Inward-rectifying potassium channels in the rabbit superior lacrimal gland.
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Animals↗
Biomedical subjects
D K Martin
Publications and source records attributed to D K Martin.
Can raters consistently evaluate the content of focus groups?
Focus groups are increasingly being used to provide insights to researchers and policy makers. These data complement quantitative approaches to understanding the world. Unfortunately, quantitative and qualitative methodologies have often been viewed as antithetical, rather than complementary, strategies. While focus groups can clearly generate rich information that is unobtainable through other quantitative methods, it is important to determine the degree to which different raters can consistently extract information from transcripts. Thus, our goal was to quantify agreement in the interpretation of transcripts from patient and physician focus groups, using decision-making in ischemic heart disease as a model. We used data from focus groups with both patients and physicians that sought to identify factors affecting diagnostic and treatment decisions in ischemic heart disease. Three raters independently reviewed transcribed audiotapes from focus groups of patients with ischemic heart disease, as well as focus groups of physicians who care for these patients. We found that raters could not distinguish between major and minor factors reliably. More troubling, however, is that consistency regarding the apparently straightforward judgment as to the mere presence or absence of a factor was difficult to achieve. In particular, the three raters of each transcript failed to agree on between one third and one half of the factors. This reasonably high level of disagreement occurred despite the raters: (1) having generated the individual factors themselves based upon their reading a random sample of actual transcripts and (2) being trained in the use of rating forms (including standard definitions of themes). These data suggest that if a single rater evaluates focus group transcripts, as is commonly done, judgments may not be reproducible by other raters. Moreover, a single rater may not extract all important information contained in the transcripts.
Female↗
Characterization of an inwardly rectifying potassium channel in the rabbit superior lacrimal gland.
PURPOSE: To characterize the properties of an inwardly rectifying K+ (KIR) current in fresh, enzymatically isolated acinar cells from the rabbit superior lacrimal gland. METHODS: New Zealand White rabbits of both sexes were killed by injecting 45 mg/kg pentobarbital sodium, and the glands were excised. Single acinar cells were isolated enzymatically from these glands. Standard patch-clamp techniques were used to record ion currents. RESULTS: Hyperpolarizing voltages evoked KIR currents that had a conductance of 2.7 +/- 0.16 nS (n = 6) in the range -50 mV to -160 mV. The KIR current was activated with steep voltage dependence on hyperpolarization, and the conductance was approximately dependent on the square root of the external K+ concentration. Increasing the pipette Ca2+ concentration from 10(-9) M to 10(-6) M increased the conductance to 5.3 +/- 0.45 nS (n = 7). Internal substitution of K+ with various cations gave the following permeability sequence: K+ (1.0) > Rb+ (0.83) > Li+ (0.15). The KIR current was inhibited by Ba2+ (100 microns), tetraethylammonium (10 mM), and Cs+ (5 mM) but was insensitive to 4-aminopyridine (5 mM). The single-channel conductance was 43 +/- 2.7 pS (n = 11), and the relationship between between single-channel conductance (gamma) and external K+ concentration ([K]o) is given by: gamma = 7.04[K]o0.37 (pS, r2 = 0.99, P < 0.05). The relationship between [K]o and zero current potential (Erev) is given by: Erev = 35.5 log[K]o - 77.8 (mV; r2 = 0.99, P < 0.05). CONCLUSIONS: The KIR current identified in these lacrimal acini has a similar dependence on [K]o as other inward rectifiers of excitable tissues and exocrine glands. However, this study highlights that there are interspecies variations and similarities between KIR channels that could be related to their individual physiological roles. The authors' investigations suggest that one role of the KIR channel in the rabbit superior lacrimal gland acinar cells is to set and stabilize the resting membrane potential. However, this KIR channel may also be involved in secretion, as has been shown to occur in the sheep parotid gland.
Animals↗
In vitro transesterification of cocaethylene (ethylcocaine) in the presence of ethanol. esterase-mediated ethyl ester exchange esterase-mediated ethyl ester exchange.
