Induction of B cell hyperplasia in simian immunodeficiency virus-infected rhesus macaques with the simian homologue of Kaposi's sarcoma-associated herpesvirus.

A simian homologue of Kaposi's sarcoma-associated herpesvirus (KSHV), the eighth human herpesvirus (HHV8), was isolated from a simian immunodeficiency virus (SIV)-infected rhesus macaque (Macaca mulatta) that developed a multicentric lymphoproliferative disorder (LPD). This simian rhadinovirus is genetically similar to a recently described rhesus rhadinovirus (RRV) (Desrosiers, R.C., V.G. Sasseville, S.C. Czajak, X. Zhang, K.G. Mansfield, A. Kaur, R.P. Johnson, A.A. Lackner, and J.U. Jung. 1997. J. Virol. 71:9764-9769) and is designated RRV 17577. RRV 17577 was experimentally inoculated into rhesus macaques with and without SIV(mac239) infection to determine if RRV played a role in development of the LPD observed in the index case. In contrast to control animals inoculated with SIV(mac239) or RRV alone, two animals coinfected with SIV(mac239) and RRV 17577 developed hyperplastic LPD resembling the multicentric plasma cell variant of Castleman's disease, characterized by persistent angiofollicular lymphadenopathy, hepatomegaly, splenomegaly, and hypergammaglobulinemia. Hypergammaglobulinemia was associated with severe immune-mediated hemolytic anemia in one RRV/SIV-infected macaque. Both RRV/SIV-infected macaques exhibited persistent RRV viremia with little or no RRV-specific antibody response. The macaques inoculated with RRV alone displayed transient viremia followed by a vigorous anti-RRV antibody response and lacked evidence of LPD in peripheral blood and lymph nodes. Infectious RRV and RRV DNA were present in hyperplastic lymphoid tissues of the RRV/SIV-infected macaques, suggesting that lymphoid hyperplasia is associated with the high levels of replication. Thus, experimental RRV 17577 infection of SIV-infected rhesus macaques induces some of the hyperplastic B cell LPDs manifested in AIDS patients coinfected with KSHV.

Live birth after ovarian tissue transplant.

Radiation and high-dose chemotherapy may render women with cancer prematurely sterile, a side-effect that would be avoided if ovarian tissue that had been removed before treatment could be made to function afterwards. Live offspring have been produced from transplanted ovarian tissue in mice and sheep but not in monkeys or humans, although sex steroid hormones are still secreted. Here we describe the successful transplantation of fresh ovarian tissue to a different site in a monkey, which has led to the birth of a healthy female after oocyte production, fertilization and transfer to a surrogate mother. The ectopically grafted tissue functions without surgical connection to major blood vessels and sets the stage for the transplantation of cryopreserved ovarian tissue in humans.

Radiation-induced endometriosis in Macaca mulatta.

Female rhesus monkeys received whole-body doses of ionizing radiation in the form of single-energy protons, mixed-energy protons, X rays, and electrons. Endometriosis developed in 53% of the monkeys during a 17-year period after exposure. Incidence rates for endometriosis related to radiation type were: single-energy protons, 54%; mixed-energy protons, 73%; X rays, 71%; and electrons, 57%. The incidence of endometriosis in nonirradiated control monkeys was 26%. Monkeys exposed to single-energy protons, mixed-energy protons, and X rays developed endometriosis at a significantly higher rate than control monkeys (chi 2, P less than 0.05). Severity of endometriosis was staged as massive, moderate, and minimal. The incidence of these stages were 65, 16, and 19%, respectively. Observations of clinical disease included weight loss in 43% of the monkeys, anorexia in 35%, space-occupying masses detected by abdominal palpation in 55%, abnormal ovarian/uterine anatomy on rectal examination in 89%, and radiographic evidence of abdominal masses in 38%. Pathological lesions were endometrial cyst formation in 69% of the monkeys, adhesions of the colon in 66%, urinary bladder in 50%, ovaries in 86%, and ureters in 44%, focal nodules of endometrial tissue throughout the omentum in 59%, and metastasis in 9%. Clinical management of endometriosis consisted of debulking surgery and bilateral salpingo-oophorectomy combined in some cases with total abdominal hysterectomy. Postoperative survival rates at 1 and 5 years for monkeys recovering from surgery were 48 and 36%, respectively.

