Prophylactic low-dose vancomycin treatment in very-low-birth-weight infants. For the prophylaxis of septicemia with coagulase-negative staphylococci in a high-risk very-low-birth-weight population, we administered 5 mg/kg of vancomycin every 12 h. Distribution volume and half-life of vancomycin were determined. Serum peak and trough levels were obtained on day 3 of treatment. With this low-dose regimen, serum concentrations in the therapeutic range were achieved in 35 of the 45 patients. Distribution volume and half-life were 0.692 liters/kg and 7.4 h, respectively. The distribution volume was not related to the gestational age; the half-life in the group of patients with a gestational age < 30 weeks was considerably higher. The 10 small-for-gestational-age children had a significantly smaller distribution volume. The vancomycin trough levels correlated with the serum creatinine concentrations and, therefore, with the gestational age. Our study indicates that this low vancomycin dose is sufficient in very-low-birth-weight infants to achieve therapeutic serum levels, being suitable for both prophylaxis and sepsis therapy.


Methamphetamine detection from meconium and amniotic fluid in guinea pigs depends on gestational age and metabolism. Significant adverse perinatal effects of maternal methamphetamine use have been reported, but little is known about factors influencing methamphetamine screening test results during the perinatal period. We tested the hypothesis that gestational age would affect quantitative recovery of methamphetamine in meconium and amniotic fluid. Time-bred guinea pigs received an intraperitoneal (i.p.) injection of 1 mg/kg methamphetamine at either 44 days (0.65 of term, n = 5), 50 days (0.74, n = 8), 56 days (0.82, n = 9) or 63 days (0.93, n = 4) gestation. At 1 or 7 days after i.p. methamphetamine, meconium and amniotic fluid were collected for quantitative methamphetamine assay by gas chromatography-mass spectrometry. Recovery from amniotic fluid and meconium 1 day after injection was influenced by gestational age. Greater values in amniotic fluid and meconium and a higher percentage of positive samples were seen in older fetuses. Collectively at all gestational ages, combined testing of amniotic fluid and meconium yielded detectable methamphetamine or its metabolites in 87% of guinea pigs 1 day after injection. However, methamphetamine was not detectable 1 week after injection in any sample (n = 63) at either 0.74 or 0.82 of term except for one positive amniotic fluid sample. Finally, demethylation of methamphetamine to amphetamine was higher in older fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)


The valproic acid metabolite E-2-n-propyl-2-pentenoic acid does not induce spina bifida in the mouse. The antiepileptic drug valproic acid (VPA) has been implicated as a human teratogen causing spina bifida aperta. Recently we developed a mouse model inducing spina bifida aperta and occulta with VPA. In a search for novel antiepileptic agents the VPA metabolite E-2-n-propyl-2-pentenoic acid (2-en-VPA) had been developed. In the mouse, 2-en-VPA exhibits anticonvulsive potency similar to VPA but very low teratogenic potency (induction of exencephaly). We have now compared VPA and its metabolite 2-en-VPA in regard to induction of spina bifida in our mouse model. 2-en-VPA was administered 3 times during the period of gestation most sensitive for the induction of spina bifida aperta with VPA: on day 9 of gestation at 0, 6 and 12 h. The following doses were injected (in mmol 2-en-VPA-Na/kg): (a) 3 x 2.1, (b) 3 x 2.7 (the equimolar dose of VPA is the threshold dose for induction of spina bifida aperta) and (c) 3 x 3.0 (the equimolar VPA dose produced spina bifida aperta). 2-en-VPA did not induce spina bifida aperta in the mouse in any of these groups. We then investigated the induction of spina bifida occulta in the three dose groups. Spina bifida occulta is a less serious form of spina bifida and may provide a sensitive method to estimate the potency of a compound to induce more severe forms of spina bifida. This malformation was demonstrated in alcian-blue- and alizarin-red-stained fetal skeletons by measurements of the distance between the cartilaginous ends of each vertebral arch.(ABSTRACT TRUNCATED AT 250 WORDS)


[The functional significance of coronary collateral circulation during sudden coronary occlusion]. The functional significance of the coronary collateral circulation remains controversial. It has been suggested that collateral circulation possibly helps prevent myocardial ischemia. Seventeen target lesions in 15 patients were studied to determine the relationship between the extent of the coronary collateral circulation and the degree of ventricular dysfunction during percutaneous transluminal coronary angioplasty (PTCA). During the first balloon inflation, diastolic indices such as left ventricular end-diastolic pressure, max negative dP/dt and the time constant of early relaxation were measured immediately before and at 60 sec following balloon inflation. During the second inflation, the contralateral and ipsilateral collateral circulations were evaluated. The latter was graded as follows: 0 = none; I = filling of side branches only; II = partial filling of the epicardial segment; and III = complete filling of the epicardial segment. Following balloon inflation, a significant increase was noted in the time constant of early relaxation in patients with grade 0 collateral circulation (40 +/- 7 to 47 +/- 7 msec: p < 0.01) and grade II collateral circulation (52 +/- 12 to 56 +/- 13 msec: p < 0.05). The percent increase in the time constant of early relaxation of patients with grade 0 and I collateral circulations exceeded that of patients with grade II (p < 0.05) or grade III collateral circulation (p < 0.05). Left ventricular end-diastolic pressure was elevated in all groups during PTCA. There was no significant difference in the percent increase of left ventricular end-diastolic pressure (LVEDP) between the 4 groups. However, LVEDP before PTCA was higher in patients with grade III collateral circulation than in patients in the other groups. Max negative dP/dt did not change significantly in any group. In conclusion, collateral circulation helps prevent myocardial ischemia during acute coronary occlusion, which is most precisely shown by the time constant of early relaxation. The degree of this protective function of collateral circulation seems to vary.


[Clinical characteristics of pulmonary edema in patients with unstable angina]. To elucidate the clinical characteristics of pulmonary edema in unstable angina, 120 patients with unstable angina who admitted to the hospital within 6 hours after the onset of chest pain were studied. The criteria for the diagnosis of pulmonary edema included interstitial pulmonary edema and diffuse alveolar edema. Pulmonary edema was present in 24 patients. In these patients, the duration of chest pain was relatively longer, and the incidences of diabetes mellitus, emergency coronary revascularization and multiple-vessel coronary artery disease were higher than in those without pulmonary edema. In addition, in-hospital mortality rate in patients with pulmonary edema was higher than in those without it (21 vs 1%, p < 0.001), which is probably due to a large area of myocardial ischemia. For these patients, therefore, early diagnosis and appropriate therapy to save viable segments of the myocardium are mandatory.


Left atrial function in ischemic heart disease assessed by intravenous digital subtraction angiography. To investigate changes in left atrial morphology and dimensions during the cardiac cycle, the atrium was visualized by intravenous digital subtraction angiography (DSA). The study subjects consisted of 22 male patients whose average age was 54.5 +/- 8.6 years. They had ischemic heart disease without mitral valve disease and were in sinus rhythm. They were 11 patients with old myocardial infarction (OMI group) and 11 who had chest pain without evidence of infarction (AP group). DSA was performed in the continuous mode. Contrast material (35 ml) was injected at a rate of 18 ml/sec via a catheter in the superior vena cava and subtraction images were obtained at a speed of 30 frames/sec in the right anterior oblique projection. The left atrial and left ventricular margins were traced manually, their areas were calculated, and fractional changes in area were analyzed. The left ventricular ejection fraction (LVEF) was calculated by densitometry. Cardiac catheterization was performed in 16 patients and the left ventricular end-diastolic pressure (LVEDP) and mean pulmonary arterial wedge pressure (PAWP) were measured. The entire left atrium was clearly imaged using DSA. Phase analysis of the time-area curves in the right anterior oblique projection revealed that the left atrial area was maximal during left ventricular end-systole (%LA1 = 100%), it decreased during early left ventricular diastole (%LA2), and then increased slightly again during mid-diastole (%LA3). After left atrial contraction, the minimum area was obtained (%LA4). The left atrium showed a two-stage decrease in the area due to passive emptying and active contraction during left ventricular diastole. Passive emptying (%LA1-%LA2) was significantly less in the OMI group than in the AP group (6.3 +/- 3.6 vs 13.3 +/- 4.8%, p < 0.01, respectively). In all 22 subjects, passive emptying correlated with LVEF (r = 0.70, p < 0.001) and LVEDP (r = -0.58, p < 0.05). There was no difference in active contraction (%LA3-%LA4) between the 2 groups (26.0 +/- 5.7% in the OMI group, 28.2 +/- 8.4% in the AP group), and it did not correlate with LVEF or LVEDP. The ratio of passive emptying to active contraction [(%LA1-%LA2)/(%LA3-%LA4)] correlated with LVEF (r = 0.63, p < 0.01). These findings suggested that impaired left ventricular diastolic function and a relative increase in atrial contraction were present in patients with a lower LVEF. The %LA4 correlated with LVEDP and PAWP (r = 0.65, r = 0.63, p < 0.01, respectively). In conclusion, DSA proved to be a useful method for investigating left atrial morphology and function.


[Efficacy of intracoronary thrombolysis versus percutaneous transluminal coronary angioplasty for treating acute myocardial infarction]. The usefulness of percutaneous transluminal coronary angioplasty (PTCA) in patients with evolving myocardial infarction remains controversial. We retrospectively assessed the efficacy of PTCA on myocardial salvage in acute myocardial infarction in comparison with the efficacy of intracoronary thrombolysis (ICT). Sixty-two patients with initial anteroseptal myocardial infarction who had been treated within 6 hrs after the onset of chest pain were categorized into 4 groups: 1) spontaneous recanalization: n = 14, 2) successful PTCA: n = 25 (this group was further subdivided into 2 groups: direct PTCA group, primary PTCA without prior ICT: n = 19; and rescue PTCA group, PTCA after unsuccessful ICT: n = 6), 3) successful ICT group (n = 12), and 4) unsuccessful recanalization group (n = 11). Left ventricular function in the chronic phase was assessed by contrast ventriculography using the global ejection fraction (EF) and regional wall motion (RWM) was assessed by the centerline method. Patients with recanalization had a significantly higher EF than did those without (62 +/- 12 vs 50 +/- 13%, p < 0.01). The mean EFs for groups with successful reperfusion were as follows: 65 +/- 8% for the spontaneous recanalization group, 61 +/- 14% for PTCA group (64 +/- 13% for direct PTCA group, 51 +/- 13% for rescue PTCA group) and 60 +/- 12% for the ICT group. The EFs for the spontaneous recanalization group and the direct PTCA group were significantly greater than that for the rescue PTCA group. The time to reperfusion and the thrombolysis in myocardial infarction (TIMI) flow grade before reperfusion did not affect the preservation of global left ventricular function. RWM of the infarcted area in patients with recanalization were less hypokinetic than that in patients without (p < 0.01). The mean RWM (SD/chord) in the successfully reperfused groups were -2.3 +/- 1.2 for the spontaneous recanalization group, -2.6 +/- 1.2 for the PTCA group (-2.3 +/- 1.1 for the direct PTCA group, -3.3 +/- 1.0 for rescue PTCA group) and -3.0 +/- 0.5 for the ICT group. Hypokinesis of the infarcted area was more severe in the rescue PTCA group than in the spontaneous recanalization group and the direct PTCA group (multiple comparison test p < 0.01, respectively), and hypokinesis was more severe in the ICT group than in the direct PTCA group (Student's t-test, p < 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)


