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Biomedical subjects

J H Johnson

Publications and source records attributed to J H Johnson.

At least 73 records · Page 4Linked to original sources
PubMed1993

GLUT-2 function in glucose-unresponsive beta cells of dexamethasone-induced diabetes in rats.

Spontaneous and dexamethasone-induced noninsulin-dependent diabetes mellitus (NIDDM) in rats is associated with loss of glucose-stimulated insulin secretion (GSIS) and a reduction in both GLUT-2-positive beta cells and high Km glucose transport. To determine if the chronology and correlation of these abnormalities is consistent with a causal relationship, Zucker (fa/fa) rats were studied longitudinally before and during 10 d of dexamethasone-induced (0.4 mg/kg per d i.p.) NIDDM. Within 24 h of dexamethasone treatment blood glucose rose and GSIS declined, becoming paradoxically negative (-87 +/- 12 microU/ml per min) on day 10. Blood glucose was negatively correlated with GSIS (r = -0.92; P < 0.001). 3-0-methyl-D-glucose (3MG) transport was unchanged at 12 h, 23% below normal on day 1, and declined further to a nadir 59% below normal. The GLUT-2-positive beta cell area did not decline until 48 h, reaching a nadir of 35% of normal at 10 d. The area of GLUT-2-positive beta cells was correlated with GSIS (r = 0.77; P < 0.005). We conclude that the chronology and correlation between GSIS loss and hyperglycemia is consistent with a cause-effect relationship, but that the subtotal impairment in glucose transport by itself cannot explain the total loss of GSIS if one assumes that normal beta cells are functionally homogenous.

3-O-Methylglucose↗
PubMed1993

GLUT2 expression and function in beta-cells of GK rats with NIDDM. Dissociation between reductions in glucose transport and glucose-stimulated insulin secretion.

GLUT2 underexpression has been reported in the beta-cells of Zucker diabetic fatty rats and db/db mice, models of spontaneously occurring NIDDM with antecedent obesity. To determine whether the beta-cells of a nonobese rodent model of NIDDM exhibit the same abnormalities in GLUT2, we studied Goto-Kakizaki rats. In these mildly diabetic animals glucose-stimulated insulin secretion was reduced at all ages examined from 8 to 48 wk. In normal control Wistar rats, immunostainable GLUT2 was present on all insulin-positive cells in the pancreatic islets. Only 85% of beta-cells were GLUT2-positive in GK rats at 12 wk of age, and only 34% were positive at 48 wk of age. GLUT2 mRNA was 50% of normal in 12-wk-old GK rats. In the latter age-group, glucose-stimulated insulin secretion was only 28% of normal at a time when 85% of beta-cells were GLUT2-positive and initial 3-O-methyl-D-glucose transport rate was 77% of the control value. We conclude that although GLUT2 is underexpressed, neither the magnitude of the underexpression of GLUT2 nor of the reduction in GLUT2 transport function in islets of GK rats is sufficient by itself to explain the profound reduction in glucose-stimulated insulin secretion.

Aging↗
PubMed1993

Characterization of particle- and vapor-phase organic fraction emissions from a heavy-duty diesel engine equipped with a particle trap and regeneration controls.