This study reports that cocaethylene undergoes an esterase-mediated ethyl ester exchange with ethanol, resulting in an increase in the apparent in vitro t1/2, compared with control conditions. Homogenized liver from male Sprague Dawley rats in pH 7.4 phosphate buffer was centrifuged at 9000g, and the resulting supernatant (S9) fraction was collected. Tubes containing the rat S9 fraction and 50 microM cocaethylene plus aqueous buffer (control), 50 mM ethanol, or 51. 3 mM 2H6-ethanol were incubated at 37 degrees C for 4 hr. Samples were collected from the incubation tubes at various times, extracted with a solid-phase extraction system, and assayed for cocaethylene and 2H5-cocaethylene by GC/MS. Concentration-time profiles were constructed and kinetic parameters were determined. The experiment was repeated in the presence of specific and nonspecific esterase inhibitors. Enzyme kinetic parameters were also determined. Cocaethylene underwent ethyl ester exchange, being converted to 2H5-cocaethylene in the presence of 2H6-ethanol. The average apparent in vitro t1/2 value for cocaethylene (13.0 +/- 1.4 min) incubated with the S9 fraction and buffer only was increased approximately 5-fold (67.8 +/- 0.3 min) in the presence of ethanol. Formation of 2H5-cocaethylene was totally blocked with the addition of bis-(p-nitrophenyl)phosphate but was unaffected by physostigmine. The intrinsic metabolite formation clearance of 2H5-cocaethylene from cocaethylene and 2H6-ethanol (1.92 +/- 0.03 microl/min.mg protein) was several times greater than the corresponding value for cocaethylene formation from cocaine and ethanol (0.94 +/- 0.01 microl/min.mg protein) or 2H6-ethanol (0.87 +/- 0.04 microl/min.mg protein).
Animals↗
California's county hospitals and the University of California graduate medical education system. Current issues and future directions.
California's county hospitals train 45% of the state's graduate medical residents, including 33% of residents in the University of California system. This paper describes the interrelationships of California's county hospitals and the University of California (UC) graduate medical education (GME) programs, highlighting key challenges facing both systems. The mission of California's county health care systems is to serve all who need health care services regardless of ability to pay. Locating UC GME programs in county hospitals helps serve the public missions of both institutions. Such partnerships enhance the GME experience of UC residents, provide key primary care training opportunities, and ensure continued health care access for indigent and uninsured populations. Only through affiliation with university training programs have county hospitals been able to run the cost-effective, quality programs that constitute an acceptable safety net for the poor. Financial stress, however, has led county hospitals and UC's GME programs to advocate for reform in both GME financing and indigent care funding. County hospitals must participate in constructing strategies for GME reform to assure that GME funding mechanisms provide for equitable compensation of county hospitals' essential role. Joint advocacy will also be essential in achieving significant indigent care policy reform.
California↗
Molecular cloning and expression of a chloride ion channel of cell nuclei.
Ion channels are known to be present on the plasma membrane of virtually all cells and have been found on the membranes of various intracellular organelles. However, until recently they were believed not to occur at the nuclear membrane. In this study we describe the molecular cloning and characterization of a nuclear ion channel protein, designated nuclear chloride channel-27 (NCC27), from the human myelomonocytic cell line, U937. NCC27 is a novel chloride ion channel protein that was found to localize principally to the cell nucleus. Its only known homologue is a bovine chloride ion channel protein (p64) believed to localize to internal organelles. NCC27 therefore represents the first human member of a new class of organellar chloride ion channel proteins.
Amino Acid Sequence↗
Leptin is a four-helix bundle: secondary structure by NMR.
Leptin is a signaling protein that in its mutant forms has been associated with obesity and Type II diabetes. The lack of sequence similarity has precluded analogies based on structural resemblance to known systems. Backbone NMR signals for mouse leptin (13C/15N -labeled) have been assigned and its secondary structure reveals it to be a four-helix bundle cytokine. Helix lengths and disulfide pattern are in agreement with leptin as a member of the short-helix cytokine family. A three-dimensional model was built verifying the mechanical consistency of the identified elements with a short-helix cytokine core.
Amino Acid Sequence↗
Examination of racial differences in management of cardiovascular disease.
