Genome‐wide mosaicism in divergence between zoonotic malaria parasite subpopulations with separate sympatric transmission cycles

Plasmodium knowlesi is a significant cause of human malaria transmitted as a zoonosis from macaque reservoir hosts in South‐East Asia. Microsatellite genotyping has indicated that human infections in Malaysian Borneo are an admixture of two highly divergent sympatric parasite subpopulations that are, respectively, associated with long‐tailed macaques (Cluster 1) and pig‐tailed macaques (Cluster 2). Whole‐genome sequences of clinical isolates subsequently confirmed the separate clusters, although fewer of the less common Cluster 2 type were sequenced. Here, to analyse population structure and genomic divergence in subpopulation samples of comparable depth, genome sequences were generated from 21 new clinical infections identified as Cluster 2 by microsatellite analysis, yielding a cumulative sample size for this subpopulation similar to that for Cluster 1. Profound heterogeneity in the level of intercluster divergence was distributed across the genome, with long contiguous chromosomal blocks having high or low divergence. Different mitochondrial genome clades were associated with the two major subpopulations, but limited exchange of haplotypes from one to the other was evident, as was also the case for the maternally inherited apicoplast genome. These findings indicate deep divergence of the two sympatric P. knowlesi subpopulations, with introgression likely to have occurred recently. There is no evidence yet of specific adaptation at any introgressed locus, but the recombinant mosaic types offer enhanced diversity on which selection may operate in a currently changing landscape and human environment. Loci responsible for maintaining genetic isolation of the sympatric subpopulations need to be identified in the chromosomal regions showing fixed differences.     

Diet-induced obesity suppresses ghrelin in rat gastrointestinal tract and serum

The aims of the present study were to examine ghrelin expression in serum and gastrointestinal tract (GIT) tissues, and to measure tissue ghrelin levels and obesity-related alterations in some serum biochemical variables in rats with diet-induced obesity (DIO). The study included 12 male rats, 60 days old. The rats were randomly allocated to two groups (n = 6). Rats in the DIO group were fed a cafeteria-style diet to induce obesity, while those in the control group were fed on standard rat pellets. After a 12 week diet program including an adaptation period all rats were decapitated, tissues were individually fixed, ghrelin expression was examined by immunohistochemistry , and tissue and serum ghrelin levels were measured by radioimmunoassay. Serum biochemical variables were measured using an autoanalyzer. When the baseline and week 12 body mass index and GIT ghrelin expression were compared between DIO and control rats, BMI had increased and ghrelin expression decreased due to obesity. The RIA results were consistent with these findings. Serum glucose, LDL cholesterol, and total cholesterol levels were elevated and HDL cholesterol significantly decreased in the DIO group. A comparison of GIT tissues between the control and obese groups demonstrated that ghrelin was decreased in all tissues of the latter. This decrease was brought about a decline in the circulating ghrelin pool. This suggests that rather than being associated with a change in a single tissue, obesity is a pathological condition in which ghrelin expression is changed in all tissues.     

Effects of Genotype on Testis, Semen Quality, and Mineral Composition of Semen in Various Ram Breeds

This study was carried out on nine 3–5-year-old rams of different species: four Norduz, two Karakas, and three Ile de France?×?Akkaraman rams. Sperm of each ram was collected by means of an artificial vagina once per month from June to October 2007. In each case, the sperm volume, pH, viscosity, mass activity, motility, concentration, abnormal spermatozoa ratio, and the sperm death/live ratio were recorder along with the animals’ testis diameter and scrotum circumferences. The concentrations of calcium, potassium, copper, magnesium, iron, and zinc levels in seminal plasma were determined. There were changes in the measured parameters according to genotype and time of sampling, which can be useful for better planning of anestrous mating programs for these ram species.     

