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Biomedical subjects

D H Ryan

Publications and source records attributed to D H Ryan.

At least 55 records · Page 3Linked to original sources

Failure to change exploration or saccharin preference in rats exposed to chronic mild stress.

Chronic mild stress (CMS) exposes animals to unpredictable stressors. Reduced consumption of sucrose or saccharin solutions by CMS rats has been used as a putative measure of anhedonia, typical of depression. Our objective was to determine whether saccharin consumption and preference and suppression of exploratory and rearing behaviors in the open field were reliable indicators of CMS-induced behavioral depression. In Experiment 1, male Wistar rats subjected to 6 weeks of CMS consumed significantly less food and gained less weight than controls. CMS did not effect saccharin intake, or preference, measured in a two-bottle test with water. CMS rats exposed to a novel open field showed increased exploration and rearing. In a second test, performed immediately after a novel stress of restraint, there were no differences in exploratory or rearing behavior of CMS and control rats. In Experiment 2, CMS was reduced to 3 weeks and rats were single or group housed in their home cages. Open field activity of CMS rats was similar to that in Experiment 1. Saccharin preference of CMS rats was significantly suppressed when tested after 24 hours of water deprivation, but was not different from controls after 5 hours of water deprivation. In the final experiment Sprague Dawley rats behaved the same as Wistar rats in the CMS paradigm. Therefore, the CMS protocol used in these experiments did not induce behaviors indicative of depression but did cause a mild anorexia and weight loss. Saccharin intake of CMS rats was dependent upon their dehydration state and could not be attributed to stress-induced anhedonia.

Animals↗

Telomerase is up-regulated in human germinal center B cells in vivo and can be re-expressed in memory B cells activated in vitro.

The extensive proliferation that B lymphocytes undergo in germinal centers could compromise generation of long term B cell memory if there occurs shortening of the telomeres of germinal center B cells with cell division. Telomere length, which is thought to act as a "mitotic clock" for somatic cells that dictates cellular senescence, can be preserved by the enzyme telomerase. Human tonsil germinal center B cells consistently expressed 100- to 1000-fold higher levels of telomerase activity than naive or memory B cells, which had no or very low detectable activity, as analyzed by the telomere repeat amplification protocol assay. In vitro stimulation of human memory B cells through surface Ig or CD40 was capable of up-regulating telomerase. The findings suggest that longevity of B cell memory is maintained, despite multiple cell divisions in the generation of a memory B cell, by up-regulation of telomerase in germinal center and activated memory B cells.

ADP-ribosyl Cyclase↗

TGF-beta down-regulates stromal IL-7 secretion and inhibits proliferation of human B cell precursors.

Development of lymphoid progenitors in vivo requires interaction with a bone marrow stromal microenvironment containing multiple cytokines involved in the development of nonlymphoid hemopoietic lineages. We tested the effect of one such cytokine, TGF-beta, on the proliferation of early human clonogenic lymphoid progenitors using a stroma-dependent in vitro culture system. TGF-beta caused a dose-dependent inhibition of lymphoid progenitor colonies that was reversible at low TGF-beta doses by addition of exogenous IL-7 to the cultures. IL-7 was unable to reverse the inhibitory effect of higher TGF-beta concentrations or inhibition caused by IL-1alpha, IL-4, or TNF-alpha. Stromal IL-7 mRNA expression and protein secretion were markedly down-regulated by TGF-beta, suggesting that inhibition of stromal IL-7 secretion partially accounts for the inhibitory effect of TGF-beta on lymphopoiesis in this culture system. It is likely that higher TGF-beta concentrations do not inhibit lymphopoiesis by down-regulating IL-7 receptor expression, since this cytokine did not reduce IL-7R alpha or gamma c mRNA levels in normal B cell precursors. Since direct stromal contact is required for in vitro lymphopoiesis, the potential regulation of the IL-7 pathway by cell adhesion was examined. Adhesion of human B cell precursors to stroma did not alter stromal IL-7 expression or expression of IL-7R alpha or gamma c-chains by B cell precursors. These results indicate that TGF-beta is a significant negative regulator of stroma-dependent proliferation of early human lymphoid progenitors and acts in part by down-regulating stromal IL-7 secretion.

B-Lymphocytes↗

Expression of interleukin-7 receptor by lineage-negative human bone marrow progenitors with enhanced lymphoid proliferative potential and B-lineage differentiation capacity.

