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Biomedical subjects

L R Johnson

Publications and source records attributed to L R Johnson.

At least 127 records · Page 7Linked to original sources

Spirometry in the Lung Health Study. 1. Methods and quality control.

The Lung Health Study is a randomized clinical trial designed to determine whether a smoking intervention program and prescription of an inhaled bronchodilator can reduce the annual rate of decline of FEV1 (FEV1 slope) in cigarette smokers with mild to moderate airflow limitation. During the recruitment period, spirometry measurements were obtained during three screening visits from 5,887 cigarette smokers 35 to 60 yr of age. To improve the statistical power of the study to detect significant differences in the FEV1 slopes among study groups, the spirometry testing protocol carefully controls for multiple factors that are known to increase intraindividual variability of FEV1 and FVC results. These factors include participant preparation, maneuver acceptability and reproducibility, technician training and performance monitoring, equipment design and calibration, and result processing. The objective of this report is to describe the success of the spirometry quality control program in minimizing the short-term intraindividual FEV1 variability. The mean difference between the FEV1 values obtained at the second and third screening visits (mean 25 days apart) was +5.7 ml in women and -14.7 ml in men, with a mean intraindividual variability of 119 ml in women and 162 ml in men, with a coefficient of variation of 5.8% for both. During only 2.1% of the test sessions were participants unable to produce three acceptable FVC maneuvers with the two best FEV1 values matching within 5% or 100 ml. Institution of site visits and technician performance monitoring was associated with improved test session quality grades and maintenance of high quality grades thereafter.

Adult↗

Activation of thymocyte deoxyribonucleic acid degradation by endogenous glucocorticoids.

Immature lymphocytes in the thymus gland are killed by treatment with exogenous glucocorticoids. This steroid-mediated lymphocytolysis is preceded by numerous alterations in lymphocyte metabolism, including a DNA-degrading process in which the genome is cleaved at internucleosomal intervals. To date, this process has only been characterized by treating lymphocytes in vitro with glucocorticoids or by exogenous treatment of whole animals with adrenal steroids. To determine whether thymocyte DNA degradation could be activated by endogenous glucocorticoids, 4-wk-old chicks were treated with porcine adrenocorticotropic hormone (ACTH). This procedure elevated serum corticosterone levels approximately 80-fold within 2 h of hormone treatment. Following ACTH administration, thymocyte DNA was isolated and analyzed by agarose gel electrophoresis. The ACTH activated a DNA-degrading process that generated internucleosomal fragments of DNA identical in size to those observed following exogenous treatment with synthetic or naturally occurring glucocorticoids. Furthermore, this response could be inhibited by the glucocorticoid antagonist RU486 (17 beta-hydroxy-11 beta, 4-dimethylaminophenyl-17 alpha-propynl-estra-4,9,diene-3-one), indicating that adrenal steroids activate this process via the glucocorticoid receptor. These results demonstrate that lymphocyte DNA degradation does not result solely from exogenous glucocorticoid treatment; moreover, endogenous glucocorticoids can mediate this process and may thereby play an important role in thymic gland function.

Adrenocorticotropic Hormone↗

Selective induction of c-jun and jun-B but not c-fos or c-myc during mitogenesis in SV40-transformed cells at the predifferentiation growth arrest state.

Insulin has recently been reported to function as a complete mitogen for SV40 large T antigen-transformed 3T3 T-cells, designated CSV3-1, but not for nontransformed 3T3 T-cells (H. Wang and R. E. Scott, J. Cell. Physiol., 147: 102-110, 1991). It is now reported that sodium orthovanadate mimics this effect of insulin. For example, when exposed to 1-5 microM vanadate, most predifferentiation growth-arrested CSV3-1 cells undergo DNA synthesis within 24 h, but neither vanadate nor insulin induces mitogenesis in nontransformed 3T3 T-cells. To investigate the possible mechanisms by which mitogenesis is induced in CSV3-1 cells, the effects of insulin and vanadate on the expression of growth-related genes were examined. Whereas insulin and vanadate had no effect on the expression of c-fos, c-myc, c-jun, jun-B, or ornithine decarboxylase activity in nontransformed 3T3 T-cells, insulin and vanadate showed different effects on the expression of these genes in CSV3-1 cells. Insulin induced a rapid and transient accumulation of c-fos mRNA followed by induction of c-myc, c-jun, jun-B, and ornithine decarboxylase. In contrast, vanadate induced the expression of c-jun, jun-B, and ornithine decarboxylase without inducing c-fos and c-myc. These observations suggest that SV40 large T antigen may play an important role in insulin- and vanadate-induced mitogenesis and that insulin and vanadate may mediate their mitogenic effects by different signal transduction pathways.

