Review on flavivirus vaccine development. Proceedings of a meeting jointly organised by the World Health Organization and the Thai Ministry of Public Health, 26-27 April 2004, Bangkok, Thailand.

In light of the continuous spread of human pathogenic flaviviruses, in particular the mosquito-transmitted species, vaccine development remains a high priority on the public health agenda. On 26-27 April 2004, a conference was held in Bangkok, Thailand, to review current status of flavivirus vaccine development and related issues, focussing on dengue (DEN) and Japanese encephalitis (JE). This event, co-sponsored by the World Health Organization (WHO) and the Thai Ministry of Public Health, reviewed the progress made with vaccine development, sero-epidemiological studies and other accompanying activities critical for vaccine development and vaccination. The considerable interest in and awareness of the flavivirus diseases and their prevention by public health decision makers, as well as the establishment of two dedicated programmes for dengue and Japanese encephalitis vaccine development raise hopes that new or improved vaccines will become available in the coming years.

Effects of rice powder salt solution and milk-rice mixture on acute watery diarrhea in young children.

A randomized pilot study was carried out to compare the safety and effectiveness of rice powder salt solution (RPSS) in combination with milk-rice mixture (RPSS-MR group, n = 17) with other two regimens, glucose-based oral rehydration solution (ORS) combined with MR (ORS-MR group, n = 17) and ORS combined with formula milk (ORS-milk group, n = 14) in the treatment of acute watery diarrhea with mild to moderate dehydration in 48 boys younger than 2 years. Results showed that in the first 24 hours patients in the RPSS-MR group had significantly smaller amounts of stool weight (32.7 g/kg) than those in the ORS-MR group (67.5 g/kg) and ORS-milk group (59.2 g/kg) (p< 0.05 for both measurements). Patients in the RPSS-MR group also had significantly shorter duration of diarrhea (29.6 hours) than the other two groups (43.8 hours and 49.6 hours, respectively) (p < 0.05 for both measurements). The stool weight and duration of diarrhea between the ORS-MR group and the ORS-milk group were not significantly different. The positive effect of milk rice mixture was not demonstrated in the study due to the significantly more severe diarrhea in the ORS-MR group. The effectiveness of the RPSS-MR is therefore likely due to mainly RPSS.

A new Vero cell rabies vaccine: results of a comparative trial with human diploid cell rabies vaccine in children.

We evaluated the immunogenicity and safety of a chromatographically purified rabies vaccine (CPRV) compared with human diploid cell rabies vaccine (HDCV) after pre-exposure immunizations (both primary and booster). Intramuscular doses of either 0.5 mL of CPRV or 1.0 mL of HDCV were given to 400 schoolchildren on days 0, 7, 28, and 365 (booster). Adequate titers of antibody (> or = 0.15 IU/mL, as defined by the Centers for Disease Control and Prevention) were observed in serum samples from all children 14 days after primary immunization with CPRV and HDCV; the antibodies persisted in all but one child up until 1 year. Fourteen days after the primary immunization series (day 42) and 7 days after booster immunization (day 372), all children had antibody titers of > or = 0.5 IU/mL. Local and systemic reactions after primary and booster immunizations occurred significantly less frequently in the CPRV group. A severe allergic reaction (angioedema) was reported in only one child after booster immunization with HDCV. CPRV has adequate immunogenicity for primary and booster pre-exposure immunizations in children and has a better safety profile than does HDCV.

Persistence of antibodies in children after intradermal or intramuscular administration of preexposure primary and booster immunizations with purified Vero cell rabies vaccine.

