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Biomedical subjects

R M Smith

Publications and source records attributed to R M Smith.

At least 253 records · Page 14Linked to original sources

The transverse plane deformity of structural scoliosis.

An examination of the transverse plane aspect of deformed vertebrae from specimens of both human and animal scoliosis identified a consistent pattern of intravertebral deformity. In the animal model, dynamic bone growth studies illustrated bone drift in the opposite direction to the rotation of the scoliosis, suggesting that the bone growth in the transverse plane was attempting to correct rather than produce the deformity. The observed pattern of vertebral deformity can be explained in simple terms because it obeys the standard laws of bone growth and remodeling. This observed growth pattern is consistent only with the production of idiopathic scoliosis by the rotation of a primary lordosis.

Animals↗

One hour in 1 ATA oxygen enhances rat alveolar macrophage chemiluminescence and fungal cytotoxicity.

The purpose of this study was to determine if 100% O2 would enhance rat pulmonary alveolar macrophage (PAM) oxidative killing of conidia of the fungus Neurospora crassa. First, we found that incubation in 100% O2 had no effect on conidia viability in the absence of PAM. We obtained resident PAM from nonpretreated anesthetized male Sprague-Dawley rats by bronchoalveolar lavage. Compared with similar air exposures we found that 1 h in vitro exposure of PAM to 100% O2 (1.0 atmosphere absolute) increased their killing of opsonized phagocytized conidia by 52%, without altering phagocytosis. A reactive oxygen killing mechanism is suggested because 1) 100% O2 increased PAM chemiluminescence (CL) by 60% both at rest and during 1 h of phagocytosis, and 2) PAM in 100% O2 killed albino conidia (lacking free radical-quenching carotenoids) 2.9 times more readily than they killed wild-type conidia. Compared with air, 100% O2 delayed the PAM respiratory burst time to peak by 18% but did not alter the burst maximal acceleration. The latter and the similar 60% increase in PAM CL in 100% O2 both at rest and during 1 h of phagocytosis suggests a nonenzymatic O2 enhancement of the respiratory burst. Changed timing of burst events in 100% O2 suggests early O2 toxicity. We conclude that 1 h in 100% O2 increases PAM free radical production and fungicidal activity.

Animals↗

Respiratory burst oxidase and three of four oxidase-related polypeptides are associated with the cytoskeleton of human neutrophils.

Resting and phorbol-activated human neutrophils were separated by treatment with Triton X-100 into detergent-extractable and cytoskeleton fractions. Respiratory burst oxidase activity was restricted entirely to the cytoskeleton. The cytoskeleton also contained approximately 15% of the neutrophil cytochrome b558, an oxidase-associated heme protein, as well as most of the oxidase-related cytosolic polypeptide p67phox. In contrast, the components of the oxidase-associated phosphoprotein family p47phox were found almost exclusively in the detergent extract, suggesting that p47phox is needed for oxidase activation but not for O2- production by the activated oxidase. Activation of the oxidase had no apparent effect on the distribution of any of these species between the cytoskeleton and the detergent extract. Our results support earlier studies implying that the cytoskeleton participates in an important way in regulating the activity of the O2(-)-forming respiratory burst oxidase of neutrophils.

Blotting, Western↗

Effects of dietary cadmium chloride throughout gestation on blood and tissue metabolites of primigravid and neonatal dairy cattle.

Thirty-six postpubertal Holstein heifers were allocated to three groups and fed the same diet, which differed only in the concentration of Cd: control group (.25 ppm of Cd), low-Cd group (1 ppm of Cd), and high-Cd group (5 ppm of Cd). Cadmium was supplemented to the low-Cd and high-Cd groups using CdCl2. Liver, kidney cortex, and abdominal muscle were biopsied for mineral analysis from one-half of the heifers of each group before Cd supplementation and again from the same animals within 5 d after parturition, 394 d later. Blood, liver, and muscle were collected from each calf within 5 h after birth. In the dam, 5 ppm of dietary Cd caused a 62-, 27-, and 4-fold increase in Cd of the kidney, liver, and muscle, respectively; kidney Zn and Fe increased (76%) and decreased (33%), respectively, whereas the serum Cu was reduced (31%). Liver Cu was reduced to 40 and 17% by dietary Cd of 1 and 5 ppm, respectively, in the dams. Calves from dams consuming 5 ppm of Cd had a 29 and 43% reduction in liver Cu and Zn, respectively. In these same calves, packed cell volume, hemoglobin concentration, and serum Cu were decreased by 17, 18, and 25%, respectively, whereas serum Zn was increased (55%). Serum sodium and potassium were reduced by 4 and 13%, respectively, and blood urea nitrogen was increased by 63% in calves from dams consuming 5 ppm of Cd. Feeding primigravid dairy cattle up to 5 ppm of Cd as CdCl2 throughout gestation did not influence the concentration of Cd in the neonate but caused reductions in liver Cu and Zn; teratogenesis was not apparent.