Antigens, Viral, Tumor↗

Cocarcinogenesis between cadmium and Aroclor 1254 in planarians is enhanced by inhibition of glutathione synthesis.

Asexual Dugesia dorotocephala planaria continuously exposed to 50 micrograms/beaker of Aroclor 1254 (A1254) and cadmium (Cd) developed tumors after 11 days of continuous exposure to 0.65 or 1.3 mg/liter Cd and after 23 days to 0.13 mg/liter Cd. The tumor rate at 14 days averaged 53% for the survivors in both 0.65 and 1.3 mg/liter Cd + A1254 and 40% at 0.13 mg/liter Cd + A1254. Other groups were either preexposed or coexposed to L-buthionine-S,R-sulfoximine (BSO), a specific inhibitor of glutathione synthesis. In the presence of A1254 (50 micrograms/beaker), animals continuously exposed to BSO and 0.13 mg/liter Cd first developed tumors at 18 days, with a tumor yield equal to 41% of the survivors. Tumors developed by 6 days at 0.65 mg/liter Cd, with a yield equal to 75% of survivors. At 1.3 mg/liter Cd, BSO did not change the rate or frequency of planarian tumor production. Continuous BSO always produced high mortality over 20 days. In contrast, a 24-h preexposure to BSO caused little mortality. In these groups, tumor yields increased with cadmium concentration (0.13, 0.25, 0.65 mg/liter), and the large cocarcinogenic effect of A1254 was clearly evident. No tumors developed in animals exposed only to 50 micrograms/beaker of Aroclor 1254 or continuously only to 1 mM BSO.

Animals↗

Tricyclic antidepressant toxicosis.

Accidental ingestions of TCAs by companion animals often occur. During the past 4 years, over 450 cases have been reported to the IAPIC. At least 7% of the animals that displayed clinical signs of toxicosis eventually died. Overdoses of TCAs adversely affect the cardiovascular, parasympathetic, and central nervous systems. The cardiovascular system is involved most seriously and ventricular arrhythmias with severe hypotension are believed to be the primary cause of death. Animals that ingest a potentially lethal dose (over 15 mg/kg) may die within 1 to 2 hours if appropriate treatment is not administered. Treatment involves the use of initial life-supportive measures (control of seizures, maintenance of an airway, ventilation, and so on), detoxification of the animal (enterogastric lavage, activated charcoal, etc.), and the intravenous use of sodium bicarbonate (2-3 mEq/kg) to control signs of acidosis, hypotension, tachycardia, bradycardia, and other cardiac conduction abnormalities. The animal must then be monitored closely for the return of the clinical signs and sodium bicarbonate therapy should be repeated as needed. In addition, to enhance removal of the TCAs from the gastrointestinal tract and, ultimately, from the body, activated charcoal should be repeated at 3-hour intervals until the animal is asymptomatic.

Animals↗

Assessment of occupational exposure to inorganic arsenic based on urinary concentrations and speciation of arsenic.

An analytical speciation method, capable of separating inorganic arsenic (As (V), As (III] and its methylated metabolites (MMAA, DMAA) from common, inert, dietary organoarsenicals, was applied to the determination of arsenic in urine from a variety of workers occupationally exposed to inorganic arsenic compounds. Mean urinary arsenic (As (V) + As (III) + MMAA + DMAA) concentrations ranged from 4.4 micrograms/g creatinine for controls to less than 10 micrograms/g for those in the electronics industry, 47.9 micrograms/g for timber treatment workers applying arsenical wood preservatives, 79.4 micrograms/g for a group of glassworkers using arsenic trioxide, and 245 micrograms/g for chemical workers engaged in manufacturing and handling inorganic arsenicals. The maximum recorded concentration was 956 micrograms/g. For the most exposed groups, the ranges in the average urinary arsenic speciation pattern were 1-6% As (V), 11-14% As (III), 14-18% MMAA, and 63-70% DMAA. The highly raised urinary arsenic concentrations for the chemical workers, in particular, and some glassworkers are shown to correspond to possible atmospheric concentrations in the workplace and intakes in excess of, or close to, recommended and statutory limits and those associated with inorganic arsenic related diseases.