BACKGROUND: The use of intradermal (i.d.) injections of purified Vero cell rabies vaccine (PVRV) for preexposure prophylaxis has not been well-established. We studied the safety and immunogenicity of i.d. and intramuscular (i.m.) PVRV injections for primary and booster preexposure immunizations. METHODS: One of two rabies preexposure PVRV regimens comprising three doses of either 0.1 ml i.d. or 0.5 ml i.m. administered during 28 days was assigned at random to 190 school children. One booster dose was given 1 year later either i.d. or i.m., according to their initial randomization group. Serologic results were available from 155 (82%) children at 1 year after primary immunization and 118 (62%) children at 2 years after booster. RESULTS: Although children vaccinated i.d. had significantly lower rabies-neutralizing antibody titers after primary immunization as well as after booster than children vaccinated i.m. (P< 0.001 for all time points), there were no significant differences in the percentages of children with adequate titers (> or =0.15 IU/ml) between the i.d. and i.m. groups after both primary and booster immunizations. Mild local reactions were more frequent after i.d. vaccination. Mild or moderate systemic reactions were infrequent and similar after i.d. and i.m. vaccinations. Fever and headache were reported by < or =6%. The reactions after booster were not different from those of post-primary immunization. CONCLUSIONS: Purified Vero cell rabies vaccine appears to be safe and immunogenic for primary and booster preexposure immunizations. An i.d. PVRV preexposure regimen should be useful especially for rabies-endemic countries with low per capita income.

Evaluation of the safety, immunogenicity, and pharmacokinetic profile of a new, highly purified, heat-treated equine rabies immunoglobulin, administered either alone or in association with a purified, Vero-cell rabies vaccine.

A clinical evaluation of a new, purified, heat-treated equine rabies immunoglobulin (PHT-Erig), F(ab')2 preparation, was carried out in Thailand and in the Philippines-two countries where rabies is endemic. An initial prospective, randomised, controlled trial (Study 1), compared the safety and pharmacokinetics (serum concentrations of rabies antibodies) after administration either of PHT-Erig or of a commercially-available, equine rabies immune globulin (Erig PMC). A second trial (Study 2) simulated post-exposure rabies prophylaxis by using a reference cell culture vaccine, the purified Vero-cell rabies vaccine (PVRV), administered in association with either Erig PMC or PHT-Erig. In Study 1, 27 healthy, Thai adults received a 40 IU kg(-1) dose of either Erig PMC (n = 12) or PHT-Erig (n = 15) via the intramuscular (i.m.) route; half of the dose was injected into the deltoid area and the other half into the buttocks. Serum for rabies antibody determination and F(ab')2 concentration was collected at hours (H) 0, 6 and 12, and on day (D) 2, 3, 4, 6, 8, 10, 12 and 15. Both products were safe, with no serious adverse events, and in particular, no anaphylactic reactions or serum sickness was reported. A statistical comparison of the pharmacokinetic parameters did not demonstrate bioequivalence of the two products. Nonetheless, the relative bioavailability of 93% and the similar absorption rates suggest the pharmacokinetic profiles of Erig and PHT-Erig are similar. The antibody level in either group were low throughout the 15-day study period. The geometric mean titer (GMT) values ranged from group 0.027-0.117 IU ml(-1) in the Erig group and from 0.029 to 0.072 IU ml(-1) in the PHT-Erig. There was no significant difference between the evolution of GMT values for the two groups. In Study 2, 71 healthy volunteers received 40 IU kg(-1) via the intramuscular route of either Erig PMC (n = 36) or PHT-Erig (n = 35) on D0, in association with five doses of PVRV on D0, D3, D7, D14 and D28. The safety evaluation was performed during the 28-day follow-up and serum samples for anti-rabies antibody titration were collected on D0 (before injection) D3, D7, D14 and D28. No serious reactions were reported in either group. In particular, no immediate (anaphylactic type) or delayed (serum sickness) allergic reactions were observed. Over the 28-day follow-up period, GMT profiles of the two groups were statistically equivalent. On D14, 100% of the subjects had protective antibody titers (anti-rabies antibodies > or = 0.5 IU ml(-1), which is the WHO-recommended level of seroconversion), and Erig PMC and PHT-Erig were indistinguishable according to the clinical definition chosen. On D28, the GMT values were 33.2 IU ml(-1) (95% CI, 23.8-46.1 IU ml(-1)) in the Erig PMC/PVRV group and 31.4 IU ml(-1) (95% confidence interval, CI, 23.4-42.2 IU ml(-1)) in the PHT-Erig/PVRV group, showing evidence of adequate vaccine-induced antibody responses in both groups. The increased purity, the heat-treatment step introduced in the manufacturing process of PHT-Erig, and the good clinical results substantiate the use of this new generation, purified equine F(ab')2 preparation in the post-exposure prophylaxis of rabies.