The effect of percutaneous transluminal coronary angioplasty on anaerobic threshold in patients with angina pectoris. The anaerobic threshold (AT) is regarded an objective parameter for evaluating exercise tolerance, but its relationship to the improvement of myocardial ischemia remains uncertain. To investigate this relationship, submaximal treadmill exercise tests were performed for 15 consecutive patients with angina pectoris who had undergone successful percutaneous transluminal coronary angioplasty (PTCA). Before and after PTCA, the AT was determined using cardiorespiratory monitoring, while the patients were receiving their usual vasodilator medications. 1) Before PTCA, the minute oxygen uptake (VO2) at the AT correlated well with the peak VO2 (r = 0.92, p < 0.002). The VO2 at the AT, however, showed less correlation (r = 0.71, p < 0.002) with the VO2 at ST segment depression, while the latter parameter correlated closely with the peak VO2 (r = 0.91, p < 0.002). 2) After PTCA, exercise time, peak VO2, and the double product at peak exercise increased significantly (from 640.1 +/- 212.2 to 772.9 +/- 230.0 sec, p < 0.001, from 19.1 +/- 5.2 to 22.4 +/- 4.9 ml/min/kg, p < 0.05, and from 19.7 +/- 5.0 x 10(3) to 23.7 +/- 4.5 x 10(3), p < 0.001, respectively). However, the VO2 at the AT did not increase significantly (from 15.8 +/- 4.1 to 16.6 +/- 3.5 ml/min/kg, p = NS). The heart rate, systolic blood pressure, and double product at the AT did not change significantly. In conclusion, in patients with angina pectoris, the AT is apparently related to the onset of myocardial ischemia. However, the AT does not necessarily reflect acute improvement of myocardial ischemia immediately after PTCA.


[Assessment of coronary artery bypass grafts by X-ray computed tomography with and without contrast enhancement]. To assess the patency of coronary artery bypass grafts, we tested the capability of X-ray computed tomography (CT) with and without contrast enhancement. This procedure was used on 63 grafts (30 in the LAD; 20 in the LCX; 13 in the RCA) in 32 patients with a mean age of 56 +/- 8 years, all of whom were referred to our department for postoperative management or evaluation. The CT scanner used was Toshiba TCT-60A with a scan time of 3 sec, 5 mm thick slices, and 512 x 512 pixels. CT scans without contrast enhancement were obtained from the level of the aortic arch to the left ventricle. Eight sec after 30 ml of contrast media was injected at a rate of 3 ml/sec into an antecubital vein, 5 scans were made at the same level of the pulmonary artery truncus. After positioning the regions of interests on the ascending aorta and grafts, we obtained time-density curves (TDCs) and compared the data with those recorded from an intraoperative electromagnetic flow meter (EMF). Fifty grafts were angiographically patent. The appearance time, build-up time, peak time, disappearance time and peak densities obtained from TDCs of grafts did not correlate with the flow volumes measured by EMF. Patent grafts were easily identified visually, without contrast enhancement (sensitivity; 88%, specificity; 100%, accuracy; 91%). Occluded grafts were not imaged on CT, either with or without contrast enhancement. We concluded that the TDCs of grafts obtained by CT are of no value for predicting the graft flow, and that plain CT without contrast enhancement is sufficiently useful for assessing the patency of such grafts.


[Distribution of myocardial damage in patients with hypertrophic cardiomyopathy: evaluation by exercise thallium-201 scintigraphy]. The characteristic of myocardial damage in hypertrophic cardiomyopathy (HCM) was evaluated as to whether the damage is limited to the hypertrophied wall or extends throughout the entire wall. The myocardial damage was detected by exercise thallium-201 (Tl-201) scintigraphy and was evaluated using circumferential profile analysis, calculation of initial uptake and washout rate. Eleven patients with asymmetrical hypertrophy (ASH), whose septal and posterior wall thickness ratio exceeded 1.3 on left ventriculography and biventriculography, and 13 age-matched control subjects without heart disease were studied. The mean values of initial uptake in both groups did not differ significantly, but the washout rate for the entire heart was significantly decreased only in the HCM group (p < 0.05). All of the regional washout rates (antero-septal, apical and postero-lateral) were significantly decreased in the HCM group (p < 0.05), without any difference between the hypertrophied wall and the non-thickened free wall being noted. These results demonstrated that the analysis of myocardial damage by exercise Tl-201 scintigraphy using calculation of the washout rate is a very sensitive means of detecting myocardial damage in HCM, and that such myocardial damage is not restricted to the hypertrophied wall, but rather extends to the entire wall, including the free wall which is not thickened.


[Left ventricular blood filling in patients with severe mitral stenosis: comparisons before and soon after percutaneous transluminal mitral commissurotomy]. To clarify the effects of mitral obstruction on left ventricular (LV) diastolic blood filling, 15 patients with tight mitral stenosis (each mitral valve area was less than 1.5 cm2) were studied. Each selected patient underwent successful percutaneous transluminal mitral commissurotomy (PTMC), which resulted in a 1.5 fold increase in each mitral valve area. LV pressure, left atrial (LA) pressure and cardiac output were measured before and immediately after PTMC. Left ventriculography was performed before and immediately after PTMC. The ventriculogram was traced frame by frame for one cardiac cycle. The LV volume curve was obtained from the traced image using a computer. The LV end-diastolic and end-systolic volumes (EDVI, ESVI), and ejection fraction in the subsequent cardiac cycle were calculated. The diastolic filling period was divided into 3 equal parts: namely, early, mid-, and late diastole. The blood volume entering the LV during early, mid-, and late diastole, which indicated the filling properties of each part, were calculated. After successful PTMC, both the mitral valve area (1.1 +/- 0.3 cm2 to 1.9 +/- 0.6 cm2, p < 0.01) and the cardiac index (3.2 +/- 0.8 l/min/m2 to 3.6 +/- 1.1 l/min/m2, p < 0.05) increased with the decreases in the mean diastolic pressure gradients between the LA and LV (13.4 +/- 4.5 mmHg to 5.9 +/- 2.6 mmHg, p < 0.01). The blood volume entering the LV during early diastole increased significantly without significant change in the blood volume entering the LV during mid- and late diastole.(ABSTRACT TRUNCATED AT 250 WORDS)


[Clinical and hemodynamic sequelae of mitral prostheses evaluated by Doppler echocardiography]. To determine the relative superiority of a prosthesis in the mitral position, in vivo hemodynamics were examined by Doppler echocardiography, and the results were compared with other types of mechanical mitral valves including 63 Björk-Shiley convexo-concave (BS) values, 30 Duromedics (DM) valves, and 58 Medtronic Hall (MH) valves. For this comparison, the following indices were evaluated: peak velocity of mitral flow (PV), mitral valve orifice area (MVA), mitral valvular regurgitation, New York Heart Association (NYHA) classification, pulmonary capillary wedge pressure (PC), cardiac index (CI) and valve-related complications. On Doppler echocardiograms, PV ranged from 1.2 to 2.0 m/sec with a mean of 1.6 m/sec. There was no evident relationship between the PV and the valve size in each type of prosthesis, and no significant difference in the PVs among the valves. The mean MVA was 2.6 cm2 (25 mm DM, 25 mm MH), which was regarded satisfactory from a clinical standpoint. MVA increased with the increase in the valve size in all types of valves, and of all sizes, MVA was larger in the DM and MH groups than in the BS group. Similarly, the incidence of valvular regurgitation was relatively low in all groups, and the degree of regurgitation proved to be grade II or less in all cases. As for the clinical results, clinical symptoms (NYHA) and hemodynamic states (PC, CI) improved postoperatively, with the differences among the types of prosthetic valves being insignificant. The incidences of thromboembolism, valvular thrombosis, valve failure and prosthetic endocarditis were relatively low in all groups.(ABSTRACT TRUNCATED AT 250 WORDS)


[Surgical decisions for active infective endocarditis in patients with acute neurological complications]. The surgical management of 7 patients with active infective endocarditis and recent (within 16 days) neurological injury was presented. All patients had preoperative computed tomographic scans which revealed no evidence of intracranial hemorrhage and underwent successful corrective cardiac surgery. In the early postoperative period, 4 patients died of cerebral hemorrhage, subarachnoid hemorrhage, or progression of cerebral edema. Two of the 3 surviving patients showed no aggravation of cerebral infarcts postoperatively. In the remaining surviving patient, intracerebral mycotic aneurysms were resolved spontaneously after postoperative antibiotic therapy, although new cerebral hemorrhage, a complication of emboli, occurred after open heart surgery. The results of this study indicated that 1) cerebrovascular complications were the causes of the 4 deaths in this series, and 2) although heparinization during open heart surgery may result in intracerebral hemorrhage from mycotic aneurysm or infarction, early surgical intervention after recent cardiogenic embolic strokes may save patients with minor cerebral infarcts.


[Magnetic resonance imaging: evaluation of the Blalock-Taussig shunts and anatomy of the pulmonary artery]. The morphology and circulation of the pulmonary arteries and shunt vessels were evaluated by magnetic resonance imaging (MRI) in 8 patients with cyanotic heart disease after a Blalock-Taussig shunt operation. Their ages ranged from one month to 17 years. MRI permitted assessment of the size and patency of the Blalock-Taussig shunts, as well as the size and morphology of the pulmonary arteries in all patients. Measurements of the vessel diameters on MRI correlated well with the angiographic measurements (main pulmonary artery, r = 0.98; right pulmonary artery, r = 0.98; left pulmonary artery, r = 0.98; and Blalock-Taussig shunt, r = 0.97). MRI successfully imaged 3 of 4 shunt obstructions and 3 of 4 pulmonary stenoses with high resolution. In assessing peripheral pulmonary stenosis or obstruction, MRI was superior to echocardiography, the latter being unable to image peripheral pulmonary arteries satisfactorily. We concluded that MRI is an excellent noninvasive method for serially evaluating the anatomy and function of Blalock-Taussig shunts and pulmonary arteries, which is particularly useful for children with cyanotic congenital heart disease.