Animals↗

Effects of long-term dietary cadmium chloride on tissue, milk, and urine mineral concentrations of lactating dairy cows.

The effects of long-term consumption of 1 and 5 ppm of Cd on Cd, Cu, Fe, and Zn concentrations in milk and a variety of tissues of first-lactation dairy cows was investigated. Thirty-six Holstein heifers were allocated to three groups and fed similar diets differing only in the concentration of Cd (.25, 1, and 5 ppm) for a 394-d period before calving. One- and 5-ppm Cd concentrations were achieved using CdCl2. Liver, kidney cortex, and muscle were biopsied from one-half of the heifers of each group before Cd supplementation and again from the same heifers within 5 d after parturition. Colostrum and milk were sampled throughout the 150 d of lactation. Urine was sampled after an average of 450 d of Cd exposure. At slaughter (after an average of 554 d), 11 tissues were sampled from 12 cows representing all treatment groups. During the first 394 d, Cd accumulated in kidney and liver with increasing dietary concentrations of Cd but did not further increase by 554 d. However, by 554 d Cd had also accumulated in the adrenal glands, ovaries, spleen, and uteri of cows consuming 5 ppm of Cd. Dietary Cd did not influence the concentration of Cd, Cu, Fe, or Zn in colostrum or milk. However, urine pH, Zn, and K were lower in cows consuming 5 ppm of Cd. Liver Cu was reduced by 1 and 5 ppm of Cd at both 394 and 554 d.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Hmong folk remedies: limited acetylation of opium by aspirin and acetaminophen.

The traditional folk medicine of the Hmong and other Southeast Asian refugees has accompanied them during their immigration in this country over the past two decades. In two recent cases involving Hmong defendants, unknown solids, resembling charcoal in consistency and purported to be "backache remedies," were analyzed and found to be complex mixtures of aspirin, acetaminophen, caffeine and partly acetylated opium. In particular, significant amounts of acetylacetaminophen, 3-O-acetylmorphine, 6-O-acetylcodeine, 6-O-acetylmorphine, and heroin were identified by gas chromatography/mass spectrometry. Heating approximately equal weights of solid opium, aspirin, and acetaminophen at 130 degrees C for several hours produced a mixture of compounds showing a similar acetylation pattern.

Acetaminophen↗

Embryonic chicken lens cells cultured in reconstituted basement membrane: an experimental model to maintain the epithelial phenotype in culture.

The action of a reconstituted basement membrane has been studied on primary cultures of embryonic lens cells. When a solution of this matrix (Matrigel) was included in the culture medium, a high percentage of cells maintained the epithelial phenotype, judged by electron microscopy criteria, in contrast to the differentiated state induced by serum. Complete matrix stimulated by 6-fold the incorporation of 3H-thymidine into the cells, while one of its defined components, laminin, only had a 2-fold stimulatory effect. Thus, the basement membrane may stimulate mitogenesis and play a role complementary to that of growth factors in development.

Animals↗

Bird attack on milk bottles: possible mode of transmission of Campylobacter jejuni to man.

A case-control study was carried out to test the hypothesis that the rise in the rate of Campylobacter jejuni infection in the Brigend area of South Wales during May was due to the consumption or handling of milk from bottles that had been attacked by birds. 32 of 36 cases meeting the case definition were interviewed, along with 2 controls per case, matched for age, sex, and area of residence. There were strong associations between campylobacter infection and doorstep delivery of milk bottles, a history of milk bottle attack by birds, milk bottle attack by birds during the week before illness, and consumption of milk from attacked bottles during the week before illness. There was a very strong dose-response relation between frequency of bird attack and illness. Controls with a history of milk bottle attack by birds were more likely than cases to have taken preventive measures against bird attack and consumption of contaminated milk. Although few people witnessed the attacks, the likely culprits are magpies (Pica pica) and jackdaws (Corvus monedula).

Adolescent↗

The p67-phox cytosolic peptide of the respiratory burst oxidase from human neutrophils. Functional aspects.