Relatively little is known about the relationship of lymphoid-associated gene expression to the proliferation and differentiation potential of early human bone marrow lymphoid progenitors. Surface expression of interleukin-7 (IL-7) receptor-alpha (IL-7R alpha), a component of the high-affinity receptor for the lymphoid precursor growth factor IL-7, defined a CD34+ progenitor subset lacking the CD19+ pro-B phenotype but demonstrating markedly enhanced lymphoid clonogenic capacity and the ability to differentiate into pro-B cells in short-term culture. These progenitors expressed mRNA for the lymphoid-associated genes Ig beta, RAG-1, and PAX-5, and were uniformly TdT-positive (TdT+). In contrast, IL-7R alpha-/CD19-/ CD34+ progenitors had a 50-fold reduced lymphoid clonogenic capacity and did not differentiate into pro-B cells in short-term culture. Expression of TdT and the lymphoid-associated genes Ig beta and RAG-1, but not PAX-5, was detected in this fraction, although at lower levels than in the IL-7R alpha+ progenitors. In contrast to IL-7R alpha, loss of the stem cell factor receptor c-kit was associated with enhanced lymphoid clonogenic potential and increased B-lineage differentiation potential. These results indicate that IL-7R alpha expression defines entry into a developmental stage characterized by upregulation of multiple lymphoid-associated genes and enhanced fitness for B-lymphoid differentiation. The onset of IL-7R alpha and PAX-5 expression immediately before acquisition of CD19 is consistent with evidence suggesting upregulation of CD19 through pathways involving PAX-5 and IL-7.

Adult↗

Prevalence of binge eating disorder in obese adults seeking weight loss treatment.

Binge eating has been identified as a common problem in samples of obese persons. Earlier studies found that approximately 30% of participants presenting for weight loss treatment could be diagnosed with Binge Eating Disorder (BED). This study investigated the prevalence of BED using the Questionnaire on Eating and Weight Patterns (QEWP) and the Interview for the Diagnosis of Eating Disorders (IDED) in a sample of 468 obese adults seeking weight loss treatment at two research facilities. The study found that only a small percentage of the participants met Diagnostic and Statistical Manual for Mental Disorders, 4th Revision (DSM-IV) diagnostic criteria for BED using either the IDED (1.3%) or QEWP (7.3%). A larger percentage of the sample (10.7% based on the IDED and 20.5% based on the QEWP) reported binge eating, but did not endorse all criteria necessary to warrant a diagnosis of BED. The primary finding of the study was that the prevalence of BED in treatment seeking obese adults was much lower than was reported in previous studies. Also, there was significant discrepancy in prevalence rates of BED as defined by self-report and interview assessment methods, with the interview method yielding lower estimates of prevalence. These findings suggest that the prevalence of BED may be lower than estimates of earlier reports. We recommend that future studies of BED use reliable and valid interview methods and that this research focus on more diverse populations, including men and a variety of racial and ethnic groups.

Adult↗

Sleep deprivation by the "flower pot" technique and spatial reference memory.

This study investigated whether paradoxical, or rapid eye movement (REM), sleep deprivation (SD) affected spatial memory. SD was induced in male Wistar rats by housing them on small platforms over water. They fell into the water if they lost muscle tone. Controls were either housed in tanks with large platforms (TC) or in normal cages (CC). All rats had free access to food and water. Each day they were tested in a place-learning set task using a Morris water maze. The rats were released from 6 different starting points (sets) and allowed 2 min to find a submerged platform. Two trials were conducted from each starting point. SD caused a significant decrement in performance in Trial 1 from Day 2. By Day 4, when distance swum to find the platform was plotted against set, area under the curve was doubled in SD compared to that in TC and CC rats, indicating a significant impairment in reference spatial memory. There was no consistent effect on working memory, indicated by Trial 2. SD caused weight loss and increased serum corticosterone compared to that in CC rats. There were no differences in concentrations of hypothalamic, hippocampal, or cortical catecholamines or their metabolites. Serotonin metabolism was elevated in the hypothalamus and hippocampus in SD rats. These results indicate that SD induced in rats housed on small platforms causes a substantial impairment of reference memory. The memory deficit may not be specific to SD because the rats are physically stressed and lose some nonREM sleep when housed in these conditions.

Animals↗

Appetitive operant behavior and free-feeding in rats exposed to acute stress.

