Tumor Necrosis Factor -α, Interleukin-10, Intercellular and Vascular Adhesion Molecules Are Possible Biomarkers of Disease Severity in Complicated Plasmodium vivax Isolates from Pakistan

Background

Cytokine-mediated endothelial activation pathway is a known mechanism of pathogenesis employed by Plasmodium falciparum to induce severe disease symptoms in human host. Though considered benign, complicated cases of Plasmodium vivax are being reported worldwide and from Pakistan. It has been hypothesized that P.vivax utilizes similar mechanism of pathogenesis, as that of P.falciparum for manifestations of severe malaria. Therefore, the main objective of this study was to characterize the role of cytokines and endothelial activation markers in complicated Plasmodium vivax isolates from Pakistan.

Methods and Principle Findings

A case control study using plasma samples from well-characterized groups suffering from P.vivax infection including uncomplicated cases (n=100), complicated cases (n=82) and healthy controls (n=100) were investigated. Base line levels of Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Interleukin-10 (IL-10), Intercellular adhesion molecule-1 (ICAM-1), Vascular adhesion molecule-1(VCAM-1) and E-selectin were measured by ELISA. Correlation of cytokines and endothelial activation markers was done using Spearman’s correlation analysis. Furthermore, significance of these biomarkers as indicators of disease severity was also analyzed. The results showed that TNF-α, IL-10, ICAM-1and VCAM-1 were 3-fold, 3.7 fold and 2 fold increased between uncomplicated and complicated cases. Comparison of healthy controls with uncomplicated cases showed no significant difference in TNF-α concentrations while IL-6, IL-10, ICAM-1, VCAM-1 and E-selectin were found to be elevated respectively. In addition, significant positive correlation was observed between TNF-α and IL-10/ ICAM-1, IL-6 and IL-10, ICAM-1 and VCAM-1.A Receiver operating curve (ROC) was generated which showed that TNF-α, IL-10, ICAM-1 and VCAM-1 were the best individual predictors of complicated P.vivax malaria.

Conclusion

The results suggest that though endothelial adhesion molecules are inducible by pro-inflammatory cytokine TNF-α, however, cytokine-mediated endothelial activation pathway is not clearly demonstrated as a mechanism of pathogenesis in complicated P.vivax malaria cases from Pakistan.     

Coinfection with <Emphasis Type="Italic">Lichtheimia corymbifera</Emphasis> and <Emphasis Type="Italic">Aspergillus flavus</Emphasis> in an Immune-Competent Patient Mimicking as Pulmonary-Renal Syndrome

Lichtheimia corymbifera and Aspergillus flavus pulmonary coinfection has been rarely reported in immune-competent patients. We report case of a young male who presented with clinical features of pulmonary-renal syndrome and was later diagnosed to have bilateral polymicrobial fungal lung infection.     

Rapid test for determination of urea hydrolysis

A urease test for the rapid determination of urea hydrolysis is described in which diluted urea agar concentrate was used in small amounts with dense inoculum of the test organisms. The method was evaluated and compared with Christensen's urea agar slants by using 728 clinical isolates of gram-negative bacteria. Of the 325 strains of urease-positive Proteus-Providencia-Morganella, 282 (87%) gave positive results within 5 min with the rapid test. Urease activity of 97% of these organisms became evident within 30 min. All 287 isolates which showed no urease activity on Christensen's urea agar also remained negative by this test.     

Adaptive radiation and ecological opportunity in Sulawesi and Philippine fanged frog (Limnonectes) communities

Because island communities are derived from mainland communities, they are often less diverse by comparison. However, reduced complexity of island communities can also present ecological opportunities. For example, amphibian diversity on Sulawesi Island is lower than it is in the Philippines, but Sulawesi supports a surprising diversity of Sulawesi fanged frogs (Limnonectes). Here we examine molecular, morphological, and geographical variation of fanged frogs from these two regions. Using genealogical concordance, morphology, and a Bayesian approach to species delimitation, we identified 13 species on Sulawesi, only four of which have been previously described. After evolutionary history is accounted for, a model with multiple body size optima in sympatric species is favored over a "random-walk" model of body size evolution. Additionally, morphological variation is higher among sympatric than nonsympatric species on Sulawesi but not in the Philippines. These findings suggest that adaptive radiation of fanged frogs on Sulawesi was driven by natural selection to infiltrate ecological niches occupied by other frog lineages in the Philippines. This supports a role of ecological opportunity in community assembly: diversification in mature communities, such as the Philippines, is limited by a dearth of unoccupied ecological niches. On Sulawesi, evolutionary novelties originated in a predictable and replicated fashion in response to opportunities presented by a depauperate ancestral community.     

