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Biomedical subjects

D B Williams

Publications and source records attributed to D B Williams.

At least 109 records · Page 6Linked to original sources
PubMed1990

Absorption of doxycycline from a controlled release pellet formulation: the influence of food on bioavailability.

A three-way crossover study was performed to compare the bioavailability of a new pelletised doxycycline product administered either with food or without food and a reference product taken without food. Four different methods were used to calculate pharmacokinetic parameters from the data. The sums of squares, Akaike's Information Criterion (AIC), and the ranges for the parameters obtained were used for comparison. Good fits to the data were obtained when all four methods were used, each with a lag time. The two compartment open model was the most efficient method for describing the data. The one compartment open model was the least efficient, particularly with respect to predicting the peak concentration of doxycycline in plasma. All the models gave similar rank order results with respect to bioavailability differences between the three treatments. Analysis of the data by different methods suggests that pelletised doxycycline is bioequivalent to the reference product when taken in the absence of food. A standardized feeding regimen affected the rate, but not extent of absorption of doxycycline from the pelletised formulation.

Absorption↗
PubMed1990

Bioavailability of a new sustained-release theophylline capsule in fasted and non-fasted healthy subjects: single and multiple dosing studies.

Eighteen healthy, non-smoking, adult volunteers participated in single and multiple dose three-way crossover studies to evaluate a sustained-release, pellet-filled capsule of theophylline, Austyn. The effect of food on the bioavailability of the sustained-release capsule was investigated by administering 300 mg single doses of Austyn, with a high-fat meal and without food and a divided 300 mg dose of the reference product Elixophyllin elixir, given after fasting. Plasma theophylline concentrations were measured by fluorescence polarization immunoassay (FPIA) which had been validated against HPLC. The single dose study data showed that there were no significant differences (n = 18, ANOVA, p greater than 0.05) between the three regimens with respect to AUC0-infinity values (mg h l-1), (mean +/- SD); Elixophyllin fasting = 97.1 +/- 33.7, Austyn with food = 90.9 +/- 31.3, Austyn fasting = 91.2 +/- 33.8. Similarly, multiple dosing with rapid-release Nuelin tablets, Austyn capsules, and sustained-release Theo-Dur tablets demonstrated that there were no significant differences between regimens with respect to AUC0-24h, AUC0-12h, and AUC12-24h values calculated from the steady-state concentrations (5th day, 24 h sampling). However, the percentage fluctuation at steady-state over the total blood sampling period was significantly less for treatment with the sustained-release capsule. Austyn, compared with the sustained-release tablet, Theo-Dur (Austyn = 36.7 +/- 13.7 per cent, Theo-Dur = 53.1 +/- 14.1 per cent). The results of the single and multiple dose studies indicate that Austyn capsules demonstrate complete bioavailability, and good controlled release characteristics not influenced by concomitant intake of a high-fat meal and with no evidence of dose dumping.

Adult↗
PubMed1990

An essential member of the HSP70 gene family of Saccharomyces cerevisiae is homologous to immunoglobulin heavy chain binding protein.

Immunoglobulin heavy chain binding protein (BiP) is present in the lumen of the mammalian endoplasmic reticulum, where it associates transiently with a variety of newly synthesized secretory and membrane proteins or permanently with mutant proteins that are incorrectly folded. We describe a unique member of the Saccharomyces cerevisiae 70-kDa heat shock protein gene family (HSP70) that encodes a protein homologous to mammalian BiP. The DNA sequence contains a 2046-nucleotide open reading frame devoid of introns, and examination of the predicted amino acid sequence reveals features not found in most other yeast HSP70 proteins but which are present in BiP. Most notable are a 42-residue sequence at the N terminus that exhibits characteristics of a cleavable signal sequence and a C-terminal sequence, -His-Asp-Glu-Leu, that is involved in determining endoplasmic reticulum localization in yeast. The 5' flanking region of this gene contains two overlapping sequences between nucleotides -146 and -169 that closely resemble consensus heat shock elements. The yeast BiP gene is strongly heat shock-inducible, whereas the BiP genes in various other species are either weakly or non-heat-inducible. We demonstrate that a functional BiP gene is essential for vegetative growth. An evolutionary comparison of amino acid sequences of 34 HSP70 proteins from 17 species suggests that BiP genes share a common ancestor, which diverged from other HSP70 genes near the time when eukaryotes first appeared.

