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.

Naturally acquired circumsporozoite antibodies and their role in protection in endemic falciparum and vivax malaria.

The role of naturally acquired circumsporozoite (CS) antibodies in protection against falciparum and vivax malaria was evaluated in a group of Thai endemic villagers using a prospective cohort and a case-control study design. There was no evidence of protection by either the presence of positive CS antibody levels at the presumed time of sporozoite exposure or in individuals who persistently had measurable levels of the antibodies. The study defined levels of CS antibodies that were not protective in natural infection.

An improved high-performance liquid chromatographic method for the simultaneous measurement of halofantrine and desbutylhalofantrine in human serum.

A rapid, accurate, and sensitive high-performance liquid chromatographic (HPLC) method, with ultraviolet detection, for simultaneous measurement of halofantrine (HAL) and desbutylhalofantrine (BHAL) in human serum is described. Sample preparation involved protein precipitation, followed by a solid-phase clean-up and a liquid-liquid extraction. Chromatographic separation was carried out on an Ultrasphere C8 column (25 cm x 4.

Characteristics of natural antibody responses to the circumsporozoite protein of Plasmodium vivax.

The antibody response to the prototype circumsporozoite (CS) protein of Plasmodium vivax (CSPV) was studied in Thai soldiers experiencing occupational malaria. Seventy-four (65%) of 114 men followed during assignment to a malaria transmission area developed blood-stage infection with P. vivax.

Acridine orange fluorescent microscopy and the detection of malaria in populations with low-density parasitemia.

Detection of low-density malaria parasites with Giemsa-stained thick smears (G-TS) requires time and experience and becomes impractical with high sample loads. Acridine orange fluorescent microscopy (AO/FM) of capillary centrifuged blood may offer an alternative technique. We compared AO/FM readings with G-TS in 290 specimens from asymptomatic people in Thai villages endemic for malaria.

Basal forebrain lesions with or without reserpine injection inhibit cortical reorganization in rat hindpaw primary somatosensory cortex following sciatic nerve section.

To test the hypothesis that cortical reorganization depends on acetylcholine and one or more of the monoamines, the hindpaw cortex was mapped in eight different groups of mature rats: (1) untreated; (2) after sciatic nerve transection; (3) after intraperitoneal injections of reserpine, to reduce the level of cortical monoamines; (4) after ibotenic acid lesion of the nucleus basalis of Meynert (NBM), to destroy cholinergic cells projecting to the cortex; (5) after reserpine treatment and transection; (6) after ibotenic acid lesion and transection; (7) after reserpine treatment and ibotenic acid lesion; and (8) after reserpine treatment, ibotenic acid lesion, and transection. Four days after transection, the cortex had reorganized in the transected group. However, this process of reorganization was prevented in transected animals with NBM lesions.

The early development of the neopallial wall and area choroidea in fetal rats. A light and electron microscopic study.

The telencephalic wall was studied by light and electron microscopy in 11-13 day old fetal rats (E11-13). A few specimens from E14-16 were also included for comparisons. Two areas were selected: the dorso-lateral convexity of the hemispheric vesicles, called the neopallial wall, and the area choroidea, the posterior part of the telencephalic roof which unites the two hemispheres.

Folate and cobalamin metabolism in Plasmodium falciparum.

During the past several years, Jerapan Krungkrai, H. Kyle Webster and Yongyuth Yuthavong have characterized the metabolic pathway of folate biosynthesis and folate-dependent reactions, including the cobalamin-dependent activity of methionine synthase, in P. falciparum grown in vitro.

Comparison of an enzyme-linked immunosorbent assay (ELISA) with a radioimmunoassay (RIA) for the measurement of rat insulin.

A recently developed competitive enzyme-linked immunosorbent assay (ELISA) was compared with a conventional competitive radioimmunoassay (RIA) for the measurement of rat insulin in culture medium. Fifty-six samples were analysed by both assays. There was a correlation coefficient of r = 0.

Macrophage activation in falciparum malaria as measured by neopterin and interferon-gamma.

Macrophage activation during acute falciparum malaria in 71 Thai adults was investigated by measuring urinary neopterin and serum interferon-gamma (IFN-gamma). Neopterin, a product of IFN-gamma-activated macrophages, was elevated in 94% of patients upon admission (day 0, prior to treatment) and in all at some time during the period of study. Neopterin levels tended to rise further (days 1-5) before falling back towards the normal range as patients recovered following effective chemotherapy (days 6-8).