Evaluation of transit-time and electromagnetic flow measurement in a chronically instrumented nonhuman primate model.

The Physiology Research Branch at Brooks AFB conducts both human and nonhuman primate experiments to determine the effects of microgravity and hypergravity on the cardiovascular system and to identify the particular mechanisms that invoke these responses. Primary investigative efforts in our nonhuman primate model require the determination of total peripheral resistance, systemic arterial compliance, and pressure-volume loop characteristics. These calculations require beat-to-beat measurement of aortic flow. This study evaluated accuracy, linearity, biocompatability, and anatomical features of commercially available electromagnetic (EMF) and transit-time flow measurement techniques. Five rhesus monkeys were instrumented with either EMF (3 subjects) or transit-time (2 subjects) flow sensors encircling the proximal ascending aorta. Cardiac outputs computed from these transducers taken over ranges of 0.5 to 2.0 L/min were compared to values obtained using thermodilution. In vivo experiments demonstrated that the EMF probe produced an average error of 15% (r = .896) and 8.6% average linearity per reading, and the transit-time flow probe produced an average error of 6% (r = .955) and 5.3% average linearity per reading. Postoperative performance and biocompatability of the probes were maintained throughout the study. The transit-time sensors provided the advantages of greater accuracy, smaller size, and lighter weight than the EMF probes. In conclusion, the characteristic features and performance of the transit-time sensors were superior to those of the EMF sensors in this study.

A conscious baboon model for evaluation of hemodynamics in altered gravity.

A model was developed for evaluation of cardiovascular parameters in conscious baboons exposed to altered gravitational environments. Baboons were trained to sit quietly in a confinement chair of unique design which allowed a range of normal physical activity. They were then instrumented with high-fidelity blood pressure transducers in the aorta and left ventricle, electromagnetic flow probes encircling the proximal ascending aorta, left and right atrial fluid catheters, left ventricular sonomicrometer crystals in a 3-axis configuration, and a hydraulic occluder cuff encircling the inferior vena cava. Catheters and transducer wires were exteriorized at the midscapular region of the back. Viability of percutaneous exit sites was enhanced by use of velour cuffs on the transducer wires, providing a scaffold for wound healing. Pressure transducers and flow probes were calibrated and balanced during postoperative cardiac catheterization procedures. This instrumentation allowed measurement of beat-to-beat stroke volume and cardiac output not reliant on thermodilution techniques. Postoperative longevity was from 1 to 10 months. Instrumentation failure included endocardial trapping of ventricular pressure transducers, corrosion of ventricular sonomicrometer crystals, and catheter tip thrombosis. Acquisition of high quality data was possible with this model in several different environments of altered gravitational stress, allowing characterization of aortic flow and ventricular performance.

A method for repeated high-fidelity micromanometer measurement of intracardiac pressures.

Dial-tipped, high-fidelity micromanometers were inserted through polyurethane catheters to acutely measure blood pressures within the chambers of the heart and the great vessels of baboons, rhesus monkeys, and goats. Repeated measurements of atrial, ventricular, aortic, and pulmonary artery pressure were possible with this method, with calibration of micromanometers accomplished immediately prior to and after pressure recordings to assure data accuracy. All attempts to pass micromanometers into the atria in all species were successful. Passage of micromanometers from the left ventricle across the aortic valve and into the aorta was successful in 97% of the attempts in baboons, 100% for rhesus monkeys, and 75% for goats; while insertions into the pulmonary artery from the right ventricle were successful in 64% of the baboons, 40% of the rhesus monkeys, and 75% of the goats. Advantages of this technique are that a permanent conduit for cardiac vascular access is available and that high-fidelity pressure signals may be acquired.

Cardiovascular responses to propofol and etomidate in long-term instrumented rhesus monkeys (Macaca mulatta).