OBJECTIVES: We sought to identify the clinical characteristics associated with, and to investigate the impact of cohort selection criteria on, interracial use of invasive cardiac procedures and to determine survival. BACKGROUND: Although interracial differences in the use of invasive cardiac procedures have been previously reported, the underlying reasons are not known. METHODS: A retrospective cohort study was conducted at a Veterans Affairs Medical Center. Study patients were evaluated for cardiovascular disease between January 1 and December 31, 1993. RESULTS: The study included 1,406 male patients (85% white, 58% married), with a mean age of 63.4 years. African Americans were less likely than whites to undergo procedures (cardiac catheterization: odds ratio [OR] 0.37, 95% confidence interval [CI] 0.24 to 0.58; coronary angioplasty: OR 0.60, 95% CI 0.25 to 1.49; coronary bypass surgery: OR 0.22, 95% CI 0.08 to 0.63; any procedure: OR 0.32, 95% CI 0.21 to 0.50). On bivariate analysis, patients who underwent cardiac procedures were more likely to be younger, married and reside nonlocally and less likely to have severe comorbid disease; however, African Americans were less likely to be married and to reside nonlocally and more likely to have severe comorbid disease. Cohorts adjusting for referral status and specified cardiac diagnoses reduced or reversed interracial treatment differences. Thirty-day and 1-year survival rates (96% and 87.6%, respectively) were equivalent. CONCLUSIONS: Racial disparity in invasive cardiac procedure use may be partially explained by clinical differences and cohort selection bias. Despite treatment differences, survival rates were equivalent in African Americans and whites.
Black or African American↗
Carboxylesterase-mediated transesterification of meperidine (Demerol) and methylphenidate (Ritalin) in the presence of [2H6]ethanol: preliminary in vitro findings using a rat liver preparation.
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Animals↗
Kainic acid blocks a TTX-sensitive sodium channel in retinal horizontal cells of the turtle (Pseudemys scripta elegans).
The aim of this study was to investigate the effects of excitatory amino acids on channels found in horizontal cell membranes using patch-clamp techniques. We unexpectedly found that the excitatory amino acid receptor agonist, kainic acid, reversibly inhibited the transient tetrodotoxin (TTX)-sensitive Na+ current in isolated horizontal cell bodies and axons from the retina of the turtle (Pseudemys scripta elegans). The effect of kainic acid was antagonized by the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione. Kainic acid activated a non-selective cation current, a finding that was consistent with previous reports, and which would account for the kainate induced depolarisation of these cells. The inhibition of the transient TTX-sensitive Na+ current by kainic acid might be important in the modification of the kinetics of responses to excitatory amino acid analogues often observed during intracellular recording from these cells.
Animals↗
Some techniques for the analysis of work sampling data.
Work sampling is an observational technique that produces counts representing the number of times that an individual has been observed performing each of several tasks. These data are collected using either systematic or random times of observation, and typically exhibit correlation between repeated observations on the same individual, with the degree of correlation being a function of the amount of time elapsed between measurements. Using several recently developed statistical techniques, we illustrate how it is possible to carry out analyses of these nominal outcomes that account for the correlation between repeated outcomes. We use description of a work sampling study to motivate the techniques and we compare empirically results from analyses based on several different underlying assumptions.
Adult↗
Chloride ion channels are associated with adherence of lymphatic endothelial cells.
We have examined the role of chloride ion channels as part of the control mechanism for adherence and growth of lymphatic endothelial cells. The chloride channel inhibitor N-phenylanthranilic acid (0.1 mM) inhibited the initial adherence of previously nonadherent cells in the period up to 3 hr following seeding onto gelatin-coated culture dishes. The potassium channel inhibitor 4-aminopyridine (1 mM) had no effect on the rate of cell adherence. N-Phenylanthranilic acid had no effect when added to confluent monolayers of cells that had been growing for 3 days. This, and the observation that N-phenylanthranilic acid did not affect the ultrastructure of the cells, suggested that plasma membrane chloride channels are involved in the initial signaling cascade triggered when lymphatic endothelial cells begin to adhere to a substrate.
Animals↗
Risk factors for nonelective hospital readmissions.
