Search PubMed⌕ Search

Biomedical subjects

R M Smith

Publications and source records attributed to R M Smith.

At least 199 records · Page 11Linked to original sources
PubMed1994

Fatigue in operational settings: examples from the aviation environment.

The need for 24-h operations creates nonstandard and altered work schedules that can lead to cumulative sleep loss and circadian disruption. These factors can lead to fatigue and sleepiness and affect performance and productivity on the job. The approach, research, and results of the NASA Ames Fatigue Countermeasures Program are described to illustrate one attempt to address these issues in the aviation environment. The scientific and operational relevance of these factors is discussed, and provocative issues for future research are presented.

Aerospace Medicine↗
PubMed1994

Acute effects of a single dose of porcine surfactant on patients with the adult respiratory distress syndrome.

In an attempt to restore functional surfactant to the lungs of patients with the adult respiratory distress syndrome (ARDS), we have treated six patients within the first 2 days of the onset of ARDS with a single dose of hydrophobic components of porcine surfactant. Surfactant (4 g in 50 ml) delivered via a bronchoscope in aliquots to each of the lobar bronchi was well tolerated and caused a modest transient improvement in gas exchange. No significant changes in chest radiograph or lung compliance were detected. Analysis of bronchoalveolar lavage (BAL) fluid showed no change in albumin, alpha-1-proteinase inhibitor specific activity, or cell count. Bronchoalveolar lavage phospholipid concentrations were elevated 3 h after surfactant administration relative to preadministration levels and fell by 24 h. In addition, in two patients we found reduced inhibition of surfactant function in BAL after surfactant replacement. These observations suggest a role for surfactant replacement in the treatment of patients with ARDS and support the need for continuing investigation.

Adult↗
PubMed1994

Anti-class II MHC antibodies prevent and treat EAE without APC depletion.

We have demonstrated that a murine IgG1 anti-class II major histocompatibility complex (MHC) antibody (OX6) is able both to prevent and treat experimental autoimmune encephalomyelitis (EAE) in the Biozzi AB/H mouse. We have confirmed that this antibody is poorly cytotoxic, allowing mechanisms of action other than depletion of antigen-presenting cells (APC) to be assessed. Despite its effect on disease, OX6 administration does not prevent priming of T cells in the draining lymph node, implying an alternative mechanism for regulation of the autoimmune disease process.

Acute Disease↗
PubMed1994

A subgroup of murine monoclonal anti-deoxyribonucleic acid antibodies traverse the cytoplasm and enter the nucleus in a time-and temperature- dependent manner.

BACKGROUND: The capacity of lupus autoantibodies to enter living cells and bind to molecules for which they have intrinsic affinity is not well appreciated. In previous studies, we identified a subgroup of three murine monoclonal IgG anti-DNA antibodies, derived from lupus-prone MRL-lpr/lpr mice, that localized within nuclei of cells in multiple organs and induced functional perturbations, in vivo, after passive transfer to normal mice. To examine the mechanisms of this phenomenon, we now extend these observations, using the same monoclonal anti-DNA antibodies and cultured cell lines. EXPERIMENTAL DESIGN: Multiple experimental approaches were utilized to track nuclear localization of anti-DNA antibodies, including direct immunofluorescence, confocal microscopy and immunoelectron microscopy. The requirements for nuclear localization were further evaluated quantitatively, in nuclei isolated from co-cultures of cells and 125I-Ig, under varying experimental conditions. RESULTS: Nuclear localization was observed with the same subset of anti-DNA antibodies that localized within nuclei in vivo; it was dependent on the antigen-binding region of the molecule; and it was not found with other anti-DNA antibodies. At progressive intervals, the Ig were observed: at the cell surface, within the cytoplasm, clustered at the nuclear pore, and within the nucleus. Nuclear localization of Ig was found to be a time- and temperature- dependent process, specific for a subset of anti-DNA antibodies and dependent on the antigen binding region of the Ig. CONCLUSIONS: This is the first demonstration that monoclonal autoantibodies can traverse both the cell and nuclear membranes to localize within the nuclei of cultured cells. Furthermore, nuclear localization of Ig was regulated in a manner analogous to that of other large cytoplasmic proteins that enter the nucleus. This confirms and extends our results using the same antibodies in whole animals, and it provides the basis to further examine the underlying mechanisms and consequences of this phenomenon.

