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H L Collins

Publications and source records attributed to H L Collins.

At least 55 records · Page 3Linked to original sources
PubMed1997

CR3-dependent phagocytosis by murine macrophages: different cytokines regulate ingestion of a defined CR3 ligand and complement-opsonized Cryptococcus neoformans.

Phagocytosis is a fundamental process in innate resistance to infection. We have used the pathogenic yeast Cryptococcus neoformans to study the interaction of this encapsulated organism with murine macrophages in vitro. In the absence of exogenous opsonins the encapsulated yeast is almost totally resistant to ingestion by murine macrophages. Owing to its ability to activate the alternative complement pathway, the anti-phagocytic properties of the polysaccharide capsule can be partially overcome following opsonization in vitro with non-immune mouse serum and subsequent phagocytosis via complement receptors. Here, we demonstrate the importance of the complement receptor type 3 (CR3) in in vitro phagocytosis of the yeast and in in vivo resistance to infection. In vitro, 70% of a population of resident murine macrophages are able to ingest C. neoformans and then only inefficiently (1-2 organisms per cell). Previously we have shown that tumour necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF) efficiently enhance ingestion of serum-opsonized encapsulated C. neoformans, and we now show that the cytokines convert a population of resident macrophages to a state where all the cells are competent for ingestion of large numbers of yeasts (6-8 per cell). We also show that these cytokines have a direct effect on CR3, as enhanced levels of complement-opsonized sheep red blood cells (EIgMC) bind to macrophages activated in this way. However, cytokines that have previously been shown to enhance phagocytosis of EIgMC have no effect on ingestion of encapsulated C. neoformans. These results demonstrate that the cytokines regulating CR3-dependent ingestion of C. neoformans are different to those regulating ingestion of EIgMC and reinforce the importance of studying pathogens rather than inert ligands in understanding the regulation of phagocytosis.

Animals↗
PubMed1997

Daily exercise attenuates the sympathetic component of the arterial baroreflex control of heart rate.

The influence of daily spontaneous running (DSR) on the sympathetic (SC) and parasympathetic components of the arterial baroreflex control of heart rate (HR) was examined in 16 female Long Evans rats [8 sedentary (SED) and 8 DSR]. After 8-9 wk of SED control or DSR, animals were chronically instrumented with arterial and venous catheters. DSR resulted in an increased heart weight-to-body weight ratio (2.71 +/- 0.11 vs. 3.09 +/- 0.09 g/kg) and a resting bradycardia (378 +/- 6 vs. 330 +/- 5 beats/min). Arterial baroreflex function was examined during ramp infusions of phenylephrine and sodium nitroprusside under the following three experimental conditions: 1) control, 2) after beta 1-adrenergic receptor blockade (beta 1-X), and 3) after muscarinic-cholinergic receptor blockade (M-X). Arterial baroreflex function parameters were compared between SED and DSR rats. In the control condition, DSR attenuated the range (182 +/- 15 vs. 124 +/- 18 beats/min), maximum HR (464 +/- 9 vs. 394 +/- 15 beats/min), and maximal gain (Gmax; 5.57 +/- 0.42 vs. 3.2 +/- 0.45 beats.min-1.mmHg-1). Similarly, after M-X, DSR attenuated the range (84 +/- 5 vs. 62 +/- 8 beats/min), maximum HR (449 +/- 11 vs. 412 +/- 11 beats/min), and Gmax (2.73 +/- 0.37 vs. 1.57 +/- 0.32 beats.min-1.mmHg-1). In contrast, after beta 1-X, DSR did not alter the range (61 +/- 13 vs. 70 +/- 7 beats/min), maximum HR (326 +/- 9 vs. 313 +/- 7 beats/min), or Gmax (3.04 +/- 0.54 vs. 3.75 +/- 0.52 beats.min-1.mmHg-1). Results demonstrate that DSR attenuated the arterial baroreflex control of HR by reducing the SC.

Animals↗
PubMed1997

Transfer of phagocytosed particles to the parasitophorous vacuole of Leishmania mexicana is a transient phenomenon preceding the acquisition of annexin I by the phagosome.