The effects of a ceramic particle trap on the chemical and biological character of the exhaust from a heavy-duty diesel engine have been studied during steady-state operation and during periods of trap regeneration. Phase I of this project involved developing and refining the methods using a Caterpillar 3208 engine, and Phase II involved more detailed experiments with a Cummins LTA10-300 engine, which met Federal 1988 particulate matter standards, and a ceramic particle trap with built-in regeneration controls. During the Phase I experiments, samples wee collected at the Environmental Protection Agency (EPA)* steady-state mode 4 (50% load at intermediate speed). Varying the dilution ratio to obtain a constant filter-face temperature resulted in less variability in total particulate matter (TPM), particle-associated soluble organic fraction (SOF), solids (SOL), and polynuclear aromatic hydrocarbon (PAH) levels than sampling with a constant dilution ratio and allowing filter-face temperature to vary. A modified microsuspension Ames assay detected mutagenicity in the SOF samples, and in the semivolatile organic fraction extracted from XAD-2 resin (XAD-2 resin organic component, XOC) with at least 10 times less sample mass than the standard plate incorporation assay. Measurement techniques for PAH and nitro-PAH in the SOF and XOC also were developed during this portion of the project. For the Phase II work, two EPA steady-state rated speed modes were selected: mode 11 (25% load) and mode 9 (75% load). With or without the trap, filter-face temperatures were kept at 45 degrees +/- 2 degrees C, nitrogen dioxide (NO2) levels less than 5 parts per million (ppm), and sampling times less than 60 minutes. Particle sizes were determined using an electrical aerosol analyzer. Similar sampling methods were used when the trap was regenerated, except that a separate dilution tunnel and sampling system was designed and built to collect all of the regeneration emissions. The SOF and XOC were extracted from their collection media with dichloromethane. Levels of 12 PAH and nitro-PAH compounds with known biological effects were determined using high-pressure liquid chromatography with fluorescence detection. Two additional dinitro-PAHs were analyzed semi-quantitatively, and several representative SOF and XOC samples were screened for three additional PAH compounds. Mutagenicity was assessed using the modified Ames microsuspension assay.(ABSTRACT TRUNCATED AT 400 WORDS)

Air Pollutants↗
PubMed1992

Cloning of the alpha 1 subunit of a voltage-dependent calcium channel expressed in pancreatic beta cells.

The isoforms of the alpha 1 subunits of voltage-dependent Ca2+ channels expressed in human pancreatic islets were identified by using a pair of degenerate oligonucleotide primers and the polymerase chain reaction (PCR) to amplify mRNAs encoding alpha 1 subunit-like sequences. The sequences of the PCR products indicate that islets express the heart-type alpha 1 subunit as well as a second isoform whose complete sequence has not been previously reported. The sequences of cloned cDNAs encoding the human beta-cell, or neuroendocrine-type, alpha 1 subunit indicate that it is composed of 2181 amino acids. It shares 68%, 64%, and 41% identity with the sequences of the alpha 1 subunits of rabbit heart, skeletal muscle, and brain, respectively, and is predicted to have a similar structure including four homologous domains composed of six membrane-spanning segments each. RNA blotting studies indicate that the beta-cell-type alpha 1 subunit is also expressed in brain as well as in the insulin-producing cell lines RINm5F and beta TC-3; however, it could not be detected by RNA blotting in a third cell line, HIT-T15. In situ hybridization studies revealed expression of beta-cell-type alpha 1 subunit mRNA in beta cells of rat pancreatic islets, implying that this protein may play a role in the regulation of insulin secretion.

Amino Acid Sequence↗
PubMed1992

Engineering of glucose-stimulated insulin secretion and biosynthesis in non-islet cells.

The high-capacity glucose transporter known as GLUT-2 and the glucose phosphorylating enzyme glucokinase are thought to be key components of the "glucose-sensing apparatus" that regulates insulin release from the beta cells of the islets of Langerhans in response to changes in external glucose concentration. AtT-20ins cells are derived from anterior pituitary cells and are like beta cells in that they express glucokinase and have been engineered to secrete correctly processed insulin in response to analogs of cAMP, but, unlike beta cells, they fail to respond to glucose and lack GLUT-2 expression. Herein we demonstrate that stable transfection of AtT-20ins cells with the GLUT-2 cDNA confers glucose-stimulated insulin secretion and glucose regulation of insulin biosynthesis and also results in glucose potentiation of the secretory response to non-glucose secretagogues. This work represents a first step toward creation of a genetically engineered "artificial beta cell."

Adrenocorticotropic Hormone↗
PubMed1992

Bronchovascular mucormycosis in the diabetic: an urgent surgical problem.