[Left ventricular systolic time intervals during paroxysmal supraventricular tachycardia: the difference between A-V nodal re-entry and A-V re-entry]. In this study, the differences in hemodynamic changes during paroxysmal supraventricular tachycardia (PSVT) between A-V nodal re-entry and A-V re-entry were evaluated. In 8 patients with A-V nodal re-entrant tachycardia and 10 with A-V re-entrant tachycardia, electrophysiological studies were performed to measure systolic time intervals (pre-ejection period: PEP, ejection time: ET, PEP/ET ratio: PEP/ET). These measurements were obtained in the control state (atrial pacing at 90/min) and during PSVT with simultaneous recordings of electrocardiogram and femoral arterial pulse tracing. During PSVT, there was no difference in the heart rate between the 2 groups, but ventriculo-atrial conduction time was shorter in A-V nodal re-entry than in A-V re-entry. There was a marked fall in the ET and an increase in PEP/ET in all the patients when PSVT was induced. PEP increased significantly in A-V nodal re-entry, but did not change in A-V re-entry. This resulted in a greater increase in the PEP/ET suggesting a greater deterioration of the hemodynamic consequences in A-V nodal re-entry than in A-V re-ent y. Thus, the hemodynamic changes of PSVT differ between these 2 types of re-entrant circuits, which are mainly influenced by the ventriculo-atrial conduction time.


[Development of computer software in ramp slope controller for treadmill ergometer]. We developed computer software to produce a treadmill ramp protocol through which oxygen uptake (VO2) is increased in a linear fashion, thus enabling subjects to walk until the end of exercise. The developed software simulates the increases in speed and grade of the treadmill displayed on a personal computer screen, and produces the ramp protocol by arbitrarily determining the variables of the following formula: 1) Increments of VO2 = a1t+a2 (ml/min/kg) [t = exercise time (min)] 2) Predicted VO2 by speed (S) and grade (G) = a3S2 + a4G2 + a5SG + a6S + a7G + a8 (ml/min/kg) [S = speed (km/hr); G = grade (%)] 3) Speed suitable for desired exercise time (S or S2) = a9t2 + a10t + a11 (km/hr) 4) Grade suitable for desired exercise time (G or G2) = a12t2 + a13t + a14 (%) In this study, the increment of VO2 was determined by considering the subject's exercise capacity (VO2 = 4t + 7 ml/min/kg), using the Ito's formula (VO2 = 0.067S2 + 0.289SG + 7.73 ml/min/kg). The formula of grade was determined following the formula arbitrarily (G2 = 25t + 5%). The formula of speed for exercise time was calculated automatically. The new ramp protocol, which was applied to 10 healthy subjects (mean age: 24.8 +/- 4.8 years old), disclosed a similar linear relationship between the predicted VO2 and the measured VO2.


[Mechanisms of vasovagal syncope elucidated by upright-tilt with isoproterenol infusion]. To elucidate the role of increased basal vagal activity in vasovagal syncope, we compared patients with bradyarrhythmia due to increased vagal tone and patients with vasovagal syncope using an upright-tilt (60 degrees) positioning test with isoproterenol infusion. Eight patients with unexplained recurrent syncope after clinical and electrophysiological investigations and 5 patients without syncope who had bradyarrhythmias due to increased vagal tone were studied. All 8 patients with recurrent syncope had some prodrome suggestive of vasovagal syncope. The upright-tilting test was considered positive if syncope developed in association with hypotension or bradycardia, or both. If 10 min of control tilting was negative, the patient was lowered to the supine position. Upright-tilting was then repeated during continuous intravenous isoproterenol infusion at successive incremental doses of 0.01 to 0.03 microgram/kg/min. During the control upright-tilting test, none of the patients had positive responses. During the upright-tilting with isoproterenol infusions, all patients with vasovagal syncope had positive responses; whereas, all patients with bradyarrhythmia due to increased vagal tone had negative responses. In patients with vasovagal syncope, the heart rate (HR) and the mean blood pressure (mBP) were higher at the time of supine positioning than at the time of syncope (HR: 109 +/- 16-->88 +/- 16 bpm, p < 0.05) (mBP: 86 +/- 5-->53 +/- 6 mmHg, p < 0.01). However, in patients with bradyarrhythmia there was no significant change in HR and mBP between the supine and 10 min of the upright-tilting with isoproterenol infusion.(ABSTRACT TRUNCATED AT 250 WORDS)


[Hemodynamic effects of alcohol ingestion on anaerobic threshold]. Hemodynamic influence of alcohol ingestion was evaluated using expiratory gas analysis in 10 healthy men whose mean age was 26.1 +/- 2.2 years. Protocol of the control study (C) was as follows: Oxygen uptake (VO2) at rest and during warm-up, anaerobic threshold (AT) and peak VO2 were measured with ramp protocol using a bicycle ergometer. AT was determined by the V-slope method. Protocol of the alcohol study (A) was as follows: On a different day, the same parameters as C were measured after the ingestion of 0.7 g ethanol per kg body weight. The following results were obtained: 1) The serum ethanol concentration was 79.1 +/- 13.5 mg/dl. 2) VO2 in the resting state was higher in A than in C (4.51 +/- 0.74 vs 3.61 +/- 0.62 ml/min/kg), (p < 0.05). 3) AT and peak VO2 in C and A were 17.15 +/- 2.76, 32.09 +/- 6.77 ml/min/kg and 15.43 +/- 2.86, 29.60 +/- 5.35 ml/min/kg, respectively. 4) Exercise time to the AT in C and A was 222.90 +/- 37.32 and 172.0 +/- 33.1 sec, respectively. 5) The heart rate was higher in A than in C. It was concluded that peak VO2 and AT decreased by alcohol ingestion.


[High incidence of left ventricular thrombosis and systemic embolism in patients with left ventricular assist system]. The purpose of this study was to determine the incidence of left ventricular (LV) thrombosis and systemic embolism in 14 patients with LV assist systems. Echocardiography was used to detect LV wall motion abnormalities, intracavitary smoke-like echoes and thrombosis, and the effect of anticoagulant therapy was serially examined. During full assist of the circulation, the aortic valve did not open in any patient. Smoke-like echoes were observed in 9 patients (64%) and thrombi in 8 (57%). The thrombus developed within the first 3 assist days. Systemic anticoagulant therapy decreased the thrombus size in only 3 patients, but there was a possibility of intracranial or mediastinal bleeding in other 3 patients. Systemic embolism was noted in 7 of 11 autopsy patients (64%). The characteristic finding was that there were multiple embolized organs, such as the brain, kidneys, spleen and liver, in all patients. Development of a thrombus is a serious complication in all patients with LV assist systems. However, the problem does not lie in the assist system but in the left ventricle of the patient's own heart. It is also noteworthy that systemic anticoagulation is not effective for an LV thrombus. A new method of assisting the failing heart, or a new anticoagulant delivery technique for the LV cavity to prevent LV thrombus development is needed.


[Acute hemodynamic effects of pimobendan and captopril: a comparative study in the same patients with chronic heart failure]. In this study, the acute hemodynamic effects of pimobendan (2.5 mg), a new drug, was compared with that of captopril (12.5 mg) in the same 8 patients with chronic heart failure (NYHA class II-III); 3 with dilated cardiomyopathy and 5 with regurgitant valvular heart disease. The hemodynamics were serially assessed before and after drug administration for at most 6 hours. Pimobendan reduced mean blood pressure, mean pulmonary artery pressure, pulmonary capillary wedge pressure, right atrial pressure, total systemic vascular resistance, and total pulmonary vascular resistance but it increased heart rate. By contrast, captopril reduced mean blood pressure and double product. No significant changes were noted in the cardiac index, stroke volume index, AV-O2 difference or the arterial oxygen pressure between the 2 drugs. In conclusion, pimobendan seems to function as a strong arterio-veno-dilator rather than as an inotropic agent in patients with chronic heart failure.


[Cardiac sarcoidosis with multinucleated giant cells detected in endomyocardial biopsy specimens: report of 3 cases]. This is a report of 3 cases of cardiac sarcoidosis. None of the 3 patients had other overt systemic involvement. Multinucleated giant cells and non-caseating epithelial cell granulomas with small lymphocytic infiltrations were detected in the endomyocardial biopsy specimen. Some focal left ventricular wall motion abnormalities were detected by echocardiography and angiography in all 3 patients. Paroxysmal ventricular tachycardia and multifocal ventricular premature complexes were observed by Holter 24-hour electrocardiography in all 3 patients. Two of the 3 patients did not receive corticosteroids, but remaining patient is on a regimen of corticosteroid.


Risk factors for Trypanosoma cruzi infection among blood donors in central Brazil. Characteristics and possible risk factors associated with Trypanosoma cruzi infection among blood donors were assessed within a routine screening programme in blood banks in an endemic area of Chagas disease. 6,172 voluntary blood donors were interviewed and tested for anti-T. cruzi antibodies by Haemagglutination and Complement Fixation tests in six blood banks in Goiânia-Central Brazil from October 1988 to April 1989. An overall prevalence of 2.3% for T. cruzi infection was obtained, being 3.3% for first-time blood donors and 1.9% for regular ones (p < 0.01). Considering this seropositivity among regular blood donors, selection of candidates relying only on the history of previous donation was found to be inadequate. The risk of infection increased inversely with the degrees of education and monthly income. There was a 9.2 risk of infection (95% CI 3.8-22.6) for those who had lived more than 21 years in an endemic area compared to subjects who had never lived in rural settings, after multivariate analysis. These informations may help to review the criteria of selection of donors in order to improve quality of blood products in endemic areas.


Evaluation of some organic compounds on bloodstream forms of Trypanosoma cruzi. Accidental transmission of Chagas' disease to man by blood transfusion is a serious problem in Latin-America. This paper describes the testing of several synthetic, semi-synthetic, and natural compounds for their activity against blood trypomastigotes in vitro at 4 degrees C. The compounds embody several types of chemical structures: benzoquinone, naphthoquinone, anthracenequinone, phenanthrenequinone, imidazole, piperazine, quinoline, xanthene, and simple benzenic and naphthalenic derivatives. Some of them are for the first time tested against Trypanosoma cruzi. The toxic effect of these compounds on this parasite was done by two quite distinct sets of experiments. In one set, the compounds were added to infected blood as ethanolic solution. In this situation the most active one was a furan-1,2-naphthoquinone, in the same range as gentian violet, a new fact to be considered in the assessment of structure-activity relationships in this class of compounds. In other set, we tentatively evaluated the biological activity of water insoluble compounds by adding them in a pure form without solvent into infected blood. In this way some appear to be very active and it was postulated that the effectiveness of such compounds must result from interactions between them and specific blood components.


