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Biomedical subjects

W A Maier

Publications and source records attributed to W A Maier.

At least 19 recordsLinked to original sources

Growth-stimulating influence of human chorionic gonadotropin (hCG) on Plasmodium falciparum in vitro.

In this study, it is reported that human Chorionic Gonadotropin (hCG), being one of the most important hormones of pregnancy, has a growth-stimulating effect on the asexual stages of Plasmodium falciparum in vitro. On the one hand, it is shown that the effect of the hormone is dose-related: The highest growth-rates of Plasmodium falciparum in vitro are achieved, when doses of 8.32 i.u./ml (= 50 i.u. hCG/6 ml) and 16.67 I.U./ml (= 100 i.u. hCG/6 ml) are added to the culture medium. These doses correspond to the physiological peak amounts of hCG between the 9th and 16th week of pregnancy, when parasitaemia also reaches its highest rate. On the other hand, it is shown, that any growth-stimulating effect disappears after inactivation of the hormone by heating at 120 degrees C for 20 minutes. These data support the hypothesis, that hCG does not only possess immunosuppressive properties acting on the response of T-lymphocytes, but also increases the growth of Plasmodium falciparum in vitro. The combination of both effects may explain why malaria still remains one of the most serious complications of pregnancy.

Animals

Morphologic changes in Nosema algerae (Microspora) during extrusion.

As a member of the phylum Microspora, Nosema algerae is a small obligate intracellular parasite. Its free invasive stage is a spore with a characteristic cellular organization, including an apically anchored polar tube that serves as a tool for the transmission of genetic material into the host cell. By detailed electron micrographic documentation of the spore ultrastructure we present the aspects related to the biologic process of spore extrusion. Our ultrastructure findings confirm that the extrusion process of microsporidian spores is based on extreme changes in their organization. This study is the first complete ultrastructural documentation of N. algerae concerning the extrusion process, which can be subdivided into different stages: the breakdown of the microsporidian cellular compartmentation; the filling of a preformed polar tube with modified sporoplasm; the uncoiling of the polar tube, which in this stage has reached its final length; and, finally, its extrusion and screw-like movement.

Animals

Seasonal density, sporozoite rates and entomological inoculation rates of Anopheles gambiae and Anopheles funestus in a high-altitude sugarcane growing zone in Western Kenya.

An entomological study was conducted on vectors of malaria and their relative contribution to Plasmodium falciparum transmission in Mumias, a high-altitude site and large-scale sugarcane growing zone in Kakamega district, western Kenya. Anopheles gambiae s.l., the predominant vector species, represented 84% (n=2667) of the total Anopheles mosquitoes collected with An. funestus comprising only 16%. Polymerase chain reaction (PCR) identified all 600 specimens of the An. gambiae complex tested as An. gambiae sensu stricto, an indication that it is the only sibling species represented in the high-altitude sites in western Kenya. Plasmodium falciparum sporozoite rates of 6.3% (133/2118) for An. gambiae s.l. and 9.5% (38/402) for An. funestus by ELISA were obtained in Mumias. None of 1600 mosquitoes tested for P. malariae sporozoites was positive. ELISA tests of mosquito blood meals indicated a high tendency of anthropophagy, a behaviour contributing significantly to malaria transmission by the vector species, with 95.9%, 4.86% and 0.2% having taken at least one blood meal on human, bovine and avian hosts, respectively Malaria transmission intensity was low as revealed by the low entomological inoculation rates (EIR) recorded. The EIR values for An. gambiae s.l. were 29.2 infective bites per person per year (ib/p/year) and 17.5 ib/p/year for An. funestus in Mumias. The highest inoculation rate for both vector species was 7.0 ib/p/month in July. Plasmodium falciparum parasite rate among asymptomatic children was 55.4% and 44% in the wet (July-September) and dry (December-February) seasons, respectively. These results indicate that malaria transmission intensity in the high-altitude site is low but perennial, with transmission being maintained by An. gambiae s.s. and An. funestus.

Agriculture

Occurrence of an unusual polymerase chain reaction product during identification of Anopheles gambiae sibling species (Diptera: Culicidae).

Using the polymerase chain reaction (PCR) for species differentiation within the Anopheles gambiae complex, we examined several hundred Cameroonian mosquito specimens. Applying an approved routine protocol for DNA extraction and PCR conditions, apart from the indubitable identification of most of the specimens, we came across ten PCR products that did not correspond in length to any of the hitherto known amplification products of the sibling species. Sequencing experiments showed the ten products to be of identical length (117 bp) and nucleotide sequence. The total sequence of the novel product is included in the PCR product specific for A. melas, which is known to occur in the same collection area as the ten unidentifiable mosquito specimens. On alignment of the novel PCR product sequence and the A. melas one starting at the 3'-end primer annealing site, the last 20 nucleotides of the novel product, reflecting the sequence of the 2nd PCR primer, showed only 60% homology with the then-corresponding A. melas DNA site. Explanations for the occurrence of the unusual PCR products are given and discussed.

