Glycosylphosphatidylinositol anchors of Plasmodium falciparum: molecular characterization and naturally elicited antibody response that may provide immunity to malaria pathogenesis.

Induction of proinflammatory cytokine responses by glycosylphosphatidylinositols (GPIs) of intraerythrocytic Plasmodium falciparum is believed to contribute to malaria pathogenesis. In this study, we purified the GPIs of P. falciparum to homogeneity and determined their structures by biochemical degradations and mass spectrometry. The parasite GPIs differ from those of the host in that they contain palmitic (major) and myristic (minor) acids at C-2 of inositol, predominantly C18:0 and C18:1 at sn-1 and sn-2, respectively, and do not contain additional phosphoethanolamine substitution in their core glycan structures. The purified parasite GPIs can induce tumor necrosis factor alpha release from macrophages. We also report a new finding that adults who have resistance to clinical malaria contain high levels of persistent anti-GPI antibodies, whereas susceptible children lack or have low levels of short-lived antibody response. Individuals who were not exposed to the malaria parasite completely lack anti-GPI antibodies. Absence of a persistent anti-GPI antibody response correlated with malaria-specific anemia and fever, suggesting that anti-GPI antibodies provide protection against clinical malaria. The antibodies are mainly directed against the acylated phosphoinositol portion of GPIs. These results are likely to be valuable in studies aimed at the evaluation of chemically defined structures for toxicity versus immunogenicity with implications for the development of GPI-based therapies or vaccines.

Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro.

Curcumin, a widely used spice and colouring agent in food has been shown to have a broad spectrum of biological activities such as anti-inflammatory, anti-neoplastic, antimutagenic and antioxidant. We have used liver slice culture model to demonstrate hepatoprotective activity of curcumin in vitro. Ethanol has been used as a hepatotoxin and the cytotoxicity of ethanol is estimated by quantitating the release of LDH. Ethanol induces 3.5 times more release of LDH from the liver cells and twice the amount of lipid peroxidation as compared to the cells from untreated liver tissue and this was significantly reduced in presence of curcumin (5 microM). We measured the activity of antioxidant enzymes (AOEs) namely superoxide dismutase, catalase and peroxidase and found that in ethanol treated cells activity of all three enzymes was elevated. However, when curcumin was added along with ethanol their levels were kept low. The fact that release of LDH is significantly reduced along with lipid peroxidation and the activity of AOEs is kept low indicates that curcumin by its antioxidant activity reduced the oxidative stress induced by ethanol and protected the liver cells in vitro.

Electromechanical coupling constant extraction of thin-film piezoelectric materials using a bulk acoustic wave resonator.

Thin-film piezoelectric materials such as ZnO and AlN have great potential for on-chip devices such as filters, actuators and sensors. The electromechanical coupling constant is an important material parameter which determines the piezoelectric response of these films. This paper presents a technique based on the Butterworth Van-Dyke (BVD) model which, together with a simple one-mask over-moded resonator, can be used to extract the bulk, one-dimensional electromechanical coupling constant K(2) of any piezoelectrically active thin-film. The BVD model is used to explicitly define the series resonance, parallel resonance, and quality factor Q of any given resonating mode. Common methods of defining the series resonance, parallel resonance, and Q are shown to be inaccurate for low coupling, lossy resonators such as the over-moded resonator. Specifically, an electromechanical coupling constant K(2) of (2.6+/-0.1)% was measured for an (002) c-axis textured AlN film with an X-ray diffraction rocking curve of 7.5 degrees using the BVD based extraction technique.

Measurements of the bulk, C-axis electromechanical coupling constant as a function of AlN film quality.

Piezoelectric thin film AlN has great potential for on-chip devices such as thin-film resonator (TFR)-based bandpass filters. The AlN electromechanical coupling constant, K(2), is an important material parameter that determines the maximum possible bandwidth for bandpass filters. Using a previously published extraction technique, the bulk c-axis electromechanical coupling constant was measured as a function of the AlN X-ray diffraction rocking curve [full width at half maximum (FWHM)]. For FWHM values of less than approximately 4 degrees , K (2) saturates at approximately 6.5%, equivalent to the value for epitaxial AlN. For FWHM values >4 degrees , K(2) gradually decreases to approximately 2.5% at a FWHM of 7.5 degrees . These results indicate that the maximum possible bandwidth for TFR-based bandpass filters using polycrystalline AlN is approximately 80 MHz and that, for 60-MHz bandwidth PCS applications, an AlN film quality of >5.5 degrees FWHM is required.

Sigmoid kidney associated with double urethra.

An unusual case of crossed renal ectopia (sigmoid kidney) in an adult, compounded with double urethra, is being presented for its rarity and mode of presentation.