Loss of dendritic cell migration and impaired resistance to Leishmania donovani infection in mice deficient in CCL19 and CCL21

The encounter between APC and T cells is crucial for initiating immune responses to infectious microorganisms. In the spleen, interaction between dendritic cells (DC) and T cells occurs in the periarteriolar lymphoid sheath (PALS) into which DC and T cells migrate from the marginal zone (MZ) along chemokine gradients. However, the importance of DC migration from the MZ into the PALS for immune responses and host resistance to microbial infection has not yet been elucidated. In this study, we report that following Leishmania donovani infection of mice, the migration of splenic DC is regulated by the CCR7 ligands CCL19/CCL21. DC in plt/plt mutant mice that lack these chemokines are less activated and produce less IL-12, compared with those in wild-type mice. Similar findings are seen when mice are treated with pertussis toxin, which blocks chemokine signaling in vivo. plt/plt mice had increased susceptibility to L. donovani infection compared with wild-type mice, as determined by spleen and liver parasite burden. Analysis of splenic cytokine profiles at day 14 postinfection demonstrated that IFN-gamma and IL-4 mRNA accumulation was comparable in wild-type and plt/plt mice. In contrast, accumulation of mRNA for IL-10 was elevated in plt/plt mice. In addition, plt/plt mice mounted a delayed hepatic granulomatous response and fewer effector T cells migrated into the liver. Taken together, we conclude that DC migration from the MZ to the PALS is necessary for full activation of DC and the optimal induction of protective immunity against L. donovani.     

Isolation and molecular characterization of causal agent of blue mold on Allium cepa L. and its control by Pennisetum flaccidum Griseb

Blue mold pathogen, isolated from infected Allium cepa L., was identified as a Penicillium species through morphological and molecular characterisation. Internal Transcribed spacer (ITS) region of ribosomal DNA (rDNA) was utilised for DNA sequencing. Basic Local Alignment Search Tool (BLAST) analysis has found the maximum similarity index of the fungus to be 82.39% with the Uncultured Penicillium clone (Accession: MF535522). So, the isolated Penicillium specie is the first reported specie of the genus that infects onion. A phylogenetic tree was constructed to establish a relationship of the isolated fungus with the most relevant species reported on GenBank. Extracts of Pennisetum flaccidum Griseb. were evaluated against the isolated fungus as a potential biocontrol agent. Among the five tested methanol concentrations (0.5%, 1%, 1.5%, 2% and 2.5%) of each plant part (root, inflorescence and foliage), 0.5% root extract showed maximum growth retardation, i.e. 89%. For bioassay-guided fractionation, the root extract was partitioned in n-hexane, chloroform, n-butanol and ethyl acetate. Ethyl acetate (1%) was proved to be the most potent one. Phytochemical screening has confirmed the occurrence of terpenoids, tannins, saponins and alkaloids. The applied molecular approach has deduced that the Penicillium specie collected from Pakistan might be novel. This study can be concluded that P. flaccidum contains potent phytochemicals which might be used as antifungal agent against Penicillium species.     