We previously reported a predictive model that identified potentially modifiable risk factors for nonelective readmission to a county hospital. The objectives of this study were to determine if those risk factors were generalizable to a different population. We found that the previously reported risk factors were generalizable, and other potentially modifiable risk factors were identified in this population of veterans. However, further research is needed to establish whether or not the risk factors can be modified and whether or not modification improves outcomes.
Aged↗
Candida albicans induces selective expansion of human T lymphocytes expressing the T-cell receptor variable region V beta 5.1.
Candida albicans is a common pathogen which can present major problems as an opportunistic skin pathogen in patients with immunodeficiency. The exact nature of the T cell responses to C. albicans is poorly understood. The purpose of this study was to determine whether C. albicans could stimulate the selective expansion of T lymphocytes expressing particular V beta gene segments. Human T lymphocytes stimulated in vitro with an extract of C. albicans were analyzed for T cell receptor V beta gene expression by using a quantitative PCR technique. We found that stimulation of peripheral blood mononuclear cells (PBMC) produced a selective increase in the expression of V beta 5.1 and 5.2 gene transcripts. Using cytofluorographic analysis with available anti-V beta monoclonal antibodies, we verified that there was a significant selective expansion (P = 0.035) of V beta 5.1 positive T lymphocytes in PBMC from six subjects stimulated in vitro with C. albicans. PCR analysis of V beta 5.1 expansion in 10 subjects showed increases in V beta 5.1 gene transcripts in 7/10 subjects. More importantly, analysis of the T cell infiltrate 48 h after intradermal injections with C. albicans also showed significant expression of V beta 5.1 in the infiltrates, along with the infiltration of V beta 8.1 + T cells. The selective expansion of V beta 5.1 bearing T lymphocytes in PBMC stimulated with C. albicans and in skin test reactions to C. albicans suggests that a restricted population of T cells react to C. albicans. Furthermore, our present data raise the provocative possibility that one or more antigens in C. albicans can act as a superantigen, producing selective expansion of a population of T lymphocytes bearing a particular V beta specificity.
Antibodies, Monoclonal↗
Immunohistochemical and histochemical characterisation of epithelial cells of rabbit lacrimal glands in tissue sections and cell cultures.
The purpose of this study was to establish conditions for isolation and long term culture of acinar cells from the Harderian gland, and superior and inferior lacrimal glands of the rabbit and to compare the in vitro growth patterns of cultured cells from these glands. In order to determine the predominant cell type in the cultures, cells and tissue sections were stained using a variety of antibodies to cytokeratins, smooth muscle actin, and neuron specific enolase. Similarly, PAS and alcian blue histochemistry were used to test for the presence of mucins. The glands were excised and cells isolated using enzymatic digestion and then established in long term culture. Different media and substrata were trialed for suitability. When cultured on uncoated Costar plastic in DMEM/10%FBS, the pattern of cell growth was similar for all glands with distinct phases involving aggregation and migration out from the aggregates before cells died between 20 to 30 days. Immunohistochemical staining indicated that the cultures were of acinar cells with a small percentage of ductal cells. The acinar cells of the lacrimal glands in situ and in vitro stained with antibody MNF116 directed against cytokeratins 5, 6, 8 and 17 but did not stain for antibodies to cytokeratin 18. The reverse staining pattern was true for the Harderian gland. Sections from the white lobe of the Harderian gland showed islets of serous secreting cells which showed positive staining when MNF116 was used. In situ, PAS positive cells were found in a small number of demilunes in the superior and inferior lacrimal glands and also in cells of the intercalated ducts. Surprisingly, in culture nearly all cells, including those isolated form the Harderian gland became PAS positive. In this study we have demonstrated that acinar cells from the Harderian and lacrimal glands of rabbit can be isolated and maintained in culture for 20 to 30 days, and that despite dramatic morphological changes, these cells retain their distinctive phenotype as indicated by antibody staining to specific cellular structural proteins such as cytokeratins and actin. However, the cultured cells also begin to produce mucins as indicated by PAS staining.
Animals↗
Characterization of the properties of cocaine in blood: blood clearance, blood to plasma ratio, and plasma protein binding.