Developmental stage-specific biosynthesis of glycosylphosphatidylinositol anchors in intraerythrocytic Plasmodium falciparum and its inhibition in a novel manner by mannosamine.

Glycosylphosphatidylinositols (GPIs) are the major glycoconjugates in intraerythrocytic stage Plasmodium falciparum. Several functional proteins including merozoite surface protein 1 are anchored to the cell surface by GPI modification, and GPIs are vital to the parasite. Here, we studied the developmental stage-specific biosynthesis of GPIs by intraerythrocytic P. falciparum. The parasite synthesizes GPIs exclusively during the maturation of early trophozoites to late trophozoites but not during the development of rings to early trophozoites or late trophozoites to schizonts and merozoites. Mannosamine, an inhibitor of GPI biosynthesis, inhibits the growth of the parasite specifically at the trophozoite stage, preventing further development to schizonts and causing death. Mannosamine has no effect on the development of either rings to early trophozoites or late trophozoites to schizonts and merozoites. The analysis of GPIs and proteins synthesized by the parasite in the presence of mannosamine demonstrates that the effect is because of the inhibition of GPI biosynthesis. The data also show that mannosamine inhibits GPI biosynthesis by interfering with the addition of mannose to an inositol-acylated GlcN-phosphatidylinositol (PI) intermediate, which is distinctively different from the pattern seen in other organisms. In other systems, mannosamine inhibits GPI biosynthesis by interfering with either the transfer of a mannose residue to the Manalpha1-6Manalpha1-4GlcN-PI intermediate or the formation of ManN-Man-GlcN-PI, an aberrant GPI intermediate, which cannot be a substrate for further addition of mannose. Thus, the parasite GPI biosynthetic pathway could be a specific target for antimalarial drug development.

Plasmodium falciparum glycosylphosphatidylinositol-induced TNF-alpha secretion by macrophages is mediated without membrane insertion or endocytosis.

The glycosylphosphatidylinositols (GPIs) of Plasmodium falciparum are believed to contribute to the pathogenesis of malaria by inducing the secretion of proinflammatory cytokines by macrophages. Previous studies have shown that P. falciparum GPIs elicit toxic immune responses by protein tyrosine kinase (PTK)- and protein kinase C (PKC)-mediated cell signaling pathways, which are activated by the carbohydrate and acyl moieties of the intact GPIs, respectively. In this study, we show that induction of TNF-alpha by P. falciparum GPIs in macrophages is mediated by the recognition of the distal fourth mannose residue. This event is critical but not sufficient for the productive cell signaling; interaction by the acylglycerol moiety of GPIs is also required. These novel interactions are coupled to previously demonstrated PTK and PKC pathways, since the specific inhibitors of these kinases effectively blocked the GPI-induced TNF-alpha production. Surprisingly, sn-2 lyso-GPIs were also able to elicit TNF-alpha secretion. Contrary to the prevailing notion, GPIs are neither inserted to the plasma membranes nor endocytosized. Thus, this study defines the GPI structural requirements and reveals a novel mechanism for the outside-in activation of cell signaling by P. falciparum GPIs in inducing proinflammatory responses.

Gravidity-dependent production of antibodies that inhibit binding of Plasmodium falciparum-infected erythrocytes to placental chondroitin sulfate proteoglycan during pregnancy.

During pregnancy, Plasmodium falciparum-infected erythrocytes sequester in the placenta by adhering to chondroitin 4-sulfate, creating a risk factor for both the mother and the fetus. The primigravidae are at higher risk for placental malaria than the multigravidae. This difference in susceptibility has been attributed to the lack of antibodies that block the adhesion of infected erythrocytes to placental chondroitin 4-sulfate in primigravid women. However, recent results show that many primigravidae at term have antibody levels similar to those of multigravidae, and thus the significance of antiadhesion antibodies in providing protection against malaria during pregnancy remains unclear. In this study, we analyzed plasma samples from women of various gravidities at different gestational stages for antiadhesion antibodies. The majority of women, regardless of gravidity, had similar levels of antibodies at term. Most primigravidae had low levels of or no antiadhesion antibodies prior to ~20 weeks of pregnancy and then produced antibodies. Multigravidae also lacked antibodies until ~12 weeks of pregnancy, but thereafter they efficiently produced antibodies. In pregnant women who had placental infection at term, higher levels of antiadhesion antibodies correlated with lower levels of placental parasitemia. The difference in kinetics of antibody production between primigravidae and multigravidae correlated with the prevalence of malaria in these groups, suggesting that antibodies are produced during pregnancy in response to placental infection. The early onset of efficient antibody response in multigravidae and the delayed production to antibodies in primigravidae appear to account for the gravidity-dependent differential susceptibilities of pregnant women to placental malaria.