Genetic Diversity and Protective Efficacy of the RTS,S/AS01 Malaria Vaccine.

BACKGROUND: The RTS,S/AS01 vaccine targets the circumsporozoite protein of Plasmodium falciparum and has partial protective efficacy against clinical and severe malaria disease in infants and children. We investigated whether the vaccine efficacy was specific to certain parasite genotypes at the circumsporozoite protein locus. METHODS: We used polymerase chain reaction-based next-generation sequencing of DNA extracted from samples from 4985 participants to survey circumsporozoite protein polymorphisms. We evaluated the effect that polymorphic positions and haplotypic regions within the circumsporozoite protein had on vaccine efficacy against first episodes of clinical malaria within 1 year after vaccination. RESULTS: In the per-protocol group of 4577 RTS,S/AS01-vaccinated participants and 2335 control-vaccinated participants who were 5 to 17 months of age, the 1-year cumulative vaccine efficacy was 50.3% (95% confidence interval [CI], 34.6 to 62.3) against clinical malaria in which parasites matched the vaccine in the entire circumsporozoite protein C-terminal (139 infections), as compared with 33.4% (95% CI, 29.3 to 37.2) against mismatched malaria (1951 infections) (P=0.04 for differential vaccine efficacy). The vaccine efficacy based on the hazard ratio was 62.7% (95% CI, 51.6 to 71.3) against matched infections versus 54.2% (95% CI, 49.9 to 58.1) against mismatched infections (P=0.06). In the group of infants 6 to 12 weeks of age, there was no evidence of differential allele-specific vaccine efficacy. CONCLUSIONS: These results suggest that among children 5 to 17 months of age, the RTS,S vaccine has greater activity against malaria parasites with the matched circumsporozoite protein allele than against mismatched malaria. The overall vaccine efficacy in this age category will depend on the proportion of matched alleles in the local parasite population; in this trial, less than 10% of parasites had matched alleles. (Funded by the National Institutes of Health and others.).

Heart rate, blood pressure, and running speed responses to mesencephalic locomotor region stimulation in anesthetized rats.

The decerebrate rat locomotor preparation described in a previous study requires extensive brain surgery with the possibility of significant blood loss. The purpose of this study was to improve on the previous model by using lightly anesthetized instead of decerebrated rats. After initial surgery consisting of boring a small hole through the parietal bone, the animals were maintained on low levels of halothane anesthetic. The mesencephalic locomotor region was then located by physiological criteria using stereotaxic coordinates from the previous study. Locomotor speed, blood pressure and heart rate responses were then measured over a wide range of stimulation currents that elicited a maximal running speed. Stimulation currents ranged from 36 microA for walking to 82 microA for fast galloping. Locomotor speeds ranged from 20 m/min for walking to 64 m/min for fast galloping. Some animals easily achieved galloping speeds beyond 100 m/min. Blood pressure and heart rate increased with increasing stimulation currents. Blood pressure also increased during stimulation after muscular paralysis. This was not due to current spread, suggesting that the mesencephalic locomotor region might be involved in central command mechanisms. Heart rate did not increase after paralysis. This supports other multi-joint dynamic studies suggesting that movement per se may be necessary to induce heart rate changes, presumably via joint mechanoreceptors. The range of locomotor patterns and cardiovascular responses were obtained under self-supported conditions. By defining the mesencephalic locomotor region via physiological criteria, and by grouping blood pressure and heart rate measurements by gait rather than by stimulation currents, the potential use of the intact model for cardiovascular control studies was demonstrated. The animals were able to run and gallop at high speeds considering they were anesthetized. The simplified preparation will be useful for more complex cardiovascular experiments requiring intact and self-supported conditions.

Cisapride. Drug interactions of clinical significance.

