Lovastatin and (+)‐geodin production by Aspergillus terreus from crude glycerol

The use of pure substrate represents a significant proportion of the cost of manufacturing a drug such as lovastatin. This study explores the production of lovastatin and (+)‐geodin by Aspergillus terreus ATCC 20542 using biodiesel‐derived crude glycerol (CG) as a feedstock. Shake flask experiments showed reduced lovastatin production and glycerol consumption in the presence of 10–50 g/L CG with respect to pure glycerol controls. At 50 g/L, lovastatin and (+)‐geodin production was significantly reduced by 82 and 73%, respectively. The lowest lovastatin inhibition was detected in 30 g/L of CG (48%), which was accompanied by a significant rise in (+)‐geodin production (338%). Further investigation was performed on three major impurities found in CG, namely methanol (MeOH), sodium chloride (NaCl), and fatty acids (oleic acid and palmitic acid (PA), soap). None was particularly inhibitory for lovastatin, except soap and PAs, which reduced its production by more than 50% at all concentrations tested. In contrast, (+)‐geodin was inhibited in the presence of MeOH and PA by up to 46 and 91%, respectively. These observations indicate that partial purification of CG would be potentially useful in improving production of lovastatin and (+)‐geodin by A. terreus.     

Plasmodium vivax Drug Resistance Genes; Pvmdr1 and Pvcrt-o Polymorphisms in Relation to Chloroquine Sensitivity from a Malaria Endemic Area of Thailand

The aim of the study was to explore the possible molecular markers of chloroquine resistance in Plasmodium vivax isolates in Thailand. A total of 30 P. vivax isolates were collected from a malaria endemic area along the Thai-Myanmar border in Mae Sot district of Thailand. Dried blood spot samples were collected for analysis of Pvmdr1 and Pvcrt-o polymorphisms. Blood samples (100 μl) were collected by finger-prick for in vitro chloroquine susceptibility testing by schizont maturation inhibition assay. Based on the cut-off IC₅₀ of 100 nM, 19 (63.3%) isolates were classified as chloroquine resistant P. vivax isolates. Seven non-synonymous mutations and 2 synonymous were identified in Pvmdr1 gene. Y976F and F1076L mutations were detected in 7 (23.3%) and 16 isolates (53.3%), respectively. Analysis of Pvcrt-o gene revealed that all isolates were wild-type. Our results suggest that chloroquine resistance gene is now spreading in this area. Monitoring of chloroquine resistant molecular markers provide a useful tool for future control of P. vivax malaria.     

Nitrogen excretion by the sheep abomasal parasite Teladorsagia circumcincta

Excretion of nitrogenous substances by Teladorsagia circumcincta was investigated during incubation of L3 in phosphate buffer for up to 30 h and adult worms for 4-6 h. Ammonia was the main excretory product, with about 20% urea. For the first 4-6 h, ammonia excretion by L3 was temperature dependent, directly proportional to the number of larvae, but independent of the pH or strength of the phosphate buffer. Later, ammonia excretion slowed markedly in L3 and adults and reversed to net uptake in L3 by 30 h. An initial external ammonia concentration of 600 μM did not alter the pattern or magnitude of excretion. Re-uptake of ammonia did not occur at extremes of pH or low buffer strength and was slightly reduced at the highest external concentrations. Ammonium transporters and enzymes of glutamate metabolism, including glutamate dehydrogenase, glutamine synthetase and possibly glutamate synthase, are worthy of further investigation as anthelmintic targets.     

Rational use of Jatropha curcas L. in food and medicine: from toxicity problems to safe applications

Jatropha curcas L. has become an important plant for biorefinery and production of biodiesel. From its ethnobotanical use, the plant is known for several activities which are associated with high toxicity. The latest development in engineering technology enables detoxification of native oil and other parts of the plant for new pharmaceutical purposes. Hence a revised look to the rich metabolic spectra of partly structurally rare secondary compound becomes an interesting field of research to be explored. In this review, we discuss recent developments in the technology of detoxification process and give insight about how this ethnomedicinal plant can be applied to new fields of experimental medicine. The review highlights recent data on biological activities and discusses concepts and strategies for turning a poison plant into a valuable crop with high pharmaceutical potential.     

Genetic isolation between the Western and Eastern Pacific populations of pronghorn spiny lobster Panulirus penicillatus

The pronghorn spiny lobster, Panulirus penicillatus, is a circumtropical species which has the widest global distribution among all the species of spiny lobster, ranging throughout the entire Indo-Pacific region. Partial nucleotide sequences of mitochondrial DNA COI (1,142-1,207 bp) and 16S rDNA (535-546 bp) regions were determined for adult and phyllosoma larval samples collected from the Eastern Pacific (EP)(Galápagos Islands and its adjacent water), Central Pacific (CP)(Hawaii and Tuamotu) and the Western Pacific (WP)(Japan, Indonesia, Fiji, New Caledonia and Australia). Phylogenetic analyses revealed two distinct large clades corresponding to the geographic origin of samples (EP and CP+WP). No haplotype was shared between the two regional samples, and average nucleotide sequence divergence (Kimura's two parameter distance) between EP and CP+WP samples was 3.8±0.5% for COI and 1.0±0.4% for 16S rDNA, both of which were much larger than those within samples. The present results indicate that the Pacific population of the pronghorn spiny lobster is subdivided into two distinct populations (Eastern Pacific and Central to Western Pacific), with no gene flow between them. Although the pronghorn spiny lobster have long-lived teleplanic larvae, the vast expanse of Pacific Ocean with no islands and no shallow substrate which is known as the East Pacific Barrier appears to have isolated these two populations for a long time (c.a. 1MY).     