Cytogenetics as a tool for triatomine species distinction (Hemiptera-Reduviidae). Several cytogenetic traits were tested as species diagnostic characters on five triatomine species: Rhodnius pictipes, R. nasutus, R. robustus, Triatoma matogrossensis and T. pseudomaculata. Four of them are described for the first time. The detailed analysis of the meiotic process and the application of C-banding allowed us to identify seven cytogenetic characters which result useful to characterize and differentiate triatomine species.


Relative susceptibility of different stages of Rhodnius prolixus to the entomopathogenic hyphomycete Beauveria bassiana. Laboratory bioassays were conducted to determine the relative susceptibility of eggs, 1st-, 3rd-, 5th-instar nymphs and adults of Rhodnius prolixus to one isolate of the entomopathogenic hyphomycete, Beauveria bassiana. Treatments consisted of directly spraying on insects of increasing doses of inoculum (3 x 10(2) to 3 x 10(5) conidia per cm2). Mortality due to all doses of conidia was very high in the five tested stages of the target insect. Experiments on eggs demonstrated that the fungal isolate was able to kill eggs before they hatched. Both time-mortality and dose-mortality responses showed that the susceptibility of R. prolixus varied according to its stage of development and increased with age. As a matter of fact, at the dose of 3 x 10(3) conidia per cm2, LD 50 varied between 11.2 days in 1st-instar nymphs and 6.4 days in both 5th-instar nymphs and adults. Comparison of LD50 permitted to estimate that 1st-instar nymphs were about 700-fold less susceptible than the two oldest stages.


Influence of mating on ovarian follicle development in Triatoma infestans (Klug, 1834). This work examines the influence of mating on ovarian follicle development in Triatoma infestans. The observations were carried out on both virgin and mated females, which were killed at various times after their emergence. There was no difference in the ovarian development of both experimental groups during the first gonadotrophic cycle. By the 7th day mated females as well as virgin females showed vitellogenic oocytes. The coriogenesis and ovulation process began on the 13th day after imaginal moulting. However we could observe that egg-laying was dependent on mating. Mated females laid eggs whereas virgin females did not lay eggs. However ovarian production was significantly greater in the mated females. It is suggested that in T. infestans mating stimulates egg-laying but it does no influence the oogenesis and ovulation process.


Ultrastructure of the ovary of Dermatobia hominis (Diptera: Cuterebridae). III. Gonial cell degeneration. We studied the ultrastructural aspects of pre-pupae and pupae ovaries of Dermatobia hominis. Physiological degeneration of gonial cells was observed: (a) after the ovarioles differentiation, in the oogonia residing in the apical region of the ovary; (b) at the beginning of vitellogenesis, in the cystoblasts close to the terminal filament. The significance of gonial cell degeneration was correlated with the physiological changes which occur in the ovary during development.


Ecology of phlebotomine sand flies (Diptera: Psychodidae) in a focus of Leishmania (Viannia) braziliensis in northeastern Colombia. The phlebotomine sand fly fauna of two coffee plantations in a Leishmania-endemic area of Norte de Santander, Colombia was studied. Regular insect collections using a variety of methods were made for three and a half years. Information was obtained on diurnal resting sites, host range and seasonal abundance for 17 species, of which five (Lutzomyia spinicrassa, Lu. serrana, Lu. shannoni, Lu. ovallesi and Lu. gomezi) were far more numerous than the others, anthropophilic and present throughout the year. The behaviour of these and the remaining 12 species is discussed in relation to their potential role in transmission of Leishmania (Viannia) braziliensis in the area.


Dispersal of phlebotomine sand flies (Diptera: Psychodidae) in a Colombian focus of Leishmania (Viannia) braziliensis. Dispersal of five species of phlebotomine sand flies was studied in a coffee plantation near Arboledas, Colombia, by mark-release-recapture studies using fluorescent powders. The estimated recapture rate for males of Lutzomyia shannoni marked and released during the day was 28.1%, significantly higher than that for all other species (p < 0.05). Recapture rate of female Lu. shannoni was 9.5%, and no females of the other four species were recovered. This suggests either that Lu. shannoni is a more sedentary species than the others, or that the large trees on which these insects were captured and recaptured function as foci of lekking behaviour in males. The high recapture rates of females of this species may indicate that oviposition occurs in close proximity to the bases of these trees. Although most marked sand flies were recaptured within 200 m of their release point, a single female Lu. gomezi was recovered 960 m away 36 h after release. This suggests that the dispersal capacity of Lutzomyia species may be greater than has been thought, an important consideration in future control programs directed against these insects in Leishmania-endemic areas.


Onchocerciasis in Ecuador: the situation in 1989. Details are given of the prevalence rates of onchocerciasis from the most recent surveys (1989) conducted in northern Ecuador. The disease has intensified and dispersed considerably due to migration of infected individuals and the presence of a highly efficient vector. Comparison of these data with those from two previous surveys carried out in 1982/83 and 1986 and correlated with entomological findings highlight the danger of the formation of new foci of onchocerciasis in areas currently free of the disease. Recommendations are made for further entomological studies in areas either recently or likely to be affected by the disease where potential vectors are unknown or different to those registered in the Santiago focus. Ivermectin treatment with local vector control in specific areas is advocated to reduce the disease to a low level of public health importance.


The feasibility of forecasting influenza epidemics in Cuba. A large influenza epidemic took place in Havana during the winter of 1988. The epidemiologic surveillance unit of the Pedro Kourí Institute of Tropical Medicine detected the beginning of the epidemic wave. The Rvachev-Baroyan mathematical model of the geographic spread of an epidemic was used to forecast this epidemic under routine conditions of the public health system. The expected number of individuals who would attend outpatient services, because of influenza-like illness, was calculated and communicated to the health authorities within enough time to permit the introduction of available control measures. The approximate date of the epidemic peak, the daily expected number of individuals attending medical services, and the approximate time of the end of the epidemic wave were estimated. The prediction error was 12%. The model was sufficiently accurate to warrant its use as a practical forecasting tool in the Cuban public health system.


Forecast of acute respiratory infections: expected nonepidemic morbidity in Cuba. A forecast of nonepidemic morbidity due to acute respiratory infections were carry out by using time series analysis. The data consisted of the weekly reports of medical patient consultation from ambulatory facilities from the whole country. A version of regression model was fitted to the data. Using this approach, we were able to detect the starting data of the epidemic under routine surveillance conditions for various age groups. It will be necessary to improve the data reporting system in order to introduce these procedures at the local health center level, as well as on the provincial level.


Molecular karyotype analysis and mapping of housekeeping genes to chromosomes of selected species complexes of Leishmania. The molecular karyotypes for 20 reference strains of species complexes of Leishmania were determined by contour-clamped homogeneous electric field (CHEF) electrophoresis. Determination of number/position of chromosome-sized bands and chromosomal DNA locations of housekeeping genes were the two criteria used for differentiating and classifying the Leishmania species. We have established two gel running conditions for optimal separation of chromosomes, which resolved DNA molecules as large as 2,500 kilobase pairs (kb). Chromosomes were polymorphic in number (22-30) and size (200-2,500 kb) of bands among members of five complexes of Leishmania. Although each stock had a distinct karyotype, in general the differences found between strains and/or species within each complex were not clear enough for parasite identification. However, each group showed a specific number of size-concordant DNA molecules, which allowed distinction among the Leishmania complex parasites. Clear differences between the Old and New world groups of parasites or among some New World Leishmania species were also apparent in relation to the chromosome locations of beta-tubulin genes. Based on these results as well as data from other published studies the potential of using DNA karyotype for identifying and classifying leishmanial field isolates is discussed.


Disseminated American muco-cutaneous leishmaniasis caused by Leishmania braziliensis braziliensis in a patient with AIDS: a case report. The authors report a case of culture-proven disseminated American muco-cutaneous leishmaniasis caused by Leishmania braziliensis braziliensis in an HIV positive patient. Lesions began in the oropharynx and nasal mucosa eventually spreading to much of the skin surface. The response to a short course of glucantime therapy was good.


Macrophage activation and histopathological findings in Calomys callosus and Swiss mice infected with several strains of Trypanosoma cruzi. Peritoneal macrophage activation as measured by H2O2 release and histopathology was compared between Swiss mice and Calomys callosus, a wild rodent, reservoir of Trypanosoma cruzi, during the course of infection with four strains of this parasite. In mice F and Y strain infections result in high parasitemia and mortality while with silvatic strains Costalimai and M226 parasitemia is sub-patent, with very low mortality. H2O2 release peaked at 33.6 and 59 nM/2 x 10(6) cells for strains Y and F, respectively, 48 and 50 nM/2 x 10(6) for strains Costalimai and M226, at different days after infection. Histopathological findings of myositis, myocarditis, necrotizing arteritis and absence of macrophage parasitism were found for strains F and Costalimai. Y strain infection presented moderate myocarditis and myositis, with parasites multiplying within macrophages. In C. callosus all four strains resulted in patent parasitemia which was eventually overcome, with scarce mortality. H2O2 release for strains Y and F was comparable to that of mice-peaks of 27 and 53 nM/2 x 10(6) cells, with lower values for strains Costalimai and M226-16.5 and 4.6 nM/2 x 10(6) cells, respectively. Histopathological lesions with Y and F strain injected animals were comparable to those of mice at the onset of infections; they subsided completely at the later stages with Y strain and partially with F strain infected C. callosus. In Costalimai infected C. callosus practically no histopathological alterations were observed.


Quantification of Trypanosoma cruzi in the heart, lymph nodes and liver of experimentally infected mice, using limiting dilution analysis. Limiting dilution analysis was used to quantify Trypanosoma cruzi in the lymph nodes, liver and heart of Swiss and C57Bl/10 mice. The results showed that, in Swiss and Bl/10 mice infected with T. cruzi Y strain, the number of parasites/mg of tissue increased during the course of the infection in both types of mice, although a greater number of parasites were observed in heart tissue from Swiss mice than from Bl/10. With regard to liver tissue, it was observed that the parasite load in the initial phase of infection was higher than in heart. In experiments using T. cruzi Colombian strain, the parasite load in the heart of Swiss and Bl/10 mice increased relatively slowly, although high levels of parasitization were nonetheless observable by the end of the infection. As for the liver and lymph nodes, the concentration of parasites was lower over the entire course of infection than in heart. Both strains thus maintained their characteristic tissue tropisms. The limiting dilution assay (LDA) proved to be an appropriate method for more precise quantification of T. cruzi, comparing favorably with other direct microscopic methods that only give approximate scores.