Amino Acid Sequence↗
PubMed1990

Respective contribution of intracellular calcium release and extracellular calcium influx for interleukin-2 synthesis in activated T-cell hybrids.

Triggering of the T-cell receptor (TcR)alpha beta/CD3 receptor complex with anti-allotypic antibodies or concanavalin A (Con A) induced a rapid release of intracellular calcium in a murine T-cell hybridoma model system. Internal calcium release preceded the influx of extracellular calcium, as judged by comparative analysis of time-dependent changes in Quin 2 fluorescence following T-cell activation in the presence and absence of extracellular calcium. The magnitude of intracellular calcium release and extracellular calcium influx depended on the degree of receptor-occupancy and cross-linking. Correlations between the concentration of stimulating ligand, cytosolic calcium increase and IL-2 synthesis indicated a positive but non-linear relationship. Our data suggest that TcR cross-linking may provide a third T-cell activation signal which, in conjunction with protein kinase C activation and cytosolic calcium elevation, together form a signal triad responsible for interleukin-2 (IL-2) synthesis.

Animals↗
PubMed1989

Different class I antigen oligosaccharides on a murine tumor and a lectin-resistant variant are not responsible for the differential recognition of the tumors by CTL.

Previous studies have shown that whereas a highly malignant mouse cell line termed MDAY-D2 (d haplotype) does not elicit a detectable response by cytotoxic T lymphocytes (CTL) in DBA/2 mice, strong anti-tumor CTL are generated against a wheat-germ-agglutinin-resistant variant, designated MDW3. Additional evidence suggests these anti-MDW3 CTL may not be a consequence of a unique antigenic determinant on the variant cells. Because MDW3 cells are expected to differ from MDAY-D2 cells in their surface carbohydrate structures (due to their lectin resistance) and Class I major histocompatibility molecules play a crucial role in CTL-mediated responses, we speculated that the Asn-linked oligosaccharides present on Class I molecules of MDAY-D2 and MDW3 might be different and could potentially influence recognition analyses and Con A-Sepharose affinity chromatography clearly demonstrated that the oligosaccharides isolated from the H-2Dd molecule of MDAY-D2 cells are larger and more highly branched than those of the MDW3 variant. Taken together with the finding that anti-MDW3 CTL are restricted by H-2Dd, these results suggested that the larger H-2Dd oligosaccharides on MDAY-D2 cells could potentially mask or perturb determinants required for recognition by these CTL. To test this postulate, the surface Class I oligosaccharides of both MDAY-D2 and MDW3 cells were converted to simpler hybrid structures by treatment with the oligosaccharide processing inhibitor, swainsonine. However, no effect was observed on the lysis or binding of either MDAY-D2 or MDW3 cells by anti-MDW3 CTL. Thus, the results do not support the possibility that the larger H-2Dd oligosaccharides on MDAY-D2 cells are, in themselves, responsible for the poor recognition of the parent tumor by anti-MDW3 CTL. Our data do indicate, however, that CTL target binding and effector functions are not dependent on the fine structure of complex Asn-linked carbohydrates present on Class I molecules and possibly on other, accessory molecules at the target cell surface, since MDW3 cells maintained their sensitivity to lysis by CTL following swainsonine treatment.

Alkaloids↗
PubMed1989

Role of beta 2-microglobulin in the intracellular transport and surface expression of murine class I histocompatibility molecules.