Most cases of cytosol-defective chronic granulomatous disease are due to the deficiency of a 47-kD protein (p47-phox) whose phosphorylation normally accompanies the activation of the respiratory burst oxidase. Recently, a form of chronic granulomatous disease was described in which the failure of O2- production was associated with the absence of a 67-kD polypeptide (p67-phox) from the cytosol of affected neutrophils. Using neutrophils obtained from a patient with this form of the disease, we examined the function of p67-phox in the activation of the oxidase. Our studies showed that in whole p67-phox-deficient neutrophils, p47-phox was phosphorylated in a normal fashion. In the cell-free oxidase-activating system, the ability of the p67-phox-deficient cytosol to support oxidase activation was partly restored by the addition of p47-phox-deficient cytosol; the p67-phox-deficient cytosol, however, was not complemented by cytosol inactivated with NADPH dialdehyde, an affinity label previously found to block the NADPH-binding component of the oxidase. Despite these differences, the kinetic properties of the p67-phox-deficient cytosol closely resembled those of the p47-phox-deficient cytosol. Taken together with earlier findings, these results suggest that (a) in the neutrophil cytosol, p67-phox is at least partly complexed to p47-phox; (b) it is in the form of this complex that p67-phox participates in oxidase activation; and (c) p47-phox appears to be translocated from the cytosol to the plasma membrane during oxidase activation, but complexation to p67-phox is not necessary for this translocation, nor for the accompanying extra protein phosphorylation.

Blood Proteins↗

Differences in adenosine triphosphate dependency of receptor-mediated endocytosis of alpha 2-macroglobulin and insulin correlate with separate routes of ligand-receptor complex internalization.

Biochemical and morphological studies compared the ATP requirements for and the internalization routes of alpha 2-macroglobulin and insulin in H35 hepatoma cells. Cellular ATP concentrations were decreased more than 94% by 1 mM 2,4-dinitrophenol or 10 mM sodium azide, potassium cyanide, or oligomycin. ATP depletion decreased total cell-associated alpha 2-macroglobulin 70-90% by inhibiting binding 67-77% and receptor-mediated internalization 90-96%. Under the same conditions, insulin binding was decreased less than 10%, and endocytosis and intracellular accumulation were not affected. Quantitative electron microscopic analysis of the distribution of occupied receptors on the surface of control and treated cells was performed using colloidal gold-labeled alpha 2-macroglobulin or insulin. alpha 2-Macroglobulin concentrated in and was internalized almost exclusively by coated pits. Insulin was rarely associated with coated pits, but was found in and internalized by noncoated invaginations. ATP depletion did not affect receptor mobility or ligand-induced aggregation of either receptor. There was an increase in the amount of alpha 2-macroglobulin found in coated pit-like structures. The coat underlying pits in ATP-depleted cells was poorly defined and may account for the inability of coated pits to form and/or internalize. These results showed that receptor-mediated internalization via coated pits was ATP dependent, whereas internalization via pinocytotic invaginations was energy independent, which explained the difference in the ATP dependency of uptake for the two ligands. These observations suggested that autophosphorylation of the insulin receptor may not be involved in either the aggregation or internalization of the insulin-receptor complex, since ATP depletion did not affect either process. This study provided evidence that specialized mechanisms exist for the internalization of insulin which may be related to some of its intracellular effects.

Adenosine Triphosphate↗

The state of differentiation of embryonic chicken lens cells determines insulin-like growth factor I internalization.

Microdissected epithelial cells from chicken embryonic lens differentiate into fiber cells during primary culture. Binding of IGF-I to both cell types has been documented biochemically. This report describes differences in the internalization of insulin-like growth factor I (IGF-I) in the two cell types using three electron microscopic approaches. Cells were incubated with a biologically active colloidal gold-labeled-IGF-I complex or [125I]IGF-I and prepared for electron microscopy. Other cells were incubated with unlabeled IGF-I and prepared for immunoelectron microscopy, using a colloidal gold-labeled anti-IGF-I antibody to detect the IGF-I. Each technique demonstrated binding of IGF-I on the surface of epithelial and differentiated fiber cells. IGF-I was internalized in epithelial cells. In contrast, the ligand exhibited little endocytosis in fiber cells during a 5 h incubation. Intracellular IGF-I was observed in the endosomes, Golgi apparatus, and lysosomes of epithelial cells, in addition, it was shown for the first time that IGF-I was translocated to the nucleus of epithelial cells. The differences between epithelial and fiber cells regarding internalization and nuclear translocation of IGF-I suggest that there are cell-specific itineraries of the hormone, depending on the differentiation stage of the cell. These differences may relate to the specific biological actions of the growth factor on each of these cells, particularly in allowing proliferation and differentiation of the epithelial cells.