This report reviews the successful surgical experience with a diabetic patient with bronchial obstruction due to Mucorales infection. A review of the reported medical and surgical experience is included. The danger of a lethal pulmonary hemorrhage makes early surgical intervention mandatory.

Airway Obstruction↗
PubMed1992

Child temperament, maternal adjustment, and changes in family life style.

Child temperament has been implicated as a relevant factor in understanding parental adjustment. In a study of 77 mothers of 3- and 4-year-old children, difficult child temperament was found to be directly related to maternal distress, discomfort in the role of parent, poor spousal relationships, and negative changes in way of life. Quality and intensity of the child's mood were most predictive of these difficulties.

Adaptation, Psychological↗
PubMed1992

Pharmacokinetics and inflammatory fluid penetration of sparfloxacin.

A single 400-mg oral dose of sparfloxacin was given to each of six healthy male volunteers, and the concentrations of the drug were measured in plasma, cantharides-induced inflammatory fluid, and urine over the subsequent 52 h. The mean peak concentration in plasma of 1.6 micrograms/ml was attained at a mean time of 2.7 h postdose. The mean peak concentration in inflammatory fluid of 1.3 micrograms/ml was attained at a mean time of 5 h postdose. The mean elimination half-life in plasma was 17.6 h, and that in inflammatory fluid was 19.7 h. The overall penetration into inflammatory fluid was 117%. Urinary recovery within the first 52 h postdose was 8.8% of the administered dose. Our results indicate that a once-daily dosage of sparfloxacin should be adequate to treat systemic infections caused by most common bacterial pathogens.

Administration, Oral↗
PubMed1992

Roles of insulin resistance and beta-cell dysfunction in dexamethasone-induced diabetes.

The roles of insulin resistance and beta-cell dysfunction in glucocorticoid-induced diabetes were determined in Wistar and Zucker (fa/fa) rats. All Wistar rats treated with 5 mg/kg per d of dexamethasone for 24 d exhibited increased beta-cell mass and basal and arginine-stimulated insulin secretion, indicating insulin resistance, but only 16% became diabetic. The insulin response to 20 mM glucose was normal in the perfused pancreas of all normoglycemic dexamethasone-treated rats but absent in every diabetic rat. Immunostainable high Km beta-cell transporter, GLUT-2, was present in approximately 100% of beta-cells of normoglycemic rats, but in only 25% of beta cells of diabetic rats. GLUT-2 mRNA was not reduced. All Zucker (fa/fa) rats treated with 0.2-0.4 mg/kg per d of dexamethasone for 24 d became diabetic and glucose-stimulated insulin secretion was absent in all. High Km glucose transport in islets was 50% below nondiabetic controls. Only 25% of beta cells of diabetic rats were GLUT-2-positive compared with approximately 100% in controls. Total pancreatic GLUT-2 mRNA was increased twofold suggesting a posttranscriptional abnormality. We conclude that dexamethasone induces insulin resistance, whether or not it induces hyperglycemia. Whenever hyperglycemia is present, GLUT-2-positive beta cells are reduced, high Km glucose transport into beta cells is attenuated and the insulin response to glucose is absent.

Animals↗
PubMed1992

The tissue distribution of tumor necrosis factor biosynthesis during endotoxemia.

Tumor necrosis factor (TNF) is a protein hormone implicated in the development of septic shock and other pathologic states. However, complexities inherent in detecting TNF synthesis by individual tissues have left the precise origins of this protein undefined. In addition, the possibility that localized TNF production may contribute to the pathogenesis of organ-specific diseases such as type I diabetes has not been explored in vivo. We have developed a transgenic mouse line bearing a reporter gene construct in which the TNF coding sequence and introns are replaced by a chloramphenicol acetyltransferase (CAT) coding sequence. In normal transgenic animals, CAT activity is expressed only in the thymus. When endotoxin is administered to the animals, CAT activity is also evident in kidney, heart, islets of Langerhans, spleen, lung, fallopian tubes, and uterus, but not in other organs. The biosynthesis of CAT in vivo correlated with tissue capacity to secrete TNF in vitro. Thus, TNF was secreted by all the tissues that expressed CAT, including lung, spleen, thymus, uterus/fallopian tubes, pancreatic islets, renal glomeruli, and cultured cardiac cells after exposure to endotoxin.