We have examined the requirement for beta 2-microglobulin (beta 2m) in the intracellular transport of murine class I histocompatibility molecules to the cell surface. R1E cells that are defective in the synthesis of beta 2m were transfected with either the class I H-2Kb or H-2Db genes alone, or together with the beta 2m gene. Kb or Db heavy chains synthesized in the presence of beta 2m were transported rapidly through the cell and expressed efficiently at the cell surface. In the absence of beta 2m, no "free" Kb heavy chains were detectable at the cell surface and their intracellular transport was blocked at an early stage. In contrast, a significant quantity of "free" Db heavy chains could be detected at the cell surface as we have reported previously. However, we have shown here that defects in intracellular transport were apparent in that the majority (approximately 70%) of newly synthesized Db heavy chains accumulated intracellularly and were degraded. Therefore, although Kb and Db heavy chains differ in their abilities to be expressed at the cell surface in the absence of beta 2m they both require association with beta 2m for efficient intracellular transport. In addition, R1E cells transfected with a deletion construct of the Kb gene expressed a truncated molecule lacking the alpha 3 extracellular domain (Kb-3) at the cell surface, but, like free Db, most newly synthesized Kb-3 molecules accumulated intracellularly. The free Kb, Kb-3, and Db heavy chains were not recognized by most mAb specific for Kb and Db, respectively. Therefore, even the transported forms of free Db and Kb-3 were not native in conformation, which is surprising given the current view that correct folding is essential for intracellular transport. Interestingly, the free Db and Kb-3 heavy chains that reached the cell surface differed in their detergent binding properties from those retained within the cell. This suggests that the transported heavy chains may have folded differently thus allowing their export to the cell surface.

Animals↗
PubMed1988

Intracellular transport of class I histocompatibility molecules. Influence of protein folding on transport to the cell surface.

To examine the structural requirements for the intracellular transport and surface expression of Class I histocompatibility molecules, we studied somatic cell variants that produce altered forms of the H-2Kb molecule. One variant, R8.10, produced a mutant Kb molecule that was expressed on the cell surface at about 15% of the wild-type level. Nucleic acid sequence analysis identified the mutation as a single nucleotide change resulting in an amino acid substitution (Trp----Arg) at residue 167 in the alpha 2 extracellular domain. A comparative kinetic analysis of the intracellular transport of the wild-type and mutant molecules revealed that transport of the mutant product was remarkably impaired. Whereas wild-type molecules arrived at the cell surface with a half-time of about 30 min, 80% of newly synthesized mutant molecules did not progress beyond the rough endoplasmic reticulum, where they were slowly degraded. Surprisingly, the remaining 20% of the mutant population was capable of reaching the plasma membrane at a rate about one-half that of wild-type Kb. The accumulation of most of the mutant molecules in the rough endoplasmic reticulum was not due to aggregation or insolubility, nor could it be attributed to a lack of association with beta 2-microglobulin. Several techniques were employed in an effort to detect structural features unique to the transport-deficient mutant population. Monoclonal antibody binding experiments revealed structural differences between wild-type and mutant molecules in the region of the mutation but failed to distinguish the transport-competent and -deficient mutant populations. Detergent partitioning studies were also ineffective in this regard. However, differences between transported and untransported molecules were readily demonstrated by their disparate susceptibilities to proteolytic digestion. The results indicated that the transport-deficient form of the mutant assumed a conformation that was altered relative to both the transport-competent form and the wild-type molecule. The fact that the ability of wild-type and mutant molecules to be transported correlated with conformational features rather than with their specific primary sequences suggests that proper folding is an important requirement for their passage through the exocytotic pathway.

Animals↗
PubMed1988

Pharmacokinetics of low-dose oral modified release, soluble and intravenous aspirin in man, and effects on platelet function.

The pharmacokinetics of low-dose aspirin and the resulting salicylic acid were studied in 6 healthy volunteers. Each received a single 50-mg dose of (1) oral modified release capsules, (2) oral solution and (3) intravenous solution. The volunteers also received 50 mg modified release capsules daily for 6 days to determine the effect on collagen, ADP and arachidonate induced platelet aggregation and thromboxane production, and to compare the pharmacokinetics after repeated dosing with the parameters obtained after the single dose. The formulation and route of administration profoundly influenced several pharmacokinetic parameters for aspirin: the maximum concentration (Cmax, ng.ml-1) was 221 and 191 after modified release for single and chronic dosing respectively, 1323 after the oral solution and 6000 after intravenous injection; the time to achieve this maximum concentration (tmax, h) was 3.42 and 3.02 after modified release for single and chronic dosing respectively, and 0.29 after the oral solution; the area under the plasma drug concentration versus time curve (AUC, microgram.h.ml-1) was 0.38 and 0.27 after modified release single and chronic dosing respectively, 0.68 after the oral solution and 1.57 after intravenous injection. The elimination of aspirin after the two solutions was at least biphasic. The terminal phase rate constant ranged from 1.52 h-1 after intravenous injection to 1.88 h-1 after the oral modified release form. The absorption of the oral forms of aspirin was complete as reflected by the total recovery of the doses as salicylic acid in urine.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1988

Familial motor neuron disease: differing penetrance in large pedigrees.