This study investigated whether appetitive operant food reinforcement or free-feeding behavior in rats, food-restricted to 85% of body weight, was disrupted by exposures to 3 h of restraint stress or by 3 h of restraint plus water immersion stress (RWI). Rats were trained under a 3-cycle 10-min time-out-10-min time-in fixed-ratio 15 (FR15) schedule of food reinforcement. Free-feeding was measured in a 3-cycle 10-min food jar-out-10-min food jar-in test conducted in the operant chambers. Three hours of restraint stress did not significantly affect response rate or food reinforcement in the operant FR15 task or in the free-feeding condition. In contrast, 3 h of RWI completely abolished operant food reinforcement and suppressed response rate, whereas free-feeding was significantly reduced but not abolished in food-restricted, hungry rats. We conclude that acute restraint stress in food-restricted, hungry rats does not affect their appetite or motor ability to lever-press for food nor did it affect their ability to feed in a free-feeding situation. In contrast, RWI stress may have induced a motor impairment, or some other aspect of motivation independent of hunger, that disrupted their performance in the operant FR15 task.

Animals↗

CRF receptor antagonist attenuates immobilization stress-induced norepinephrine release in the prefrontal cortex in rats.

Neuroanatomical, neurophysiological, and behavioral studies suggest that brain stem nucleus locus coeruleus (LC) plays an important role in stress response. The present study was designed to clarify, whether infusion of CRF antagonist, alpha hCRF, into LC could attenuate or block stress-induced changes in norepinephrine (NE) concentrations in microdialysates collected from the medial prefrontal cortex (PFM). Rats were implanted with a bilateral cannulae assembly aimed in the LC and a microdialysis probe (4 mm active membrane length) into the LC. Immobilization of animals significantly increased the concentration of NE in microdialysates from PFM to a maximum of 170.8 +/- 12.8% of the baseline ten minutes after the onset of stressor. Concentration of NE in dialysates remained significantly elevated for the next 40 min. Infusion of alpha hCRF into the LC significantly attenuated stress-induced increase in PFM NE concentration in samples collected at 10, 20, 30, and 50 min after the onset of immobilization. Infusion of alpha hCRF alone (no immobilization) did not change concentrations at any time during sample collection. These results are consistent with other studies and suggest that stress can facilitate NE release in the PFM through the activation of the CRF system in the brain.

Animals↗

Does vegetable oil attenuate the beneficial effects of fish oil in reducing risk factors for cardiovascular disease?

Contradictory reports on the protective effect of fish consumption on cardiovascular disease (CVD) risk could be due to variations in the intake of n-3 and n-6 polyunsaturated fatty acids (PUFAs). Metabolic competition between n-3 and n-6 PUFAs suggests that n-6 PUFAs in vegetable oils could attenuate the efficacy of n-3 PUFAs in fish oil to favorably alter endpoints relevant to CVD risk. We determined the effects of varying dietary amounts of fish oil on lipid and thrombotic endpoints relevant to risk factors for CVD and whether these effects were attenuated by vegetable oils. Two randomized, double-blind, placebo-controlled, parallel studies were conducted in human subjects fed varying amounts of n-3 and n-6 PUFAs; n-3 PUFA intake was varied by using fish or placebo oil capsules, and n-6 PUFA intake was modified by incorporating varying amounts of safflower oil into the diet. Endpoints included changes in membrane fatty acid composition, blood lipids, and thrombotic profile. The results indicated that absolute amounts of fish oil, and not the relative amounts of fish and vegetable oil (ratios of n-3 to n-6 PUFAs), determined the magnitude of the reduction of arachidonic acid and increase in eicosapentaenoic acid in phospholipids of plasma and platelets. The suppression of plasma triacylglycerols by fish oil was not affected by varying amounts of dietary n-6 PUFAs. Fibrinogen concentrations decreased with 15 g but not with 9 g fish oil/d fed at the same ratio of n-3 to n-6 PUFAs. The efficacy of fish oil in favorably modifying certain risk factors for CVD was not attenuated by vegetable oil.

Adolescent↗

Effect of restraint stress on food intake and body weight is determined by time of day.