BACKGROUND AND PURPOSE: Cardiac and arterial responses to prescribed doses of propofol and etomidate in rhesus monkeys were compared. METHODS: Intravenously administered induction doses of propofol (2 mg/kg of body weight) or etomidate (1 mg/kg) followed by continuous intravenous infusions of propofol (200 microg/kg/min) or etomidate (100 microg/kg/min) were administered. Left ventricular and right atrial access catheters were implanted for long-term use, along with a transit-time flow probe on the ascending aorta, and pericardial electrocardiogram leads. A dual sensor 3-F micromanometer was used to measure left ventricular pressure and aortic pressure, and an active redirectional transit-time probe measured aortic flow. Noordergraaf's four-element model was used to estimate total peripheral resistance and systemic arterial compliance. RESULTS: Significant (P < 0.01) decreases in mean arterial pressure, heart rate, and myocardial contractility were accompanied by an increase in systemic arterial compliance associated with propofol and etomidate. Only minimal changes in left ventricular diastolic pressure, cardiac output, stroke volume, and total peripheral resistance were found for both drugs. The changes associated with propofol are comparable to results in human beings, whereas the changes associated with etomidate did not agree with results of published human studies. CONCLUSION: The significant cardiovascular alterations associated with both agents were attributed to reductions in heart rate, although the possibility exists that negative inotropic effects may have had a role.

Left ventricular, systemic arterial, and baroreflex responses to ketamine and TEE in chronically instrumented monkeys.

Effects of prescribed doses of ketamine five minutes after application and influences of transesophageal echocardiography (TEE) on left ventricular, systemic arterial, and baroreflex responses were investigated to test the hypothesis that ketamine and/or TEE probe insertion alter cardiovascular function. Seven rhesus monkeys were tested under each of four randomly selected experimental conditions: (1) intravenous bolus dose of ketamine (0.5 ml), (2) continuous infusion of ketamine (500 mg/kg/min), (3) continuous infusion of ketamine (500 mg/kg/min) with TEE, and (4) control (no ketamine or TEE). Monkeys were chronically instrumented with a high fidelity, dual-sensor micromanometer to measure left ventricular and aortic pressure and a transit-time ultrasound probe to measure aortic flow. These measures were used to calculate left ventricular function. A 4-element Windkessel lumped-parameter model was used to estimate total peripheral resistance and systemic arterial compliance. Baroreflex response was calculated as the change in R-R interval divided by the change in mean aortic pressure measured during administration of graded concentrations of nitroprusside. The results indicated that five minutes after ketamine application heart rate and left ventricular diastolic compliance decreased while TEE increased aortic systolic and diastolic pressure. We conclude that ketamine may be administered as either a bolus or continuous infusion without affecting cardiovascular function 5 minutes after application while the insertion of a TEE probe will increase aortic pressure. The results for both ketamine and TEE illustrate the classic "Hawthorne Effect," where the observed values are partly a function of the measurement process. Measures of aortic pressure, heart rate, and left ventricular diastolic pressure should be viewed as relative, as opposed to absolute, when organisms are sedated with ketamine or instrumented with a TEE probe.

Evaluation of dual-tip micromanometers during 21-day implantation in goats.

Investigative research efforts using a cardiovascular model required the determination of central circulatory haemodynamic and arterial system parameters for the evaluation of cardiovascular performance. These calculations required continuous beat-to-beat measurement of pressure within the four chambers of the heart and great vessels. Sensitivity and offset drift, longevity, and sources of error for eight 3F dual-tipped micromanometers were determined during 21 days of implantation in goats. Subjects were instrumented with pairs of chronically implanted fluid-filled access catheters in the left and right ventricles, through which dual-tipped (test) micromanometers were chronically inserted and single-tip (standard) micromanometers were acutely inserted. Acutely inserted sensors were calibrated daily and measured pressures were compared in vivo to the chronically inserted sensors. Comparison of the pre- and post-gain calibration of the chronically inserted sensors showed a mean sensitivity drift of 1.0 +/- 0.4% (99% confidence, n = 9 sensors) and mean offset drift of 5.0 +/- 1.5 mmHg (99% confidence, n = 9 sensors). Potential sources of error for these drifts were identified, and included measurement system inaccuracies, temperature drift, hydrostatic column gradients, and dynamic pressure changes. Based upon these findings, we determined that these micromanometers may be chronically inserted in high-pressure chambers for up to 17 days with an acceptable error, but should be limited to acute (hours) insertions in low-pressure applications.