Trypanosomatidae codon usage and GC distribution. A study of Trypanosomatidae GC distribution and codon usage is presented. The codon usage patterns in coincidence with the phylogenetical data are similar in Crithidia and Leishmania, whereas they are more divergent in Trypanosoma brucei and T. cruzi. The analysis of the GC mutational pressure in these organisms reveals that T. brucei, and to a lesser extent T. cruzi, have evolved towards a more balanced use of all bases, whereas Leishmania and Crithidia retain features of a primeval genetic apparatus. Tables with the approximated GC mutational pressure in homologous genes, and codon usage in Trypanosomatidae are presented.


Experimental heteroxenous cycle of Lagochilascaris minor Leiper, 1909 (Nematoda: Ascarididae) in white mice and in cats. Reports of natural infections of sylvatic carnivores by adult worms of species similar to Lagochilascaris minor in the Neotropical region led to attempts to establish experimental cycles in laboratory mice and in cats. Also, larval development was seen in the skeletal muscle of an agouti (Dasyprocta leporina) infected per os with incubated eggs of the parasite obtained from a human case. In cats, adult worms develop and fertile eggs are expelled in the feces; in mice, larval stages of the parasite develop, and are encapsulate in the skeletal muscle, and in the adipose and subcutaneous connective tissue. From our observations, we conclude that the larva infective for the mouse is the early 3rd stage, while for the final host the infective form is the later 3rd stage. A single moult was seen in the mouse, giving rise to a small population of 4th stage larvae, long after the initial infection.


Trypanosoma rangeli (Tejera, 1920): observations upon pleomorphism. Meta-trypomastigotes of Trypanosoma rangeli Tejera, 1920, harvested from LIT medium, were inoculated i.p. or s.c. into 6, 16, and 26 g NMRI mice, these representing increasing degrees of immunological maturity. In all cases, similar pleomorphic patterns were observed. Four morphobiometrically differentiable types of trypanosome were encountered in an overlapping temporal sequence. These observations, taken in comparison with those on pleomorphism in this and other species of Trypanosoma by other workers, are consistent with the hypothesis that the pleomorphic types represent the natural development of the parasite, rather than the result of the immune response of the mammal host. Small, slender trypanosomes prevalent at the onset of the parasitemia either reinvade the tissue cells for relatively limited subsequent generations of tissue reproduction, or else differentiate toward the forms that are only capable of colonizing the insect vector.


Pterigodermatites (Paucipectines) spinicaudatis n. sp. (Nematoda: Rictularidae) from Dromiciops australis (Marsupialia: Microbiotheriidae) in Bariloche, Rio Negro, Argentina. Biogeographical distribution and host-parasite relationships. Pterigodermatites (P.) spinicaudatis sp. n. from Dromiciops australis is proposed and described. The simple morphology of the ovijector and the presence of a well developed spine between the two cuticular projections at the caudal extremity of the female distinguish the studied nematode from the remainder species of the genus parasitizing South American Edentata, marsupials and cricetid rodents. The distribution area of the hosts of the different species of P. (P.) are given. The studied genus does not parasitize any Australian marsupials. It was found in the endemic South American Microbiotheriidae. This fact suggests from a parasitological point of view that D. australis is not related to the Australian marsupials but to the South American ones.


Life history studies of heterophyid trematodes in the Neotropical region: Ascocotyle (Leighia) hadra sp. n. The life cycle of Ascocotyle (Leighia) hadra n. sp. was experimentally reproduced, starting from cercariae from naturally infected Littoridina parchappei, collected from Los Ranchos stream, near Mercedes city, Buenos Aires Province, Argentina. Metacercariae were found encysted in the liver and mesentery of experimentally and naturally infected fishes Cnesterodon decemmaculatus and Jenynsia lineata. Adults were obtained experimentally in chicks and mice. The natural host is unknown. The new species is compared with Ascocotyle (Leighia) mcintoshi Price 1936 as described by Leigh, 1974, differing in behavior and morphology of cercarial, metacercarial and adult stages.


Schistosomiasis mansoni in three localities of western lowland of the state of Maranháo before and after mass treatments. A cross-sectional study for schistosomiasis was carried out in the localities of Aliança, Alegre and Coroatá (districts of Cururupu, São Bento and São João Batista, respectively) in the lowland of the state of Maranhão, after respectively 13, 11 and 4 mass treatments with oxamniquine in the period of ten years (1977-1987). The study included clinical and quantitative fecal examination, skin test for Schistosoma mansoni infection, evaluation of man-water contact of the total population (829 persons) in the three localities and other epidemiological investigations such as infection rate and dynamics of the snail population. After 13 mass treatments in Aliança, the prevalence of S. mansoni infection was reduced from 57.9% to 7.4%. In Coroatá with 11 mass treatments the prevalence fell from 69.2% to 12.8% and in Alegre, with only 4 mass treatments there was practically no reduction in prevalence: 22.9% to 21%. After mass treatments the type II hepatointestinal clinical form was 10.8% in Aliança, 17.9% in Alegre and 18% in Coroatá. The hepatosplenic (type III) form was not seen in Aliança and Coroatá but unexplanably it was 7.6% in Alegre. There was no correlation between the egg load elimination and the clinical forms.


Dengue 2 virus enhancement in asthmatic and non asthmatic individual. During the 1981 dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) Cuban epidemic, bronchial asthma (BA) was frequently found as a personal or family antecedent in dengue hemorrhagic fever patients. Considering that antibody dependent enhancement (ADE) plays an important role in the etiopathogenic mechanism of DHF/DSS, we decide to study the Dengue 2 virus (D2V) capability of replication in peripheral blood leukocytes (PBL) from asthmatic patients and healthy persons. In 90% of asthmatic patients and 53.8% of control group it was obtained PBL with a significant D2V enhancing activity (X2 p < 0.01). Power enhancement was higher in asthmatic group. This is the first in vitro study relating BA and the dengue 2 virus immuno enhancement. The results obtained support the role of BA as a risk factor for DHF/DSS as already described on epidemiological data.


Replication of dengue viruses in mosquito cell cultures--a model from ultrastructural observations. Mosquito cell cultures infected with human sera from dengue-1 and dengue-2 outbreaks, started in Rio de Janerio by 1986 and 1990 respectively, were examined by electron microscopy at different times post the infection of cell cultures. More information was obtained about cell penetration of virus particles in the presence or not of antibodies, their pathway inside the cells, replication mode and exist. Infectiveness of the virus at those different stages can only be attributed to the particles appearing inside the trans-Golgi vesicles; most of all newly formed virus particles remain inside the RER-derived cell vesicles or inside lysosomes, even during cell lysis. Groups of larger particles, 65-75 nm in diameter at dengue-2 infections, persist during cell passage. The large amounts of smooth membrane structures, as vesicles or tubules inside the RER, are attributed to a cell response to viral infection.


Delayed phagocytosis and bacterial killing in Chediak-Higashi syndrome neutrophils detected by a fluorochrome assay. Ultrastructural aspects. The few studies already published about phagocyte functions in Chediak-Higashi syndrome (CHS) has stated that neutrophils present slow rate of bacterial killing but normally ingest microorganisms. In the present study, both phagocytosis and killing of Staphylococcus aureus were verified to be delayed in neutrophils from two patients with CHS when these functions were simultaneously evaluated by a fluorochrome phagocytosis assay. Electron microscopic examination showed morphologic differences among neutrophils from CHS patients and normal neutrophils regarding the cytoplasmic structures and the aspects of the phagolysosomes. It was noteworthy the presence of giant phagolysosomes enclosing bacteria in active proliferation commonly observed in CHS neutrophils after 45 min of phagocytosis, which corresponded with the impaired bactericidal activity of these leukocytes. The present results suggest that phagocytosis may also be defective in CHS, and point out to the sensitivity of the fluorochrome phagocytosis assay and its application in clinical laboratories.


Simian malaria in Brazil.

Simian malaria in Brazil. In Brazil simian malaria is widely spread, being frequent in the Amazon region (10% of primates infected) and even more in the forested coastal mountains of the Southeastern and Southern regions (35% and 18% infected, respectively), but absent in the semi-arid Northeast. Only two species of plasmodia have been found: the quartan-like Plasmodium brasilianum and the tertian-like P. simium, but the possible presence of other species is not excluded. P. brasilianum is found in all enzootic foci, but P. simium was detected only on the coast of the Southeastern and Southern regions, between paralles 20 degrees S and 30 degrees S. Nearly all hosts are monkeys (family Cebidae, 28 species harbouring plasmodia out of 46 examined), and very rarely marmosets or tamarins (family Callitrichidae, 1 especies out of 16). P. brasilianum was present in all infected species, P. simium in only two. The natural vector in the Southeastern and Southern regions was found to be Anopheles cruzi, but has not been conclusively identified in the Amazon. One natural, accidental human infection due to P. simium was observed. There is no evidence of the relation of simian to human malaria in the Southeastern and Southern regions, where human malaria was eradicated in spite of the high rates of monkeys infected, but in the Amazon recent serological studies by other workers, revealing high positivity for P. brasilianum/P. malariae antibodies in local indians, would suggest that among them malaria might possibly be regarded as a zoonosis.


Use of glass beads and CF 11 cellulose for removal of leukocytes from malaria-infected human blood in field settings. Passage of malaria-infected blood through a two-layered column composed of acid-washed glass beads and CF 11 cellulose removes white cells from parasitized blood. However, because use of glass beads and CF 11 cellulose requires filtration of infected blood separately through these two resins and the addition of ADP, the procedure is time-consuming and may be inappropriate for use in the field, especially when large numbers of blood samples are to be treated. Our modification of this process yields parasitized cells free of contaminating leukocytes, and because of its operational simplicity, large numbers of blood samples can be processed. Our procedure also compares well with those using expensive commercial Sepacell resins in its ability to separate leukocytes from whole blood. As a test of usefulness in molecular biologic investigations, the parasites obtained from the blood of malaria-infected patients using the modified procedure yield genomic DNA whose single copy gene, the circumsporozoite gene, efficiently amplifies by polymerase chain reaction.


Extrachromosomal nucleic acids in bovine Babesia. Two kinds of small extrachromosomal nucleic acid elements were found in the bovine babesias, Babesia bovis and B. bigemina. One element with an apparent size of 5.5 kilobase pairs (kbp) is a double stranded RNA related to virus like particles. Another molecule is a double stranded DNA with a molecular size of about 6.2 kbp. Southern blot comparison of restriction DNA fragments of the latter molecule, which is present in both B. bovis and B. bigemina is described.