Arsenic↗

Role of ornithine decarboxylase in repair of gastric mucosal stress ulcers.

The purpose of this study was to determine whether ornithine decarboxylase (ODC) has a role in mucosal repair during the first 24 h after stress-induced damage. Rats were fasted 22 h, placed in a restraint cage, and immersed in water to the xiphoid process for 6 h. Animals were killed either immediately after the period of stress or at 2-h intervals up to 24 h thereafter. Gastric mucosal ODC increased significantly from 0 to 12 h and peaked 4 h after the 6-h stress period. By 24 h enzyme activity in the gastric mucosa was near normal. Macroscopic lesions were regularly produced after 6 h of stress. Histologically, stress caused extensive damage to the superficial epithelial cells, extending in some cases into the mucosa and beyond the basal lamina. However, after stress the mucosa recovered quickly, returning to near normal 24 h later. The decreases in mucosal content of DNA, RNA, and protein caused by stress also were restored and reached near-normal levels 24 h after stress. alpha-Difluoromethylornithine (DFMO), a specific inhibitor of ODC, not only inhibited the ODC activity but significantly delayed the recovery from injury as well. DFMO also prevented the restoration of DNA, RNA, and protein content of the gastric mucosa. In conclusion, stress-induced gastric mucosal lesions are accompanied by significant increases in ODC activity. The increased ODC is necessary for the normal repair of the mucosa.

Animals↗

Intestinal ornithine decarboxylase: half-life and regulation by putrescine.

Ornithine decarboxylase (ODC) is the primary rate-limiting enzyme for polyamine synthesis. ODC levels are increased in most tissues, including the intestinal mucosa, by growth-promoting agents. This enzyme has a brief half-life of from 5 to 30 min in mammalian tissues and is regulated by its product; putrescine. The current study examines the turnover and regulation of ODC in the mucosa of the small intestine. With the use of scraped intestinal mucosa from cycloheximide-treated rats, the time course of the decline in ODC activity yielded a half-life of approximately 22 min. Labeling enzyme protein with [3H]difluoromethylornithine (DFMO) resulted in a nearly identical estimation of half-life. ODC activity of mucosa from isolated gut segments stimulated by luminal glycine (0.1-0.4 M) was enhanced 60-100% by 10 mM putrescine administered luminally. Putrescine alone had no effect on ODC. In contrast, 10(-7) M putrescine prevented 80% of the ODC activity stimulated by asparagine in IEC-6 cells (a rat intestinal crypt cell line). The half-life of ODC in unstimulated IEC-6 cells was 20 min and increased to 35 min in cells exposed to 10 mM asparagine. These data demonstrate that ODC of nonproliferating villous cells is regulated differently from the identical enzyme in proliferating crypt cells. Therefore, conclusions regarding mucosal growth should not be based totally on ODC activity from whole mucosa, since it is essentially a measure of only the enzyme present in the villous cells.

Animals↗

Regulation of ornithine decarboxylase activity in LoVo cells.

The role of Na+ and Na(+)-H+ exchange in the stimulation of ornithine decarboxylase (ODC) activity has been investigated in a human colon adenocarcinoma cell line, LoVo. Asparagine (Asn; 10 mM) or 10% fetal bovine serum (FBS) increased ODC activity from undetectable levels to greater than 500 pmol CO2.mg protein-1.h-1 in 4 h. This increase could be reduced 50% by concentrations of Na(+)-H+ exchange inhibitors that did not reduce protein synthesis. (approximately 0.2 mM for amiloride and 0.05 mM for hexamethyleneamiloride). Asn was able to double the uptake of 22Na+, whether an ionic (choline chloride) or nonionic (D-mannitol) substance was substituted for Na+, and the substitution of these compounds as well as N-methyl-glucamine for Na+ largely prevented the stimulation of ODC by Asn. Another factor influencing ODC activity was extracellular pH (pHo). When pHo was lowered, intracellular pH (pHi) also fell, and ODC activity was reduced. When pHo was raised, pHi also rose, and ODC activity increased. The well-known correlation between increased pHi and Na+ uptake with the stimulation of growth may be due to their influence on ODC activity.