Three experiments were conducted to investigate the effect of restraint stress applied at different times of the light-dark cycle on feeding behavior and body weight of rats. Sprague-Dawley rats were restrained for 3 h in restraining tubes either at the start or the end of the light cycle. There was a significant reduction in food intake on the day of restraint and no change in food intake during a 10-day recovery period in either experiment. Reductions of food intake on the day of restraint were about the same for both restrained groups compared with their controls. When stress was applied in the evening, eating was inhibited during the first 2 h after restraint, whereas in rats restrained in the morning, feeding was suppressed twice: during the 4 h after restraint and during the first 2 h of the dark cycle. Restraint induced a significant weight loss that was greater in the rats stressed in the morning. Neuropeptide Y (NPY) levels determined at the time of food suppression for both experiments (beginning of the dark cycle) revealed an elevation of NPY in the paraventricular nucleus of rats stressed in the morning compared with other groups, but no difference in hypothalamic NPY mRNA expression. Expression of uncoupling protein mRNA in brown adipose tissue and leptin mRNA in epididymal fat, measured at the start of the dark period, was not altered by stress. There was an elevation of dopamine turnover in the hypothalami of rats restrained at the end of light cycle, but not those restrained in the morning. These results show that restraint stress has a greater effect on metabolism and energy balance when it is applied in the morning. Additional studies are needed to elucidate mechanisms involved in the suppression of food intake 9 h after restraint.

3,4-Dihydroxyphenylacetic Acid↗

A paradigm of experimentally induced mild hyperthyroidism: effects on nitrogen balance, body composition, and energy expenditure in healthy young men.

Although T3 exerts major regulatory actions in both animals and humans, most clinical studies of T3 administration have been relatively short term. The present study examined the effects of more than 2 months (63 days) of low dose T3 treatment on nitrogen balance, body composition, 24-h energy expenditure (EE), and protein turnover in seven healthy men studied at an in-patient metabolic unit. Subjects were also randomly assigned to either high or low fat diets to determine the effects of diet composition. T3 treatment produced significant losses in both lean mass (1.5 +/- 0.3 kg) and fat mass (2.7 +/- 0.4 kg) by 6 weeks, with similar reductions in both at 9 weeks. The high fat diet somewhat attenuated the loss of body fat. Nitrogen balance was significantly negative for the first 3 weeks of T3 treatment, but tended to return to baseline thereafter. There were no significant effects of treatment on protein turnover at 9 weeks, although there was a slight increase in leucine oxidation (P = 0.07). Despite the apparent adaptation in nitrogen balance, total 24-h EE and sleeping EE were significantly increased at 9 weeks. We conclude that although healthy men are able to adapt to mild hyperthyroidism in terms of nitrogen balance, they exhibit significant and persistent changes in fat and fat-free mass as well as energy balance.

Adult↗

Corticotropin-releasing factor receptor antagonist infused into the locus coeruleus attenuates immobilization stress-induced defensive withdrawal in rats.

It has been proposed that corticotropin-releasing factor (CRF) released during stress in the region of the locus coeruleus (LC) induces changes in behavior that are typical indices of anxiety. The experiments tested the ability of a CRF antagonist, alpha hCRF9-41, to attenuate stress-induced defensive withdrawal in rats. 1 microgram of alpha hCRF in 300 nl was infused bilaterally in the LC of rats 10 min prior to 30 min immobilization. The apparatus consisted of a small chamber set on one side of a 1 m open field, into which the rat was placed to start the test. Restraint induced defensive withdrawal in rats familiar with the apparatus and significantly increased latency time to emerge from the chamber, total time and mean time spent in the chamber. Infusion of alpha hCRF into the LC prior to restraint significantly decreased total and mean time spent in the chamber comparing to stressed animals. These results are consistent with anatomical, electrophysiological and neurochemical evidence that CRF receptors located in, or close to, the LC region influence behaviors induced by stress.

Aggression↗

A double-blind randomized placebo-controlled trial of sibutramine.

Sibutramine is a beta-phenethylamine which blocks reuptake of norepinephrine and serotonin. In this clinical study, a group of 173 patients were randomized to treatment with sibutramine at doses of 1, 5, 10, 15, 20 or 30 mg/d and were compared with placebo in a 24-week double-blind trial. There was a dose-dependent reduction in body weight, with doses of 10, 15, 20 and 30 mg being significantly greater than placebo. Weight loss was still continuing in the highest three doses at the end of the study. When drugs were discontinued patients regained weight, as expected. Side effects were generally mild and were most evident in the group treated with the highest dose. These studies suggest that sibutramine may be a valuable new drug for treatment of obesity.

Adolescent↗

Medicating the obese patient.

