Quantitation of 6-mercaptopurine in biologic fluids using high-performance liquid chromatography: A selective and novel procedure

A simple, selective, sensitive and rapid procedure is described for the quantitation of 6-mercaptopurine (6-MP) in biological fluids. A sensitivity of at least 5 ng/ml is easily achieved in plasma on a reversed-phase octadecylsilane (C₁) column using a high-performance liquid chromatography system following an initial protein precipitation and a clean-up step. Mean extractability of the drug from plasma following this procedure is greater than 98% and the overall coefficient of variation for the assay is below 6%. Plasma levels of 6-MP were measured in a rhesus monkey for 12 h following an intravenous administration of a single bolus dose (4 mg/kg) of 6-MP.     

Determinants of Psychosocial Difficulties Experienced by Persons with Brain Disorders: Towards a ‘Horizontal Epidemiology’ Approach

BackgroundPersons with brain disorders experience significant psychosocial difficulties (PSD) in daily life, e.g. problems with managing daily routine or emotional lability, and the level of the PSD depends on social, physical and political environments, and psychologic-personal determinants. Our objective is to determine a brief set of environmental and psychologic-personal factors that are shared determinants of PSD among persons with different brain disorders.MethodsCross-sectional study, convenience sample of persons with either dementia, stroke, multiple sclerosis, epilepsy, migraine, depression, schizophrenia, substance dependence or Parkinson’s disease. Random forest regression and classical linear regression were used in the analyses.Results722 subjects were interviewed in four European countries. The brief set of determinants encompasses presence of comorbidities, health status appraisal, stressful life events, personality changes, adaptation, self-esteem, self-worth, built environment, weather, and health problems in the family.ConclusionsThe identified brief set of common determinants of PSD can be used to support the implementation of cross-cutting interventions, social actions and policy tools to lower PSD experienced by persons with brain disorders. This set complements a recently proposed reliable and valid direct metric of PSD for brain disorders called PARADISE24.     

Noninvasive genetic monitoring of tiger (Panthera tigris tigris) population of Orang National Park in the Brahmaputra floodplain, Assam, India

The Brahmaputra Valley of Assam, India, is one of the prime habitats for the endangered Royal Bengal tiger Panthera tigris tigris. With dwindling global population, estimation of the minimum number of tigers has always been a curiosity to wildlife researchers as well as to protected area managers. In the present study, DNA-based techniques were used for identifying individual tigers present in Orang National Park of Assam, from 57 faecal samples collected during February 2009. Orang National Park stands as an island of a single forest patch along the north bank of river Brahmaputra. The present study confirms the presence of 17 individual tigers in Orang National Park, with five male and 12 female. DNA-based capture–recapture analysis yielded minimum range estimate of 18 and 19 individuals, with possible overestimates of population size following two models of capture probability in CAPWIRE. The results of our genetic counting of tigers are compared with the estimates of 19 tigers based on pugmark analysis by the state Forest Department in 2000 and an independent capture–recapture estimate of 14 (±3.6) individuals based on photographic identity study in 2009. Looking at high mortality of tigers in the area, with 19 reported deaths during 2000 to 2009, our results indicate high individual turnover in the area. This study shows that Orang National Park harbours a healthy breeding population of tigers. However, the possibility of a source-sink dynamics operating in the landscape could not be ruled out, with possible immigration from nearby Kaziranga National Park on the south bank of Brahmaputra, which has the highest reported density of the species in the world.     

Molecular mechanism of in vitro oligomerization of Dps from Mycobacterium smegmatis: mutations of the residues identified by "interface cluster" analysis

The irreversible dodecamerization of native Dps trimers from Mycobacterium smegmatis, in vitro, is known to be directly associated with the bimodal function of this protein. Hence it is important to explore this pathway at the molecular level. Two types of trimers, Trimer A (tA) and Trimer B (tB), can be derived from the dodecamer due to the inherent 3-fold symmetry of the spherical crystal structure. These derived trimers were expressed as protein structure graphs (PSGs) using the computed interaction strength among the residues. Interface clusters which were identified from PSGs allowed us to convincingly predict E146 and F47 for further mutation studies. Various single and double mutants were constructed and characterized. We were finally able to generate a single mutant F47E impaired in dodecamerization and a double mutant E146AF47E as native monomer in solution. These two observed results suggest that the two trimers are important for dodecamerization and that the residues selected are important for the structural stability of the protein in vitro.     

Characteristics of fluorescamine modified bacteriorhodopsin

The light activated proton pump, bacteriorhodopsin was modified with varying amounts of flourescamine, the fluorescamine to protein ratio ranging from 1 to 100. The modified protein was washed free of excess of fluorescamine and reconstituted into phospholipid vesicles to check the proton pumping activity. Although the spectral investigations indicated chemical modification, the circular dichroism measurements pointed to an overall loss of the trimeric structure of the protein. The implications of the present study are that the modifying agent can interact non-specifically with the protein, altering its structural parameters, which in turn affects the function of the protein