Potential correlation between carboxylic acid metabolites in Biomphalaria alexandrina snails after exposure to Schistosoma mansoni infection

Carboxylic acids play an important role in both aerobic and anaerobic metabolic pathways of both the snail and the parasite. Monitoring the effects of infection by schistosome on Biomphalaria alexandrina carboxylic acids metabolic profiles represents a promising additional source of information about the state of metabolic system. We separated and quantified pyruvic, fumaric, malic, oxalic, and acetic acids using ion-suppression reversed-phase high performance liquid chromatography (HPLC) to detect correlations between these acids in both hemolymph and digestive gland gonad complex (DGG's) samples in a total of 300 B. alexandrina snails (150 infected and 150 controls) at different stages of infection. The results showed that the majority of metabolite pairs did not show significant correlations. However, some high correlations were found between the studied acids within the control group but not in other groups. More striking was the existence of reversed correlations between the same acids at different stages of infection. Some possible explanations of the underlying mechanisms were discussed. Ultimately, however, further data are required for resolving the responsible regulatory events. These findings highlight the potential of metabolomics as a novel approach for fundamental investigations of host-pathogen interactions as well as disease surveillance and control.     

Synthesis of diethylaminoethyl dextran hydrogel and its heavy metal ion adsorption characteristics

Epichlorohydrin-crosslinked diethylaminoethyl dextran (DEAE-D/ECH) hydrogel was synthesized by intermolecular side-chain reaction of DEAE-D hydroxyl groups with monomeric crosslinking agent, ECH. Swelling ability, adsorption capacity and metal removal of the hydrogel were profoundly determined and some structural parameters for the hydrogel such as volume of non-swollen gel, percentages of gellation, swelling ratio and equilibrium water content were evaluated in this study. The ability of removing heavy metal ions from Orontes River by the synthesized hydrogel, thoroughly characterized by photometric spectrometer and the adsorption characteristics of metal ions, was investigated as well as surface morphologies of the hydrogel before and after metal adsorption were examined by SEM. Structure of DEAE-D/ECH gel was analyzed by FTIR, TGA, and DSC. Gellation point of binary system reaction between DEAE-D and ECH was determined via monitoring viscosity changes during reaction. The order of affinity based on amount of metal ion uptake was found as follows: Zn(2+)>Mn(2+)>Pb(2+)>Cd(2+).     

Mechanism of Action of Recombinant Acc-Royalisin from Royal Jelly of Asian Honeybee against Gram-Positive Bacteria

The antibacterial activity of royalisin, an antimicrobial peptide from the royal jelly produced by honeybees, has been addressed extensively. However, its mechanism of action remains unclear. In this study, a recombinant royalisin, RAcc-royalisin from the royal jelly of Asian honeybee Apis cerana cerana, was expressed by fusing with glutathione S-transferase (GST) in Escherichia coli BL21, isolated and purified. The agar dilution assays with inhibition zone showed that RAcc-royalisin, similar to nisin, inhibits the growth of Gram-positive bacteria. The antibacterial activity of RAcc-royalisin was associated with its concentration, and was weakened by heat treatment ranging from 55°C to 85°C for 15 min. Both RAcc-royalisin and nisin exhibited the minimum inhibitory concentrations (MIC) of 62.5 µg/ml, 125 µg/ml, and 250 µg/ml against Gram-positive bacterial strains, Bacillus subtilis and Micrococcus flavus and Staphyloccocus aureus in the microplate assay, respectively. However, RAcc-royalisin did not show antimicrobial activity against tested Gram-negative bacterial and fungal strains. The antibacterial activity of RAcc-royalisin agrees well with the decrease in bacterial cell hydrophobicity, the leakage of 260-nm absorbing materials, and the observation by transmission electron microscopy, all indicating that RAcc-royalisin induced the disruption and dysfunction of cell walls and membranes. This is the first report detailing the antibacterial mechanism of royalisin against Gram-positive bacteria, and provides insight into the application of recombinant royalisin in food and pharmaceutical industries as an antimicrobial agent.     

