Functional characterization of N-terminal nucleotide binding domain (NBD-1) of a major ABC drug transporter Cdr1p of Candida albicans: uncommon but conserved Trp326 of Walker B is important for ATP binding

Using purified N-terminal NBD (NBD-512) domain of Cdr1p, a major multidrug extrusion pump of human pathogenic yeast Candida albicans, we show the relevance of the unique positioning of an atypical Trp326 residue. Similar to Cys193 in Walker A, Trp326 in the Walker B motif of Cdr1p is also a conserved feature of other fungal ATP Binding Cassette (ABC) transporters. By employing fluorescence spectroscopy, chemical modification, and site-directed mutagenesis, we demonstrate that of the five Trp residues in the NBD-512 domain, Trp326 alone is important for nucleotide binding and subsequent conformational changes within the domain. Furthermore, mutation of Trp326 to Ala results in an increased K(M) without appreciably affecting V(max) of ATPase activity. Thus, Trp326 in NBD-512 appears to be important for nucleotide binding and not for its hydrolysis. Additionally, the role of Trp326 in ATP binding is independent of the presence of the adjacent well-conserved Asp327 residue which, like Cys193, has a catalytic role in ATP hydrolysis. Considering that Trp326 of Cdr1p is a typical feature of fungal transporters alone, our study suggests that these ABC transporters may reflect mechanistic differences with regard to nucleotide binding and hydrolysis as compared to their counterparts of non-fungal origin.     

Electromyographic (EMG) Biofeedback in the Comprehensive Treatment of Central Pain and Ataxic Tremor Following Thalamic Stroke

Peripheral pain and ataxic tremor can appear suddenly following thalamic stroke and can significantly alter a patient's psychological, social, and physical functioning. The present paper reports the case of a 70-year-old Caucasian female who sustained an acute left posterior cerebral artery infarction involving the thalamus and left mesiotemporal regions. She subsequently developed Central Poststroke Pain and ataxic movement of her right arm and hand in addition to a significant right-side claudication. She was treated over 16 weeks (6 weeks of EMG biofeedback and 10 weeks of psychotherapy) with a combination of EMG biofeedback, progressive muscle relaxation, behavioral pain coping skills training, Forced Use Therapy, and Cognitive Behavioral Therap 7 years after her initial cerebral accident. The case demonstrates the utility of biofeedback when combined as part of a comprehensive treatment program to address the multiple complications associated with thalamic stroke.     

Synthesis of novel piperidinyl linker based energy transfer terminators and their potential use in DNA sequencing

Synthesis of novel piperidinyl linker based ET cassettes and terminators is described These novel terminators are evaluated in the DNA sequencing experiments using thermostable DNA polymerase.     

Genetic affinities of AB0 and Rh blood groups among populations of Andhra Pradesh, India: 2. Tribes

Genetic affinities of 21 tribal populations of Andhra Pradesh are reported in terms of genetic distance analysis with regard to AB0 and Rh loci. These tribal populations show a high degree of differentiation in the distribution of AB0 blood groups. Some tribes exhibit the monomorphism for Rh (D) locus. The genetic distance analysis reveals that the 21 tribes included in this study are genetically distant from one another and the genetic clustering pattern correlates with linguistic/ethnic affiliation and geographical propinquity of these tribal populations to a certain extent. The possible reasons for different cluster formations are discussed.     

The Journey from Traffic Offender to Severe Road Trauma Victim: Destiny or Preventive Opportunity?

Background

Road trauma is a leading cause of death and injury in young people. Traffic offences are common, but their importance as a risk indicator for subsequent road trauma is unknown. This cohort study assessed whether severe road trauma could be predicted by a history of prior traffic offences.