Cisapride is a prokinetic agent which restores motility of the gastrointestinal tract in conditions of decreased bowel transit. It may also alter the absorption of coadministered drugs. The absorption of morphine, diazepam, cyclosporin, alcohol (ethanol) and levodopa are increased. Initial absorption of cimetidine and raniditine is also increased, but overall absorption is lower due to increased bowel transit. The absorption of digoxin, propranolol and the anticoagulants warfarin and phenprocoumon appears unaffected by cisapride, although increase thrombotest values were seen with acenocoumarol (nicoumalone). Drug interactions leading to increased plasma concentrations of cisapride may produce an increase in adverse effects. The most important of these is QT interval prolongation and ventricular arrhythmias. Phenytoin does not appear to affect protein binding of cisapride. Cisapride metabolism is inhibited by the antifungals ketoconazole, fluconazole, itraconazole and miconazole, and by the antibacterials erythromycin, troleandomycin and clarithromycin. Cisapride should not be coadministered with these drugs. Cimetidine produces a small increase in cisapride plasma concentrations, which may be due to inhibition of metabolism. Cisapride absorption is unaffected by other antacids. Atropine may reverse the cisapride-induced increase in peristalsis. Prescribers should remain vigilant to the presence of these and other, as yet unreported, reactions.

Blood pressure responses to LBNP in nontrained and trained hypertensive rats.

To study the influences of 16 wk of endurance training on the reflex regulation of resting blood pressure, nontrained (NT) and trained (T) female hypertensive rats (SHR) were subjected to conditions of lower body negative pressure (LBNP). Measurements of muscle cytochrome oxidase activity and run time to exhaustion indicated that the animals were endurance trained. The rats (NT = 6, T = 7) were tranquilized with 300-600 micrograms.kg-1 diazepam (IV) before heart rates and blood pressures were measured over a range of 2.5-10.0 mm Hg of negative pressure. When subjected to conditions of LBNP, the reflex tachycardia of the T group was greater than the NT at the lower (-2.5 and -5.0 mm Hg) negative pressures. Although arterial pressure declines were similar in both groups, the T group experienced significantly less of a decline in central venous pressure than the NT animals. When chlorisondamine was used as a ganglionic blocker (2.5 mg.kg-1, IV), the fall in CVP at 10 mm Hg negative pressure was greater for the NT group while the fall in the initial systemic arterial pressure was more for the T group. From these results we concluded that training had altered the interaction between cardiopulmonary and arterial baroreflexes in these hypertensive rats and a nonneural component had been altered such as cardiac function.

A model of dynamic exercise: the decerebrate rat locomotor preparation.

The purpose of this study was to develop a dynamic exercise model in the rat that could be used to study central nervous system control of the cardiovascular system. Rats of both sexes were decerebrated under halothane anesthesia and prepared for induced locomotion on a freely turning wheel. Electrical stimulation of the mesencephalic locomotor region (MLR) elicited locomotion at different speeds and gait patterns and increased heart rate and blood pressure. Two maneuvers were performed to illustrate the potential use of the preparation. The first maneuver consisted of muscular paralysis, which prevents excitation of muscle mechanoreceptors and chemoreceptors resulting from exercise. MLR stimulation still increased blood pressure. The second maneuver was performed to determine whether the blood pressure response obtained during paralysis was an artifact of electrical stimulation of the MLR. After microinjection of gamma-aminobutyric acid into the MLR, electrical current thresholds for blood pressure and locomotion increased in parallel. gamma-Aminobutyric acid injection also reduced the pressor response to suprathreshold electrical stimulation by 76%. The injection results suggest that electrical stimulation of the MLR activates cells rather than fibers of passage. The blood pressure response of the exercise model is probably not an artifact of stimulation. The decerebrate rat locomotor preparation should offer another approach to investigate difficult problems in exercise physiology.

Arterial hemodynamics during head-up tilt in conscious dogs.

The purpose of this study was to measure the major arterial hemodynamic responses to head-up tilt in the conscious dog. After recovery from surgery for instrumentation, and after habituation to tilt, the dogs were tilted from horizontal to 75 degrees for 5 min. The arterial hemodynamic response after the initial cardiovascular adjustments to the tilt consisted of no change in heart rate and significantly increased arterial blood pressure, with significantly reduced stroke volume and cardiac output. Both renal blood flow and terminal aorta blood flow declined significantly, even more than cardiac output. Muscular exertion was not part of the tilt response because upright standing on the hindlimbs elicited a sustained increase in heart rate and a significantly smaller increase in estimated total peripheral resistance. When compared with the orthostatic response in humans, the increase in arterial pressure was exaggerated in the dogs.

Exercise training and the arterial baroreflex.