Efficacy and pharmacokinetics of atovaquone and proguanil in children with multidrug-resistant Plasmodium falciparum malaria.

A trial was conducted in 32 Thai children with uncomplicated multidrug-resistant falciparum malaria to assess the efficacy, safety and pharmacokinetics of atovaquone and proguanil; plasma concentrations of atovaquone, proguanil and its metabolite, cycloguanil, were measured in a subset of 9 children. The children received atovaquone (17 mg/kg/d for 3 d) plus proguanil (7 mg/kg/d for 3 d). Twenty-six children who had only Plasmodium falciparum infection and remained in hospital for 28 d were assessed for drug efficacy. The combination regimen produced a cure rate of 100%. Parasite and fever clearance times were 47 h (range 8-75) and 50 h (range 7-111), respectively. Atovaquone and proguanil were rapidly absorbed, with median time to peak concentrations of 6 h (range 6-24) and 6 h (range 6-12), respectively. Peak concentrations of cycloguanil were achieved between 6 and 12 h (median 6) after administration of proguanil. Mean peak plasma concentration of atovaquone on day 3 was 5.1 micrograms/mL (SD = 2.1). The day 3 mean peak plasma concentration of proguanil was 306 ng/mL (SD = 108) compared with 44.3 ng/mL (SD = 27.3) for cycloguanil. Mean values for the AUC (area under plasma concentration-time curve) were 161.8 micrograms/mL.h (SD = 126.9) for atovaquone, 4646 ng/mL.h (SD = 1226) for proguanil, and 787 ng/mL.h (SD = 397) for cycloguanil. Terminal elimination half-lives of atovaquone, proguanil and cycloguanil were estimated as 31.8 h (SD = 8.9), 14.9 h (SD = 3.3) and 14.6 h (SD = 2.6), respectively. No major adverse effect was attributable to the study drugs. Atovaquone/proguanil combination is safe and highly effective, and should be especially valuable for treatment of multidrug-resistant falciparum malaria.

Comparative clinical trial of artesunate suppositories and oral artesunate in combination with mefloquine in the treatment of children with acute falciparum malaria.

A randomized pilot study to compare the safety and efficacy of artesunate suppositories (15 mg/kg/day for three days) versus oral artesunate (6 mg/kg/day for three days), both in combination with mefloquine (25 mg/kg), was conducted in 52 Thai children with uncomplicated multidrug-resistant falciparum malaria. Forty-five patients (87%) had a full 28-day follow-up in the hospital to assess efficacy and exclude reinfection. Mean [range] times to fever clearance of the two groups were similar (42 hr [15-104] versus 42 hr [6-119]). Artesunate suppositories resulted in significantly longer times to achieve 50% and 90% reductions of the initial parasite counts (17 and 26 hr versus 9 and 15 hr; P < 0.05 and P < 0.001). Time [range] to parasite clearance was longer in the artesunate suppositories group (42 hr [14-93] versus 35 hr [16-69]), but the difference was not significant. The cure rates by days 28 were not significantly different, 92% for artesunate suppository-treated patients and 100% for oral artesunate-treated patients. Both drug regimens are safe and effective. Further studies are needed to characterize the pharmacokinetic properties and the optimum regimen of artesunate suppositories for the treatment of severe malaria.

Lactobacillus GG and acute diarrhoea in young children in the tropics.