Virological surveillance of human influenza in Indonesia, October 2008-March 2010

Despite the high prevalence of highly pathogenic H5N1 influenza A viruses in Indonesia, epidemiology information on seasonal human influenza is lacking. The present authors, therefore, conducted virologic surveillance in Surabaya, East Java from October 2008 to March 2010. Influenza viruses, including pandemic (H1N1) 2009 viruses, were isolated from 71 of 635 individuals tested. Seasonal influenza peaked in the rainy season. Compared with seasonal influenza viruses, pandemic 2009 viruses were isolated from younger patients with milder symptoms. Given the high prevalence of H5N1 infections in humans, continued influenza surveillance is essential for pandemic preparedness.     

Sequential action of ATP-dependent subunit conformational change and interaction between helical protrusions in the closure of the built-in lid of group II chaperonins

ATP drives the conformational change of the group II chaperonin from the open lid substrate-binding conformation to the closed lid conformation to encapsulate an unfolded protein in the central cavity. The detailed mechanism of this conformational change remains unknown. To elucidate the intra-ring cooperative action of subunits for the conformational change, we constructed Thermococcus chaperonin complexes containing mutant subunits in an ordered manner and examined their folding and conformational change abilities. Chaperonin complexes containing wild-type subunits and mutant subunits with impaired ATP-dependent conformational change ability or ATP hydrolysis activity, one by one, exhibited high protein refolding ability. The effects of the mutant subunits correlate with the number and order in the ring. In contrast, the use of a mutant lacking helical protrusion severely affected the function. Interestingly, these mutant chaperonin complexes also exhibited ATP-dependent conformational changes as demonstrated by small angle x-ray scattering, protease digestion, and changes in fluorescence of the fluorophore attached to the tip of the helical protrusion. However, their conformational change is likely to be transient. They captured denatured proteins even in the presence of ATP, whereas addition of ATP impaired the ability of the wild-type chaperonin to protect citrate synthase from thermal aggregation. These results suggest that ATP binding/hydrolysis causes the independent conformational change of the subunit, and further conformational change for the complete closure of the lid is induced and stabilized by the interaction between helical protrusions.     

Evaluation of scanning resolution,detector choice and detector orientation to be used for accurate and time-efficient commissioning of a 6MV clinical linear accelerator

The present study is aimed at exploring different scanning parameters, detectors and their orientations for time-efficient and accurate commissioning of a 6 MV clinical linear accelerator (LINAC). Beam profiles and percentage depth dose (PDD) curves were measured with a PTW dosimetry diode, a PTW Semiflex and a PinPoint ion chamber in different orientations. To acquire beam data, equidistant (step size of 0.5 mm, 1 mm, 2 mm and 3 mm) and fanline (step size of 2–0.5 mm, 2–1 mm, 3–0.5 mm and 3-1 mm) scanning modes were employed and data measurement time was recorded. Scan time per measurement point was also varied (0.2 s, 0.5 s and 1.0 s) to investigate its effect on the accuracy and acquisition time of beam data. Accuracy of the measured data was analyzed on the basis of the variation between measured data and data modeled by a treatment planning system. Beam profiles (particularly in penumbra region) were found to be sensitive to variation in scanning resolution and showed an improved accuracy with decrease in step size, while PDD curves were affected negligibly. The accuracy of beam data obtained with the PTW dosimetry diode and the PinPoint ion chamber was higher than those obtained with the PTW Semiflex ion chamber for small fields (2?×?2 cm² and 3?×?3 cm²). However, the response of the PTW diode and the PinPoint ion chamber was significantly indifferent in these fields. Furthermore, axial orientation of the PTW Semiflex ion chamber improved accuracy of profiles and PDDs as compared to radial orientation, while such a difference was not significant for the PinPoint ion chamber. It is concluded that a scan time of 0.2 s/point with a fanline scanning resolution of 2–1 mm for beam profiles and 3 mm for PDDs are most favorable in terms of accuracy and time efficiency. For small fields (2?×?2 cm² and 3?×?3 cm²), a PinPoint ion chamber in radial orientation or a dosimetry diode in axial orientation are recommended for both beam profiles and PDDs. If a PinPoint ion chamber and a PTW dosimetry diode are not available, a Semiflex ion chamber in axial orientation may be used for small fields.