Epidemiology and control of malaria and other arthropod-borne diseases. Malaria and other arthropod born diseases remain a serious public health problem affecting the lives and health of certain social groups when the two basic strategies to control fail due to: (1) the lack of effective chemoprophylaxis/chemotherapy or the rapid development of drug resistance of the infectious agents and (2) the ineffectiveness of pesticides or the arthropod vectors develop resistance to them. These situations enhances the need for the design and implementation of other alternatives for sustainable health programmes. The application of the epidemiological methods is essential not only for analyzing the relevant data for the understanding of the biological characteristics of the infectious agents, their reservoirs and vectors and the methods for their control, but also for the assessment of the human behaviour, the environmental, social and economic factors involved in disease transmission and the capacity of the health systems to implement interventions for both changes in human behaviour and environmental management to purpose guaranteed prevention and control of malaria and other arthropod born diseases with efficiency, efficacy and equity. This paper discuss the evolution of the malaria and arthropod diseases programmes in the American Region and the perspectives for their integration into health promotion programs and emphasis is made in the need to establish solid basis in the decision-making process for the selection of intervention strategies to remove the risk factors determining the probability to get sick or die from ABDs. The implications of the general planning and the polices to be adopted in an area should be analyzed in the light of programme feasibility at the local level, in the multisectoral context of specific social groups and taking in consideration the principles of stratification and equity.


Epidemiological risk stratification of malaria in the Americas. During the last years, malaria had a significant increase in Latin America, emerging again as one critical health problem in the Region of the Americas. More than 1.04 million new cases were reported in 1990. This resurgence of malaria needed a comprehensive strategy for its prevention and control. National malaria control programs recognized the epidemiological stratification of malaria as a valuable method to assist them in the recognition of local variations and factors that specifically contribute to the level and intensity of transmission in critical malarious areas. Also it serves as a useful instrument for the selection of needed malaria prevention and control activities. The principal feature of this approach is to provide a dynamic and ongoing process for assessing the epidemiological importance of different risk factors (socio-economic, ecological, organization of health services) in malaria transmission. Health interventions are based on this assessment and are aimed directly at the reduction or elimination of the identified risk factors operating at the local level. Intersectorial co-participation and the integration of malaria programs in local health services are also important aspects of this public health approach.


Household-based malaria control in a highly endemic area of Africa (Tanzania): determinants of transmission and disease and indicators for monitoring--Kilombero Malaria Project. The Kilombero Malaria Project (KMP) attempts to define operationally useful indicators of levels of transmission and disease and health system relevant monitoring indicators to evaluate the impact of disease control at the community or health facility level. The KMP is a longitudinal community based study (N = 1024) in rural Southern Tanzania, investigating risk factors for malarial morbidity and developing household based malaria control strategies. Biweekly morbidity and bimonthly serological, parasitological and drug consumption surveys are carried out in all study households. Mosquito densities are measured biweekly in 50 sentinel houses by timed light traps. Determinants of transmission and indicators of exposure were not strongly aggregated within households. Subjective morbidity (recalled fever), objective morbidity (elevated body temperature and high parasitaemia) and chloroquine consumption were strongly aggregated within a few households. Nested analysis of anti-NANP40 antibody suggests that only approximately 30% of the titer variance can be explained by household clustering and that the largest proportion of antibody titer variability must be explained by non-measured behavioral determinants relating to an individual's level of exposure within a household. Indicators for evaluation and monitoring and outcome measures are described within the context of health service management to describe control measure output in terms of community effectiveness.


Human babesiosis in Europe.

Human babesiosis in Europe. Human babesiosis in Europe came to medical attention in 1957 and until now 19 cases have been reported, most of them due to Babesia divergens. The onset of the disease is characterized by hemoglobinuria, high fever and renal failure ensue rapidly. The patients were generally asplenic and resident in a rural area. Intraerythrocytic pleomorphic parasites (1-3 microns) observed in stained thin blood smears are essential for Genus diagnosis. Parasitemia varied from 5 to 80% of red blood cells. Massive blood exchange transfusion (2-3 blood volumes) followed by intravenous clindamycin (3-4 times daily) and oral quinine (600 mg base, 3 times daily) were successfully used in the treatment of three recent cases. Splenectomised individuals should be aware for prevention.


The level of infestation with the vector of cattle babesiosis in Argentina. Studies were carried out to determine the differential aptitude to sustain the only vector of cattle babesiosis in Argentina, the tick Boophilus microplus, throughout the infested region of this country. Tick counts on Bos taurus cattle were used as the main criterion to classify favourable (F), intermediate (I) and unfavourable (U) areas for its development. The geographical limits of each area were set up using data of non-parasitic tick stages, temperature, water balance and map recognition of flooded and unflooded zones. The F area contained 16.5 x 10(6) ha with a cattle population of 6 x 10(6); the I and U areas had 25 x 10(6) ha with 2.7 x 10(6) cattle and 19 x 10(6) with population of 2.4 x 10(6) cattle, respectively. Research on the relationship amongst Babesia-Boophilus-cattle is needed in the F area for tick development which coincides with the best region for cattle breeding.


Transmission and diagnosis of equine babesiosis in South Africa. The transmission and prevalence of Babesia equi and B. caballi are being studied. Rhipicephalus evertsi mimeticus an ixodid tick from Namibia was identified as a new vector of B. equi, however, R. turanicus, previously reported to be a vector, failed to transmit both B. equi and B. caballi in the laboratory. The accurate diagnosis of B. caballi is being investigated because the nature of its low level parasitaemia does not allow easy detection in thin blood smears, routinely used for diagnosis, by clinicians. Consequently its role as a pathogen remains obscure. The importance of identifying infected horses, destined for export to Babesia-free countries, is also stressed. Thick and thin blood smears, serology (IFAT) and DNA probes are currently employed to study disease prevalence. To date 293 healthy, adult, thoroughbred horses have been screened by all three methods. The percentage positives are as follows: B. equi 4.4%, 70.6%, 13% and B. caballi 0.7%, 37%, 18.4% respectively. The DNA probes were more sensitive than blood smear examination for diagnosing carrier infections but are probably not sensitive enough to identify all carrier infections. A poor correlation was found between detection of the parasites' DNA and seropositivity. However, polymerase chain reaction could be used to amplify parasite DNA in a particular sample and this could result in more accurate diagnosis.


Impact of Babesia bovis and Babesia bigemina on the production of beef cattle in Uruguay. Uruguay is situated in a marginal area for the development of Boophilus microplus (30 degrees 35 degrees South Lat.) with important areas of enzootic instability for Babesia bovis and B. bigemina. The livestock products represent 70% of our exports, for which reason it is fundamental to evaluate the losses in the production that these haemoparasites cause as basic information to take future decisions. In the period 1988-1990, several works were carried out by our laboratory to know the incidence of babesiosis in the reduction of liveweight gains. The results are shown and discussed in the work. Experiment I: the weight increase of the control group (x = 0.248 kg/day), was 23% higher than that of the infected group with Babesia spp (from Uruguay), but significant statistical differences were not found (P < 0.05). These animals were kept in boxes and the food was controlled for 76 days. Experiment II: the incidence of Babesia spp (same strain) was studied for 140 days on Hereford heifers (n = 14) on natural pastures. The control group obtained x = 25.29 kg of liveweight gain and it was 45% higher than that of the infected group, significant statistical difference were found (P < 0.05). Experiments with attenuated strains III: four studies were carried out inoculating B. bovis and B. bigemina in bovines about one year old, in different growth systems, searching for the limit of application. Significant statistical differences between those groups were not found during the experiment (about 180 days) (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


On the current existence of a coccidial line of development in the malaria parasites: a theory. Most opinion favors the origin of the malaria parasites from a coccidial ancestor. It is assumed that whatever the process through which the coccidia differentiated into a Plasmodium this phenomenon very probably occurred millions of year ago, and during that differentiation process the original coccidia vanished. Therefore it has never been repeated. At the light of some experiments the existence, at the present time, of a coccidial cycle of development in the malaria parasites, is proposed. The connection routes and mechanisms through which the malaria parasite changes to a coccidial life, and the routes in reverse are exposed. Transmission of the malaria-coccidial forms is suggested.


Immunogenicity and antigenicity of the N-term repeat amino acid sequence of the Plasmodium falciparum P126 antigen. The P126 protein, a parasitophorus vacuole antigen of Plasmodium falciparum has been shown to induce protective immunity in Saimiri and Aotus monkeys. In the present work we investigated its immunogenicity. Our results suggest that the N-term of P126 is poorly immunogenic and the antibody response against the P126 could be under a MHC restricted control in C57BL/6(H-2b) mice, which could be problematic in terms of a use of the P126 in a vaccine program. However, we observed that a synthetic peptide, copying the 6 octapeptide repeat corresponding to the N-term of the P126, induces an antibody response to the native molecule in C57BL/6 non-responder mice. Moreover, the vaccine-P126 recombinant induced antibodies against the N-term of the molecule in rabbits while the unprocessed P126 did not.


A recombinant hybrid protein as antigen for an anti-blood stage malaria vaccine: a study on the conservation of a protective component. Recently we have shown that two hybrid proteins expressed in Escherichia coli confer protective immunity to Aotus monkeys against an experimental Plasmodium falciparum infection (Knapp et al., 1992). Both hybrid proteins carry a sequence containing amino acids 631 to 764 of the serine stretch protein SERP (Knapp et al., 1989b). We have studied the diversity of this SERP region in field isolates of P. falciparum. Genomic DNA was extracted from the blood of six donors from different endemic areas of Brazil and West Africa. The SERP region encoding amino acids 630 to 781 was amplified by polymerase chain reaction (PCR) and sequenced. Only conserved amino acid substitutions in maximally two positions of the analyzed SERP fragment could be detected which supports the suitability of this SERP region as a component of an anti-blood stage malaria vaccine.


Development of an immunoenzymatic assay using a monoclonal antibody against a 50-kDa catabolite from the P126 Plasmodium falciparum protein to the diagnosis of malaria infection. The WHO criterion of defering any donation of blood by a confirmed case of malaria for three years after cessation of therapy can not be applied in areas where malaria is endemic. For this reason we developed an immunoenzymatic assay for the detection of plasmodial antigens for blood screening in malarial endemic areas. So, we tested sera from 191 individuals. Among patients with active disease 100% of the cases of Plasmodium falciparum or mixed infections and 91.7% of those with P. vivax were positive for the presence of plasmodial antigens. The lower parasitaemia detected was 0.0003% for P. falciparum and 0.001% for P. vivax malária. When the frequency of positive circulating malarial antigens was evaluated among asymptomatic and symptomatic individuals with negative TBS, positive results were found in respectively 38.7% and 17.7% of the individuals studied in the 30 days after confirmed malaria attack. Data provide by these assays have shown that ELISA seemed to be more sensitive than parasitological examination for malaria diagnosis. This test by virtue of its high sensitivity and the facilities in processing a large number of specimens, can prove to be useful in endemic areas for the recognition of asymptomatic malaria and screening of blood donors.