It is likely that adult-onset motor neuron disease is inherited more frequently than has previously been believed. We have studied 9 families with familial motor neuron disease and have found apparently wide variation in penetrance. Only one has autosomal dominant inheritance with high penetrance and in this family the average age of onset is similar to that reported in reviews of familial motor neuron disease. The remaining families have a pattern consistent with autosomal dominant inheritance with diminished penetrance. The average ages of onset in these families are comparable with those in sporadic motor neuron disease. Low penetrance appears to be related to higher average age of disease onset because gene carriers have an increased likelihood of dying from other causes before developing motor neuron disease. When penetrance is low the family history may be unknown, causing affected individuals to be mistakenly regarded as having sporadic disease. We conclude that the incidence of familial motor neuron disease is likely to have been underestimated by the exclusion of apparently sporadic cases in which the familial disease has low penetrance. These results and conclusions imply that the lower average age of disease onset reported in familial motor neuron disease may be due to selective recognition of high penetrance families with lower average ages of onset.

Adult↗
PubMed1986

Transfer factor as a therapy for multiple sclerosis: a follow-up study.

The result of a two year, double blind, controlled trial of Transfer Factor (TF) in the treatment of multiple sclerosis (MS) were reported in 1980. It was demonstrated that TF significantly reduced the rate of progression of the disability but the benefit of therapy was not apparent until 18 months after its commencement. After the completion of the trial, TF treatment was offered to all the trial participants. Forty-five of these people accepted TF as treatment and have been followed for the subsequent three years. The twenty-three people who had received TF during the trial, and continued on TF after the trial, consistently had a slower rate of progression of their MS. Although the twenty-two patients initially on placebo had a significantly faster rate of progression during the trial, this slowed with commencement of TF treatment. After 3 years of TF, the rate of progression of disease was similar to that of the group receiving TF continuously for 5 years. In addition, 470 patients with clinically definite MS are being treated in New South Wales in an open study of TF. The rate of progression of the disease is being monitored by neurological assessments and appears to be similar to that of patients who had received TF in the original trial. The follow-up study of the 1980 TF trial patients and the open study of 470 MS patients confirm the original observation that TF has some effect on slowing the course of MS.

Clinical Trials as Topic↗
PubMed1985

Intracellular transport of membrane glycoproteins: two closely related histocompatibility antigens differ in their rates of transit to the cell surface.

The intracellular transport of two closely related membrane glycoproteins was studied in the murine B cell lymphoma line, AKTB-1b. Using pulse-chase radiolabeling, the kinetics of appearance of the class I histocompatibility antigens, H-2Kk and H-2Dk, at the cell surface were compared and found to be remarkably different. Newly synthesized H-2Kk is transported rapidly such that all radiolabeled molecules reach the surface within 1 h. In contrast, the H-2Dk antigen is transported slowly with a half-time of 4-5 h. The rates of surface appearance for the two antigens closely resemble the rates at which their Asn-linked oligosaccharides mature from endoglucosaminidase H (endo H)-sensitive to endo H-resistant forms, a process that occurs in the Golgi apparatus. This suggests that the rate-limiting step in the transport of H-2Dk to the cell surface occurs before the formation of endo H-resistant oligosaccharides in the Golgi apparatus. Subcellular fractionation experiments confirmed this conclusion by identifying the endoplasmic reticulum (ER) as the site where the H-2Dk antigen accumulates. The retention of this glycoprotein in the ER does not appear to be due to a lack of solubility or an inability of the H-2Dk heavy chain to associate with beta 2-microglobulin. Our data is inconsistent with a passive membrane flow mechanism for the intracellular transport of membrane glycoproteins. Rather, it suggests that one or more receptors localized to the ER membrane may mediate the selective transport of membrane glycoproteins out of the ER to the Golgi apparatus. The fact that H-2Kk and H-2Dk are highly homologous (greater than or equal to 80%) indicates that this process can be strongly influenced by limited alterations in protein structure.