Anatomical and micromorphological studies on <Emphasis Type="Italic">Teucrium</Emphasis> sect. <Emphasis Type="Italic">Isotriodon</Emphasis> (Lamiaceae) in Turkey with a taxonomic note

In this study, the anatomical features of the leaf and stem, besides the nutlet characteristics of some Teucrium sect. Isotriodon (Lamiaceae) taxa in Turkey, T. montbretii Betham subsp. montbretii, T. montbretii subsp. pamphylicum P. H. Davis, T. odontites Boiss. &; Bal., T. cavernarum P. H. Davis, T. antitauricum T. Ekim, along with an isolated population of T. montbretii (T. montbretii subsp.) were investigated. The anatomical studies revealed that the taxa share generally similar anatomical characters, such as thicker upper leaf cuticles and larger upper leaf epidermal cells compared to lower ones and diacytic to anomocytic stomata on the leaves. However, the portion of the mesophyll occupied by palisade parenchyma and the occurrence of mucilage cells in leaf epidermis shows difference among the taxa. Furthermore, the studied taxa have general stem characteristics of the Lamiaceae family, except for having poorly developed collenchyma at the corners. With the amphistomatic leaves and developed sclerenchymatic tissue in the leaf median vein, T. cavernarum is seperated from the other taxa. Trichome types on the vegetative organs and nutlet shape and sculpturing are generally the same or similar in the studied taxa, but trichomes on the nutlets are different among them. Based on nutlet characteristics and some morphological ones, it was revealed that the isolated population of T. montbretii represent a new subspecies, T. monbretii subsp. yildirimlii M.Dinç &; S.Do?u subsp. nov.     

Analysis of the Plasmodium and Anopheles transcriptional repertoire during ookinete development and midgut invasion

Plasmodium, the causative agent of malaria, has to undergo sexual differentiation and development in anopheline mosquitoes for transmission to occur. To isolate genes specifically induced in both organisms during the early stages of Plasmodium differentiation in the mosquito, two cDNA libraries were constructed, one enriched for sequences expressed in differentiating Plasmodium berghei ookinetes and another enriched for sequences expressed in Anopheles stephensi guts containing invading ookinetes and early oocysts. Sequencing of 457 ookinete library clones and 652 early oocyst clones represented 175 and 346 unique expressed sequence tags, respectively. Nine of 13 Plasmodium and four of the five Anopheles novel expressed sequence tags analyzed on Northern blots were induced during ookinete differentiation and mosquito gut invasion. Ancaspase-7, an Anopheles effector caspase, is proteolytically activated during Plasmodium invasion of the midgut. WARP, a gene encoding a Plasmodium surface protein with a von Willebrand factor A-like adhesive domain, is expressed only in ookinetes and early oocysts. An anti-WARP polyclonal antibody strongly inhibits (70-92%) Plasmodium development in the mosquito, making it a candidate antigen for transmission blocking vaccines. The present results and those of an accompanying report (Srinivasan, P., Abraham, E. G., Ghosh, A. K., Valenzuela, J., Ribeiro, J. M. C., Dimopoulos G., Kafatos, F. C., Adams, J. H., and Jacobs-Lorena, M. (2004) J. Biol. Chem. 279, 5581-5587) provide the foundation for further analysis of Plasmodium differentiation in the mosquito and of mosquito responses to the parasite.     

Temporal alterations of <Emphasis Type="Italic">Nannochloropsis</Emphasis><Emphasis Type="Italic">salina</Emphasis> (Eustigmatophyceae) grown under aqueous diesel fuel stress