Adenocarcinoma↗

Gastric and duodenal mucosal ornithine decarboxylase and damage after corticosterone.

We recently found that stress increases gastric and duodenal ornithine decarboxylase (ODC) activity and damages both tissues. The current study investigated whether corticosterone induces ODC activity in gastric and duodenal mucosa in rats and compared plasma corticosterone levels after stress and treatment with corticosterone to determine whether this hormone mediated the effects of stress on the mucosa. Rats were fasted 22 h, placed in a restraint cage, and immersed in water to the xiphoid process for 6 h. Stress markedly increased plasma corticosterone levels; the increase was 10.5 times control and lasted the duration of stress. The maximum increase was observed 1 h after a single injection of corticosterone (5 mg/kg sc) and represented 11.1 times control values. By 6 h, plasma corticosterone had returned to normal levels. A single injection of corticosterone had no effect on the gastric mucosa, but duodenal ODC was increased significantly from 4 to 8 h after injection, peaking at 6 h. Histological examination revealed no damage in either tissue. Administration of corticosterone three times daily for 3 days dramatically elevated ODC activity and produced significant microscopic damage. The surface epithelium of the stomach was disrupted, with many surface cells shed, and most villi were absent from the duodenal mucosa. Corticosterone also markedly decreased DNA and RNA content of both tissues. Inhibition of ODC with DL-alpha-difluoromethylornithine additionally decreased DNA, RNA, and protein content, exacerbating the damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Luminal polyamines stimulate repair of gastric mucosal stress ulcers.

The purpose of this study was to examine whether luminal polyamines can substitute for tissue polyamines in the healing process of gastric mucosal stress ulcers. Rats were fasted 22 h, placed in restraint cages, and immersed in water to the xiphoid process for 6 h. Animals were killed either immediately or at 4, 12, or 24 h after the period of stress. Stress significantly increased ornithine decarboxylase (ODC) activity and tissue polyamine content. Mucosal polyamine levels peaked 4 h after stress and remained significantly elevated for 12 h. The healing process, which was significant by 12 h, was inhibited by DL-alpha-difluoromethylornithine (DFMO), a specific inhibitor of ODC. DFMO totally prevented the marked increases in ODC and polyamine levels that usually followed stress. Oral administration of polyamines, putrescine, cadaverine, spermidine, or spermine, immediately after stress increased the normal rate of healing and prevented the inhibition of repair caused by DFMO. Spermidine or spermine accelerated healing better than putrescine or cadaverine. The delayed recovery of mucosal DNA, RNA, and protein content after stress in the DFMO-treated rats was also significantly prevented by exogenous polyamines. The reduced amounts of gastric mucosal spermidine and spermine in rats treated with DFMO returned toward control levels after administration of exogenous spermidine (100 mg/kg). These results show that 1) increased levels of polyamines provided by ODC are absolutely required for normal healing of gastric mucosal stress ulcers, 2) the polyamines are active from the luminal side, and 3) polyamines accelerate healing at least partly through a mechanism involving cell renewal.

Animals↗

Long-term reproducibility of tests of small airways function. Comparisons with spirometry.

We used a tracking index to measure the reproducibility of single breath nitrogen test variables (CV/VC, CC/TLC, delta N2/L) and spirometric variables (FEV1 and FEV1/FVC) and to compare the characteristics of individuals whose pulmonary function tracks well with those whose pulmonary function tracks poorly. Data were derived from two cohorts followed longitudinally over a 9-11 year period. All variables were adjusted for age, sex and height by expressing them as Z-scores. In all smoking groups and in both cohorts, the tracking index was highest for FEV1, indicating that this variable was the most reproducible over the period of follow-up; delta N2/L and FEV1/FVC had very similar but slightly lower tracking indices; CV/VC consistently had the lowest tracking indices. Tracking indices were generally higher in smokers than in nonsmokers. The reproducibility of CC/TLC increased over the period of follow-up whereas the FEV1 reproducibility remained constant. We found no significant difference between those with high tracking indices and those with low indices in terms of sex, smoking status, prevalence of respiratory symptoms, history of respiratory disease, and rate of decline of FEV1. We conclude that FEV1 is the most reproducible of the variables examined, both within and between individuals, and that poor tracking or reproducibility are not related to smoking or to the presence of respiratory symptoms or disease.