Animals↗
PubMed1992

Applicability of the Impact on Family Scale for assessing families with behaviorally difficult children.

The present paper reports a series of studies on the potential usefulness of the Stein and Riessman Impact on Family Scale for assessing the effects of behaviorally difficult children on their families. Subjects were parents of 54 clinic and 76 nonclinic children who completed a child behavior or child temperament questionnaire in addition to the Impact on Family Scale. Findings suggest that, while the scale was developed for assessing the influence of a physically ill child on the family, this general assessment approach may also have value in assessing family-related changes associated with having a child who is difficult to look after.

Adaptation, Psychological↗
PubMed1992

Lanomycin and glucolanomycin, antifungal agents produced by Pycnidiophora dispersa. II. Structure elucidation.

Two novel antifungal agents, lanomycin and glucolanomycin, as well as a biologically inactive degradation product, lanomycinol, were isolated from liquid fermentations of Pycnidiophora dispersa. All three compounds share an E,E,E-triene appended to a pyran ring. Lanomycin contains a glycine ester and glucolanomycin possesses a glucose unit attached to the glycine nitrogen. The structures, including absolute stereochemistry, were determined by spectroscopic analysis and partial chemical degradation. Both of the glycine containing compounds show activity against several pathogenic fungi in vitro.

Aminoglycosides↗
PubMed1992

Risk factors for heterotopic ossification in children and adolescents with severe traumatic brain injury.

Previous reports on the incidence of heterotopic ossification (HO) in children and adolescents with traumatic brain injury (TBI) have ranged from 3% to 20%. These studies mention HO briefly or address patient cohorts with coma more than three months. We reviewed the medical records of 90 children younger than 19 years old, who were comatose more than 24 hours after TBI. Mean followup was 33.1 months (range = 2 to 128 months). Incidence of HO was 14.4% (n = 13). One to five sites were involved, with hip and knee most frequent, followed by shoulder, elbow, and nonjoint sites. Children with HO were older than 11 years (relative risk [RR] = 18.85, confidence interval [CI] = 3.7, 95.7), had a greater length of coma (RR = 7.22, CI = 1.4, 37.1), and had a greater associated risk for poor functional outcome (RR = 2.89, CI = 1.02, 7.9) compared to those without RO. Multiple limb fractures showed a trend toward association with increased incidence of HO. Other factors, including gender, history of skull fracture, brain CAT scan findings, and presence of spasticity, were not predictive of HO. Eleven children were treated with etidronate, four with nonsteroidal antiinflammatory drugs, and one with surgery. Only three children had residual functional impairments attributed to the presence of HO.

Adolescent↗
PubMed1991

Analysis of neurotransmitter receptors mediating changes in LH release induced by electrical stimulation of the dorsal raphe nucleus in the rat.