Methodology and Principal Findings

Clinical data of all adult road trauma patients admitted to the Western Australia (WA) State Trauma Centre between 1998 and 2013 were linked to traffic offences records at the WA Department of Transport. The primary outcomes were alcohol exposure prior to road trauma, severe trauma (defined by Injury Severity Score >15), and intensive care admission (ICU) or death, analyzed by logistic regression. Traffic offences directly leading to the road trauma admissions were excluded. Of the 10,330 patients included (median age 34 years-old, 78% male), 1955 (18.9%) had alcohol-exposure before road trauma, 2415 (23.4%) had severe trauma, 1360 (13.2%) required ICU admission, and 267 (2.6%) died. Prior traffic offences were recorded in 6269 (60.7%) patients. The number of prior traffic offences was significantly associated with alcohol-related road trauma (odds ratio [OR] per offence 1.03, 95% confidence interval [CI] 1.02–1.05), severe trauma (OR 1.13, 95%CI 1.14–1.15), and ICU admission or death (OR 1.10, 95%CI 1.08–1.11). Drink-drinking, seat-belt, and use of handheld electronic device offences were specific offences strongly associated with road trauma leading to ICU admission or death—all in a ‘dose-related’ fashion. For those who recovered from road trauma after an ICU admission, there was a significant reduction in subsequent traffic offences (mean difference 1.8, 95%CI 1.5 to 2.0) and demerit points (mean difference 7.0, 95%CI 6.5 to 7.6) compared to before the trauma event.

Significance

Previous traffic offences were a significant risk factor for alcohol-related road trauma and severe road trauma leading to ICU admission or death.     

Lead Retention by Soils at Field Moisture Contents

The majority of studies pertaining to lead retention by clays and soils have examined the mechanisms, kinetics, and adsorption isotherms using the batch experiment technique that employs solid: water extracts of 1:10 and 1:20. Field soil deposits generally have much lower gravimetric water content ranging between 9 and 45%. Given the wide disparity in the solids: water ratio employed in the batch experiment technique and that prevailing in field deposits, this paper examines the lead retention characteristics of soils at field moisture contents (6%, 13%, and 25%) using artificially lead-contaminated soil specimens. A residually derived (i.e., formed by in-situ weathering of parent rock) red soil was used to prepare the artificially contaminated soil specimens. The impact of variations in clay content on lead retention was examined by diluting the residual soil with various amounts (0 to 60%) of river sand. Soil specimens remolded at 6 and 13% moisture contents produced very stiff to hard soils on compaction, while specimens remolded at 25% moisture content existed in the slurry state. The soil specimens were contaminated with low (30 mg/kg) to high (2500 mg/kg) concentrations of lead ions by remolding them with 160 ppm to 10,000 ppm ionic lead solutions. Lead retention by soils at field moisture contents was determined by extracting the lead from the soil using a water leach test. Experimental results showed that the bulk (71 to 99%) of the added lead was retained by the soil in insoluble form at the field moisture content. Correlations between the amount of lead retained and soil/solution parameters indicated that the amounts of Pb retained at field moisture content is a function of the initial Pb addition, total sand content, effective clay porosity, and soil pH.     

Remote sensing of biodiversity: what to measure and monitor from space to species?

Global biodiversity monitoring systems through remote sensing can support consistent assessment, monitoring, modelling and reporting on biodiversity which are key activities intended for sustainable management. This work presents an overview of biodiversity monitoring components, i.e. biodiversity levels, essential biodiversity variables, biodiversity indicators, scale, biodiversity inventory, biodiversity models, habitat, ecosystem services, vegetation health and biogeochemical heterogeneity and discusses what remote sensing through Earth Observations has contributed to the study of biodiversity. The technological advancements in remote sensing have enabled information-rich data on biodiversity. Remote sensing data are making a strong contribution in providing unique information relevant to various biodiversity research and conservation applications. The extensive use of Earth observation data are not yet realized in biodiversity assessment, monitoring and conservation. The development of direct remote sensing approaches and the techniques for quantifying biodiversity at the community to species level is likely to be a great challenge for comprehensive earth observation-based monitoring strategy.