A prospective, placebo controlled, triple blind clinical trial was undertaken in Thailand to determine the effect of Lactobacillus GG on recovery from acute diarrhoea in children. Thirty-nine children (mean age = 8 months) were enrolled and following rehydration received either oral Lactobacillus GG (n = 20) as a freeze-dried preparation or placebo (n = 19) twice daily for 2 days. The clinical characteristics of the study groups were similar. There was no significant difference overall in clinical response detected between the study groups. When only those with acute non-bloody diarrhoea (n = 26) were considered, the mean duration of diarrhoea was significantly shorter in the lactobacillus group (1.9 days) than in the placebo group (3.3 days) (P < 0.055). Stool frequency was less on the second day in the lactobacillus group (P < 0.05). The results suggest that Lactobacillus GG accelerates recovery from acute watery diarrhoea in young children in a tropical setting.

Salivary sampling for hepatitis B surface antigen carriage: a sensitive technique suitable for epidemiological studies.

Vertical transmission from mothers to infants of hepatitis B infection result in a high incidence of carriage in children with long-term consequences, and is the focus of current immunization strategies. The use of saliva for hepatitis B surface antigen (HBsAg) screening in a country with an intermediate to high prevalence of chronic carriage was investigated. We recruited 88 Thai women with known HBsAg status (44 positive, 44 negative) who were attending an antenatal clinic. The collection of saliva was acceptable to all patients. Aspects of technique regarding collection and handling in a tropical setting are discussed. Comparing the detection of HBsAg in saliva to serum using a commercially available serological test kit without modification, the sensitivity was 92.0% (95% CI 84.8-99.5) and the specificity was 86.8% (95% CI 76.0-97.6). Population acceptability and safety advantages along with a high sensitivity strongly support the use of salivary sampling for epidemiological surveillance of hepatitis B virus.

Tepid sponging to reduce temperature in febrile children in a tropical climate.

The effectiveness of tepid sponging, in addition to antipyretic medication, in the reduction of temperature in febrile children living in a tropical environment, was assessed in a prospective, randomized, open trial. Seventy-five children aged between 6 and 53 months who attended the casualty department of the Children's Hospital, Bangkok, Thailand, with fever (rectal temperature > or = 38.5 degrees C) of presumed viral origin were randomized to received either tepid sponging and oral paracetamol (sponged group) or paracetamol alone (control group). Rectal temperature and the occurrence of crying, irritability, and shivering were recorded over the following 2 hours. A greater and more rapid fall in mean rectal temperature occurred in the sponged group than in the control group. Temperature fell below 38.5 degrees C sooner in children in the sponged group than in control children (P < 0.001). At 60 minutes, 38 (95.0%) of the controls still had a temperature of 38.5 degrees C or greater, compared with only 15 children (42.9%) in the sponged group (P < 1 x 10(-5). Crying was associated with sponging, but shivering and irritability occurred in only one child who was being sponged. It is concluded that tepid sponging, in addition to antipyretic medication, is clearly more effective than antipyretic medication alone in reducing temperature in febrile children living in a tropical climate.

Pharmacokinetic study of mefloquine in Thai children aged 5-12 years suffering from uncomplicated falciparum malaria treated with MSP or MSP plus primaquine.

A triple combination of mefloquine, sulfadoxine and pyrimethamine (MSP) was used with primaquine for the radical treatment of falciparum malaria in Thailand. Primaquine was reported to inhibit hepatic microsomal enzymes and drug metabolism in animal and man. 23 children hospitalized in the Hospital for Tropical Diseases, Faculty of Tropical Medicine, Bangkok, Thailand, were randomly allocated into two groups, group I received MSP equivalent to 20 mg base of mefloquine/kg body weight and group II received MSP plus primaquine (0.75 mg/kg). No statistical difference was noted for clinical response except the gametocyte clearance time was shorter in children given MSP plus primaquine (7 +/- 2.7 days) than the children given MSP alone (21.9 +/- 4.4 days) (P < 0.01). No serious side effects were recorded in this study. The plasma samples were obtained for kinetic calculations by HPLC in 18 children (11 in group I, 7 in group II). Mean Cmax was 7.4 +/- 5.2 h in group I and 6.6 +/- 7.0 h in group II. Mean t1/2, Cl/f and Vd/f were 9.8 +/- 1.6 days, 0.43 +/- 0.16 ml/min/kg and 8.84 +/- 4.21 l/kg in group I, 9.3 +/- 1.4 days, 0.41 +/- 0.12 ml/min/kg and 8.91 +/- 3.00 l/kg group II, respectively. The comparison of kinetic parameters in groups I and II revealed no significant difference (P > 0.05) suggesting no drug interaction. These kinetic values in children given MSP suspension were considerably different to those in adult patients with shorter tmax, t1/2 and MRT. The coadministration of MSP and primaquine in children would benefit by reducing the transmission of malaria in endemic areas.