Animals

Serum-free cultivation of several Plasmodium falciparum strains.

Plasmodium falciparum strains from different malaria-endemic regions were grown in continuous culture without human serum. The medium was complemented with Nutridoma-SR instead of serum. All parasite strains developed well in serum-free medium. Two strains were thawed without serum and showed good multiplication in subsequent continuous culture.

Animals

Serum-free cultivation of Plasmodium falciparum gametocytes in vitro.

Several components were tested for their ability to replace human serum in cultures of Plasmodium falciparum set up for the development of gametocytes. Besides a serum-free medium, A*I*M*V (Gibco BRL), RPMI medium supplemented with commercially available serum substitutes was used to culture gametocytes. The following substances served as serum replacements: Basal Medium Supplement (Biochrom), Ultroser G (Gibco BRL) and Nutridoma-SR (Boehringer Mannheim). All serum-free additives supported some parasite growth, but only in RPMI supplemented with Nutridoma-SR were morphologically mature gametocytes obtained. The asexual forms developed almost as well as in RPMI with human serum added.

Animals

The effects of hormones on the gametocytogenesis of Plasmodium falciparum in vitro.

To examine possible effects of hormones on the gametocytogenesis of Plasmodium falciparum in vitro, insulin, progesterone, 17-beta-estradiol and testosterone were added in different concentrations to the culture medium. The medium containing RPMI was free of human serum but complemented with BMS (Basal-Medium-Supplement, Biochrom), a semidefined serum replacement. As controls, cultures were grown either with BMS supplemented RPMI without hormones, or with human serum complemented RPMI. Only addition of insulin increased the parasitemia. All tested steroid hormones added to the medium enhanced the number of gametocytes.

Animals

[Drinking water and parasites].

Entamoeba histolytica, Giardia lamblia, Cryptosporidium parvum, Isospora belli, Balantidium coli, and Microsporidia spp. are cosmopolitan parasites. They often cause diarrheal diseases. The waterborn transmission of all these parasites is possible (41). Surface water supplies used for drinking water are potential sources of contamination. Giardia lamblia and Cryptosporidium spp. have received great attention in industrialized countries during the last years because they are the etiological agents of waterborne diseases. The life cycles of Giardia lamblia and Cryptosporidium are described with a special reference to drinking water technologies aimed at removing these parasites.

Animals

The effect of nosematosis on the development of Plasmodium falciparum in Anopheles stephensi.

To quantify the effect of Nosema algerae (Microsporidia, Nosematidae) on the development of Plasmodium falciparum in Anopheles stephensi (Diptera, Culicidae), we carried out infection experiments under standardized laboratory conditions. Apart from a mean reduction of 69% in oocyst development, smaller numbers of oocysts and fewer sporozoites were found in the Nosema-infected mosquitoes. In addition, nosematosis resulted in higher mortality. The potential role of Nosema algerae as a biological control agent is discussed.

Animals

The effects of Nosema algerae on the development of Plasmodium yoelii nigeriensis in Anopheles stephensi.

Experimental simultaneous infections of Anopheles stephensi (Diptera: Culicidae) with Nosema algerae (Microsporida: Nosematidae) and Plasmodium yoelii nigeriensis under standardized laboratory conditions showed partial suppression of the malaria parasite. At 9 days after an infective bloodmeal, the oocysts in the midgut were counted; 12.1%-66.6% of the double-infected mosquitoes exhibited no oocysts, whereas only 4.5%-12% of the control group showed no oocysts. The mean reduction in oocyst numbers under the influence of Nosema was 84.68%. At 14 days after infection with Plasmodium, the amount of sporozoites was examined; their mean reduction in eight experiments was 70%.

Animals

Plasmodium falciparum: effect of chloroquine, halofantrine and pyrimethamine on the infectivity of gametocytes for Anopheles stephensi mosquitoes.