Animals↗
PubMed1985

Oral verapamil for prophylaxis of supraventricular tachycardia after myocardial revascularization. A randomized trial.

To determine if the prophylactic administration of oral verapamil is effective in reducing the incidence of supraventricular tachycardia after myocardial revascularization, 141 patients were entered into a double-blind randomized trial. Seventy patients received verapamil 80 mg every 8 hours beginning immediately after operation and continuing for 5 days, and 71 patients received placebo. Patients were monitored during the study period and observed for the occurrence of supraventricular tachycardia. The verapamil and placebo groups were similar with regard to sex, age, preoperative antianginal drugs, number of bypass grafts, average bypass time, and average occlusion time. A total of 39 patients had supraventricular tachycardia, 20 were receiving verapamil, and 19 were receiving placebo. The average ventricular rate was 127 +/- 13 beats/min for the verapamil group with supraventricular tachycardia compared with 140 +/- 12 beats/min for the placebo group with the mean differences not significant (p = 0.10). On the basis of these data, it is concluded that verapamil 80 mg every 8 hours beginning early after myocardial revascularization is not effective in reducing the incidence of supraventricular tachycardia.

Administration, Oral↗
PubMed1984

Control of asparagine-linked oligosaccharide chain processing: studies on bovine pancreatic ribonuclease B. An in vitro system for the processing of exogenous glycoproteins.

To gain an understanding of why the polymannose-type oligosaccharide chain of bovine pancreatic ribonuclease B is not processed to a complex-type chain in vivo, the processing of this glycoprotein was studied in two cell-free systems. Addition of native 125I-ribonuclease B to rat liver Golgi membranes in the presence of UDP-GlcNAc resulted in the conversion of the high mannose chain to a complex type as evidenced by the acquisition of resistance to digestion with endoglucosaminidase H. Processing was linearly dependent on time and on the amount of Golgi membranes. Omission of UDP-GlcNAc from the reaction mixtures completely abolished processing of the glycoprotein. Product identification studies confirmed that the formation of ribonuclease that was resistant to digestion with endoglucosaminidase H was accompanied by the appearance of a complex-type oligosaccharide that contained one or more terminal beta-GlcNAc residues. In vitro processing of 125I-ribonuclease B that had been denatured by reduction and alkylation revealed that the rate of complex chain formation was only slightly greater than that observed with the native enzyme. In contrast to the results obtained with the heterologous rat liver system, Golgi membranes from bovine pancreas failed to process native ribonuclease B to the complex form. However, bovine pancreas Golgi membranes did readily process the denatured form of the enzyme. The presence of a factor in bovine pancreas that binds only to native ribonuclease B and thereby prevents its oligosaccharide chain from being processed was considered to be unlikely on the basis of gel filtration studies and mixing experiments. These findings indicate that some aspect of the conformation of native ribonuclease B prevents one or more of the processing enzymes of bovine pancreas from acting on the oligosaccharide chain. In addition, the substrate specificity of this processing enzyme(s) differs markedly from its counterpart in rat liver. These two factors, conformation of the substrate and specificity of the processing enzymes, apparently combine to produce the high mannose oligosaccharide chain of ribonuclease B observed in vivo.

Animals↗
PubMed1984

Platelet activating factor: regulation by mast cells and aspirin.

We have investigated some aspects of the regulation of production of rat platelet activating factor (PAF)2 in vitro. Suspensions of unseparated (PLC1), mast cell-depleted (PLC2), or mast cell (MC)-enriched rat peritoneal lavage cells (PLC) were analyzed for PAF content by extraction at alkaline pH. PAF activity extracted from PLC1 varied inversely with viable cell concentration: at 1 X 10(6) cells/ml, 32 +/- 9.3 PAF units, decreasing to 11.2 +/- 9.5 units at 10 X 10(6) cells/ml, and no activity at higher concentrations. Incubation of PLC1 in Tyrode's buffer or acetylsalicylic acid (ASA), but not salicylate, resulted in a time-dependent loss of PAF activity. Mean PAF activity of PLC2 was similar to that in PLC1, while no PAF activity was extractable from MC. Co-incubation with MC extracts inhibited PAF activity of PLC1 extracts in a dose-dependent fashion. Ultracentrifugation of PAF-containing samples led to a loss of all PAF activity in PLC1 extracts, suggesting the association of PAF activity with subcellular components. PAF appears to be derived from a non-MC population of rat PLC, is not extractable from rat PLC in the presence of ASA and is inhibited by MC extracts. These studies suggest that ASA regulates PAF availability unrelated to its effect on cyclooxygenase and that MC membrane products directly inhibit PAF activity from rat PLC.

