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Biomedical subjects

C Miller

Publications and source records attributed to C Miller.

At least 199 records · Page 11Linked to original sources

Tetrameric stoichiometry of a prokaryotic K+ channel.

Genes with sequences reminiscent of neuronal K+ channels have recently been identified in prokaryotes. These putative K+ channels appear to be integral membrane proteins, with multiple transmembrane sequences identified by hydrophobicity analysis and a sequence strikingly similar to the pore-lining "P-region" motif found in all known eukaryotic K+ channels. This study examines the oligomeric state and stability in detergent micelles of SliK, a K+ channel homologue from Streptomyces lividans. A synthetic gene for SliK was expressed at high levels in Escherichia coli, and the protein was purified. The predominant form of the protein runs in SDS-PAGE gels as an oligomer of the 19-kDa polypeptide, but harsh treatments such as heat or high pH convert this slowly-migrating material into monomeric form. A "mass-tagging" strategy developed to examine subunit stoichiometry shows that SliK is a homotetramer in SDS and dodecyl maltoside micelles. The tetrameric structure can be disrupted by P-region mutations known to prevent the functional expression of neuronal K+ channels. The tetramer is remarkably stable, showing no conversion to the monomeric form after 14 days at room temperature. Although SliK-mediated cation flux activity was not observed, the tetrameric behavior of the protein argues that SliK may provide a system for a direct attack on the structure of a K+ channel P-region sequence.

Amino Acid Sequence↗

A controlled trial of isoniazid in persons with anergy and human immunodeficiency virus infection who are at high risk for tuberculosis. Terry Beirn Community Programs for Clinical Research on AIDS.

BACKGROUND: Patients with human immunodeficiency virus (HIV) infection and latent tuberculosis are at substantial risk for the development of active tuberculosis. As a public health measure, prophylactic treatment with isoniazid has been suggested for HIV-infected persons who have anergy and are in groups with a high prevalence of tuberculosis. METHODS: We conducted a multicenter, randomized, double-blind, placebo-controlled trial of six months of prophylactic isoniazid treatment in HIV-infected patients with anergy who have risk factors for tuberculosis infection. The primary end point was culture-confirmed tuberculosis. RESULTS: The study was conducted from November 1991 through June 1996. Over 90 percent of the patients had two or more risk factors for tuberculosis infection, and nearly 75 percent of patients were from greater New York City. After a mean follow-up of 33 months, tuberculosis was diagnosed in only 6 of 257 patients in the placebo group and 3 of 260 patients in the isoniazid group (risk ratio, 0.48; 95 percent confidence interval, 0.12 to 1.91; P=0.30). There were no significant differences between the two groups with regard to death, death or the progression of HIV disease, or adverse events. CONCLUSIONS: Even in HIV-infected patients with anergy and multiple risk factors for latent tuberculosis infection, the rate of development of active tuberculosis is low. This finding does not support the use of isoniazid prophylaxis in high-risk patients with HIV infection and anergy unless they have been exposed to active tuberculosis.

AIDS-Related Opportunistic Infections↗

Infestation of sheep dung by nematophagous fungi and implications for the control of free-living stages of gastro-intestinal nematodes.

A field trial was conducted to assess the rate at which dung becomes infested by fungi which parasitise nematodes (nematophagous fungi) after deposition. Sheep dung was placed on field plots of bare ground, ryegrass (Lolium perenne), browntop (Agrostis capillaris) and white clover (Trifolium repens) in summer (February) and autumn (April), and subsamples were examined at intervals for the presence of nematophagous fungi. Nematophagous fungi occurred in 71% of 129 samples recovered in February and 57% of 58 samples recovered in April. Arthrobotrys oligospora, Monacrosporium candidum and Nematoctonus spp. were the most frequently isolated nematode-trapping fungi in both seasons. The endoparasitic nematophagous fungus Harposporium leptospira also occurred frequently in dung deposited in February, but not April. Fungi entered dung quickly, with 83% and 58% of dung samples containing nematophagous fungi at 3 days after deposition in February and April, respectively. The percentage of dung infested by nematophagous fungi on plots of bare ground, ryegrass, white clover and browntop was 76%, 75%, 61% and 55%, respectively. Results suggest that a number of species of nematophagous fungi are able to enter dung soon after deposition on a variety of types of ground cover.

Animals↗

Norepinephrine transmitter metabolite induces apoptosis in differentiated rat pheochromocytoma cells.