Non-immunologic methods of diagnosis of babesiosis. The diagnosis of tick-borne diseases such as babesiosis still depends on observing the parasite in the infected erythrocyte. Microscopic observation is tedious and often problematic in both early and carrier infections. Better diagnostic methods are needed to prevent clinical disease, especially when susceptible cattle are being moved into disease enzootic areas. This study evaluates two techniques for early diagnosis of Babesia bovis infections in cattle, DNA probes specific for the organism and fluorescent probes specific for nucleic acid. The radioisotopically labeled DNA probes are used in slot blot hybridizations with lysed blood samples, not purified DNA. Thusfar, the probe is specific for B. bovis and can detect as few as 1000 B. bovis parasites in 10 microliters of blood. The specificity of the fluorescent probe depends on the characteristic morphology of the babesia in whole blood samples, as determined microscopically. The fluorescent probe detects as few as 10,000 B. bovis parasites in 10 microliters os blood. The application of each method for laboratory and field use is discussed.


New methods for the diagnosis of Babesia bigemina infection. Accurate diagnosis of Babesia bigemina infection, an economically important tick-transmitted protozoan parasite of cattle, is essential in the management of disease control and in epidemiological studies. The currently used methods of diagnosis are blood smear examination and serological tests which include agglutination and immunofluorescence tests. These tests have been used in the fild but because they lack sensitivity and specificity, newer and improved methods of diagnosis are being developed. The quantitative buffy coat (QBC) method, using microhaematocrit tubes and acridine orange staining allows rapid and quicker diagnosis of B. bigemina and other blood parasites compared to light microscopic examination of stained smears. Parasite specific monoclonal antibodies have been used in antigen/antibody capture enzymelinked immunosorbent assays with greater sensitivity and specificity than previously described serological tests. Similarly, DNA probes, derived from a repetitive sequence of the B. bigemina genome, offer a method of detecting very small numbers of parasites which are undetectable by conventional microscopy. An extrachromosomal DNA element, present in all the tick-borne protozoan parasites so far tested, provides an accurate means of differentiating mixed parasite populations in infected animals. These improved methods will greatly facilitate epidemiological studies.


Detection of Babesia bigemina infection: use of a DNA probe--a review. The development of a repetitive DNA probe for Babesia bigemina was reviewed. The original plasmid (p(Bbi)16) contained an insert of B. bigemina DNA of approximately 6.3 kb. This probe has been evaluated for specificity and analytical sensitivity by dot blot hybridization with isolates from Mexico, the Caribbean region and Kenya. A partial restriction map has been constructed and insert fragments have been subcloned and utilized as specific DNA probes. A comparison of 32P labelled and non-radioactive DNA probes was presented. Non-radioactive detection systems that have been used include digoxigenin dUTP incorporation, and detection by colorimetric substrate methods. Derivatives from the original DNA probe have been utilized to detect B. bigemina infection in a) experimentally inoculated cattle, b) field exposed cattle, c) infected Boophilus microplus ticks, and d) the development of a PCR amplification system.


Polypeptides reactive with antibodies eluted from the surface of Babesia bovis-infected erythrocytes. A technique was sought that would enable identification of surface-exposed parasite antigens on Babesia bovis-infected erythrocytes (BbIE) that are not detectable by surface-specific immunoprecipitations. Antibodies which bind to the surface of BbIE were recovered from intact cells using a low pH wash procedure. The eluted antibodies were then used in conventional immunoprecipitation assays to identify parasite-synthesized polypeptides carrying epitopes that are exposed on the surface or are cross-reactive with such epitopes. The results of these experiments support our previous data, obtained using a surface-specific immunoprecipitation technique, in the identification of a repertoire of parasite-derived antigens on the surface of infected erythrocytes (Allred et al., 1991). In addition, two polypeptides of M(r) 68,000 and 185,000 were identified which react strongly with the eluted antibodies but which are not detected by surface-immunoprecipitation. These data illustrate the potential of this approach for identification of parasite polypeptides which carry epitopes exposed on, or cross-reactive with exposed epitopes of the infected erythrocyte surface.


Evaluation of a colorimetric Babesia bigemina-DNA probe within an epidemiological survey. An epidemiological survey was conducted in southeast Mexico, in an effort to establish the serological reactivity and carrier status to Babesia bigemina of an indigenous cattle population. The prevalence was obtained through the Indirect Fluorescent Antibody Test (IFAT), using an in vitro culture-derived B. bigemina antigen. A specific, digoxigenin-coupled, approximately 6 Kb B. bigemina-DNA probe (BBDP), was used to indicate the presence of the parasite. Serum samples from 925 animals of all ages, were obtained within the three regions (I, II, III) of the state of Yucatan and tested by IFAT. In addition, whole blood samples drawn from 136 of the same animals of region II were analyzed using the BBDP. Positive IFAT (IFAT+) reactions were observed in 531 sera for a 57% overall prevalence. Regional values were: I = 157+ (56%), II = 266+ (68%) and III 108+ (42%). Only 32 (23%) of the blood samples tested with BBDP showed distinctive hybridization signal, in contrast with 100 (73%) IFAT+ animals. The response distribution for IFAT vs. BBDP was: +/+ 23, +/- 77, -/+ 9 and -/- 27 respectively. It was found that the analytical sensitivity of BBDP appears to be low for its utilization in widespread epidemiological surveys. It was considered, however, that the colorimetric probe might be useful to safely detect transmission prone carriers, since it is able to detect parasitemias as low as 0.001%.


Antibodies in falciparum malaria: what matters most, quantity or quality? In view of the recent demonstration that antibodies that are protective against Plasmodium falciparum malaria may act in collaboration with blood monocytes, we have investigated the isotype content of sera from individuals with defined clinical states of resistance or susceptibility to malaria. Profound differences in the distribution of each Ig subclass and particularly in the ratio of cytophilic versus noncytophilic antibodies were found. In protected subjects, two cytophilic isotypes, IgG1 and IgG3 were found to predominate. In non-protected subjects, i.e. children and primary attack adults, three different situations were encountered: a) an imbalance in which IgG2, a non-cytophilic class, predominated (mostly seen in primary attacks); b) an imbalance in which mostly IgM antibodies predominated (a frequent event in children) or c) less frequently, an overall low level of antimalarial antibodies. Of 33 non immune subjects studied all, except one, had one of the above defects. The function of total Ig presenting such an isotype imbalance was studied in vitro in Antibody-Dependent-Cellular-Inhibition assays. Not only did IgG from protected subjects cooperate efficiently with blood monocytes, whilst IgG from non-protected groups did not, but moreover the latter inhibit the in vitro effect of the former: in competition assays whole IgG from primary attack cases with increased IgG2 content, competed with IgG from immune adults, thus suggesting that non-protected subjects had antibodies to epitopes critical for protection, but that these antibodies are non functional.


A rapid, reliable method of evaluating growth and viability of intraerythrocytic protozoan hemoparasites using fluorescence flow cytometry. Fluorescence flow cytometry was employed to assess the potential of a vital dye, hydroethidine, for use in the detection and monitoring of the viability of hemoparasites in infected erythrocytes, using Babesia bovis as a model parasite. The studies demonstrated that hydroethidine is taken up by B. bovis and metabolically converted to the DNA binding fluorochrome, ethidium. Following uptake of the dye, erythrocytes containing viable parasites were readily distinguished and quantitated. Timed studies with the parasiticidal drug, Ganaseg, showed that it is possible to use the fluorochrome assay to monitor the effects of the drug on the rate of replication and viability of B. bovis in culture. The assay provides a rapid method for evaluation of the in vitro effect of drugs on hemoparasites and for analysis of the effect of various components of the immune response, such as lymphokines, monocyte products, antibodies, and effector cells (T, NK, LAK, ADCC) on the growth and viability of intraerythrocytic parasites.


Basic biochemical investigations as rationale for the design of original antimalarial drugs. An example of phospholipid metabolism. The future of antimalarial chemotherapy is particularly alarming in view of the spread of parasite cross-resistances to drugs that are not even structurally related. Only the availability of new pharmacological models will make it possible to select molecules with novel mechanisms of action, thus delaying resistance and allowing the development of new chemotherapeutic strategies. We reached this objective in mice. Our approach is hunged on fundamental and applied research begun in 1980 to investigate the phospholipid (PL) metabolism of intraerythrocytic Plasmodium. This metabolism is abundant, specific and indispensable for the production of Plasmodium membranes. Any drug able to interfere with this Plasmodium membranes. Any drug able to interfere with this metabolism blocks parasitic development. The most effective interference yet found involves blockage of the choline transporter, which supplies Plasmodium with choline for the synthesis of phosphatidylcholine, its major PL, this is a limiting step in the pathway. The drug sensitivity threshold is much lower for the parasite, which is more dependent on this metabolism than host cells. The compounds show in vitro activity against P. falciparum at 1 to 10 nM. They show a very low toxicity against a lymphoblastoid cell line, demonstrating a total absence of correlation between growth inhibition of parasites and lymphoblastoid cells. They show antimalarial activity in vivo, in the P. berghei or P. chabaudi/mouse system, at doses 20- to 100-fold lower than their acute toxicity limit. The bioavailability of a radiolabeled form of the product seemed to be advantageous (slow blood clearance and no significant concentration in tissues). Lastly, the compounds are inexpensive to produce. They are stable and water-soluble.