Endometriosis: clinical and pathologic findings in 70 rhesus monkeys.

Endometriosis was histologically confirmed in 70 female rhesus monkeys (Macaca mulatta). Severity of endometriosis was staged as minimal (14% of the monkeys), moderate (20%), or massive (66%). Clinical examination indicated masses in the abdomen and/or pelvic cavity palpable abdominally (64% of the monkeys) or rectally (93%), constipation (29%), anorexia (39%), irregular menses (17%), and radiographic evidence of abdominal soft tissue masses (38%). Pathologic findings were of intra-abdominal endometrial cyst formation (73% of the monkeys), and adhesions involving the ureters (51%), colon (66%), urinary bladder (50%), or ovaries (81%). Metastatic growth of endometrial tissue was found in 4 (6%) of the monkeys.

Surgical treatment of endometriosis in 50 rhesus monkeys.

Surgical treatment of endometriosis was attempted in 50 female rhesus monkeys (Macaca mulatta). Endometriosis was clinically staged as minimal (8% of the monkeys), moderate (18%), or massive (74%). Elective euthanasia was performed in 15 (30%) of the monkeys. Total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed in 30 (60%) of the monkeys, and bilateral salpingo-oophoretomy alone was performed in 5 (10%) monkeys. Statistical differences were not found in Kaplin-Meier survival curves between the different stages of endometriosis (P = 0.6297).

Central hemodynamics in a baboon model during microgravity induced by parabolic flight.

We developed a chronically instrumented nonhuman primate model (baboon) to evaluate the central cardiovascular responses to transient microgravity induced by parabolic flight. Instrumentation provided simultaneous recording of high fidelity (Ao) and pulmonary artery (PA) pressures, right and left ventricular and atrial pressures, Ao and PA blood flow velocities and vessel dimensions, ECG and pleural pressures. Four daily flights in 1991 and five in 1992 were flown with forty parabola per flight. Animals flown in 1991 were not controlled for volume status. Animals flown in 1992 were studied in one of three conditions: 1) volume depleted by furosemide (DH), 2) volume expanded by saline infusion (VE), and 3) euvolemic (EU, no intervention, used for echo only). Mean right atrial pressures (RAP) during 1991 flights had a variable early microgravity response: increases in n=3 and decrease in n=3 (supine) and increases in n=5, decreases in n=2 (upright). In 1992 flights, DH, upright and supine, changed -10 +/- 4.1 mmHg, -3.2 +/- 2.2 mmHg, respectively (p < .05) compared to the pull-up phase. In contrast, VE changed (from pull-up to microgravity) +13 +/- 1.5 mmHg and +4.25 +/- 2.9 mmHg (upright and supine, respectively, p < .05). EU increased with microgravity +6.9 +/- .9 mmHg (upright only). LAP responses were similar, but more variable. Finally, heart chamber areas paralleled pressure changes. Thus, right and left heart filling pressure changes with sudden entry into microgravity conditions were dependent on initial circulatory volume status and somewhat modified by position (supine vs upright).

Renal medullary solute depletion resulting from psychogenic polydipsia in a rhesus monkey.

A rhesus monkey developed psychogenic polydipsia after a change in feeding regimen was instituted. Hyposthenuria, hyponatremia, hypochloremia, and an abnormal calculated plasma osmolality were found. Water deprivation testing and administration of antidiuretic hormone indicated that renal medullary solute depletion had resulted from the prolonged polyuria. Restricted water consumption resulted in resolution of the electrolyte abnormalities and restored ability to excrete concentrated urine.

Effects of dipyridamole on the cardiovascular response to +Gz stress in miniature swine.