The activity of chloroquine, halofantrine and pyrimethamine against the gametocytes and sporogonic stages of Plasmodium falciparum (strain NF54) was tested. Five-day-old gametocytes (stages I and II) from in vitro cultures were exposed to the drugs for 48 hours. The effect of the drugs on gametocyte development was assessed by counting gametocytes on days nine and 15 of culture and determining the infectivity of the drug-treated gametocytes to mosquitoes. Gametocytogenesis was partially inhibited by all three drugs; there were 71% of the number of gametocytes in drug-free control cultures in cultures with 3 x 10(-8) M chloroquine, 51% with 5 x 10(-8) M chloroquine, 78% with 5-7 x 10(-9) M halofantrine, and 48% with 10(-7) M pyrimethamine. Halofantrine- and pyrimethamine-treated gametocytes were found to be more infective to Anopheles stephensi than untreated controls. The three drugs were also administered to the mosquitoes, either in the first bloodmeal, which contained gametocytes from in vitro cultures, or in the second, parasite-free bloodmeal, given four days after infection. The sporontocidal activity of the drugs was evaluated by counting the number of oocysts on the midgut seven or eight days after infection, or the number of sporozoites in the salivary glands 15 days after infection. A sporontocidal effect was observed only when pyrimethamine was administered with the infective bloodmeal. Neither chloroquine nor halofantrine had any marked effect on sporogony at the concentrations tested.

Animals

A new model for testing gametocytocidal effects of some antimalarial drugs on Plasmodium falciparum in vitro.

A technique is described for obtaining pure gametocyte cultures of Plasmodium falciparum, using pyrimethamine at the minimum concentration for inhibition of asexual parasites. Routine cultures producing sexual stages were exposed to pyrimethamine on days 5 and 6. These cultures grew synchronously and contained gametocytes of stages II, III and V on day 7, 9 and 15 of the cultures respectively. The pyrimethamine-treated gametocytes were more infective to mosquitoes than were untreated controls. This model for the culture of pure gametocytes was used to observe the activity of chloroquine, halofantrine, pyrimethamine and quinine on the gametocyte stage III of Plasmodium falciparum strain NF54 in vitro. NF54 was shown to be sensitive to chloroquine, quinine and pyrimethamine, but the results showed that halofantrine was the most effective drug in reducing the number of gametocytes. A concentration of 3 x 10(-9) M halofantrine was lethal to both asexual parasites and gametocytes. The gametocytocidal EC90 of chloroquine (1 x 10(-6) M) and that of quinine (9 x 10(-7) M) were equal to the minimum inhibitory concentration of asexual stages of isolates of P. falciparum considered as highly resistant to these drugs. A high concentration of pyrimethamine (1 x 10(-4) M) had, in contrast, little effect on gametocytes.

Animals

Trials to infect Anopheles stephensi with Plasmodium yoelii nigeriensis by the membrane feeding technique.

The aim of this study was to find optimal conditions for the membrane feeding technique to obtain maximum infection rates of mosquitoes with Plasmodium yoelii nigeriensis. The results show that the malaria parasite Plasmodium yoelii nigeriensis is most infective to Anopheles stephensi mosquitoes on day 3 of the infection in the mice, 1 day before the peak of parasitaemia. The mortality rate of the mosquitoes fed on mice on day 3 after infection was the highest as compared to mosquitoes fed on other days after infection. Gametocytes from mice 3 days after infection were fed to mosquitoes by three different membrane feeding methods. The results indicate that feeding during the first 10 min after blood collection gave the highest infection rates. Keeping the blood meal at a pH of 7.2 yields higher infection rates than keeping it at pH of 8.5. Stirring of the blood and supplying it with CO2 is not necessary when feeding of the mosquitoes is completed within the first 10 min after collection of the blood.

Animals

Experiments on cryopreservation of Plasmodium falciparum.

A comparative study was carried out in order to assess the effectivity of the deep-freezing conditions for P. falciparum cultures: 1. by plunging them into liquid nitrogen, 2. by use of an established freezing program for lymphocytes. After thawing in a waterbath at 37 degrees C, samples were diluted with equal volumes of 3.5% sodium chloride solution or 15% phosphate-buffered saline supplemented glucose solution. Other samples were not diluted after thawing, but the medium was replaced twice during the first day in culture. Survival was evaluated by counting the parasites in Giemsa-stained smears taken on day 3 after thawing. We obtained the best survival rate using NaCl solution, whereas replacing the medium twice resulted in the lowest level of survival. In synchronized cultures we studied survival rates of the different developmental stages of the intraerythrocytic parasites: young trophozoites (ring stages) and schizonts. The results indicate that irrespective of the freezing technique used, the survival level of young trophozoites was high (40 and 60%). The more sensitive stages (late trophozoites and schizonts) were better protected by using the freezing program designed for lymphocytes.

Animals