The influence of an aqueous extract of diesel fuel was tested on growth of the marine eustigmatophyte Nannochloropsis (Monallantus) salina Hibberd. An increase in the concentration of the pollutant led to a decrease in growth rate as measured by optical density, with maximum effect observed (33% of control) at 100% aqueous pollutant. Spectrophotometric examination of cell viability (using Evan’s blue dye) showed a significant negative effect of the diesel extract (p ≤ 0.05, r = −0.92). Infrared spectra showed a slight change in the absorbance of contaminated compared with controlled cells. Proteome analysis (sodium dodecyl sulfate polyacrylamide gel electrophoresis – “SDS-PAGE”) indicated that cell protein profiles depended on the pollutant concentration. Some of the resultant bands were characteristic to the pollutant concentration applied, indicating a distinct effect of the pollutant on the proteome structure. Iodine and toluidine blue dyes were applied using light microscopy to detect starch and mucilage, respectively. This indicated the presence of starch during all treatments, while the mucilage has been reduced. Transmission electron microscopy showed alterations to cell walls and membranes with different degrees of plasmolysis leading to a gradual increase in cell volume. However, the nucleus, the nucleolus and the pyrenoid remained unaffected. Similar results were obtained when the alga was cultured for 25 days in the 100% aqueous diesel extract indicating that long-term culture does not affect the degree of pollutant stress. Further, these cells recovered their normal appearance and characteristics within two days of being transferred to culture medium free of extract, indicating that N. salina shows a high tolerance to aqueous diesel fuel pollution.     

Gene diversity in some Muslim populations of North India

North Indian Muslim populations have historical, linguistic, and socioreligious significance to the Indian subcontinent. Although sociocultural and political dimensions of their demography are well documented, no detailed genetic structure of the populations is available. We have undertaken a survey of the gene frequencies of the ABO, Rh, PTC taste ability, sickling, and G6PD systems for different endogamous groups: Sheikh, Syed, Pathan, Ansari, Saifi, and Hindu Bania. All the groups at most loci showed statistically nonsignificant differences, except for ABO and PTC traits, for which interpopulational differences were seen. Heterozygosity ranged from 0.048 to 0.617 among the Sheikh, 0.149 to 0.599 among the Pathan, 0.105 to 0.585 among the Ansari, 0.25 to 0.869 among the Syed, 0.107 to 0.565 among the Saifi, and 0.100 to 0.492 among the Hindu Bania. The average D(ST) and G(ST) values for the five marker loci were 0.0625 +/- 0.098 and 0.1072 +/- 0.041, respectively. A dendrogram was constructed using the UPGMA clustering method. Our results revealed that the Pathan and the Sheikh form one cluster, the Syed and the Hindu Bania form another cluster, and the two clusters join together (the so-called higher caste); also, the Saifi and the Ansari form a separate cluster (lower caste). The results of the genetic distance analysis are useful for understanding the pattern of genetic relationships between different endogamous groups of Muslims.     

Gastroretentive Microsponge as a Promising Tool for Prolonging the Release of Mitiglinide Calcium in Type-2 Diabetes Mellitus: Optimization and Pharmacokinetics Study

Diabetes mellitus is one of the leading causes of death due to the persistent hyperglycemia that leads to potential complications. Lack of patients’ adherence to their prescribed medication regimens, due to the requirement of frequent dosing, leads to failure of 40–50% of patients to manage their disease. Thus, microsponges of the novel short half-life mitiglinide calcium (MTG) were formulated using Quasi-emulsion solvent diffusion method, employing Eudragit RS100, ethyl cellulose, and polyvinyl alcohol, then characterized in terms of production yield, entrapment efficiency, particle size, in vitro buoyancy, in vitro drug release, and in vivo pharmacokinetics in rabbits. Optimization was done using response surface methodology; the optimized formulation was investigated by FTIR, DSC, and SEM. Results revealed that the optimized MTG microsponge was successfully formulated with high production yield (61.61%?±?0.6), entrapment efficiency (77.7% ±1.37), and particle size of 192.76 μm and it remained buoyant over simulated gastric fluid for 24 h with high percentage of in vitro buoyancy (91.01%?±?2.5). Moreover, it sustained the in vitro drug release with cumulative % release of 83.74?±?1.5 after 24 h. This microsponge was highly porous in nature with interconnected pores where MTG was entrapped with good compatibility as confirmed by SEM, DSC, and FTIR analysis; Pharmacokinetic studies showed improvement in C_max and AUC_0-∞ (1.92- and 20.68-fold, respectively) with marked prolongation in MRT and t_1/2 (7.22- and 7.97-fold, respectively) than the marketed tablet. Thus, it is a promising approach to improve diabetic patients’ compliance by eliminating the necessity of frequent dosing thus attaining better diabetes control.