The treatment of falciparum malaria in children with halofantrine suspension. Malaria treatment of children is particularly difficult because of the absence of palatable suspensions for young children. Halofantrine hydrochloride is available as a suspension which is both palatable and simple to administer, and has been studied in a number of trials in the past 5 years. Children (331) ranging from 4 months to 17 years of age (mean 4.7 years) were treated with the 5% suspension using various dose regimens and 364 children ranging from 4 months to 14 years of age (mean 5.7 years) were treated with the 2% suspension 6 hourly for 3 doses. Using the 3-dose regimen there were only 2/462 (0.4%) who failed to clear the initial parasitaemia. Recrudescence occurred in 28/367 (7.6%) children with evaluable follow up data. The mean parasite clearance time in this group was 57.1 h (n = 417) and the mean fever clearance time was 50.9 h (n = 325). Symptoms related to malaria cleared rapidly following treatment generally by 24-48 h post treatment. Side effects possibly related to treatment were uncommon but were similar to those reported in adults. The frequency of diarrhoea and abdominal pain was lower than that seen in adults and was also less frequent following multiple doses and the use of the more dilute suspension. Since there was evidence that the majority of recrudescences were seen in younger children or those living in areas with low or seasonal transmission it is recommended that a further course of treatment 7 days later is given to these patients to prevent recrudescence.(ABSTRACT TRUNCATED AT 250 WORDS)


Prevalence and control of babesiosis in the Americas. This review presents up-to-date information on the distribution and control measures of babesiosis in Latin America. Bovine babesiosis caused by Babesia bovis and B. bigemia will be emphasized. The disease is endemic in most countries and poses a serious economic burden on livestock production in the region (U.S. $1365 million/year, FAO, 1989). Of the estimated 250 million cattle in Central and South America, approximately 175 million (70%) are in tick-infested regions. Humid, tropical and subtropical areas favor development of the main vector, the one-host tick Boophilus microplus. In many regions bovine babesiosis is enzootically stable as a consequence of a balanced host-parasite relationship. However, Latin America offers a wide range of epidemiologic conditions that are influenced by variations from tropical to cool climates and by susceptible purebred cattle that are regularly imported to upgrade local stocks. The control measures employed in most countries for babesiosis essentially rely on chemotherapy, use of acaricides for B. microplus, and to a lesser degree, on immunization methods. In general, these measures are expensive, time consuming, and in many cases, provide limited success. Finally, the zoonotic potential of babesiosis will be addressed, with special emphasis on the situation in the United States. Even though bovine babesiosis has long been eradicated from the U.S.A., human babesiosis is endemic in the northeastern region of the country.


Mefloquine resistant malaria in Cameroon and correlation with resistance to quinine. Based on the results of in vitro sensitivity of Plasmodium falciparum to chloroquine, quinine and mefloquine, and evaluation of drug consumption conducted in 1987-1988 in four areas in the north and south-west of Cameroon, two opposite situations were encountered in this country. In northern Cameroon where mefloquine resistance is prevalent a close correlation was found between the responses of P. falciparum to mefloquine and to quinine, but not between mefloquine and chloroquine. In the south, where chloroquine resistance is highly prevalent, no correlation was found neither between mefloquine and chloroquine nor mefloquine and quinine, but the responses to quinine and chloroquine appear partly correlated. These results lead to formulate the hypothesis of a "southern" type of P. falciparum submitted to a high chloroquine drug pressure inducing a secondary cross resistance, whilst a "northern" type submitted to a relatively high and abortive quinine drug pressure inducing a primary quinine resistance and a secondary cross resistance with mefloquine.


Babesia divergens vaccine.

Babesia divergens vaccine. A vaccine strategy against Babesia divergens bovine babesiosis was successfully developed after perfecting of an efficient in vitro culture. Crude supernatants and purified fractions were able to induce a vaccine protection in gerbils against B. divergens infection. More, supernatants induced an effective vaccine protection in cattle. The role of B. divergens exoantigens of 17, 37, 46, 70 and 90 kDa in the development of the immune response was clearly demonstrated in gerbils, cattle, and man.


Field evaluation of an exoantigen-containing Babesia vaccine in Venezuela. Bovine babesiosis is endemic in Venezuela, causing significant losses in highly susceptible imported cattle. Current immunoprophylactic methods include the less desirable use of live parasites. Inactivated vaccines derived from exoantigen-containing supernatant fluids of in vitro Babesia bovis and B. bigemina cultures have been developed and constitute a major improvement in vaccine safety, stability and ease of handling. Vaccination trials conducted under field conditions provide the final evaluation of a culture-derived B. bovis-B. bigemina vaccine. During a 5-year period, approximately 8,000 cattle were vaccinated and 16 clinical trials carried out in 7 states of Venezuela. Clinical, serologic and parasitologic data were collected monthly from 10% of the animals over a 2-year period. Data were also collected from a similar number of nonvaccinated control cattle. Analysis of results from these trials demonstrated a reduction in the incidence of clinical disease among vaccinated animals and complete protection against mortality caused by babesiosis. Vaccine efficacy was measured calculating the incidence rates of disease and mortality among vaccinated and nonvaccinated cattle. Use of this inactivated vaccine offers the best combination of safety, potency and efficacy for the effective immunoprophylactic control of bovine babesiosis.


Successful vaccination against Boophilus microplus and Babesia bovis using recombinant antigens. Current methods for the control of the cattle tick Boophilus microplus and the agent of bovine babesiosis, Babesia bovis are unsatisfactory. Effective immunological control of both parasites would have great advantages. However, naturally acquired immunity to the tick is generally unable to prevent serious production losses. A vaccine against the tick, based on a novel form of immunization, is being developed. A protective antigen has been isolated from the tick, characterized and produced as an effective, recombinant protein. A vaccine incorporating this antigen is currently undergoing field trials. In the Australian situation, improved tick control will probably increase endemic instability with respect to B. bovis. Fortunately, a trivalent, recombinant B. bovis vaccine has also been developed. This too is now undergoing pre-registration field trials.


A study on the pathogenesis of human cerebral malaria and cerebral babesiosis. Cerebral complications are important, but poorly understood pathological features of infections caused by some species of Plasmodium and Babesia. Patients dying from P. falciparum were classified as cerebral or non-cerebral cases according to the cerebral malaria coma scale. Light microscopy revealed that cerebral microvessels of cerebral malaria patients were filled with a mixture of parasitized and unparasitized erythrocytes, with 94% of the vessels showing parasitized red blood cell (PRBC) sequestration. Some degree of PRBC sequestration was also found in non-cerebral malaria patients, but the percentage of microvessels with sequestered PRBC was only 13%. Electron microscopy demonstrated knobs on the membrane of PRBC that formed focal junctions with the capillary endothelium. A number of host cell molecules such as CD36, thrombospondin (TSP) and intercellular adhesion molecule I (ICAM-1) may function as endothelial cell surface receptors for P. falciparum-infected erythrocytes. Affinity labeling of CD36 and TSP to the PRBC surface showed these molecules specifically bind to the knobs. Babesia bovis infected erythrocytes produce projections of the erythrocyte membrane that are similar to knobs. When brain tissue from B. bovis-infected cattle was examined, cerebral capillaries were packed with PRBC. Infected erythrocytes formed focal attachments with cerebral endothelial cells at the site of these knob-like projections. These findings indicate that cerebral pathology caused by B. bovis is similar to human cerebral malaria. A search for cytoadherence proteins in the endothelial cells of cattle may lead to a better understanding of the pathogenesis of cerebral babesiosis.


Optimization and inhibition of the adherent ability of Plasmodium falciparum-infected erythrocytes. The vast majority of the 1-2 million malaria associated deaths that occur each year are due to anemia and cerebral malaria (the attachment of erythrocytes containing mature forms of Plasmodium falciparum to the endothelial cells that line the vascular beds of the brain). A "model system" for the study of cerebral malaria employs amelanotic melanoma cells as the "target" cells in an in vitro cytoadherence assay. Using this model system we determined that the optimum pH for adherence is 6.6 to 6.8, that high concentrations of Ca2+ (50mM) result in increased levels of binding, and that the type of buffer used influences adherence (Bis Tris > MOPS > HEPES > PIPES). We also observed that the ability of infected erythrocytes to cytoadhere varied from (erythrocyte) donor to donor. We have produced murine monoclonal antibodies against P. falciparum-infected red cells which recognize modified forms of human band 3; these inhibit the adherence of infected erythrocytes to melanoma cells in a dose-responsive fashion. Antimalarials (chloroquine, quinacrine, mefloquine, artemisinin), on the other hand, affected adherence in an indirect fashion i.e. since cytoadherence is due, in part, to the presence of knobs on the surface of the infected erythrocyte, and knob formation is dependent on intracellular parasite growth, when plasmodial development is inhibited so is knob production, and consequently adherence is ablated.


Variation in the cytoadherence characteristics of malaria parasites: is this a true virulence factor? The sequestration of Plasmodium falciparum-infected erythrocytes to the endothelial cells of brain capillaries is believed to represent one of the determining factors in the pathogenesis of cerebral malaria. In vitro studies of cytoadherence provide an experimental approach to understand the mechanism of sequestration and the respective roles played by parasite and host components in this interaction. This paper critically reviews current studies on cytoadherence, with particular emphasis on the nature of the information provided by such studies and their limitations. The paper also describes how cytoadherence studies using the patient's own monocytes can provide original information on the level of receptor up-regulation in the course of malarial infection.


Cytokines and dysregulation of the immune response in human malaria. The dysregulation of the immune response by malaria parasite has been considered as a possible constraint to the effectiveness of malaria vaccination. In spite of the important role interleukin-1 (IL-1) plays on the immunoregulation, and its ability to mimic various features of clinical malaria, reports on IL-1 in malaria are lacking. We found that only 2 out of 35 subjects with acute malaria showed increased levels of serum IL-1 alpha by enzyme immunoassay. To assess whether IL-1 could interfere with T-lymphocyte responses, blood mononuclear cells from patients infected with Plasmodium falciparum, P. vivax, or healthy subjects were cultured with phytohemagglutinin, and lymphocyte proliferation measured 72 h later by 3H-thymidine incorporation. Our data showed that T-lymphocyte responses are depressed both in P. falciparum (10,500 +/- 2,900) and P. vivax malaria (13,000 +/- 3,300), as compared to that of healthy individuals (27,000 +/- 3,000). Addition of IL-1 partially reversed depression of malaria lymphocytes, but had no effect on normal cells. On the other hand, T-lymphocytes from malaria infected-subjects presented a minimal decrease in proliferation, when cultured in the presence of exogenous PGE2. These data indicate the occurrence of two defects of immunoregulation in malaria: a deficiency of IL-1 production by monocytes/macrophages, and an increased resistance of lymphocytes to the antiproliferative effect of PGE2.


Epidemiological distribution of Plasmodium falciparum drug resistance in Brazil and its relevance to the treatment and control of malaria. With the use of a simple formulary, filled by health agents was established a monitoring programme for responses of P. falciparum to the antimalarial drugs. This monitoring programme is emphasized for the knowledge of the epidemiology of the drug resistance and the control of malaria falciparum in Amazon Basin where occurs more than 95% of Brazilian malaria cases every year. It was demonstrated that still now 4-aminoquinolines have a great importance for the mortality control in areas where just SUCAM (National Health Foundation--Health Ministry) agents are present without any medical assistance. The results obtained permitted the simplification of malaria treatment in Brazil. Important conclusions were established in the field of malaria drug resistance.