Animals↗
PubMed1994

Sodium transport activity in cheek epithelial cells from adolescents at increased risk of hypertension.

Sodium transport including amiloride-sensitive Na+/H+ antiporter activity was measured in cheek epithelial cells of adolescents displaying either high or low BP tracking characteristics and in a subgroup of high BP tracking adolescents exhibiting a positive family history of hypertension. From the BP tracking behaviour of over 500 adolescents measured over a period of three years, 24 low BP tracking and 29 high BP tracking adolescents were recruited for the study. Cheek cells were collected from these subjects and proton-dependent, amiloride-sensitive Na+/H+ antiporter activity and the response of this antiporter to a proton gradient were measured. Cheek cell Na+/H+ antiporter activity was 50% lower (P = 0.0004) in the high BP tracking group (1.02 +/- 0.15 nmol Na+/mg protein/5 min (mean +/- SEM) compared with the activity in the low BP tracking group (2.05 +/- 0.24). A significantly lower Na+/H+ antiporter activity (69%; P < 0.01) was also apparent in the high BP tracking adolescents with family history of hypertension (n = 7) compared with the low BP tracking group. The graded response of cheek cell Na+/H+ antiporter activity to the proton gradient was 58% lower (P = 0.0039) for adolescents in the high BP tracking group compared with the low BP tracking group. Passive Na+ influx was also significantly lower in the cheek cells of the high BP tracking group. Our results therefore show that the activity of the Na+/H+ antiporter in cheek cells and the passive Na+ transport activity are lower in those adolescents considered at greatest risk of future development of essential hypertension.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

Combining multileaf fields to modulate fluence distributions.

PURPOSE: A method for modulating beam fluence from a linear accelerator is discussed. The beam modulation is accomplished remotely using a multileaf collimator and does not require entering the treatment room. METHODS AND MATERIALS: The multileaf collimator is used to define a series of field shapes that are superimposed at a fixed gantry angle to produce any desired fluence pattern. A heuristic technique for deriving the field shapes and corresponding monitor unit settings is described. The technique has been tested on randomly generated fluence distributions and on distributions with a limited number of peaks and valleys. The second type of distribution more closely simulates fluence patterns obtained with dose optimization software. Estimates of the time required to use this approach to treat a four-field plan are given and compared to the technique of placing a physical compensator in each beam. RESULTS: It has been demonstrated that complex fluence patterns within a 15 x 15 cm2 field can be achieved with less than 20 fields. Estimates show that this technique is faster than entering the treatment room to change physical compensators. Some limitations of the method are discussed. CONCLUSION: Optimized distributions that conform the dose to irregularly shaped target volumes that wrap around critical structures are possible using superimposed multileaf fields. A method for defining the field shapes is presented.

Humans↗
PubMed1993

Functions of the IGFs in early mammalian development.

The identification of growth factors and/or receptors produced by mammalian embryos or present in the maternal reproductive tract is of basic interest, as well as having practical application. Early studies established that receptors binding insulin and the insulin-like growth factors (IGFs) are expressed by preimplantation mouse embryos. These studies have been confirmed at the molecular level using RT-PCR techniques. In addition, high resolution electron microscopy has shown that insulin is internalized by the cells of the blastocyst stage mouse embryo, and that immunologically intact insulin is detectable in the cells of the trophectoderm and inner cell mass. Similar studies with gold labelled IGF-I have shown that this ligand is also bound and internalized by mouse blastocysts. However, although all blastocysts express receptors that bind IGF-I on the basolateral cell surface of the trophectoderm, only 30% exhibit apically located receptors. In order to elucidate the functions of IGFs in early mouse development, we are in the process of constructing protein data-bases for embryos at the eight-cell and blastocyst stage. By the use of the database, it should prove possible to elucidate targets of growth factor action.

Animals↗
PubMed1993

Genistein inhibits insulin-stimulated glucose transport and decreases immunocytochemical labeling of GLUT4 carboxyl-terminus without affecting translocation of GLUT4 in isolated rat adipocytes: additional evidence of GLUT4 activation by insulin.