3,4-Dihydroxyphenylglycolaldehyde (DOPEGAL) is the monoamine oxidase A metabolite of norepinephrine and epinephrine. DOPEGAL, but not other metabolites, kills differentiated PC-12 cells. However, the type of DOPEGAL induced cell death, whether necrosis or apoptosis, is not known. To determine the type of cell death triggered by DOPEGAL, PC-12 cells cultured in the presence or absence of 30 microM DOPEGAL were examined by electron microscopy and DNA agarose gel electrophoresis for characteristic features of apoptosis. Results indicate that DOPEGAL induces apoptosis in these cells. Implications for degenerative diseases are discussed.

Adrenergic alpha-Agonists↗

Fertilization-independent seed development in Arabidopsis thaliana.

We report mutants in Arabidopsis thaliana (fertilization-independent seed:fis) in which certain processes of seed development are uncoupled from the double fertilization event that occurs after pollination. These mutants were isolated as ethyl methanesulfonate-induced pseudo-revertants of the pistillata phenotype. Although the pistillata (pi) mutant has short siliques devoid of seed, the fis mutants in the pi background have long siliques containing developing seeds, even though the flowers remain free of pollen. The three fis mutations map to loci on three different chromosomes. In fis1 and fis2 seeds, the autonomous endosperm nuclei are diploid and the endosperm develops to the point of cellularization; the partially developed seeds then atrophy. In these two mutants, proembryos are formed in a low proportion of seeds and do not develop beyond the globular stage. When FIS/fis plants are pollinated by pollen from FIS/FIS plants, approximately 50% of the resulting seeds contain fully developed embryos; these seeds germinate and form viable seedlings (FIS/FIS). The other 50% of seeds shrivel and do not germinate; they contain embryos arrested at the torpedo stage (FIS/fis). In normal sexual reproduction, the products of the FIS genes are likely to play important regulatory roles in the development of seed after fertilization.

Arabidopsis↗

Hematopoietic stem cells: inferences from in vivo assays.

Mice and humans both contain a population in their marrow which can permanently regenerate all of the hematopoietic lineages. This developmental potential was first demonstrated in myeloablated mice transplanted with genetically marked marrow obtained from congenic donors. More recently, this approach has been used to devise an in vivo limiting dilution assay for "competitive (lymphomyeloid) repopulating units" (CRU) that allows murine hematopoietic stem cells to be quantitated. Measurements of murine CRU have shown that this population expands concomitantly with the total hematopoietic system during ontogeny and to some extent post-transplant. During these periods of expansion, defective c-kit function can be seen to preferentially compromise CRU self-renewal more than early CRU detection (which requires differentiation and amplification of the progeny of CRU, but may not require extensive CRU self-renewal). In humans, a similar cell type with transplantable lymphomyeloid differentiation potential can be identified in cord blood on the basis of its ability to engraft sublethally irradiated immunodeficient nonobese diabetic/severe combined immunodeficient mice. Quantitation of these human CRU by limiting dilution analysis of unseparated, highly purified (CD34+CD38-) and cultured (CD34+CD38-) human cord blood cells indicates that their numbers (like the long-term culture-initiating cell [LTC-IC] population) can be slightly expanded in cytokine-supplemented serum-free media, although not as extensively as anticipated from analogous studies of human marrow LTC-IC cultured under the same conditions. Taken together, the results of our studies suggest that the self-renewal of mitotically activated hematopoietic stem cells can be enhanced by their interactions with particular cytokine combinations whose effectiveness in this regard may change during ontogeny.

Animals↗

Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles.

We previously applied the Poisson-Boltzmann equation to atomic models of phospholipid bilayers and basic peptides to calculate their electrostatic interactions from first principles (Ben-Tal, N., B. Honig, R. M. Peitzsch, G. Denisov, and S. McLaughlan. 1996. Binding of small basic peptides to membranes containing acidic lipids. Theoretical models and experimental results. Biophys. J. 71:561-575). Specifically, we calculated the molar partition coefficient, K (the reciprocal of the lipid concentration at which 1/2 the peptide is bound), of simple basic peptides (e.g., pentalysine) with phospholipid vesicles. The theoretical predictions agreed well with experimental measurements of the binding, but the agreement could have been fortuitous because the structure(s) of these flexible peptides is not known. Here we use the same theoretical approach to calculate the membrane binding of two small proteins of known structure: charybdotoxin (CTx) and iberiotoxin (IbTx); we also measure the binding of these proteins to phospholipid vesicles. The theoretical model describes accurately the dependence of K on the ionic strength and mol % acidic lipid in the membrane for both CTx (net charge +4) and IbTx (net charge +2). For example, the theory correctly predicts that the value of K for the binding of CTx to a membrane containing 33% acidic lipid should decrease by a factor of 10(5) when the salt concentration increases from 10 to 200 mM. We discuss the limitations of the theoretical approach and also consider a simple extension of the theory that incorporates nonpolar interactions.