Maternal practices and risk factors for dehydration from diarrhoea in young children: a case-control study in central Thailand slums.

To determine factors related to dehydration from diarrhoea, we conducted a hospital-based, case-control study in children aged 24 months or younger who had acute watery diarrhoea and attended Chonburi Regional Hospital in central Thailand during November 1988 through May 1989. The study compared 48 cases who had moderate or severe dehydration with 48 controls who had no dehydration. Both cases and controls belonged to low socioeconomic families and were living in urban slum areas. They had adequate health care facilities and access to ORS packets. Overall, 56% of the mothers used ORS solution at home. None of the mothers knew how to administer ORS, i.e. the fluid was not given at the onset of diarrhoea to prevent dehydration, and they gave no more than 60 ml over a 24-hour period to their dehydrated children. They also did not use home fluids. Multivariate analysis of data showed two factors significantly associated with dehydration: children's dirty fingernails that indicated inadequate maternal hygiene-related behaviour (Odds Ratio 6.4; 95% Confidence Intervals 1.5-27.6, p < 0.01), and frequency of vomiting in the 24 hours before rehydration (Odds Ratio 1.3; 95% Confidence Intervals 1.1-1.6, p < 0.001). Cases and controls had similar aetiologic agents and nutritional status. Providing proper education to mothers about oral rehydration therapy with special emphasis on the volume of ORS to be given, along with guidance to improve their personal hygiene should be considered important interventions in reducing the risk of dehydration and deaths from diarrhoea in these children.

Rice-powder salt solution in the treatment of acute diarrhea in young children.

Dehydration is the most common cause of death in diarrheal patients. Early oral rehydration therapy (ORT) can prevent or reverse dehydration from diarrhea in almost almost all cases. Shortages of oral rehydration salt (ORS) packets in certain areas remain a major problem of the Diarrheal Diseases Control Program of Thailand. To find an effective solution that can be prepared locally, a randomized trial of oral rehydration solutions was conducted. A rice-powder salt solution containing rice-power 30 g/l and salt 3.5 g/l (RPSS) was evaluated in a group (n = 23) of infants and young children aged between 4 months and 5 years with mild or moderate dehydration from acute watery diarrhea, and the results were compared with those who received WHO recommended glucose electrolyte solution (WHO-ORS) (n = 21), and glycine supplemented WHO-ORS (G-ORS) (n = 20). The efficacies of WHO ORS and G-ORS were found to be similar. The RPSS was found to be more effective than WHO-ORS and G-ORS as shown by a significantly lower stool frequency, lower rate of stool output, a significantly shorter duration of diarrhea, and a smaller intake of rehydration fluid. Promotion of the effective rice-salt solution could increase early implementation of ORT in many rural communities.

Parasitologic and clinical human response to immunoglobulin administration in falciparum malaria.