A recent study from this laboratory (Abler et al., J. Biol. Chem. 267, 18172-18179, 1992) showed genistein blocked insulin-stimulated glucose oxidation without affecting receptor autophosphorylation or tyrosine kinase activity. The mechanism by which genistein inhibited insulin-stimulated glucose metabolism was investigated in the present study. Insulin caused a approximately 12-fold increase in 3-O-methyl-D-glucose (3OMG) uptake compared to that of control cells. Basal and insulin-stimulated 3OMG transport was inhibited 40-60% by genistein in a concentration-dependent manner (10-100 micrograms/ml). Genistein had no effect on insulin-stimulated GLUT4 translocation from low density microsomes to plasma membranes as determined by Western blotting. These results suggested that genistein inhibited glucose transport in adipocytes by decreasing the intrinsic activity, rather than the number, of the plasma membrane-associated glucose transporters. We also previously reported that insulin treatment of adipocytes resulted in the immunocytochemically visualized unmasking of the carboxyl-terminus of plasma membrane-associated GLUT4 and suggested the unmasking might be related to an insulin-induced increase in the intrinsic activity of the glucose transporter (Smith et al., Proc. Natl. Acad. Sci. USA 88, 6893-6897, 1991). In the present study, genistein decreased immunocytochemical labeling of plasma membrane-associated GLUT4 by approximately 50% in control and insulin-treated adipocytes by carboxyl-terminus antibodies but had no effect on labeling observed in an amino-terminus antibody. Since genistein did not affect the number of plasma membrane-associated GLUT4 transporters, this result supports the hypothesis that conformational changes in the glucose transporter, reflected by the ability of anti-carboxyl-terminus antibodies to bind to the transporter, may be an indication of the intrinsic activity of the plasma membrane-associated transporter. We therefore conclude that conformational changes in and activation of glucose transporters, in addition to insulin-stimulated GLUT4 translocation, play an important role in insulin-regulated glucose transport in adipocytes.

3-O-Methylglucose↗
PubMed1993

Insulin induces an unmasking of the carboxyl terminus of G(i) proteins in rat adipocytes.

Several groups have shown a relationship between the insulin receptor and inhibitory G proteins, G(i). An antisera, 8729, to a peptide sequence (KNNLKDCGLF) corresponding to the carboxyl termini of G(i)alpha subunits was used to investigate this relationship by immunoelectron microscopy. Rat adipocytes were incubated in the absence or presence of 100 ng/ml insulin for 1 h and fixed for immunoelectron microscopy. Insulin-treated adipocytes stained with 8729 were labeled at the cell surface at a much higher density than control adipocytes. Subcellular fractionation of insulin-treated and control cells was followed by PAGE and Western blots of the plasma membrane and low-density microsomes with 8729. The density of the bands did not change in response to insulin treatment. Antibodies to noncarboxyl terminus sequences of the alpha subunit were used for immunoelectron microscopy and no difference was noted between insulin-treated and control adipocytes. These results indicated that 8729 was detecting a conformational change in the structure of G(i)alpha subunit in the plasma membrane in response to insulin. This unmasking of the carboxyl terminus was also seen in response to treatment with phenylisopropyladenosine and prostaglandin E2. Pertussis toxin-catalyzed ADP ribosylation also unmasked the carboxyl terminus. In contrast, isoproterenol, an agonist of stimulatory G proteins (Gs), did not induce an unmasking of the carboxyl terminus. These results support the hypothesis that some of insulin's effects are mediated through G(i) proteins in adipocytes.

Adipose Tissue↗
PubMed1993

Comparisons of specific and nonspecific bronchoprovocation in subjects with asthma, rhinitis, and healthy subjects.

BACKGROUND: We studied subjects with atopic asthma, atopic rhinitis, and nonatopic healthy subjects to evaluate responsiveness to bronchoprovocation with both methacholine and allergen. METHODS: Subjects with a demonstrable FEV1 PD20 to methacholine or allergen (responders) were further analyzed for putative sensitivity (PD20 FEV1) and reactivity (dose-response slopes) to determine whether any characteristics could distinguish individuals with asthma from other responders. Subjects were recruited without sex restrictions and were between the ages of 18 and 45 years old. They were nonsmokers, had no other medical problems, and were free of upper respiratory infection for at least 6 weeks before challenge. All had a history taken, physical examination, limited laboratory screening, chest radiography, pulmonary function testing, and intradermal skin testing before admission to the study. RESULTS: Although the groups were significantly different in both sensitivity and reactivity to methacholine, responses to allergen bronchoprovocation were sufficiently similar between responders with asthma and those with rhinitis to prevent separation on the basis of either sensitivity or reactivity. The fall in FEV1 at the nadir of the late response, which was greater in the asthma group, was significantly correlated with sensitivity and reactivity of the immediate response to allergen but not to methacholine. Regression analysis demonstrated a stronger association between allergen and methacholine responsiveness in subjects with rhinitis than in subjects with asthma. CONCLUSION: We concluded that (1) nonspecific bronchial hyperresponsiveness fails to explain why patients with allergic asthma have clinical asthma as a result of allergen exposure and patients with allergic rhinitis do not; (2) hyperresponsiveness to allergen does not simply reflect quantitative or qualitative airway nonspecific hyperresponsiveness; and (3) clinical asthma may involve mechanisms difficult to elucidate by laboratory bronchoprovocation techniques.