Adult↗

Reliability of crevicular fluid measurements taken in the presence of supragingival plaque.

The measurement of gingival crevicular fluid (GCF) is commonly used in clinical studies to quantitate the severity of gingival inflammation. This study examined the influence of the presence of supragingival plaque on GCF readings. Bilateral pairs of maxillary premolars received replicate GCF measurements. One of the teeth was randomly selected to be the test tooth. Supragingival plaque was removed from the test tooth between the first and second GCF measurements. Plaque was not removed from the control tooth. The crevicular fluid was collected on filter paper strips which were placed just subgingivally. The quantity of crevicular fluid was determined with the aid of the Periotron 6000. The mean difference between the first and second GCF measurement for the control teeth was -1.5 +/- 5.7 (+/- SD). This change was not significant (P = 0.21). The mean difference between the first and second GCF measurements for the experimental teeth (-21.3 +/- 16.5) was significant (P less than or equal to .001). The mean difference between the control and the experimental GCF differences were found to be highly significant (P less than or equal to .001). The data indicate that the presence of supragingival plaque can significantly elevate the GCF measurements when determined with the Periotron 6000.

Adult↗

Glucocorticoid activation of deoxyribonucleic acid degradation in bursal lymphocytes.

Treatment of animals with exogenous adrenal steroids or elevation of endogenous glucocorticoids results in a profound involution of lymphoid tissue. In rodent species, this lymphoinvolution is accompanied by lymphocyte cell death and extensive degradation of the genome prior to lymphocytolysis. In the present study, this process was investigated in the bursa of Fabricius of domestic fowl. Four-wk-old chicks were treated with a single injection of dexamethasone, and bursal regression and cell viability were monitored over a 72-h period. Following hormone treatment, DNA was extracted from bursal lymphocytes and analyzed by agarose gel electrophoresis. Dexamethasone treatment resulted in a rapid regression of bursal tissue that could be detected as soon as 6 h posttreatment, but lymphocyte viability was not altered until 24 h afterward. The DNA isolated from bursal lymphocytes of glucocorticoid-treated birds appeared to be degraded at internucleosomal sites and generated a "ladder" of discrete DNA fragments when analyzed by agarose gel electrophoresis. This form of hormone-induced cell death, referred to as programmed cell death, may play a key role in glucocorticoid-mediated immunosuppression.

Animals↗

Respiratory symptoms, lung function, and mortality in a screening center cohort.

Numerous studies have documented the effects of smoking and reduced pulmonary function on all-cause mortality. The effects of respiratory symptoms are less well studied. This paper examines the joint effects of respiratory symptoms, lung function, and smoking using 11-year mortality data on 698 subjects aged 25 years and older. Copies of death certificates were obtained for all 120 confirmed deaths, and cause of death was coded by a nosologist using the rules of the International Classification of Diseases, Ninth Revision. Symptoms of cough/phlegm, wheeze, and dyspnea were significantly associated with all-cause mortality in separate univariate analyses. On a cause-specific basis, these associations appeared to hold for chronic obstructive pulmonary disease, lung cancer, and vascular disease. Further analysis indicated that, for both smokers and nonsmokers, the presence of chronic cough and/or sputum production was related to mortality only in the presence of wheezing. In addition, among smokers, the presence of both cough/phlegm and wheeze. In addition, among smokers, the presence of both cough/phlegm and wheeze was significantly associated with mortality only among subjects with low initial lung function. Although the limited number of deaths and the nonrandom nature of the cohort limit the generalizability of our findings, it seems clear, based on these results and other published studies, that symptoms of cough, phlegm, and/or wheeze have important adverse health implications even in the absence of smoking and reduced lung function. More studies using common methodological approaches are needed.