Bacterial ferritin contains 24 haem groups

Pseudomonas aeruginosa bacterioferritin, also known as cytochrome b1 or cytochrome b557, has been isolated with 9 haems per 24 subunits. Various forms of the protein have been prepared including the completely haem-free protein and the fully haem-loaded protein with 24 haems per 24 subunits. The presence of the core does not significantly affect haem addition or removal. The absorbance ratio of the non-haem-iron-loaded protein, 278 nm:417 nm (oxidised), can be used to estimate the haem loading.     

Selenium Levels in First-Degree Relatives of Diabetic Patients

The present study was conducted to evaluate the serum selenium levels in first-degree relatives of diabetic patients (FDR) according to controls. Insulin resistance, serum lipid levels, inflammation markers, and blood pressure were also studied in these patients. Serum levels of selenium in FDR were significantly lower than control group (74.65 ± 5.9 vs 88.7 ± 8.7 μg/dl, p < 0.0001). HsCRP, HOMA-IR, insulin, homocysteine levels were significantly higher in FDR according to the control group (1.32 ± 0.9 vs 0.63 ± 0.4 mg/dL, p < 0.0001; 2.07 ± 0.84 vs 1.51 ± 0.69, p < 0.0001; 9.26 ± 3.8 vs 6.8 ± 2.98 μU/MI, p < 0.0001; 15.7 ± 7.4 vs 11.5 ± 5.1 μmol/L, p < 0.0001, respectively). There was significant correlation between selenium levels and hsCRP (r = − 0.450, p < 0.0001). There was also weak significant correlation also between HOMA-IR and selenium levels (r = −0.227, p = 0.003). There was a correlation between systolic blood pressure and BMI (r = 0.365, p < 0.0001). But there was no correlation between selenium levels and blood pressure or other parameters. HsCRP, HOMA-IR, homocysteine levels in individuals with selenium levels < 80 μg/L (n = 78) was significantly higher than hsCRP HOMA-IR, homocysteine levels in individuals with selenium levels ≥ 80 (n = 91; 1.23 ± 0.98 vs 0.81 ± 0.76 mg/dL, p < 0.003; 1.99 ± 0.88 vs 1.64 ± 0.74, p < 0.005; 15.0 ± 7.6 vs 12.9 ± 5.7 μmol/L, p < 0.049, respectively). Selenium deficiency may contribute to cardiovascular disease risk in FDR.     

Efficacy of selected adjuvants for protecting Spodoptera litura nucleopolyhedrovirus from sunlight inactivation

Efficacy of selected adjuvants for protecting Spodoptera litura nucleopolyhedrovirus (SpltNPV) from sunlight inactivation was tested in direct sunlight exposure bioassays and in the field on virus-treated Brassica chinensis var parachinensis plants. All adjuvants, except palm oil, gave significant protection to the virus after 16 h of sunlight exposure, as shown by the larval mortalities. Cumulative mortalities after 14 days show that the virus + Tinopal treatment and the virus not to sunlight treatment attained 100% mortality, while the virus in riboflavin treatment and the crude brown sugar treatment caused mortality of less than 30%. However, the LT₅₀ value of virus + Tinopal was 1.2 times lower than the unexposed virus. All virus-treated plants in the field trials had higher yields and lower damage indices than the control plants. Unlike the results from direct sunlight exposure bioassays, adjuvants, including virus + Tinopal LPW, did not significantly enhance efficacy of the virus against early instars of S. litura in the field. The unformulated virus was as effective as virus + Tinopal LPW and Nurelle-D in protecting the plants from infestation. Thus, in a well-planned pest management program, SpltNPV, without any adjuvant, can be used effectively for controlling S. litura on B. chinensis var parachinensis.     