The acceptance of medication as a legitimate adjunct to diet and behavior modification in the treatment for obesity is an emerging phenomenon spurred by advances in understanding the biologic basis of body weight regulation and by the demonstration of safe and effective chronic maintenance of weight loss using a pharmacobehavioral approach. The decision to medicate for obesity depends on good clinical judgment based on such considerations as body mass index; body composition; body fat dissociation; age; sex; and comorbid conditions, such as diabetes and hypertension. Several nonadrenergic agents and a serotonergic agent have FDA-approved indications for weight loss. Phenylpropanolamine is available over the counter. Clinical trials support the efficacy of fluoxetine and ephedrine or caffeine in producing weight loss, although these agents do not have FDA-approved indications for treatment for obesity. In addition, new agents are being developed or are anticipated for approval. The use of existing agents in combination and their use adjunctive to diet and behavioral approaches to obesity treatment are fertile areas for research. The expectant attention to this subject is demanded by the imperative that the health in one three people in the United States is adversely affected by obesity.

Appetite Depressants↗

The association of body weight, dietary intake, and energy expenditure with dietary restraint and disinhibition.

The hypotheses that dieting and/or overeating are associated with adiposity, eating disturbances, and lowered energy expenditure were tested in this study. A sample of 44 premenopausal women scoring high and low on measures of dietary restraint and disinhibition of dietary control, as measured by the Three Factor Eating Questionnaire, was studied. A 2 x 2 factorial design was employed (High/Low restraint x High/Low Disinhibition). Dependent variables were: body composition, dietary intake, activity, resting metabolic rate, and thermic effect of food. Unrestrained overeaters (Low Restraint/High Disinhibition group) were very obese. High Dietary Restraint was associated with intent to diet and controlled eating. High scores on the Disinhibition Scale were associated with episodic overeating. Groups did not differ in resting metabolic rate (controlled for fat-free mass). Lower thermic effect of food was found to be associated with the obesity found in High Disinhibition subjects. Thus, Dietary Restraint was not associated with significant adverse effects upon physical or psychological health. High Disinhibition, however, was associated with adiposity and significant disturbances of eating.

Adipose Tissue↗

Sibutramine: a novel new agent for obesity treatment.

Sibutramine is a novel new pharmacologic agent which is a specific reuptake inhibitor for norepinephrine and serotonin. Preclinical data show that sibutramine and its two metabolites reduce food intake of animals eating either high or low carbohydrate diets and of obese Zucker rats. An 8-week clinical trial showed a dose-dependent decrease on body weight. Sibutramine, 5 and 20 mg/day, produced a dose-related weight loss in obese subjects compared to placebo in an 8-week trial. In doses varying from 1 to 30 mg, sibutramine also produced a dose-dependent decrease in weight in the healthy obese population when used in 6-,8-,12-24- and 52-week trials. Although the majority of the weight loss occurred during the first 12 weeks of treatment, weight loss had not plateaued in by 24 weeks in the higher doses. Side effects were mild. This drug shows promise as an antiobesity drug.

Adolescent↗

Association of body mass with dietary restraint and disinhibition.

The relationship of disinhibition and dietary restraint with body mass was studied in a sample of 293 women. Results suggested that higher body mass was associated with an interaction of disinhibition and dietary restraint. The association of disinhibition with higher body mass was moderated by increased dietary restraint. Symptoms of an eating disorder were more strongly associated with disinhibition than with dietary restraint. These results suggest that dieting may moderate the increased body mass associated with overeating. Psychological and eating problems associated with dietary restraint were found to be of less significance than those associated with disinhibition.

Adolescent↗

Regulation of human B cell lymphopoiesis by adhesion molecules and cytokines.

Recent advances in the ability to culture normal human B cell precursors have emphasized the supportive relationship between these cells and stromal cells in the bone marrow microenvironment. It is now possible to examine the role of adhesion molecules and cytokines in the regulation of different stages of human lymphopoiesis using these culture systems. Direct cell-cell adhesion mediated by the integrin adhesion molecule VLA-4 plays a critical role in supporting stromal dependent proliferation of human B cell precursors. In addition, human B precursor cell lines migrate underneath the stromal layer. This transmigration is VLA-4 dependent but not inhibitable by antibody to known VLA-4 ligands. IL-7 is secreted by the stromal layer, and is necessary for stromal-dependent proliferation of early human B cell precursors. Proliferation of early human B cell precursors or mature B cells. Since the bone marrow stroma is a source of cytokines with B cell precursor growth stimulatory activity, it is possible that adhesion interactions may play a co-stimulatory role with respect to cytokine secretion or response. As the cytokine requirements for human B cell lymphopoiesis become more completely defined, it will be important to uncover the cell-cell signals that regulate lymphopoiesis either directly or through modulation of cytokine secretion by supporting cells in the bone marrow microenvironment. The dependent relationship between human B cell precursors and the bone marrow microenvironment provides a model system for these cell-cell interactions which may be applicable to progenitor development in other lineages.

Animals↗