The eukaryotic intracellular pathogen Leishmania mexicana resides inside macrophages contained within a membrane bound parasitophorous vacuole which, as it matures, acquires the characteristics of a late endosomal compartment. This study reports the selectivity of fusion of this compartment with other particle containing vacuoles. Phagosomes containing zymosan or live Listeria monocytogenes rapidly fused with L. mexicana parasitophorous vacuoles, while those containing latex beads or heat killed L. monocytogenes failed to do so. Fusigenicity of phagosomes was not primarily dependent on the receptor utilized for ingestion, as opsonization with defined ligands could not overcome the exclusion of either latex beads or heat killed organisms. However modulation of intracellular pH by pharmacological agents such as chloroquine and ammonium chloride increased delivery of live Listeria and also induced transfer of previously excluded particles. The absence of fusion correlated with the acquisition of annexin I, a putative lysosomal targeting, molecule, on the phagosome membrane. We propose that the acquisition of cellular membrane constituents such as annexin I during phagosome maturation can ultimately direct the fusion pathway of the vesicles formed and have described a model system to further document changes in vesicle fusigenicity within cells.

Animals↗
PubMed1996

Diabetes reduces growth and body composition more in male than in female rats.

Food restriction and/or starvation has a consistently greater and more permanent effect on physical growth in males than in females. Because diabetes may be viewed as being analogous to starvation, we tested the hypothesis that diabetes would reduce growth more in male than in female rats. Diabetes was induced with streptozotocin (65-125 mg/kg IP) at 3 weeks of age in 7 female and 10 male Lewis rats. Body weight (BW) and blood glucose (bGlc) were measured over the following 8 weeks. Subsequently, animals were assessed for body (ano-nasal; ANL) and bone length (tibia; TBL) and chemically analyzed for body composition. Results were compared to age-matched controls (male = 11; female = 9). A 2-way factorial analysis of covariance (ANCOVA), with body weight as the covariate, was used to test for statistical significance for the effects of gender and diabetes on body composition (fat and protein mass) and linear growth because control males and females had significantly different body weights. There were no significant differences in bGlc between genders. However, males had a greater decrease from controls in BW (-45% vs. -13%), protein (-48% vs. -11%), fat (-89% vs. -65%), TBL (-13% vs. 0%), and ANL (-17% vs. -5%) compared to females. In addition, males had a greater absolute decrease from controls in protein (-40 g vs. -5 g) and fat (-39 g vs. -23 g) mass. These results suggest that male rats are more susceptible than females to the deleterious effects of diabetes on linear growth and body composition.

Aging↗
PubMed1996

Onset of exercise increases lumbar sympathetic nerve activity in rats.

We hypothesized that lumbar sympathetic nerve activity (LSNA) increases at the onset of whole-body dynamic exercise in the rat. To test this hypothesis, we recorded LSNA, heart rate (HR), and arterial pressure (AP) at rest and during a graded exercise test in six adult rats. Rats were instrumented with arterial and venous catheters and recording electrodes around the lumbar sympathetic trunk. Following recovery, each rat ran continuously on a hand-driven or motorized treadmill at 6 m.min-1, 12 m.min-1, and 18 m.min-1 on a 10% grade for approximately 3 min at each workload. Before exercise, mean arterial pressure (MAP), HR, and LSNA averaged 108 +/- 4 mm Hg, 385 +/- 20 bpm, and 100%, respectively. As hypothesized, all variables increased abruptly and dramatically at the onset of treadmill exercise. For example, MAP (117 +/- 5 mm Hg), HR (450 +/- 15 bpm), and LSNA (225 +/- 19%) all increased significantly within the first 25 s of treadmill running at 6 m.min-1. As the exercise continued, there was a progressive increase in HR; however, MAP plateaued at 6 m.min-1 and LSNA plateaued at 12 m.min-1. Since LSNA increased at the onset of whole-body dynamic exercise in the rat, we suggest that the increase in LSNA at the onset of exercise is mediated by a central (feed forward) mechanism.

Animals↗
PubMed1995

Local modulation of adrenergic responses in the hindlimb vasculature of the intact conscious rat.