Biophysical Phenomena↗

Limited success using the "flare" protocol in poor responders in cycles with low basal follicle-stimulating hormone levels during in vitro fertilization.

OBJECTIVE: To assess an alternate protocol for stimulating poor responders. DESIGN: Prospective clinical study. SETTING: University-affiliated infertility clinic. PATIENT(S): Eighty poor responders. INTERVENTION(S): Stimulation was withheld until patients fulfilled the following criteria: basal FSH < or = 12 mIU/mL (conversion factor to SI unit, 1.00) with concurrent E2 level < or = 100 pg/mL (conversion factor to SI unit, 3.671), and P level < or = 1.0 ng/mL (conversion factor to SI unit, 3.467). They then started leuprolide acetate on cycle day 2 and a high dose of gonadotropins on cycle day 3. MAIN OUTCOME MEASURE(S): Stimulation response and pregnancy rates (PRs). RESULT(S): Nineteen cycles (23.8%) were canceled because of poor ovarian response. The mean E2 level on day of hCG was 2,578 +/- 1,339 pg/mL (9,464 +/- 4,915 pmol/L) (range, 789 to 5934 pg/mL [2,896 to 21,784 pmol/L]). The number of oocytes retrieved was 10 +/- 6.6 (range, 1 to 37). Nine patients did not have ET (failed fertilization [n = 5], no cleavage of preembryos [n = 3], freeze all [n = 1]). The number of pre-embryos transferred was 3.9 +/- 1.6 (range, 1 to 8). The clinical pregnancy rate was 7 of 61 (11.5%) per retrieval, and 7 of 52 (13.4%) per transfer with an implantation rate of 7 of 201 (3.5%). Three patients miscarried with an ongoing PR of 4 of 61 (6.5%) per retrieval and 4 of 52 (7.6%) per transfer. CONCLUSION(S): Poor responders undergoing controlled ovarian hyperstimulation with the "flare" protocol in cycles with low basal FSH will often show adequate ovarian response and reach oocyte retrieval and ET. The PRs, however, remain low.

Adult↗

Food purchase decision-making typologies of women with non-insulin-dependent diabetes mellitus.

Food selection is a key factor in the nutritional management of diabetes mellitus. Since up to 80% of food purchase decisions are made at the supermarket, the purpose of this study was to identify the criteria which influence point-of-purchase decision-making in women with NIDDM aged 40-60 years. A qualitative approach with individual interviews and in-store observations was used. Analysis of the interviews identified four decision-making typologies based on the extent nutrition, price and family needs were emphasized. The four typologies included (1) the Overloaded Shopper, (2) the Budget Shopper, (3) the Nutrition Savvy Shopper, and (4) the Out-of-Touch Shopper. Cluster analysis confirmed the typologies for 71% of the sample. Educators should classify shoppers according to a typology to determine their clients' personal needs and interests. Then, educators can tailor the educational or counseling message to meet those specific needs.

Adult↗

Divorcing and building a new life.

The purpose of this study was to provide a qualitative description of women's experiences of divorcing and building a new life. Interviews with 10 divorced women were conducted, transcribed, and analyzed using the constant comparative method. Four phases were identified in the process of divorcing and building a new life: the emotional divorce, making the decision, pulling apart, and moving beyond. Feelings and coping strategies reported by the participants are described.

Adaptation, Psychological↗

Infectious ocular complications in orthotopic liver transplant patients.

We report the frequency and type of infectious ocular complications following orthotopic liver transplantation (OLT) and review diagnostic and therapeutic strategies. During the period September 1988 through November 1994, 684 patients underwent OLT at Mount Sinai Hospital (New York). Nine orthotopic liver transplant patients (1.3%) developed ocular infections: Candida albicans endophthalmitis (2), Aspergillus fumigatus endophthalmitis (1), cytomegalovirus retinitis (4), herpes simplex virus keratitis (1), and varicella-zoster virus panophthalmitis (1). The mean time from OLT to ocular symptoms was 42 days for patients with fungal infections and 128 days for patients with viral infections. Blurred vision was the commonest symptom (five of nine cases). The mean duration of follow-up was 2 years (range, 33 days to 5 years). Permanent loss of vision occurred in three patients, five had improvement in visual acuity, and one died of disseminated aspergillosis 33 days after OLT. Infectious ocular complications following OLT may occur as isolated events or with disseminated disease. Fungal infections occur earlier (mean, 42 days after OLT) than viral infections (mean, 4 months after OLT). The clinical presentation may be atypical; aggressive vitreoretinal procedures and serial examinations may be required to establish the diagnosis. Cytomegalovirus retinitis in orthotopic liver transplant patients may not require life-long maintenance therapy with antiviral agents.