Polyesters Based on Linoleic Acid for Biolubricant Basestocks: Low-Temperature,Tribological and Rheological Properties

Presently, plant oils which contain high percentage of linoleic acid 1 are perceived to be a viable alternative to mineral oil for biolubricant applications due to their biodegradability and technical properties. In order to get biodegradable lubricant, triester derivatives compounds (1–5) were synthesized and characterized. The processes involved were monoepoxidation of linoleic acid 2, oxirane ring opening 3, esterification 4 and acylation 5. The structures of the products were confirmed by FTIR, ¹H and ¹³C-NMR and LC-MS. The results that showed lowest temperature properties were obtained for triester 5, with a pour point value (PP) of -73°C, highest onset temperature (260°C) and lowest volatility at 0.30%. Viscosity index (VI) increased for the ester’s synthetic compounds (2, 3, 4, 5), while the PP decreased. This behavior is the result of the increase of the chain length of the branching agents. Triester based linoleic acid has improved properties such as low-temperature and tribological properties. These results will make it feasible for plant oil to be used for biolubricants, fuels in chain saws, transmission oil and brake fluid.     

The pick-up of C14-DDT at different temperatures

Mosquito larvae, Aedes aegypti, picked up progressively greater amounts of C¹⁴ labeled DDT with ascending temperatures. The pick-up relationship contrasted with per cent mortality which showed a negative temperature coefficient. The exposure temperatures were 10°, 20° and 30° C. A similar relationship of pick-up of DDT occurred when heads, thoraces, or abdomens of the larvae were compared separately. The thorax contained greater concentrations than heads or abdomens at 30° and 20° C. At 10° C the heads contain more than thoraces and abdomens, but the amount was still less than that picked up at 20° and 30° C.The results show a positive coefficient of pick-up of DDT but a negative temperature coefficient for per cent mortality provided the concentration of DDT is not too high. The latter relationship agrees with considerable earlier research. The explanation for the negative temperature effect of DDT is still not understood, but we have evidence that the effect is not positively related to pick-up by whole larvae or portions of whole larvae.

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Zusammenfassung Mückenlarven (Aedes aegypti) nahmen um so größere Mengen von ¹⁴C-markiertem DDT auf, je mehr die Temperatur anstieg. Im Gegensatz dazu zeigte die prozentuale Mortalität einen negativen Temperaturkoeffiziente. Die Untersuchungstemperaturen betrugen 10, 20 und 30°. Eine ähnliche DDT-Aufnahme-Beziehung ergab sich, wenn Köpfe, Brustabschnitte oder Abdomina der Larven getrennt verglichen wurden. Bei 30 und 20° wiesen die Brustabschnitte größere Konzentrationen auf als die Köpfe und Abdomina. Bei 10° enthielten die Köpfe mehr als die Abdomina und Brustabschnitte, aber die aufgenommene Menge war doch geringer als bei 20 und 30°.Die Ergebnisse zeigen einen positiven Temperaturkoeffizienten für die DDT-Aufnahme, aber einen negativen für die prozentuale Mortalität unter der Voraussetzung, daß die DDT-konzentration nicht zu hoch ist. Die zweite Beziehung stimmt mit beträchtlich früheren Untersuchungen überein. Eine Erklärung für diesen negativen Temperatureffekt des DDT ist noch nicht gefunden, aber es scheint, daß diese Wirkung keine positive Korrelation zur DDT-Aufnahme ganzer Larven oder bei Teilen ganzer Larven aufweist.

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Paper No. 5188 Scientific Journal Series, Minnesota Agricultural Experiment Station, St. Paul 1, Minnesota.     

Effect of pyruvate and oleate on respiration of frog skeletal and heart muscle.

The authors studied the effect of pyruvate and oleate on O2 consumption of the frog sartorius and heart ventricle. 10 mM pyruvate raised O2 consumption of both tissues by over 100%, but only in the winter. Raised O2 consumption was only partly associated with glycogen synthesis from pyruvate. 0.1 mM oleate reduced O2 consumption in both tissues. A marked drop in O2 consumption was observed in the ventricle (up to 50%). Lecithin had a similar effect on O2 consumption. The addition of pyruvate plus oleate led to an 8-fold increase in O2 consumption of the ventricle, i.e. to maximum oxidation capacity of the tissue, but the addition of lecithin inhibited the pyruvate-induced increase in O2 consumption. It is assumed that both pyruvate and oleate influence resting metabolism in a specific manner which cannot be attributed solely to raised availability of substrate for resting energy metabolism requirements.