1. Local modulation of adrenergic responses was examined in the hindlimb vasculature of chronically instrumented intact conscious rats. Sprague-Dawley rats (n = 22) were instrumented with a Doppler flow probe around the right common iliac artery, a polyethylene catheter inserted just distal to the flow probe and a left carotid arterial catheter. 2. The effects of various concentrations of the alpha 1-adrenergic receptor agonist phenylephrine (0.005-0.075 microgram kg-1), the alpha 2-adrenergic receptor agonist clonidine (0.1-0.7 microgram kg-1), and the endogenous adrenergic receptor agonist adrenaline (0.02-0.08 microgram kg-1), were investigated under control conditions, and in the presence of the nitric oxide (NO) synthase inhibitor N omega-nitro-L-arginine methyl ester hydrochoride (L-NAME) (NO-X, 0.2 mg kg-1) and the cyclo-oxygenase inhibitor indomethacin (CO-X, 10 mg kg-1). Results were presented as dose-response curves. 3. Heart rate and arterial pressure were not altered by any of the agents because all were locally injected into the hindlimb vasculature and the selected doses were lower than those which elicited systemic responses. 4. Maximal vasoconstrictor responses to phenylephrine were enhanced in the presence of NO-X (50 +/- 6%) and CO-X (70 +/- 9%). Maximal vasoconstrictor responses to clonidine were also enhanced in the presence of NO-X (75.3 +/- 4.8%) and CO-X (50.6 +/- 5.7%). 5. The responses to adrenaline were biphasic; NO-X significantly attenuated the vasodilator response (87 +/- 6%), and enhanced the vasoconstrictor response (51 +/- 7%). CO-X attenuated the vasoconstrictor response (71 +/- 6%). 6. These results demonstrate local modulation of responses to alpha 1- and beta-adrenergic receptor agonists by receptor-mediated dose-dependent release of NO and prostaglandins.

Adrenergic Agonists↗
PubMed1995

An educational tool for understanding the cardiovascular changes associated with diabetes.

Diabetes, a syndrome characterized by high plasma glucose and low plasma insulin concentrations, is associated with somatic and autonomic neuropadiabetes as well as cardiac and vascular disorders. These consequences of diabetes significantly affect the organism's ability to maintain homeostasis. To understand the changes associated with diabetes, we developed a laboratory exercise that compares and contrasts the cardiovascular responses to exercise in an individual with diabetes and in an individual without diabetes. This exercise provides a unique opportunity to analyze, integrate, and interpret the changes associated with diabetes, since more is learned about how a system operated when the system is forced to perform than when it is idle. In this laboratory, anatomical and physiological data concerning diabetes are provided. Subsequently, a figure that illustrates the response of a specific cardiovascular variable during exercise (e.g., heart rate, cardiac output, blood pressure) is presented. Students are challenged to analyze and assimilate information from figures, answer questions, make calculations, fill in tables, and plot graphs. The laboratory does not require equipment or software, only rulers and pencils. The answers to the questions and tables are provided in the APPENDIX. Students obtain experience in evaluating and understanding diabetes as well as applying basic cardiovascular concepts. The emphasis is on the application of basic cardiovascular principles, interpretation of pictorial or tabular material, and problem-solving skills.

Blood Circulation↗
PubMed1995

Enhanced cardiopulmonary reflex inhibition of heart rate during exercise.

We tested the hypothesis that the reflex inhibition of heart rate (HR) during mechanical (acute bolus injection of 0.5% and 2% of estimated blood volume) and chemical (phenylbiguanide, PBG, 2.5 and 5 micrograms.kg-1) stimulation of cardiopulmonary receptors would be enhanced during exercise. Rats were instrumented with arterial and venous catheters. The reflex response to mechanical (N = 7) and chemical (N = 8) stimulation of cardiopulmonary receptors was examined at rest and during exercise (6 m.min-1, 10% grade). A two-way analysis of covariance (ANCOVA) with repeated measures was used to test for differences in the reflex regulation of HR at rest vs exercise. HR was used as the covariate because exercise significantly increased baseline HR. There was no significant treatment effect (rest vs exercise) for the reflex inhibition of HR during mechanical stimulation. However, the two-way ANCOVA revealed a significant treatment effect (rest vs exercise) for the reflex inhibition of HR during chemical stimulation. The reflex decreases in HR were enhanced (-delta 23 +/- 8 vs -delta 133 +/- 47 and -delta 208 +/- 40 vs -delta 374 +/- 10 bpm at 2.5 and 5 micrograms.kg-1, respectively). These data suggest that factors associated with exercise enhanced the cardiopulmonary reflex inhibition of heart rate during chemical stimulation.