To test the hypothesis that endurance training would attenuate the carotid sinus baroreflex in rats, studies were undertaken with 25 nontrained (NT) and 22 trained (T) male Sprague-Dawley rats that were exercised for 11-14 wk. Maximal O2 consumption was significantly increased 10% after training. The left carotid sinus region was functionally isolated in anesthetized animals. Subsequently, static carotid sinus pressure was raised in 20-Torr increments from 95 Torr until a maximal response in systemic arterial pressure and regional blood flows was recorded. Compared with the NT group, baroreflex control of blood pressure and calculated regional resistance of the T animals was less responsive to changes in carotid sinus pressure. Resting blood pressure, heart rate, and changes in peripheral blood flow velocity were similar for the two groups. Peripheral sensitivity to phenylephrine-HCl and hexamethonium bromide were also similar in the T and NT groups. It was concluded that the arterial baroreflex control of blood pressure was attenuated by exercise training. These findings support the concept that the trained individual is at disadvantage during hypotensive episodes and that endurance training will attenuate the sympathetic component of the arterial baroreflex.

Fastigial nucleus modulation of medullary parasolitary neurons.

Input from the cerebellar fastigial nuclei to neurons at the lateral margin of the nuclei of the solitary tract, particularly to the area identified as the nucleus parasolitarius was investigated in acutely prepared, anesthetized dogs. Fastigial nucleus stimulation led to short latency excitation of nucleus parasolitarius units often followed by prolonged inhibition of spontaneous activity. Excitation from deep skeletal muscle afferents, converged on 25% of the spontaneously active units excited from the fastigial nuclei; these afferents originated primarily from the ipsilateral forelimb muscles. This study provides electrophysiological evidence for fastigial modulation of neurons previously demonstrated autoradiographically to receive presumed monosynaptic fastigial nucleus efferents. The convergence of forelimb muscle afferent information tentatively identified as being from Group Ia or Group II pressure stretch receptors suggests that the nucleus parasolitarius may be an integrative area for cerebellar, sensorimotor and/or autonomic information.

Lower body negative pressure in the tranquilized rat.

The application of lower body negative pressure (LBNP) to tranquilized rats was assessed as an experimental technique to evaluate the response of the cardiovascular system to hypotension. After pilot studies had demonstrated that diazepam (600 micrograms X kg-1, i.v.) had no significant influence on the pressor response to unilateral carotid occlusion in unanesthetized and unrestrained rats, subsequent rats were tranquilized. When LBNP was applied, the decline in central venous pressure was linearly related to the level of negative pressure as was the initial fall in mean arterial pressure (MAP). Pulse-interval was highly correlated with the initial fall of MAP. The results indicate that the application of LBNP in the tranquilized rat can effectively produce systemic hypotension and elicit cardiovascular reflexes similar to those reported for other animals in response to LBNP, including humans.

Acute cardiorespiratory responses of hypertensive rats to swimming and treadmill exercise.

The acute cardiorespiratory responses of spontaneously hypertensive rats (SHR) to swimming and running exercise was investigated because SHR populations are hyperresponsive to external stimuli, of the paucity of existing data, and of the uncertainty on the role of exercise stimuli for training adaptations to occur. Male rats were assigned to one of five groups (n = 5-6/group) and designated as controls (C), inexperienced or naive free swimmers (NFS), experienced free swimmers (FS), experienced weighted swimmers (WS) (attached weights equal to 2% of their body weight) or experienced runners (R) who ran at an intensity of 75% of their VO2max. After 75 min in the water, all groups were acidotic and hypercapnic with the WS experiencing the greatest changes. Heart rate (HR) was increased in all swimmers during the initial 10 min, but declined thereafter, and after 75 min, the HR of WS (348 +/- 1 beats/min) was significantly lower than the C group (416 +/- 22 beats/min). At the same time interval, mean arterial blood pressure (MAP) was decreased in all swimming groups to values lower than the C animals. In addition, an exaggerated diving reflex was frequently noted when the rats were submerged. When the magnitudes of the changes were evaluated in the swimming animals they were directly associated with their submergence times, i.e., during 65-75 min of the swim, NFS, FS, and WS were submerged for 43, 46, and 66% of their total swim time, respectively. In sharp contrast to the swimmers, the runners exhibited increases in HR and MAP with their blood gas measurements being indicative of hyperventilation. We concluded that swimming as an exercise mode for hypertensive rats is best served to study the combined effects of excitement, prolonged submergence, and the consequences of the diving reflex.

An in situ study of the influence of a sclerosing solution in rabbit medial collateral ligaments and its junction strength.