Adolescent↗
PubMed1993

Mouse preimplantation embryos exhibit receptor-mediated binding and transcytosis of maternal insulin-like growth factor I.

High-resolution microscopy in conjunction with colloidal gold-labeled insulin-like growth factor I (IGF-I) has been used to provide evidence that the IGF-I receptor is first detected in 8-cell-stage mouse embryos, confirming the results of previous reverse transcriptase polymerase chain reaction (RT-PCR) studies. Specificity for the IGF-I receptor was demonstrated by displacement with unlabeled IGF-I and dual-labeling experiments with colloidal gold-labeled or unlabeled insulin. Labeled IGF-I ligand is internalized by means of receptor-mediated endocytosis following its concentration in coated pits, and it can be visualized within cytoplasmic organelles. Immunocytochemical analyses at the blastocyst stage, using gold-labeled antibodies to the receptor, confirmed the expression of IGF-I receptors on all cells of the embryo. Similar studies with antibodies directed against the ligand demonstrated that IGF-I internalized by the embryo in vivo is maternally derived. Approximately 40% of blastocysts showed apical plasma membrane binding of gold-labeled ligand ("responders"), while approximately 60% did not demonstrate binding ("nonresponders"); however, both classes of embryo expressed receptors on basolateral membranes of trophectoderm cells and on the surface of inner masses. Functional studies show that incubating embryos in physiological levels of IGF-I (40 ng/ml) results in increased numbers of cells in the inner cell mass (p < 0.05), but not the trophectoderm, as compared to controls.

Animals↗
PubMed1993

The triple-jump examination as an assessment tool in the problem-based medical curriculum at the University of Hawaii.

BACKGROUND: The three-step triple jump (TJ) examination aims to assess students' clinical problem-solving processes predominantly by means of subjective assessments administered by faculty. But training TJ administrators to ensure interrater reliability is both time- and cost-intensive, and difficult at best--hence the desire to test a more objective system of scoring students' TJ performances. METHOD: The sample was the 58 first-year students of the class of 1995, who in March 1992 were finishing the second 13-week unit in the problem-based curriculum at the University of Hawaii John A. Burns School of Medicine. To determine how well the school had succeeded in standardizing and objectifying its TJ examination (used for all units in the first two years), scores were correlated for various objective examinations independent of the TJ, the TJ administrators' subjective assessments, and the TJ objective assessments (regarding number of problem-based hypotheses generated and number of hypothesis-testing clinical database items elicited). The statistical methods used were linear regression, Student's unpaired t-test, chi-square, and the z-test. RESULTS: The TJ scores and independent objective examinations did not correlate significantly (suggesting that they assess different aspects of student achievement), but the TJ subjective and objective scores did correlate significantly. There were large standard deviations on the TJ objective scores, largely because one problem was significantly more difficult than the others (each student works on only one of several problems). However, the administrators' subjective scores for all problems were comparable. CONCLUSION: Because problems vary in difficulty, objective scores cannot be used across problems as a major component of all students' grades; but when a student has received an unsatisfactory score, an external reviewer can evaluate the appropriateness of the subjective score by comparing the student's objective performance with those of students who had the same problem and received higher subjective scores. That the administrators' subjective assessments for all problems were comparable not only suggests that the administrators were able to adjust for problem variability but also reinforces the appropriateness of using subjective assessments for the TJ examination.

Educational Measurement↗
PubMed1993

A truncated human insulin receptor missing the COOH-terminal 365 amino acid residues does not undergo insulin-mediated receptor migration or aggregation.