Analysis of Variance↗
PubMed1994

Lack of acidification in Mycobacterium phagosomes produced by exclusion of the vesicular proton-ATPase.

The success of Mycobacterium species as pathogens depends on their ability to maintain an infection inside the phagocytic vacuole of the macrophage. Although the bacteria are reported to modulate maturation of their intracellular vacuoles, the nature of such modifications is unknown. In this study, vacuoles formed around Mycobacterium avium failed to acidify below pH 6.3 to 6.5. Immunoelectron microscopy of infected macrophages and immunoblotting of isolated phagosomes showed that Mycobacterium vacuoles acquire the lysosomal membrane protein LAMP-1, but not the vesicular proton-adenosine triphosphatase (ATPase) responsible for phagosomal acidification. This suggests either a selective inhibition of fusion with proton-ATPase-containing vesicles or a rapid removal of the complex from Mycobacterium phagosomes.

Animals↗
PubMed1994

Vagal afferents reflexly inhibit exercise in conscious rats.

Activation of vagal afferents reflexly inhibited locomotion induced by stimulation of the mesencephalic locomotor region in decerebrate cats. However, this reflex has not been tested in intact mammals. Therefore, the purpose of this study was to test the hypothesis that vagal afferent stimulation would inhibit somatomotor activity in the intact conscious rat. Six Sprague-Dawley rats were chronically instrumented with carotid arterial and femoral venous catheters and electromyogram (EMG) electrodes inserted into the biceps femoris muscle. Cardiac autonomic efferent blockade [atropine methyl bromide (14 mg.kg-1, i.v.) and metoprolol (14 mg.kg-1, i.v.)] and alpha-adrenergic receptor blockade [phenoxybenzamine (5 mg.kg-1, i.v.)] was achieved to prevent bradycardia and hypotension. Vagal afferents were stimulated (phenyl-biguanide 2.5 and 5.0 micrograms.kg-1 i.v.) during steady state exercise (9.0 m.min-1, 10% grade). Phenyl-biguanide decreased exercise EMG activity 30 +/- 6% and 54 +/- 10% in a dose dependent manner without significantly altering mean arterial pressure or heart rate. We speculate that this reflex may serve as a negative feedback mechanism to indirectly reduce myocardial oxygen demands during exercise.

Action Potentials↗
PubMed1993

Physiology laboratory experience for high school students.

Recently, we were charged with providing a learning opportunity for high school students participating in the Upward Bound Regional Math and Science Program, a program designed to stimulate interest in mathematics and science for students from disadvantaged environments. Our challenge was to introduce students to the joys, excitement, and mystery of physiology and to stimulate their interest for future study. To this end, we developed a laboratory experience that examined basic physiological concepts in an animal model. This opportunity introduced students to how their bodies work and the importance of the use of animals in research. The students left the experience confident, motivated, and excited about learning.

Adolescent↗
PubMed1993

Attenuation of postexertional hypotension by cardiac afferent blockade.

Naloxone eliminates postexertional hypotension (PEH) in human and animal models. The effect of naloxone on sympathetic activity during hemorrhage and generation of arterial baroreflex function curves can be stimulated by blockade of cardiac afferent receptors. We tested the hypothesis that cardiac afferent blockade would eliminate PEH in eight spontaneously hypertensive rats (SHR). Rats were instrumented with a Silastic-tipped catheter inserted into the pericardial space. Four weeks later, a Teflon catheter was placed in the descending aorta via the left common carotid artery for measurement of mean arterial pressure (MAP) and heart rate (HR). MAP and HR were examined before (preexercise) and after (postexercise) a single bout of dynamic treadmill exercise (9-12.0 m/min, 10-18% grade for 30-40 min) under three experimental conditions: control, cardiac efferent blockade, and combined cardiac efferent and afferent blockade. MAP significantly decreased (29 +/- 5 and 25.6 +/- 4 mmHg) in the control and cardiac efferent blockade conditions after exercise. However, when cardiac afferents were blocked, the hypotensive response to mild dynamic exercise was significantly attenuated (-6 +/- 3 mmHg). Thus blockade of cardiac afferents eliminated PEH in the SHR. These data suggest that inhibitory influence of cardiac afferents on the circulation may be enhanced after exercise.