The protective effect of African IgG antibodies against Plasmodium falciparum malaria was investigated by passive transfer in Thai patients. Sera from 333 African adults were collected in the Cote d'Ivoire and subjected to extensive screening. One hundred fifty-three samples were discarded for safety reasons, and IgG was extracted from those remaining under conditions allowing their use by the intravenous (iv) route. Eight Thai patients with P. falciparum parasitemia were treated by iv inoculation of the IgG: six with a 100 mg/kg dose given over three days, one with a single 20 mg/kg dose, and one with a single 200 mg/kg dose. To ensure a safety margin of at least 48 hours, subjects were chosen among patients having a recrudescent parasitemia following quinine treatment failure at the RI level. At that stage, symptoms were mild or absent and parasitemia was low but increasing (range 4, 200-9,000/microliters). The IgG pool exerted a profound, stage-specific, but non-sterilizing effect on each of the strains tested, and proved to be safe. Asexual parasitemia decreased by a mean 728-fold (range 46-1,086), while gametocytes were unaffected. Clearance of parasites and symptoms was as fast or faster than with drugs, and was consistent in the eight patients treated, suggesting that target antigens were equally expressed in geographically remote isolates. In peripheral blood smears, no mature forms were seen at any time during the followup, which does not support the hypothesis that reversal of cytoadherence occurred. After the disappearance of the transferred antibodies, recrudescent parasites from three patients were found to be susceptible to the same extent (mean decrease of 1,310-fold) to the same IgG preparation, indicating that selection of parasites able to escape the effect of antibodies had not occurred. No adverse side-effects were detected during the followup, which lasted one year.

Quantitation of Fansimef components (mefloquine + sulfadoxine + pyrimethamine) in human plasma by two high-performance liquid chromatographic methods.

Two simple, precise, and selective high-performance liquid chromatographic methods are described for the simultaneous quantitation of mefloquine (MQ) plus pyrimethamine (PYR) or sulfadoxine (SDX) plus its principal metabolite N4-acetylsulfadoxine (N4SDX) in human plasma. After a single-step extraction, MQ plus PYR and SDX plus N4SDX including internal standards were separated using ion-paired and ion-suppression chromatography. Total run times for the assays were less than 12 min. Intraassay and interassay precision of the methods expressed as the coefficients of variation were less than 9% in plasma for the four compounds. The extraction recovery averaged 98% for MQ, 97% for PYR, 96% for SDX, and 81% for N4SDX. Plasma concentrations of the four compounds in a pediatric patient after a single oral dose of Fansimef (MQ + SDX + PYR) were determined to demonstrate the clinical application of the methods.

Red cell and plasma concentrations of combined quinine-quinidine and quinine in falciparum malaria.

Red cell and plasma quinine-quinidine, and quinine concentrations in children with uncomplicated falciparum malaria who were treated with a combination of quinine/quinidine/cinchonine (combined drug) and quinine alone, respectively, were measured, using the extraction fluorescence method. The cure rates obtained with the high dose regimen of the combined drug (100%) were significantly higher than in the low dose regimen group (37.5%) (p less than 0.05), and the quinine regimen produced a 50% cure rate. Similar mild and transient ECG effects were noted in both the combined drug group and the quinine group. In patients treated with the combined drug, quinine-quinidine concentrations in both red cell and plasma of the high dose regimen group were significantly higher than those in the low dose regimen group (p less than 0.001, p less than 0.001). In quinine-treated patients, red cell quinine concentration in those with RII failure was significantly lower than that in patients with cure or RI failure (p less than 0.05). Both red cell and plasma levels of quinine-quinidine were higher than quinine levels. The red cell:plasma quinine-quinidine concentration ratios rose steadily to the high level from day 3 to day 6, while the ratio of quinine alone fluctuated around the low level and then gradually fell. The evidence suggests that red cell drug concentrations are more closely related to the outcome of treatment than to plasma concentrations and that the combined drug may be very useful for treatment of multi-drug-resistant P. falciparum infections. Further study is needed.

Antibodies that protect humans against Plasmodium falciparum blood stages do not on their own inhibit parasite growth and invasion in vitro, but act in cooperation with monocytes.