Eight conscious female miniature swine experienced acceleration levels of 3, 5, and 7 +Gz before and after infusion of dipyridamole (1-2 mg.kg-1). Each animal was instrumented to measure ECG, heart level arterial pressure (AP), eye level arterial pressure (ELBP), left arterial pressure (LAP), heart rate (HR), and regional tissue blood flows. Each was also fitted with an abdominal anti-G suit which automatically inflated. Dipyridamole infusion had no direct effect on HR or LAP but AP was significantly reduced. All cardiovascular responses to +Gz were qualitatively similar before and after dipyridamole. Tachycardia always occurred. AP and CNS blood flow were maintained better prior to dipyridamole and AP always fell in proportion to acceleration intensity. +Gz was generally associated with increased blood flow to respiratory muscles and heart, decreased blood flow throughout the viscera and to the eyes. ELBP paralleled AP, but was always lower in direct proportion to the +Gz level. We conclude that dipyridamole reduces arterial pressure thus compromising the ability of the animal to sustain cerebral perfusion pressure (ELBP) during +Gz.

Cerebral and spinal cord blood flow dynamics during high sustained +Gz.

This study had two purposes. First, the use of Transonic flowprobes placed on the common carotid and internal carotid arteries of seven male baboons was evaluated for measuring cerebral blood flow (BF) during +Gz stress. The approach was to compare BF's obtained with these flowprobes to microsphere measurements of total cerebral BF. The second purpose was to measure regional variations in cerebral and spinal cord BF during +Gz to test the hypothesis that +Gz produces a differential perfusion deficit throughout the central nervous system so that BF's at the superior portion of the brain are decreased more than in areas of the brain that are nearer to the heart. The results indicate that internal carotid artery and microsphere measurements of total brain BF were related so that the relative decrease in internal carotid artery BF was consistently comparable to that measured with the labeled microsphere technique. Thus, Transonic flowprobes placed on the internal carotid artery of the baboon give reliable estimates of cerebral BF during +Gz stress. The microsphere BF data demonstrated that there were no regional differences in the relative decrease in BF measured in the brain or spinal cord during +Gz. We conclude that our results do not support the hypothesis of a gradient of BF deficit within the brain or spinal cord during +Gz.

Performance recovery following +Gz-induced loss of consciousness.

Seven male baboons (average weight, 20.6 kg) were trained in a simple shock-avoidance performance task on the USAFSAM centrifuge. A red light was presented to the baboon at approximate 2-s intervals. The animal was allowed 1 s to turn off the light or receive a 1-s shock. The shock could be abbreviated by a late trigger pull. Thus, the animal could avoid, escape, or accept the full shock. EEG was monitored from three transcranial stainless steel electrodes. Loss of consciousness (LOC) was induced by a rapid onset (4 or 6/s) exposure to 8 +Gz and sustained until LOC was identified by a near isoelectric EEG signal. Performance recovery time was measured from the return of EEG activity to the time when the animal resumed the performance task. These data were compared with previously obtained human data and found to be very similar. Also, it was found that time of performance recovery became significantly shorter after multiple LOC exposures and the performance recovery time significantly increased with increased time of unconsciousness. This study demonstrates the utility of the baboon as an animal model for G-induced LOC (G-LOC) research.

Diazepam as a treatment for nerve agent poisoning in primates.

Diazepam is known to decrease organophosphate nerve agent-induced convulsions, neuropathology, and lethality in rhesus monkeys. We report that, when added to the previously fielded treatment regimen (pyridostigmine, atropine, and pralidoxime chloride), diazepam significantly protects the performance of rhesus monkeys trained in an equilibrium task, the Primate Equilibrium Platform (PEP), against deficits induced by the nerve-agent soman. If administered soon enough to minimize the occurrence of convulsions, diazepam should increase the probability of mission accomplishment and eventual complete recovery from nerve agent poisoning. Diazepam was fielded by the United States military services during Operations Desert Shield and Storm for the treatment of possible organophosphate toxicity.

Total and regional cerebral blood flow during recovery from G-LOC.