Animals↗

Cell surface vitamin D-binding protein (GC-globulin) is acquired from plasma.

Vitamin D-binding protein (DBP) is an abundant plasma protein. The observation of immunodetectable, cell-associated DBP on peripheral blood mononuclear cells and placental cytotrophoblasts had presented the question of the origin, function, and precise subcellular localization of cell-associated DBP. Using anti-human DBP F(ab')2 with fluorescence-activated cytometric analysis and immunogold electron microscopy, we detected DBP on the plasmalemma of U937 cells, a monoblastic, histiocytic cell line grown in media supplemented with fetal calf serum (FCS). DBP was then removed from FCS by actin affinity chromatography followed by anti-DBP immunoaffinity chromatography. U937 cells in this DBP-free medium exhibited nearly identical growth rates to cells grown in medium containing native FCS. However, in contrast to cells grown with native FCS, those grown for seven to eight generations with DBP-free FCS exhibited less cell-surface DBP as quantified by fluorescence-activated cytometric analysis (73% decrease) and immunoelectron microscopy (88% decrease). DBP mRNA could not be detected in U937 cells, placental tissues, freshly prepared resting and stimulated B and T lymphocytes, or lymphocyte-derived cell lines by Northern analysis. In addition, using the sensitive reverse transcriptase/polymerase chain reaction assay no DBP fragments were detectable in U937 cells. We conclude that U937 cell-associated DBP is exogenously derived from plasma and is located on the plasmalemma. Based upon this conclusion, we postulate that specific binding sites for DBP may exist on the plasma membranes of certain cell types.

Blotting, Northern↗

Partial characterization of mechanism of insulin accumulation in H35 hepatoma cell nuclei.

The mechanism controlling insulin accumulation in nuclei of H35 hepatoma cells was investigated by incubating intact cells with 125I-labeled insulin in the presence or absence of agents that perturb different intracellular sites involved in the processing of ligand-receptor complexes. Purified nuclei were isolated, and nuclear-associated 125I-insulin was determined. Insulin accumulation in the nuclei was time and temperature dependent. Nuclear accumulation was linear and insulin-concentration dependent between 5 and 50 ng insulin/ml. However, pharmacological concentrations of insulin increased the amount of insulin translocated to the nucleus to a far greater extent than it increased total cell-associated insulin. Chloroquine, an acidotrophic agent, increased total cell-associated and intracellular insulin but had no effect on nuclear accumulation. The monovalent ionophores monensin and nigericin inhibited nuclear accumulation of insulin at low concentrations (0.5-5.0 microM) without affecting total insulin binding or intracellular accumulation. At 10 or 25 microM, monensin and nigericin also acted as acidotrophic agents and increased total insulin binding and intracellular accumulation but inhibited nuclear accumulation by a maximum of 50%. Low concentrations of monensin and nigericin were additive; maximal concentrations were not. A 23187 and valinomycin did not affect insulin binding or intracellular and nuclear accumulation of insulin. Neither depletion of ATP by sodium azide, 2,4-dinitrophenol, sodium cyanide, or oligomycin nor disruption of cytoskeletal elements by cytochalasin D or colchicine had any effect on nuclear accumulation of insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

Incidence of measurement disturbances in the Dental Admission Quantitative Reasoning Test.

This paper describes the incidence of inconsistent person-response patterns in the Quantitative Reasoning Test (QRT), which is part of the Dental Admission Test (DAT) battery. using the Rasch person analysis approach to estimate the likelihood of a response pattern resulting from only two factors (the difficulty of the items and the ability of the person), a statistical profile of inconsistent response patterns was developed for a sample of students taking the DAT. The results of two test dates, April 1987 and October 1988, were included in this study. A random sample of 1,000 persons was drawn from each of these two test administrations. For the Fall 1988 sample, the person analysis was supplemented by a questionnaire designed to check the correspondence between a student's self-reported response behavior and the statistical profile provided by the person analysis. The analysis of the two samples of response patterns indicates a high incidence (50 percent+) of atypical response patterns on the test. This has serious implications for the admissions procedure since many of the atypical response patterns invalidate the standard scores reported for those students.

Education, Dental↗