We report studies of neurotransmitter and receptor species mediating the inhibition of LH release in ovariectomized (OVX) rats and stimulation in OVX estrogen-primed rats induced by electrical stimulation of the dorsal raphe nucleus (DRN). Attempts were made to block effects of stimulation of the DRN in groups of four to nine ovariectomized rats with or without priming with estradiol benzoate (EB) by pretreatment with the 5HT receptor antagonists ketanserin, methysergide, or metergoline; the alpha-adrenergic antagonist phenoxybenzamine; or the opiate antagonist naltrexone. Blood samples were collected via jugular cannulae from the unanesthetized rats at 10-min intervals before and during electrical stimulation of the DRN, and assayed by double antibody radioimmunoassay for LH. To test the effect of stimulation, data from animals in each treatment group were subjected to analysis of variance to obtain the contrast matrix including prestimulation and stimulation time points of interest. The contrast matrix was then used to construct an F ratio and thereby to evaluate the level of significance. In unprimed OVX rats the sustained decrease in plasma LH concentration during stimulation was prevented in rats pretreated with ketanserin or phenoxybenzamine. Methysergide pretreatment delayed the inhibitory effect of DRN electrical stimulation for 30 min, whereas metergoline and naltrexone were ineffective. In EB-primed animals the increase in plasma LH observed during stimulation was prevented in rats pretreated with metergoline, and reversed in those receiving naltrexone. Ketanserin limited the duration of the increase to 10 min, while phenoxybenzamine and methysergide had no significant effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Screening of murine monoclonal antibodies against living schistosomula of Schistosoma mansoni by radioimmunoassay.

A radioimmunoassay was developed to screen supernatants of murine monoclonal antibodies against surface antigens of living schistosomula of Schistosoma mansoni. Of 196 clones screened, 10% bound schistosomula. Of these, 74% bound only schistosomula. The remaining molecules also reacted with soluble adult worm antigens and soluble egg antigens as determined by enzyme-linked immunosorbent assay. Immunoblot analysis demonstrated that monoclonal antibody 204-3E4 reacted with a 68 kDa protein, a glycoprotein that induces substantial resistance against S. mansoni infection. Recognition of an 18 kDa antigen by 204-3F1 antibody was stage-specific with the antigen being expressed in cercariae, 3- and 24-h-old parasites but not 4-day, lung stage or adult worms. Monoclonal antibody 204-4E3 reacted with purified S. mansoni paramyosin. These data indicate that radioimmunoassay using living schistosomula is a rapid alternative method to identify murine hybridomas that secrete antibodies which react with surface antigens of S. mansoni.

Animals↗
PubMed1991

Prediction of maximal heart rate during a submaximal work test.

A multiple regression equation was developed utilizing submaximal ratings of perceived exertion and heart rate collected during a treadmill walking test to predict maximal heart rate (MHR). One hundred subjects were administered a modification of the Balke Treadmill test during which time submaximal ratings of perceived exertion (Borg Scale) and heart rate measurements were recorded. Subjects worked until volitional fatigue and maximal heart was recorded. A multiple regression equation was developed which did not require the subject to work at greater than 85% of MHR. The multiple regression equation is MHR = 108.461 + 0.5108 (RPE 15)-0.6570 (8HR) + 0.6075 (10HR)-0.2641 (age in years) with RPE 15 equal to the heart rate at a perceived exertion rating of 15 on the Borg Scale, 8HR equal to the heart rate at 8 minutes and 10HR equal to the heart rate at 10 minutes. It was found that the regression equation significantly improved the ability to predict MHR and represented a 28% improvement in the commonly used equation MHR = 220-age.

Adult↗
PubMed1990

Underexpression of beta cell high Km glucose transporters in noninsulin-dependent diabetes.

The role of defective glucose transport in the pathogenesis of noninsulin-dependent diabetes (NIDDM) was examined in Zucker diabetic fatty rats, a model of NIDDM. As in human NIDDM, insulin secretion was unresponsive to 20 mM glucose. Uptake of 3-O-methylglucose by islet cells was less than 19% of controls. The beta cell glucose transporter (GLUT-2) immunoreactivity and amount of GLUT-2 messenger RNA were profoundly reduced. Whenever fewer than 60% of beta cells were GLUT-2-positive, the response to glucose was absent and hyperglycemia exceeded 11 mM plasma glucose. We conclude that in NIDDM underexpression of GLUT-2 messenger RNA lowers high Km glucose transport in beta cells, and thereby impairs glucose-stimulated insulin secretion and prevents correction of hyperglycemia.

3-O-Methylglucose↗