Animals↗
PubMed1984

On the use of ionization cross sections in analytical electron microscopy.

There are two approaches to the utilization of the ionization cross section, Q, for use in the determination of kappa AB factors for quantitative microanalysis in the analytical electron microscope. The first approach is to interpolate a value of Q from experimentally determined kappa AB factors at a fixed accelerating voltage (kV). The second approach uses a theoretical parameterization of Q generated by fitting the fundamental Bethe expression to selected experimental values of Q over a wide range of kV. This paper discusses the relative merits of the two approaches.

Electron Probe Microanalysis↗
PubMed1984

The effect of prostacyclin infusion on platelet hemostatic function in patients undergoing cardiopulmonary bypass.

Prostacyclin (PGI2) infusion was studied in patients during cardiopulmonary bypass for coronary artery bypass grafting to assess its capacity to protect platelet hemostatic function. Twelve patients received PGI2 at doses ranging from 20 to 70 ng/kg/min by continuous infusion directly into the bubble oxygenator. Nine control patients were also studied. Platelet counts were not different in the two groups; the platelet count at the completion of the operation was decreased by a mean of 29% in treated patients and 40% in control patients (p greater than 0.15). Bleeding times were uniformly greater than 30 minutes following 30 minutes on bypass in both groups (p greater than 0.5). Also there was no difference in the bleeding times between the treated and untreated groups in the immediate postbypass period (p greater than 0.05). In addition, no difference in transfusion requirements was observed between the treated and control groups. However, plasma levels of platelet factor 4 were lower in the treated patients (p less than 0.05), and, more significantly, blood pressure was lower at all time points in the treated patients (p less than 0.001) despite the greater use of vasopressors. We conclude that PGI2 infusion in this study was of no hemostatic benefit but produced significant hypotension, a potential source of morbidity.

Adult↗
PubMed1984

Hemodynamic response to positive end-expiratory pressure following right atrium-pulmonary artery bypass (Fontan procedure).

Thirteen patients were studied in the early postoperative period to determine the hemodynamic response to increasing levels of positive end-expiratory pressure (PEEP) following right atrium-pulmonary artery bypass (Fontan procedure). Hemodynamic data and arterial oxygen and carbon dioxide tensions were measured without PEEP and with PEEP = 3, 6, 9, and 12 cm H2O. Cardiac index decreased progressively with increasing levels of PEEP compared to PEEP = 0 (cardiac index = 2.7 +/- 1.2 L/min/m2), and the decrease was significant at PEEP = 9 (cardiac index = 2.2 +/- 0.8 L/min/m2, p less than 0.05) and 12 cm H2O (cardiac index = 2.0 +/- 0.7 L/min/m2, p less than 0.05). Both arterial oxygen tension and pulmonary vascular resistance index increased significantly at all levels of PEEP studied compared to PEEP = 0. Significant positive trends were demonstrated for arterial oxygen tension and pulmonary vascular resistance index and a significant negative trend was shown for cardiac index with increasing PEEP. Heart rate, right atrial pressure, left atrial pressure, mean arterial blood pressure, and arterial carbon dioxide tension did not change significantly nor consistently with increasing PEEP. From these data it appears that PEEP is an effective means of raising arterial oxygen tension after right atrium-pulmonary artery bypass. A progressive fall in cardiac index occurs with increasing PEEP, and the fall becomes significant at PEEP greater than 6 cm H2O. The fall in cardiac index appears to be mediated by a significant rise in pulmonary vascular resistance index.

Adolescent↗