants to react with the water-soluble, sulfhydryl-specific reagent pCMBS(-) was assayed electrophysiologically. Cysteines substituted for alpha(1)Ala(291) and alpha(1)Tyr(294) reacted with pCMBS(-) applied both in the presence and in the absence of GABA. Cysteines substituted for alpha(1)Phe(298), alpha(1)Ala(300), alpha(1)Leu(301), and alpha(1)Glu(303) only reacted with pCMBS(-) applied in the presence of GABA. We infer that the pCMBS(-) reactive residues are on the water-accessible surface of the protein and that GABA induces a conformational change that increases the water accessibility of the four M3 residues, possibly by inducing the formation of water-filled crevices that extend into the interior of the protein. Others have shown that mutations of alpha(1)Ala(291), a water-accessible residue, alter volatile anesthetic and ethanol potentiation of GABA-induced currents. Water-filled crevices penetrating into the interior of the membrane-spanning domain may allow anesthetics and alcohol to reach their binding sites and thus may have implications for the mechanisms of action of these agents.

4-Chloromercuribenzenesulfonate↗
PubMed1999

Calnexin discriminates between protein conformational states and functions as a molecular chaperone in vitro.

Although calnexin is thought to function as a molecular chaperone for glycoproteins, a prevalent view is that it cannot distinguish between protein conformational states, binding solely through its lectin site to monoglucosylated oligosaccharides. Using purified components in vitro, calnexin effectively prevented the aggregation not only of glycoproteins bearing monoglucosylated oligosaccharides but also proteins lacking N-glycans, an effect enhanced by ATP. It also suppressed the thermal denaturation of nonglycosylated proteins and enhanced their refolding in conjunction with other cellular components. Calnexin formed stable complexes with unfolded conformers of these proteins but not with the native molecules. Therefore, in addition to being a lectin, calnexin functions as a bona fide molecular chaperone capable of interacting with polypeptide segments of folding glycoproteins.

Adenosine Triphosphate↗
PubMed1999

The effect of large solid angles of collection on quantitative X-ray microanalysis in the AEM.

Increasing the solid angle of X-ray collection is a major factor in improving the analytical sensitivity of X-ray energy-dispersive spectrometry (XEDS) in the analytical electron microscope (AEM). A new scanning transmission electron microscope, the VG HB 603, is equipped with two XEDS detectors with the largest collection angles (0.30 and 0.17 sr) available in commercial AEMs. However, large collection angles result in a large range of take-off angles, from approximately 4 degrees to 36 degrees, and the low angles can cause strong X-ray absorption. In order to investigate possible detrimental effects of the low (and of the range of) take-off angles on quantitative microanalysis of specimens exhibiting significant absorption, a stoichiometric Ni3Al thin-film, in which the Al Kalpha line is significantly absorbed, was analysed. Furthermore, the effect of different values of the collection angle on X-ray intensities was theoretically evaluated by numerical calculations and spectral simulation. These theoretical approaches permitted correlation of changes in the X-ray take-off angle (and hence X-ray absorption) with changes in the collection angle. It is demonstrated that approximately 0.30 sr detectors, with minimum take-off angles as small as 4 degrees, only result in maximum errors of 4% in the quantification of Al in Ni3Al and, therefore, further increases in collection angle can be pursued while maintaining current levels of accuracy of quantification.

Electron Probe Microanalysis↗
PubMed1999

Endoscopic ultrasonography with fine-needle aspiration: an accurate and simple diagnostic modality for sarcoidosis.

BACKGROUND AND STUDY AIMS: Sarcoidosis is a chronic multisystem granulomatous disease that is often diagnosed after a finding of hilar and mediastinal lymphadenopathy on a chest radiograph. This often requires further evaluation by transbronchial biopsy or other clinical parameters. The present study is a descriptive, retrospective one using endoscopic ultrasound with fine-needle aspiration (EUS-FNA) of mediastinal lymph nodes in seven patients with sarcoidosis. PATIENTS AND METHODS: Among 108 consecutive patients who underwent EUS-FNA of mediastinal lymph nodes for various clinical indications between July 1994 and October 1997, seven patients were found to have sarcoidosis on EUS-FNA, and the EUS morphology was studied in these patients. RESULTS: Sarcoidosis was diagnosed in seven patients using endosonographic characteristics and clinical follow-up. EUS with FNA showed cytological evidence of sarcoidosis in six patients. Seven patients were found to have subcarinal lymph nodes, and six patients had abnormally enlarged aortopulmonary (AP) window lymph nodes. The nodes in all patients had three endosonographic criteria for malignancy. The long axis of the largest mediastinal lymph nodes measured 3.44+/-1.42 cm (range 1.8-6.0 cm). The short axis measured 2.50+/-0.69 (range 1.0-4.0 cm). The average number of nodes seen in each patient was 2.80+/-0.75 (range 2-4). The nodes in all seven patients were discrete and well demarcated. A central hyperechoic strand was evident in these nodes in four patients (57%). There were no complications. CONCLUSIONS: Mediastinal lymph nodes in patients with sarcoidosis appear to have specific echo characteristics, and EUS-FNA can be used for confirmatory tissue diagnosis.