Afferent Pathways↗
PubMed1993

Acute exercise enhances nitric oxide modulation of vascular response to phenylephrine.

The influence of the release of endothelium-derived nitric oxide (NO) on the vasoconstrictor response to phenylephrine (PE) was evaluated before and after a single bout of dynamic exercise. Each rat ran on a motor-driven treadmill at 12-18 m/min, 10-18% grade until exhaustion (avg time 45 min). Sprague-Dawley rats (n = 6) were instrumented with a Doppler ultrasonic flow probe around the right common iliac artery. Just distal to the flow probe, a catheter was placed into the right iliac artery for local infusions. A Teflon catheter was placed in the descending aorta to measure mean arterial blood pressure (MAP) and heart rate (HR). PE (0.005-0.075 microgram/kg) and NO inhibitor N omega-nitro-L-arginine methyl ester hydrochloride (L-NAME, 0.2-0.25 mg/kg) were injected into the functionally isolated hindlimb. HR and MAP were not altered by any of the injections because we selected doses below those which elicited systemic responses. Dose-response curves to PE were generated in the control and postexercise condition, with and without the NO synthase inhibitor L-NAME. Exercise significantly attenuated the maximal vasoconstrictor response to PE (45.6 +/- 1.6%). L-NAME enhanced the maximal vasoconstrictor response to PE 49.8 +/- 4.5% in the control condition and 121.4 +/- 5.9% in the postexercise conditions. Thus, although NO inhibition enhanced the vasoconstrictor response to PE in the control and postexercise conditions, the enhanced vasoconstrictor response to PE after L-NAME was significantly greater in the postexercise condition. Results suggest that NO contributes to the exercise induced attenuation of alpha 1-adrenergic receptor stimulation.

Animals↗
PubMed1993

Daily spontaneous running did not alter vagal afferent reactivity.

Exercise training alters the cardiopulmonary baroreflex regulation of the circulation; however, the mechanisms responsible are unknown. One possibility is an enhanced afferent response to cardiopulmonary stimulation. We therefore tested the hypothesis that daily spontaneous running (DSR) would enhance cardiopulmonary vagal afferent responses to mechanical (increase in left atrial pressure, LAP) and chemical (phenyl biguanide, PBG) stimulation. Reactivity of single-fiber cardiopulmonary vagal afferents was evaluated in 16 control and 12 DSR anesthetized Sprague-Dawley rats. Rats were weaned at 3 wk of age and randomly assigned to a control or DSR group. Eight to twelve weeks of DSR was associated with a 27% increase in heart weight-to-body weight ratio (3.27 +/- 0.08 vs. 2.56 +/- 0.05 g/kg, P < 0.001) and resting bradycardia (394 +/- 10 vs. 421 +/- 8 beats/min, P = 0.036). However, DSR did not alter the stimulus-response curves to increases in LAP (frequency of discharge vs. LAP) for either the high-frequency (maximum response, sedentary 59.6 +/- 3.2, DSR 60.1 +/- 5.0 spikes/s) or low-frequency (maximum response, sedentary 20.0 +/- 2.9 DSR 20.6 +/- 3.9 spikes/s) receptors. Dose-response curves to chemical stimulation (spikes/s vs. PBG dose) were also not altered by DSR. Thus DSR did not change vagal afferent reactivity to mechanical or chemical stimulation.

Afferent Pathways↗
PubMed1993

Cardiac afferents attenuate the muscle metaboreflex in the rat.