Adult↗

Mucosal ornithine decarboxylase in the small intestine: localization and stimulation.

In most tissues increases in ornithine decarboxylase (ODC) are associated with growth. Refeeding fasted rats. a potent stimulus for mucosal growth, strongly increases ODC in both small and large intestinal mucosa. In the small bowel, almost all of this increase occurs in the mature villus cells rather than the proliferative crypt cells. Nevertheless, inhibition of ODC with difluoromethylornithine blocks the growth response. Using a highly specific, polyclonal antiserum to ODC, we have determined that in the fasted rat ODC is localized almost exclusively to the villus cells. Using antiserum dilution techniques, we have shown that, within 2 h, refeeding increases the amount of immunoreactive ODC in both villus and crypt cells. Furthermore, the trophic hormone gastrin also increases ODC, but only in the crypt cells. Epidermal growth factor increased ODC to a greater extent than gastrin, but stimulation was more general, including both crypt and villus cells. Perfusing an isolated segment of small bowel in situ with glycine for 2 h also increased immunoreactive ODC but only in the villus cells. Thus in the small intestine the effect of refeeding on ODC activity appears to be mediated by different types of stimuli: luminal nutrients increase enzyme levels in the absorbing villus cells, while trophic peptides stimulate ODC synthesis in the proliferative crypt cells.

Animals↗

Putrescine uptake and release by colon cancer cells.

We have investigated the uptake and release of [3H]putrescine by a human colon adenocarcinoma cell line (LoVo) maintained on filter inserts. This culture system permits the cells to develop morphological polarity and provides separate access to the basolateral and apical surfaces of the cells. [3H]putrescine was taken up more readily by the basolateral than by the apical side of the cells. [3H]putrescine uptake could be stimulated greater than 300 times by either 10 mM asparagine or 10% fetal bovine serum. [3H]putrescine was accumulated to a concentration gradient of approximately 300-fold; uptake could be inhibited 50% by 7.5 microM unlabeled putrescine and was not dependent on Na+. The release of [3H]putrescine into the apical medium was inhibited by asparagine or fetal bovine serum. Usually, less than one-thousandth of the [3H]putrescine taken up into the cells was released into the apical medium. Release of [3H]putrescine did not correspond to the accumulation of [14C]-inulin in the apical medium. For these reasons we concluded that putrescine release was not simply passive leakage but was responsive to intracellular demand. The [3H]putrescine taken up by the cells as well as that released into the apical medium was greater than 90% unmetabolized at 4h.

Amiloride↗

Characteristics of spermidine uptake by isolated rat enterocytes.

Eukaryotic cells require polyamines for growth. The supply of polyamines to growing cells may be increased either by new synthesis or increased uptake. We have recently shown that putrescine uptake by isolated rat enterocytes is energy dependent, saturable, and ouabain insensitive. Although putrescine uptake was inhibited by putrescine and cadaverine, it was not inhibited by equal concentrations of spermine and spermidine. These data indicated that a carrier mechanism separate from that putrescine existed for spermidine and spermine. In the current study spermidine uptake by isolated enterocytes was saturable, temperature dependent, and inhibited by 1 mM KCN. Kinetic analysis resulted in a Km = 2.51 x 10(-6) M and a Vmax = 3.57 x 10(-12) mol.10(6) cells-1.15 min-1. Spermidine uptake was 70% inhibited by 1 mM ouabain. Replacement of sodium by choline, lithium, tetramethylammonium, or N-methyl-D-glucamine also inhibited spermidine uptake. Replacement of Na+ by mannitol or sucrose, however, depressed uptake but not significantly. Spermidine uptake was inhibited by 1 mM ouabain. Spermidine uptake was inhibited by relatively low concentrations of spermine and high concentrations of putrescine; while putrescine uptake was inhibited by relatively high concentrations of both spermine and spermidine. Kinetic data indicated that spermidine and spermine share a carrier that is distinct from the one mediating the uptake of putrescine. While spermidine uptake does not appear to depend on Na+ cotransport, it may be dependent on the electrical gradient established by the Na+-K+-ATPase.

Amino Acids↗