Adult↗
PubMed1999

Endoscopic ultrasound guided fine needle aspiration biopsy: a large single centre experience.

BACKGROUND: Endoscopic ultrasound guided fine needle aspiration biopsy (EUS-FNA) is a recent innovation in the evaluation of gastrointestinal and pulmonary malignancies. AIMS: To review the experience with EUS-FNA of a large single centre. METHODS: 333 consecutive patients underwent EUS-FNA. Follow up data were available on 327 lesions in 317 patients, including 160 lymph nodes, 144 pancreatic lesions, 15 extraintestinal masses, and eight intramural tumours. RESULTS: A primary diagnosis of malignancy was obtained by EUS-FNA in 62% of patients with clinically suspicious lesions. The overall accuracy of EUS-FNA for the diagnosis of malignancy was 86%, with sensitivity of 84% and specificity of 96%. With respect to lesion types, the sensitivity, specificity, and accuracy were 85%, 100%, and 89% for lymph nodes; 82%, 100%, and 85% for pancreatic lesions; 88%, 100%, and 90% for perirectal masses; and 50%, 25%, and 38% for intramural lesions, respectively. Compared with size and sonographic criteria, EUS-FNA in the evaluation of lymph nodes provided superior accuracy and specificity, without compromising sensitivity. Inadequate specimens were obtained from only six patients, including 3/5 with stromal tumors. Only one complication occurred. CONCLUSIONS: EUS-FNA is safe and can readily obtain tissue specimens adequate for cytopathological diagnoses. Compared with size and sonographic criteria, it is a superior modality for the detection of nodal metastases. While providing accurate diagnosis of pancreatic and perirectal malignancies, results suggest the technique is less useful for intramural lesions.

Adolescent↗
PubMed1998

Oligosaccharide binding characteristics of the molecular chaperones calnexin and calreticulin.

Calnexin and calreticulin are homologous molecular chaperones of the endoplasmic reticulum. Their binding to newly synthesized glycoproteins is mediated, at least in part, by a lectin site that recognizes the early N-linked oligosaccharide processing intermediate, Glc1Man9GlcNAc2. We compared the oligosaccharide binding specificities of calnexin and calreticulin in an effort to determine the basis for reported differences in their association with various glycoproteins. Using mono-, di-, and oligosaccharides to inhibit the binding of Glc1Man9GlcNAc2 to calreticulin and to a truncated, soluble form of calnexin, we show that the entire Glc alpha 1-3Man alpha 1-2Man alpha 1-2Man structure, extending from the alpha 1-3 branch point of the oligosaccharide core, is recognized by both proteins. Furthermore, analysis of the binding of monoglucosylated oligosaccharides containing progressively fewer mannose residues suggests that for both proteins the alpha 1-6 mannose branch point of the oligosaccharide core is also essential for recognition. Consistent with their essentially identical substrate specificities, calnexin and calreticulin exhibited the same relative affinities when competing for binding to the Glc1Man9GlcNAc2 oligosaccharide. Thus, differential glycoprotein binding cannot be attributed to differences in the lectin specificities or binding affinities of calnexin and calreticulin. We also examined the effects of ATP, calcium, and disulfide reduction on the lectin properties of calnexin and calreticulin. Whereas oligosaccharide binding was only slightly enhanced for both proteins in the presence of high concentrations of a number of adenosine nucleotides, removal of bound calcium abrogated oligosaccharide binding, an effect that was largely reversible upon readdition of calcium. Disulfide reduction had no effect on oligosaccharide binding by calnexin, but binding by calreticulin was inhibited by 70%. Finally, deletion mutagenesis of calnexin and calreticulin identified a central proline-rich region characterized by two tandem repeat motifs as a segment capable of binding oligosaccharide. This segment bears no sequence homology to the carbohydrate recognition domains of other lectins.

Adenosine Triphosphate↗
PubMed1998

Delirium tremens.

BACKGROUND: The hyperadrenergic syndrome, delirium tremens, as contrasted with tile milder alcohol withdrawal syndromes, is a medical emergency. The clinical features of delirium tremens were carefully described almost 200 years ago. Since then, many therapies have been suggested as superior to preceding therapeutic approaches. Unfortunately, morbidity and mortality remain relatively unchanged for the last 100 years. METHODS: Using a literature review, we review the history of delirium tremens, including the suggested therapeutic approaches. RESULTS/CONCLUSIONS: We recommend a rational approach to management and therapy, based on present knowledge.

Alcohol Withdrawal Delirium↗