The influence of cardiac afferents on the muscle metaboreflex was examined in 16 rats instrumented with a Silastic-tipped catheter in the pericardial space and right atrium, Doppler ultrasonic flow probe and a pneumatic vascular occluder around the terminal aorta, and a Teflon catheter in the thoracic aorta. In protocol I (cardiac efferent and afferent blockade), the muscle metaboreflex was examined under three experimental conditions: 1) control, 2) cardiac autonomic efferent blockade [intrapericardial methylscopolamine (10 micrograms/kg) and propranolol (50 micrograms/kg)], and 3) combined cardiac autonomic efferent and afferent blockade (intrapericardial procainamide, 2%). In protocol II (blood volume expansion), the muscle metaboreflex was examined before and after 15% blood volume expansion. Mild treadmill exercise (9 m/min, 10% grade) increased heart rate (71 +/- 9.4 beats/min), mean arterial pressure (12 +/- 2.0 mmHg), and terminal aortic blood flow velocity (6 +/- 1.0 kHz). During exercise, a reduction of terminal aortic blood flow velocity (10.5 +/- 1.1%) reduced mixed venous PO2 18 +/- 6%. The gain of the muscle metaboreflex in the control condition was 14.6 +/- 2.9 mmHg/kHz. Efferent blockade reduced the gain 51 +/- 7%. However, combined cardiac efferent and afferent blockade increased the gain 207 +/- 64% above the efferent blocked condition and restored the gain to levels above those obtained in the control condition (18.3 +/- 4.6 mmHg/kHz). In addition, 15% blood volume expansion reduced the gain of the muscle metaboreflex regulation of mean arterial pressure and heart rate (44 +/- 9.5% and 41 +/- 12.0%, respectively). Thus cardiac afferents tonically inhibit the pressor response to a reduction in terminal aortic blood flow velocity during exercise.

Animals↗
PubMed1992

Cytokine enhancement of complement-dependent phagocytosis by macrophages: synergy of tumor necrosis factor-alpha and granulocyte-macrophage colony-stimulating factor for phagocytosis of Cryptococcus neoformans.

We have examined the regulation of complement dependent phagocytosis by macrophage-activating cytokines. Tumor necrosis factor (TNF)-alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF), but not interferon-gamma, interleukin-4 or macrophage-CSF, stimulated ingestion of the encapsulated fungal pathogen Cryptococcus neoformans by resident peritoneal macrophages in vitro. This was dependent upon opsonization of the yeasts with complement, 72 h of incubation with the cytokines for maximum effect, and the obligate involvement of the macrophage CR3 receptor. TNF-alpha and GM-CSF synergized at low concentrations, resulting in dramatic up-regulation of phagocytosis when compared to either cytokine alone. Supernatants from C. neoformans-specific T cells also increased macrophage phagocytic efficiency. Finally, the administration of neutralizing mAb specific for TNF-alpha and GM-CSF increased mortality in C. neoformans-infected mice, and induced the rapid progression of disease with involvement of the brain and meninges. We conclude that TNF-alpha and GM-CSF are potent regulators of complement-dependent phagocytosis by murine macrophages. Macrophage activation with these two cytokines can completely overcome the anti-phagocytic properties of the virulent yeasts. Our results, therefore, implicate TNF-alpha and GM-CSF as important mediators of resistance to encapsulated pathogens such as C. neoformans where ingestion of the organism is a critical process in host resistance.

Animals↗
PubMed1991

Encapsulation of Cryptococcus neoformans impairs antigen-specific T-cell responses.

The encapsulated yeast Cryptococcus neoformans is a significant cause of opportunistic infection in patients with impaired cell-mediated immunity. The major virulence determinant of the organism is an antiphagocytic polysaccharide capsule synthesized after entry into the host. Using both an encapsulated virulent strain and an acapsular avirulent mutant, we have demonstrated the reduced ability of the encapsulated strain to stimulate specific T-cell responses in vitro. This reduction was mediated by the antiphagocytic action of the capsule rather than by direct inhibition of antigen processing and presentation, since prior opsonization with complement enhanced the ingestion of encapsulated yeast cells by purified antigen-presenting cells and allowed significant T-cell activation. Once ingestion had occurred, cryptococci were efficiently processed by activated macrophages via a chloroquine-sensitive pathway. Cryptococcal antigens were available for T-cell recognition within 1 to 2 h of interaction with macrophages and presented in a major histocompatibility complex-restricted manner. Our results suggest that the antiphagocytic action of the polysaccharide capsule is an important determinant for the development of T-cell immunity to C. neoformans.

Animals↗