Adult↗

Recombinant bacille Calmette-Guérin expressing the measles virus nucleoprotein protects infant rhesus macaques from measles virus pneumonia.

Measles virus infection continues to be a major cause of infant mortality. There is a need for a measles vaccine that can be administered at birth in the presence of maternal neutralizing antibody. Infant rhesus monkeys were immunized with recombinant bacille Calmette-Guérin expressing the full-length measles virus nucleoprotein (BCG-N) and subsequently challenged with measles virus. Nucleoprotein-specific lymphocyte proliferative responses were detected in the absence of anti-N antibody after vaccination. Vaccination with BCG-N did not prevent systemic measles virus infection; however, there was a significant reduction of lung inflammation after challenge. Virus titers in lymph nodes were significantly lower, and the duration of nasopharyngeal viral shedding was shorter in some vaccinated monkeys after challenge. These results suggest that measles virus-specific T cells were primed by BCG-N vaccination and that they prevented virus-induced lung pathology.

Animals↗

The challenges of clinical nursing research: strategies for successful conduct.

Although the literature is replete with stringent methodological guidelines for designing experimental and quasi-experimental studies, there is a paucity of literature addressing the challenges of conducting these studies in the "real world" of the clinical setting. This article describes the challenges experienced by a team of researchers conducting a study in the acute care setting, including those associated with navigating organizational restructuring and managing within a climate of uncertainty. The study focused on the evaluation of a nurse-delivered smoking cessation intervention for hospitalized cardiac patients. As each particular challenge emerged, strategies were developed. By explicating these challenges and strategies, we hope to help clinical researchers deal with challenges proactively rather than reactively.

Clinical Nursing Research↗

Effects of crystalloid solutions on circulating lactate concentrations: Part 1. Implications for the proper handling of blood specimens obtained from critically ill patients.

OBJECTIVES: a) To test the hypothesis that circulating lactate concentrations are the same in simultaneously collected arterial and central venous blood specimens; b) to test the hypothesis that even small amounts of crystalloid solutions, which are inadequately "cleared" from these indwelling arterial and venous catheters, can lead to clinically important and misleading changes in the measured lactate values. DESIGN: A prospective, multiexperiment study. SETTING: A critical care research laboratory and a 20-bed intensive care unit (ICU). PATIENTS: Three hundred fifty-five patients. INTERVENTIONS: Blood samples were collected. MEASUREMENTS AND MAIN RESULTS: Experiment 1: Simultaneously collected arterial and central venous blood specimens were obtained on 148 occasions from 48 medical ICU patients receiving no lactated Ringer's solution (RL). Arterial and central venous lactate values were nearly identical in these patients. The correlation between the arterial and central venous lactate concentrations was excellent (r2 = .85; p < .0001) and the agreement between the arterial and central venous lactate concentrations was also excellent (bias and precision = 0.04 mmol/L and +/- 0.38 mmol/L, respectively). Experiment 2: Arterial and mixed venous blood samples were obtained from 100 percutaneous transluminal coronary angioplasty (PTCA) and 75 cardiac surgical patients immediately before the performance of these cardiac procedures. We found the central venous lactate concentrations to be higher than arterial lactate values in the cardiac surgical group, and there was a very poor correlation (r2 = .07) between arterial and central venous lactate values in the cardiac surgical group. The correlation between central venous and arterial lactate concentrations in the PTCA patients was excellent (r2 = .84) and similar to the findings of experiment 1. Since the cardiac surgical patients received RL and the PTCA patients received no RL, we speculated that the intravenous infusion of RL in the cardiac surgical group accounted for these discordant findings. To test this speculation, we performed experiments 3 and 4. Experiment 3: In a large bench study, blood specimens were divided into multiple 1-mL aliquot portions, to which 0.01, 0.05, 0.10, 0.50, or 1.0 mL of various crystalloid solutions, containing or not containing RL, were added. In a volume-dependent and linear manner, solutions containing RL increased the circulating lactate concentration from 10% to > 400% of the baseline lactate value. In a volume-dependent and linear fashion, the non-RL crystalloid solutions decreased the lactate concentration by 0 to 66% of the baseline nondiluted lactate concentration. Experiment 4: In 30 different cardiac surgical patients, we simultaneously obtained central venous and arterial blood specimens. Patients this time received no RL, and catheter lines were adequately cleared (removal > 5 mL) of crystalloid solutions. We found a correlation (r2 = .82; p < .0001) that was virtually identical to the findings of experiment 1 and to the findings in the PTCA group of experiment 2. CONCLUSIONS: a) Arterial and central venous lactate concentrations are similar in hemodynamically stable critically ill patients, b) Even small amounts of RL-containing solutions in catheters used for blood sampling may cause false increases in the circulating lactate concentration. c) Even small amounts of non-RL crystalloid solutions in catheters used for blood sampling may falsely decrease circulating lactate values. d) When blood specimens are drawn from indwelling catheters, all crystalloid solutions must be cleared from the line.