P0 glycoprotein mRNA distribution in myelin-forming Schwann cells of the developing rat trigeminal ganglion.

A biotinylated P0 cDNA was hybridized in situ to aldehyde-fixed vibratome sections of trigeminal ganglia from day 2, day 7, day 15, day 30 and adult rats. Nickel-enhanced horseradish peroxidase (HRP) was used in an antibody sandwich method to detect hybridization. After postfixation in osmium tetroxide, the sections were dehydrated in ethanol and embedded in epon.

Prevalence of antibody to heterologous circumsporozoite protein of Plasmodium vivax in Thailand.

The distribution in Thailand of antibody to a recently discovered variant of circumsporozoite proteins of Plasmodium vivax was determined by enzyme-linked immunosorbent assay (ELISA). The ELISA capture antigens were a synthetic peptide of the principal variant sequence ANGAGNQPG and a candidate P vivax vaccine that contained the predominant repeat sequence GDRAA/DGQPA. Serological evidence of recent inoculation with the variant was found throughout Thailand and in migrants from Cambodia, Malaysia, and Burma.

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.

Occupational malaria in Thai Rangers: epidemiological, clinical and immunological features.

Malaria continues to be a serious disease threat to soldiers deployed in tropical environments. Distinct features of the disease and the host immune response require that malaria occurring in occupational groups be investigated separately from the infection as found in endemic populations. For the first time since the Vietnam War, malaria has been systematically studied in soldiers under combat conditions.

Preparation of fetal rat brains for light and electron microscopy.

To study cellular shapes, growth patterns, and fine structure during early stages of CNS development in rat embryos, preparative procedures were evaluated and modified to meet two criteria: 1) Coronal semithin sections should reveal undeformed telencephalic hemispheres that were symmetrically expanded on both sides of midline structures and were surrounded by contiguous mesenchyme. 2) In electron micrographs, cells should have intact, undistorted surface membranes, evenly distributed nucleoplasm and well preserved cytoplasmic organelles. To meet these criteria, 378 fetuses with a gestational age of 11-20 days (E11-E20) were used to test and modify procedures for anesthesia, embryo removal and handling, dissection, fixation, dehydration, and embedding of the embryonic CNS.

T cell responses in acute falciparum malaria.

T lymphocyte responses to malaria-specific antigens during acute falciparum malaria were studied to determine host-parasite interaction and its relation to the manifestations of the disease. The results indicate that while there is antigen-specific immunodepression, markedly elevated levels of soluble factors such as IL2 receptor, CD8 antigen and IFN-gamma suggest that there is intense concurrent cellular activation which however does not seem to be effective in controlling the infection. It is proposed that the cellular activation is to a large extent non-specific and polyclonal, and leads to the exaggerated production of cytokines and eventually immunopathology.

Increased gamma delta T cells in acute Plasmodium falciparum malaria.

The T cell receptor of gamma delta is normally expressed on a small percentage of peripheral lymphocytes. Although the role of gamma delta T cells in the physiologic immune response is still unknown, there is accumulating evidence that gamma delta T cells may participate in the immune response to mycobacterial and other infectious organisms. In this study, we have quantitated the number of circulating gamma delta T cells during acute Plasmodium falciparum malaria.

Immunolabeling of JC virus large T antigen in neonatal hamster brain before tumor formation.

Using immunolabeling methods, the JC virus (JCV) early or regulatory protein, large T antigen, was demonstrated in frozen sections of neonatal hamster brains before tumor formation. Three days after intracerebral inoculation of 2500 hemagglutinating units of JCV, T antigen was expressed predominantly in nuclei of cells in the external granular layer and newly forming internal granular layer of the cerebellum and also in cell nuclei located in the hippocampus, periventricular areas, and the olfactory bulb. At 7 days postinoculation (p.

Stereotaxic preparation of circumscribed cortical areas from rat brain for biochemical studies.

A method for the rapid dissection of circumscribed areas of rat cortex is described. The technique does not depend on skull-derived landmarks but uses for stereotaxic orientation the cross-point of the interhemispheric gap with the caudal margin of the cortex. An application of this dissection method to the biochemical analysis of cholinergic markers within the hindlimb representation of the primary somatosensory cortex revealed that both the activity of the enzyme choline acetyltransferase as well as the binding of [3H] quinuclidinyl benzilate to muscarinic cholinergic receptors do not seem to be affected drastically three days after unilateral transection of the sciatic nerve.