A previous study of tyrosine kinase-defective insulin receptors demonstrated that receptor autophosphorylation or tyrosine kinase activity was required for concentrating insulin receptors in coated pits, but not for their migration or aggregation on the cell surface. Furthermore, receptor migration and aggregation on the cell surface were not sufficient to cause internalization of the occupied receptors in coated pits. In the present study, biochemical and ultrastructural techniques were used to compare insulin receptor mobility and internalization in Rat 1 fibroblasts expressing wild-type human insulin receptors (HIRc) with those in cells expressing receptors truncated at residues 978 (HIR delta 978) or 1301 of the carboxyl-terminus (HIR delta CT). There were no significant differences in the mobility or internalization of insulin receptors on HIR delta CT cells compared to those of insulin receptors on HIRc cells. Ultrastructural analysis revealed that truncated insulin receptors on HIR delta 978 cells failed to migrate from their initial location on the microvilli, move to the plasma membrane, and aggregate in coated pits. Receptor-mediated insulin internalization in HIR delta 978 cells was markedly decreased due entirely to a decrease in ATP-dependent, coated pit-mediated internalization. ATP-independent endocytosis in non-coated pinocytotic invaginations was not affected by receptor truncations. These results provide evidence of the roles that regions of the beta-subunit play in the processing of occupied insulin receptors. 1) The carboxyl-terminus of the insulin receptor is not involved in the events leading to receptor internalization, i.e. migration, aggregation, and concentration in coated pits. 2) Internalization of insulin receptors by the ATP-independent noncoated invagination pathway is not regulated by residues in the insulin receptor beta-subunit distal to 978. 3) Sequences in the beta-subunit between 978-1300, but not the autophosphorylation and kinase domains, are involved in insulin-induced receptor migration and aggregation.

Amino Acid Sequence↗
PubMed1993

Inhibition of insulin-degrading enzyme increases translocation of insulin to the nucleus in H35 rat hepatoma cells: evidence of a cytosolic pathway.

We previously demonstrated the translocation of insulin to the nucleus in several cell types and partially characterized the uptake mechanisms and pathways in H35 rat hepatoma cells. Nuclear accumulation of insulin was energy independent, time and temperature dependent, and apparently was not saturable at insulin concentrations which resulted in full receptor occupancy. We also have shown insulin could be internalized by both receptor-mediated and fluid-phase endocytosis. This study investigated subsequent steps involved in the nuclear accumulation of insulin following internalization. We examined the effects of inhibiting insulin degrading enzyme (IDE) with 1,10-phenanthroline on the nuclear accumulation of insulin in H35 cells. 1,10-phenanthroline (2 mM) which markedly inhibited insulin degradation, significantly increased nuclear accumulation of insulin without having any effects on total cell-associated and intracellular insulin. This reagent increased 125I-insulin on the cellular membrane and decreased 125iodine (125I-insulin and 125I-insulin degradation products) in the cytosolic fractions. Chemical extraction and Sephadex G-50 chromatography revealed the insulin associated with the nucleus in 1,10-phenanthroline-treated cells formed the same complex(es) with the nuclear matrix as in control cells. These results suggested that inhibition of cytosolic IDE activity resulted in increased insulin translocation from the cytosol to the nucleus. Furthermore, when IDE activity was inhibited by high cytosolic insulin concentrations, the amount of 125I-insulin in the nucleus was significantly increased. Our study suggests internalized insulin is probably released from endosomes into the cytosol where modulation of IDE activity could have significant effects on the accumulation of insulin, or insulin-cytoplasmic protein complexes, in nuclei. The IDE regulatory mechanism, by controlling the translocation of insulin to the cell nucleus, could play a crucial role in insulin's regulation of gene expression and cell proliferation.

Animals↗
PubMed1993

Frontoethmoidal meningoencephaloceles: a review of 14 consecutive patients.

Over a period of 4.5 years, 14 patients with frontoethmoidal meningoencephaloceles were treated. Most patients came from Northern Namibia. Precise delineation of all cranial abnormalities was obtained by modern imaging techniques, and specific patterns of cerebral abnormality were found. The malformation was corrected in a single stage, and significant modifications have been developed to render the procedure simpler and safer. Information from our series favors delayed neural tube closure as the primary pathogenesis of the defect and suggests a common teratogen as the most probable etiological agent. Our experience leads us to advocate early correction of even small defects.

Encephalocele↗