As part of a study to examine cocaine disposition and interaction with ethanol, it was necessary to characterize various properties of cocaine in the blood of the experimental animal. All studies were conducted using blood from healthy adult male Sprague-Dawley rats. Cocaine was incubated in whole blood at 37 degrees C at concentrations of 500-4000 ng/mL. The apparent first-order rate constant for cocaine loss was independent of concentration. Blood clearance, calculated assuming blood volume to be 64 mL/kg, was estimated to be 0.056 +/- 0.003 mL/(min.kg); a value considerably smaller than estimates of systemic clearance. The addition of NaF increased the rate of loss to form benzoylecgonine, as a result of increased chemical degradation and as a consequence of increased pH (to pH 8.0 over 30 h). This NaF-enhanced degradation was abolished when NaF was added to blood buffered to pH 7.4. Ethanol had no influence on cocaine degradation, and there was no evidence of cocaethylene (ethylcocaine) formation. Blood to plasma ratios determined in spiked and authentic samples were constant (0.94-1.05 and 0.99-1.03, respectively) and independent of concentration (100-1500 ng/mL) and pH (7.2-7.6). This ratio was not influenced by NaF or ethanol. The unbound fraction (fu) of cocaine determined in spiked plasma varied from 0.62 to 0.63 over the concentration range (75-2025 ng/mL). Ethanol had no effect on binding. The values for fu determined from authentic blood samples taken from rats dosed intravenously with cocaine (10 mg/kg) ranged from 0.67 to 0.69 (over the concentration range 300-1500 ng/mL). Cocaine plasma protein binding was independent of concentration but depended upon plasma pH (fu, 0.765 and 0.486, at pHs 7.0 and 7.8, respectively.
Animals↗
An intervention on discharge polypharmacy.
OBJECTIVE: To determine if providing a way to cancel pre-admission prescriptions would reduce the number of active drug prescriptions (RXs) at discharge. DESIGN: A randomized non-blinded clinical trial. SETTING: Inpatient acute medical service of a university affiliated Veterans Administration medical center. PARTICIPANTS: Twelve medicine ward teams were randomized to control and intervention groups. Patients controlled had been discharged from these teams during 12 weeks and were receiving outpatient medications from this facility at hospital admission; control = 180, intervention = 168. INTERVENTION: At discharge, intervention teams used a computer-generated drug list to cancel or renew previous outpatient RXs or to prescribe new medications. Control teams could not cancel outpatient drugs and wrote all medications on individual prescriptions. MEASUREMENTS: The difference between admission and discharge RXs. RESULTS: There were no significant differences in patients' age, sex, race, Charlson Index (CI), or LOS between patient groups at discharge. The intervention group had fewer RXs on admission (5.4 vs 6.2, P < .05) and at discharge was not significantly different (2.9 vs 2.9, P = .87) from the control group. CONCLUSIONS: Providing a method for canceling pre-admission medications did not reduce the number of RXs at discharge. Further research is needed to evaluate the appropriateness of the large increase in RXs from admission to discharge for patients in acute hospital settings.
Analysis of Variance↗
Effect of the class III antiarrhythmic agent E-4031 on the ATP-sensitive potassium channel in rabbit ventricular myocytes.
The class III antiarrhythmic drug E-4031, a known blocker of the delayed rectifier potassium channel (IK), might also be capable of blocking the ATP-sensitive potassium channel (IKATP). We examined this possibility by studying the effect of E-4031 on single IKATP channels in membrane patches excised from ventricular myocytes that were obtained by standard enzymatic dissociation techniques from New Zealand white rabbits. In inside-out patches, E-4031 caused a dose-dependent block of IKATP with an EC50 of 31 +/- 1 microM, Hill coefficient of 0.89 +/- 0.24 and no effect on channel conductance. Open dwell-time kinetics were fitted by two exponential components, with E-4031 causing reduction of the longer time constant. In outside-out patches, the concentration of E-4031 required to produce blockade was much higher. We conclude that E-4031 blocks the ATP-sensitive potassium channel and that it does so from within the cytoplasm, with one-to-one channel binding stoichiometry. Single channel conductance is unchanged, but the longer time constant for the open state is reduced, which suggests that E-4031 may be an open channel blocker of intermediate to slow time course.
Adenosine Triphosphate↗