IgG extracted from the sera of African adults immune to malaria were injected intravenously into eight Plasmodium falciparum-infected nonimmune Thai patients. Clinical and parasitological improvement was reproducibly obtained in each case. After the disappearance of the transferred Ig, recrudescent parasites were equally susceptible to the same Ig preparation. High levels of antibodies to most parasite proteins were detected by Western blots in the receivers' sera (taken before transfer) as in the donors' Ig, thus indicating that the difference was qualitative rather than quantitative between donors and receivers. In vitro, the clinically effective Ig had no detectable inhibitory effect on either penetration or intra-erythrocytic development of the parasite. On the contrary, they sometimes increased parasite growth. In contrast, these IgG, as the receivers' Ig collected 4 d after transfer, but not those collected before transfer, proved able to exert an antibody-dependent cellular inhibitory (ADCI) effect in cooperation with normal blood monocytes. Results were consistent among the seven isolates studied in vitro, as with the recrudescent parasites. Thus, the results obtained in the ADCI assay correlate closely with clinical and parasitological observations.

Simultaneous measurement of quinine and quinidine in human plasma, whole blood, and erythrocytes by high-performance liquid chromatography with fluorescence detection.

A high-performance liquid chromatographic method with fluorescence detection is described for the simultaneous measurement of quinine and quinidine in plasma, whole blood, and erythrocytes. The compounds were separated on an Ultrasphere C18 reversed-phase column (25 cm x 4.6 mm inside diameter, 5 microns particle size) using a mobile phase of acetonitrile/water/triethylamine (11:88:1, vol/vol) at pH 2.5. The method, simple, accurate, and selective, requires only a single-step liquid-liquid extraction and uses the structurally similar alkaloid, cinchonine, as the internal standard. The commercial impurities, dihydroquinine and dihydroquinidine, and unknown metabolites were well resolved from the parent drugs. The assay is precise, with interassay coefficients of variation less than or equal to 7.0% and an accuracy of less than or equal to 7.3% over a concentration range of 0.125 to 4.0 micrograms/0.25 ml. The extraction recoveries of the two drugs were similar, averaging 82.9% for quinine and 79.3% for quinidine from the three biological fluids. The clinical application of the method for routine drug monitoring and for estimating the pharmacokinetics of quinine and quinidine in man are discussed.

Assessment of putative tests for protective immunity to falciparum malaria.

Sera from clinically immune individuals comprising 10 hospitalised patients (Group I), 30 persons residing in a malaria endemic area in Thailand (Group II) and 8 persons from a hyperendemic area in Ivory Coast (Group III) were tested by the parasite growth inhibition (PGI), indirect fluorescent antibody test of ring-infected erythrocyte surface antigen (RESA-IFA), urease-ELISA and Western blot. Paired sera from patients recovering from malaria (Group IV) as well as sera from blood donors were also tested. In the PGI test, sera were tested against three uncloned isolates of P. falciparum comprising SO, I4 and AE9 (PGI-SO, PGI-I4 and PGI-AE9 respectively). When growth inhibition of greater than or equal to 30% against any one of the three isolates was considered positive, the positive rate for the combined Groups I, II and III was 78.7%. Further analysis showed that the positive rates for PGI-SO, PGI-I4 and PGI-AE9 were 63.8%, 59.5% and 59.5% respectively and were not significantly different (p greater than 0.05). Comparison between PGI-SO, PGI-I4 and PGI-AE9 activities of Groups I, II and III sera showed no significant differences in any comparison groups except with PGI-AE9 in which Group III sera were more frequently positive than Group II sera (p = 0.004). Follow-up of PGI-SO and PGI-AE9 activities in Group IV patients showed mostly a decrease or no change in the activities of the convalescent sera taken 63 days later. RESA-IFA positive rate in the combined Groups I, II and III sera was 91.7%. There were no significant differences either in the seropositive rates or in the geometric mean antibody titers (GMT) between Groups I, II and III sera. Follow-up in Group IV patients showed no change in antibody titers in 64% of cases, decrease and increase in titers in 29% and 7% of cases respectively. The urease-ELISA seropositive rate in the combined Groups (I, II and III) was 89.5% which is not significantly different from that of RESA-IFA (p greater than 0.05). Comparison between individual Groups (I, II and III) likewise showed no significant differences in both GMT and seropositive rates. Follow-up in Group IV sera showed either no change or a decrease in antibody titers in 55.6% and 44.4% of cases respectively.(ABSTRACT TRUNCATED AT 400 WORDS)