INTRODUCTION: This study measured total and regional cerebral blood flow (BF) in baboons during +Gz-induced loss of consciousness (G-LOC) and during recovery from G-LOC. METHODS: Flowprobes (Transonic Inc., T201, Ithaca, NY) were placed on the common carotid and internal carotid arteries of five male baboons for continuous measurement of total cephalic and cerebral BF, respectively. Radiolabeled microspheres were used to measure regional central nervous system BF at discrete timepoints. G-LOC was determined from visual observations of the animals and from EEG recordings. RESULTS: Cerebral blood flow was maintained and animals remained conscious during 60 s exposure to +4 Gz. In contrast, G-LOC was observed during the first 16-25 s (mean = 20.3 +/- 3.7 s) of exposure to +8 Gz in all five animals. Internal and common carotid artery BF decreased rapidly to zero during the first few seconds of +8 Gz. BF always appeared to cease prior to the occurrence of G-LOC. During early recovery from G-LOC there was no hyperemic response recorded with flowprobes, whereas a hyperemic response was recorded following 60 s exposures to +4 Gz in which the animals did not experience G-LOC. Microsphere measurements of the regional distribution of BF are consistent with the hypothesis of a +Gz-induced differential perfusion deficit throughout the brain and central nervous system during G-LOC. CONCLUSIONS: We conclude that G-LOC is preceded by cessation of cerebral BF. The fact that the hyperemic response following +Gz exposure is less when G-LOC occurs than when G-LOC does not occur suggests CNS energy conservation during G-LOC.

Effect of hyperbaric oxygenation on peripheral nerve regeneration in adult male rabbits.

Oxygen environments were used to study the regenerative effects of hyperbaric oxygen on crushed sciatic nerves in 30 adult male rabbits. Six different oxygen environments were used, and treatments were initiated 4 days post injury. Transmission electron microscopy and light microscopy were used to evaluate the regenerative morphology of crushed nerves. The morphology of crushed nerves after 7 wk of treatment with compressed oxygen at 202, 242, and 303 kPa resembled normal uncrushed nerves, with nerve fibers uniformly distributed throughout the section. The treatment groups receiving 202 kPa compressed air, 100% normobaric oxygen, or ambient air did not display morphologies similar to normal uncrushed nerve. The nerves in these animals were edematous and contained disarrayed nerve fibers. Myelination in the animals receiving 100% O2 at high pressures resembled undamaged nerves. Collagen and blood vessels were more evident in the lower pressure/oxygen tension treatments than in the animals receiving 100% O2 at higher pressures. The neurofilamentous material inside the crushed control axons was dense, whereas in the axons of animals treated with compressed O2 it was loosely packed. These differences in morphology suggest that treatments consisting of 100% O2 under pressure can accelerate a peripheral nerve's recovery from a crush injury.

Effects of 12 days exposure to simulated microgravity on central circulatory hemodynamics in the rhesus monkey.

Central circulatory hemodynamic responses were measured before and during the initial 9 days of a 12-day 10 degrees head-down tilt (HDT) in 4 flight-sized juvenile rhesus monkeys who were surgically instrumented with a variety of intrathoracic catheters and blood flow sensors to assess the effects of simulated microgravity on central circulatory hemodynamics. Each subject underwent measurements of aortic and left ventricular pressures, and aortic flow before and during HDT as well as during a passive head-up postural test before and after HDT. Heart rate, stroke volume, cardiac output, and left ventricular end-diastolic pressure were measured, and dP/dt and left ventricular elastance was calculated from hemodynamic measurements. The postural test consisted of 5 min of supine baseline control followed by 5 minutes of 90 degrees upright tilt (HUT). Heart rate, stroke volume, cardiac output, and left ventricular end-diastolic pressure showed no consistent alterations during HDT. Left ventricular elastance was reduced in all animals throughout HDT, indicating that cardiac compliance was increased. HDT did not consistently alter left ventricular +dP/dt, indicating no change in cardiac contractility. Heart rate during the post-HDT HUT postural test was elevated compared to pre-HDT while post-HDT cardiac output was decreased by 52% as a result of a 54% reduction in stroke volume throughout HUT. Results from this study using an instrumented rhesus monkey suggest that exposure to microgravity may increase ventricular compliance without alternating cardiac contractility. Our project supported the notion that an invasively-instrumented animal model should be viable for use in spaceflight cardiovascular experiments to assess potential changes in myocardial function and cardiac compliance.