Interaction of a tyrosine kinase inhibitor,vandetanib with human serum albumin as studied by fluorescence quenching and molecular docking

Interaction of a tyrosine kinase inhibitor, vandetanib (VDB), with the major transport protein in the human blood circulation, human serum albumin (HSA), was investigated using fluorescence spectroscopy, circular dichroism (CD) spectroscopy, and molecular docking analysis. The binding constant of the VDB–HSA system, as determined by fluorescence quenching titration method was found in the range, 8.92–6.89?×?10^3?M^?1 at three different temperatures, suggesting moderate binding affinity. Furthermore, decrease in the binding constant with increasing temperature revealed involvement of static quenching mechanism, thus affirming the formation of the VDB–HSA complex. Thermodynamic analysis of the binding reaction between VDB and HSA yielded positive ΔS (52.76 J?mol^?1 K^?1) and negative ΔH (?6.57?kJ?mol^?1) values, which suggested involvement of hydrophobic interactions and hydrogen bonding in stabilizing the VDB–HSA complex. Far-UV and near-UV CD spectral results suggested alterations in both secondary and tertiary structures of HSA upon VDB-binding. Three-dimensional fluorescence spectral results also showed significant microenvironmental changes around the Trp residue of HSA consequent to the complex formation. Use of site-specific marker ligands, such as phenylbutazone (site I marker) and diazepam (site II marker) in competitive ligand displacement experiments indicated location of the VDB binding site on HSA as Sudlow’s site I (subdomain IIA), which was further established by molecular docking results. Presence of some common metal ions, such as Ca²⁺, Zn²⁺, Cu²⁺, Ba²⁺, Mg²⁺, and Mn²⁺ in the reaction mixture produced smaller but significant alterations in the binding affinity of VDB to HSA.     

Comparative analysis of realistic CT-scan and simplified human airway models in airflow simulation

Efforts to model the human upper respiratory system have undergone many phases. Geometrical proximity to the realistic shape has been the subject of many research projects. In this study, three different geometries of the trachea and main bronchus were modelled, which were reconstructed from computed tomography (CT) scan images. The geometrical variations were named realistic, simplified and oversimplified. Realistic refers to the lifelike image taken from digital imaging and communications in medicine format CT scan images, simplified refers to the reconstructed image based on natural images without realistic details pertaining to the rough surfaces, and oversimplified describes the straight wall geometry of the airway. The characteristics of steady state flows with different flow rates were investigated, simulating three varied physical activities and passing through each model. The results agree with previous studies where simplified models are sufficient for providing comparable results for airflow in human airways. This work further suggests that, under most exercise conditions, the idealised oversimplified model is not favourable for simulating either airflow regimes or airflow with particle depositions. However, in terms of immediate analysis for the prediction of abnormalities of various dimensions of human airways, the oversimplified techniques may be used.     

Cryopreservation of protocorm-like bodies (PLBs) of <Emphasis Type="Italic">Phalaenopsis bellina</Emphasis> (Rchb.f.) christenson by encapsulation-dehydration

Protocorm-like bodies (PLBs) of Phalaenopsis bellina were successfully cryopreserved by the encapsulation-dehydration approach. Various stages in obtaining successful cryopreservation using this method were optimized. Encapsulated PLBs precultured in half-strength MS medium supplemented with 0.75 M sucrose for 3 days exhibited the highest viability in terms of 2,3,5-triphenyltetrazoliumchloride (TTC) reduction. The amount of sucrose in the PLBs after incubation in different concentrations of sucrose for different periods of time determined by HPLC. The highest sucrose concentration was 7 mg/g of PLBs for the PLBs treated with 0.75 M sucrose for 3 days as compared to the control which had only 1 mg/g sucrose. After sucrose preculture, the PLBs were subjected to desiccation using one of two methods. Desiccation using silica gel was more efficient in reducing PLBs moisture content. After 6 h of desiccation, PLBs desiccated using laminar air flow had 43.5% moisture content while for those desiccated using silica gel had 32% moisture content. PLBs desiccated to different moisture contents were plunged into LN. After storage in LN the encapsulated PLBs were re-warmed. Two weeks after re-warming PLBs viability was determined by TTC reduction and re-growth assessed. Encapsulated PLBs precultured with 0.75 M sucrose for 3 days followed by desiccated using silica gel for 5 h resulting in a moisture content of 39% lead to the highest post re-warming viability in terms of TTC reduction (46.6% of control PLBs) and 30% re-growth.