Blood Specimen Collection↗

Hepatotoxicity of clozapine.

Two hundred thirty-eight patients treated with either haloperidol or clozapine were investigated to shed more light on the incidence and severity of antipsychotic-induced liver enzyme increase. Serum glutamic-pyruvic transaminase (SGPT) increase was most frequently seen in both treatment groups. When analyzing the incidence rates for patients with increased liver enzyme values (serum glutamic-oxaloacetic transaminase, SGPT, gamma-glutamyl transpeptidase) that were higher than twice the upper limit of the normal range, clozapine-treated patients showed an SGPT increase (37.3%) significantly more frequently than patients treated with haloperidol (16.6%). Both patients with higher clozapine plasma levels and male patients were at a higher risk for an SGPT increase. At least 60% of the increase of the different enzymes remitted within the first 13 weeks of treatment. In general, the authors conclude that clozapine-induced liver enzyme elevation seems to be a common and mostly transient phenomenon.

Adult↗

The combination of epidermal growth factor and transforming growth factor-beta induces novel phenotypic changes in mouse liver stem cell lines.

Mouse liver stem cell (oval cell) lines were investigated in order to determine the role which two families of growth and differentiation factors (GDFs), epidermal growth factor (EGF) family and transforming growth factor beta (TGF-beta) family, play in liver regeneration. EGF family members, including EGF, amphiregulin, betacellulin, heparin-binding epidermal growth factor, and TGF-alpha, were mitogenic for oval cell lines while TGF-beta family members, including TGF-beta1, TGF-beta2 and TGF-beta3, inhibited mitogenesis and induced apoptosis in oval cell lines. Surprisingly, the combination of EGF family members and TGF-ss family members resulted in neither proliferation nor apoptosis but instead in a novel cellular response, cellular scattering in tissue culture and morphological differentiation in Matrigel. Analysis of the signal transduction pathways activated by exposure of oval cell lines to either EGF, EGF+TGF-beta, or TGF-beta indicated that novel combinations of intracellular signals result following stimulation of the cells with the combination of EGF+TGF-beta. These data reveal that the dynamics of synergistic GDF action following tissue injury and regeneration results in a new level of complexity not obvious from the study of individual GDFs.

Animals↗

Empirical approaches for the investigation of toxicant-induced loss of tolerance.

It has been hypothesized that sensitivity to low-level chemical exposures develops in two steps: initiation by an acute or chronic chemical exposure, followed by triggering of symptoms by low levels of previously tolerated chemical inhalants, foods, or drugs. The Working Group on Toxicant-induced Loss of Tolerance has formulated a series of research questions to test this hypothesis: Do some individuals experience sensitivity to chemicals at levels of exposure unexplained by classical toxicological thresholds and dose-response relationships, and outside normally expected variation in the population? Do chemically sensitive subjects exhibit masking that may interfere with the reproducibility of their responses to chemical challenges? Does chemical sensitivity develop because of acute, intermittent, or continuous exposure to certain substances? If so, what substances are most likely to initiate this process? An experimental approach for testing directly the relationship between patients' reported symptoms and specific exposures was outlined in response to the first question, which was felt to be a key question. Double-blind, placebo-controlled challenges performed in an environmentally controlled hospital facility (environmental medical unit) coupled with rigorous documentation of both objective and subjective responses are necessary to answer this question and to help elucidate the nature and origins of chemical sensitivity.

Controlled Clinical Trials as Topic↗