     

Differential roles for the interferon-inducible IFI16 and AIM2 innate immune sensors for cytosolic DNA in cellular senescence of human fibroblasts

The IFN-inducible IFI16 and AIM2 proteins act as innate immune sensors for cytosolic double-stranded DNA (dsDNA). On sensing dsDNA, the IFI16 protein induces the expression of IFN-β whereas the AIM2 protein forms an inflammasome, which promotes the secretion of IL-1β. Given that the knockdown of IFI16 expression in human diploid fibroblasts (HDF) delays the onset of cellular senescence, we investigated the potential roles for the IFI16 and AIM2 proteins in cellular senescence. We found that increased IFI16 protein levels in old (vs. young) HDFs were associated with the induction of IFN-β. In contrast, increased levels of the AIM2 protein in the senescent (vs. old) HDFs were associated with increased production of IL-1β. The knockdown of type I IFN-α receptor subunit, which reduced the basal levels of the IFI16 but not of the AIM2, protein delayed the onset of cellular senescence. Accordingly, increased constitutive levels of IFI16 and AIM2 proteins in ataxia telangiectasia mutated (ATM) HDFs were associated with the activation of the IFN signaling and increased levels of IL-1β. The IFN-β treatment of the young HDFs, which induced the expression of IFI16 and AIM2 proteins, activated a DNA damage response and also increased basal levels of IL-1β. Interestingly, the knockdown of AIM2 expression in HDFs increased the basal levels of IFI16 protein and activated the IFN signaling. In contrast, the knockdown of the IFI16 expression in HDFs decreased the basal and dsDNA-induced activation of the IFN signaling. Collectively, our observations show differential roles for the IFI16 and AIM2 proteins in cellular senescence and associated secretory phenotype.     

Jatropha curcas: a review on biotechnological status and challenges

Plant tissue culture and molecular biology techniques are powerful tools of biotechnology that can complement conventional breeding, expedite crop improvement and meet the demand for availability of uniform clones in large numbers. Jatropha curcas Linn., a non-edible, eco-friendly, non-toxic, biodegradable fuel-producing plant has attracted worldwide attention as an alternate sustainable energy source for the future. This review presents a consolidated account of biotechnological interventions made in J. curcas over the decades and focuses on contemporary information and trends of future research.     

Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice

Background

Type 1 Diabetes Mellitus is caused by auto immune destruction of insulin producing beta cells in the pancreas. Currently available treatments include transplantation of isolated islets from donor pancreas to the patient. However, this method is limited by inadequate means of immuno-suppression to prevent islet rejection and importantly, limited supply of islets for transplantation. Autologous adult stem cells are now considered for cell replacement therapy in diabetes as it has the potential to generate neo-islets which are genetically part of the treated individual. Adopting methods of islet encapsulation in immuno-isolatory devices would eliminate the need for immuno-suppressants.

Methodology/Principal Findings

In the present study we explore the potential of human adipose tissue derived adult stem cells (h-ASCs) to differentiate into functional islet like cell aggregates (ICAs). Our stage specific differentiation protocol permit the conversion of mesodermic h-ASCs to definitive endoderm (Hnf3β, TCF2 and Sox17) and to PDX1, Ngn3, NeuroD, Pax4 positive pancreatic endoderm which further matures in vitro to secrete insulin. These ICAs are shown to produce human C-peptide in a glucose dependent manner exhibiting in-vitro functionality. Transplantation of mature ICAs, packed in immuno-isolatory biocompatible capsules to STZ induced diabetic mice restored near normoglycemia within 3–4 weeks. The detection of human C-peptide, 1155±165 pM in blood serum of experimental mice demonstrate the efficacy of our differentiation approach.

Conclusions

h-ASC is an ideal population of personal stem cells for cell replacement therapy, given that they are abundant, easily available and autologous in origin. Our findings present evidence that h-ASCs could be induced to differentiate into physiologically competent functional islet like cell aggregates, which may provide as a source of alternative islets for cell replacement therapy in type 1 diabetes.