A double-blind study was conducted to assess the influence of a sclerosing solution on rabbit medial collateral ligaments (MCL) in situ. It was shown that repeated injections of 5% sodium morrhuate into the MCL and its bony attachments significantly increased its bone-ligament-bone junction strength, ligament mass and thickness when compared to saline-injected controls. Morphometric analysis of electron micrographs showed a highly significant corresponding increase of the collagen fibril diameters in the experimental ligament compared against the control MCL. These composite findings suggested that the sclerosing solution had a significant influence on dense connective tissue at the insertion sites. The mechanisms for these changes are uncertain and are the basis for future investigations.

Metabolic and cardiovascular adaptations in trained hypophysectomized rats.

Metabolic and cardiovascular changes resulting from acute and chronic exercise were examined in male Sprague-Dawley rats assigned to sham-control or hypophysectomized groups. Two weeks after surgery, the hypophysectomized rats had decreased their maximum oxygen consumption (VO2 max) and heart rate values by 4 ml X min-1 X kg-1 and 142 beats X min-1, respectively. Twenty weeks later, hypophysectomy was associated with a 22 ml X min-1 X kg-1 decrease in VO2 max and a 215 beat X min-1 decline in their maximal heart rates when compared with sham-control means. Endurance training was responsible for the significantly higher O2 consumption values. Additionally, trained animals exhibited longer run times, higher muscle cytochrome oxidase activity, and reduced food consumption. Measurements of right atrial choline acetyltransferase (CAT) activity and [3H]quinuclidinyl benzilate (QNB) binding revealed significantly higher CAT values and fewer muscarinic receptors. However, training had no significant effect on resting blood pressure, blood pressure changes with conditions of lower body negative pressure, muscle glycogen concentrations, CAT levels and QNB binding of the left atrium and ventricular regions, or receptor density. These results indicated that many of the adaptations that are characteristic of normal populations can occur in the absence of the hormones from the pituitary gland.

Influence of training on the blood pressure changes during lower body negative pressure in rats.

The responses of non-trained and endurance-trained rats to conditions of lower body negative pressure (LBNP) was evaluated in normotensive, borderline hypertensive, and genetic hypertensive groups, as well as in sub-groups subjected to conditions of ventilation with 100% oxygen, systematic hemorrhaging, or sino-aortic denervations. Compared to their non-trained controls, normotensive trained rats exhibited significantly greater and faster falls in arterial blood pressure. This finding suggested a change in baroreceptor sensitivity. Related, but not statistically significant trends were observed with the hypertensive groups. Borderline hypertensive rats (DOCA injections) did not demonstrate any of these differences. Measurements of blood changes during the LBNP procedure and the effects of inspiring 100% oxygen indicated that the aortic and carotid chemoreceptors were not responsible for this training effect. After baroreceptor denervation, the group differences were abolished. In addition, the training effects were generally absent when hemorrhaging was performed, a result suggesting a difference in compliance. We have concluded from these results that endurance training will be associated with greater decreases in arterial blood pressure during LBNP than will be experienced by non-trained populations. However, the responsible mechanisms are unclear and will require further investigation.

Maximum oxygen consumption of rats and its changes with various experimental procedures.

A ten-stage treadmill test was developed and standardized to secure the VO2max of male and female rats assigned to various cross-sectional and longitudinal experimental groups. Repeated measurements indicated that the test procedure was reliable and could be used for research purposes. When the test was used with different strains, the untrained Sprague-Dawley rats had significantly higher VO2max values than animals of the Wistar-Kyoto (WKY) or the Okamoto-Aoki (SHR) strains. Exercise schedules were evaluated that were similar to those previously used by various investigators and it was found that most were exercising their rats at levels exceeding 75% VO2max. After 6--10 wk of chronic exercise, significant increases in VO2max occurred that ranged between 12 and 26%. Longitudinal studies (1 yr) with hypertensive (SHR) rats revealed that it was more desirable to logarithmically evaluate the relationship between VO2max and body mass than by the conventional method of ml . kg-1 . min-1. When this approach was used with SHR animals, the VO2max differences between the sexes were not apparent until the animals were 1 yr of age. On the other hand, training by male SHR rats caused significant increases in VO2max regardless of the method used to express the results. It is recommended that future studies designed to elucidate exercise mechanisms in rats should include a standardized VO2max test.