A long-range pseudoknot is required for activity of the Neurospora VS ribozyme

Four small RNA self-cleaving domains, the hammerhead, hairpin, hepatitis delta virus and Neurospora VS ribozymes, have been identified previously in naturally occurring RNAs. The secondary structures of these ribozymes are reasonably well understood, but little is known about long-range interactions that form the catalytically active tertiary conformations. Our previous work, which identified several secondary structure elements of the VS ribozyme, also showed that many additional bases were protected by magnesium-dependent interactions, implying that several tertiary contacts remained to be identified. Here we have used site-directed mutagenesis and chemical modification to characterize the first long-range interaction identified in VS RNA. This interaction contains a 3 bp pseudoknot helix that is required for tertiary folding and self-cleavage activity of the VS ribozyme.     

Molecular characterization of Mycobacterium avium complex isolates giving discordant results in AccuProbe tests by PCR-restriction enzyme analysis, 16S rRNA gene sequencing, and DT1-DT6 PCR

Based on cultural and biochemical tests, a total of 84 strains (72 clinical and 12 environmental isolates from the Caribbean Isles, Europe, and the Indian subcontinent) were identified as members of the Mycobacterium avium complex (MAC). They were further characterized with MAC, M. avium, and M. intracellulare probes of the AccuProbe system, and this was followed by selective amplification of DT6 and DT1 sequences. Seventy isolates gave concordant results; 63 were identified as M. avium, 5 were identified as M. intracellulare, and 24 remained untypeable by both methods. Fourteen isolates gave discrepant results, as they were DT1 positive but gave negative results by the M. intracellulare AccuProbe test. Consequently, a detailed molecular analysis of all DT1-positive isolates (14 discrepant strains plus 5 M. intracellulare strains) was performed by PCR-restriction analysis (PRA) of the hsp65 gene and 16S rRNA gene sequencing. The results confirmed the reported heterogeneity of M. intracellulare, as only 6 of 19 isolates (32%) gave PRA results compatible with published M. intracellulare profiles while the rest of the isolates were grouped in four previously unpublished profiles. 16S rRNA gene sequencing showed that only 8 of 19 isolates (42%) were related to M. intracellulare IWGMT 90247 (EMBL accession no. X88917), the rest being related to MCRO19 (EMBL accession no. X93030) and MIWGTMR10 (EMBL accession no. X88915). In conclusion, we have characterized a significant number of MAC isolates which were not identified by the AccuProbe test, PRA, or 16S rRNA sequencing. However, all of them were identifiable by DT1-DT6 PCR (they were DT6 negative and DT1 positive) and could be tentatively identified as M. intracellulare based on previously published observations. It is noteworthy that the majority of such isolates (14 of 19) were from the Indian subcontinent, with 12 of 14 being environmental isolates. Our study confirms the marked heterogeneity of M. intracellulare isolates and shows the utility of in-house DT1 PCR to detect this group of isolates, which would otherwise have been missed by the AccuProbe system in a routine clinical microbiology laboratory.     

Pulsed electrodeposition of CdTe thin films: effect of pulse parameters over structure, stoichiometry and optical absorption

CdTe thin films were electrochemically deposited using unipolar current pulses of high magnitude between 2.5 and 30 mA/cm² in an aqueous solution. Parametric study of the effect of periodic current pulse magnitude, average current and ON and OFF duration was undertaken to understand the effect of pulse variables on CdTe film properties. Increasing pulse deposition current modifies crystalline growth phase from single cubic to mixed cubic and hexagonal growth phases. In addition to the modification in CdTe growth phases, there is an increasing tendency of the oxide formation particularly CdTeO₃. Increase in pulse current density or average current yields Cd rich CdTe films. The optical absorption coefficient decreases with the decrease in pulse current density, whereas an increase is observed as the OFF time decreases. The optical energy gap is found to increase with OFF time. A systematic study on the effect of pulse variables over the structure, compositional and optical properties of CdTe film is described.     

Speckle interferometry measurements of the conductor displacementsin the CERN-LHC main dipole

Magnetic field quality in superconducting magnets strongly depends on conductor position in operational conditions. Simulations based on finite-element models (FEM) provide the field of stresses and displacements inside the magnet. Due to the complex mechanical behavior of the coil and to the different materials composing the magnet, it is not trivial to build a reliable mechanical model. We present an experimental method based on optical measurements to validate the accuracy of the FEM in evaluating differential displacements at room temperature. A short sample of dipole coils is loaded through a small press and speckle interferometry measurements detect differential displacements with an accuracy of 1.5 μm. Comparison with the numerical results allows the testing of the most critical features of the model, i.e., a pressure-dependent coil elasticity and the interfaces between different materials. A good agreement between measurements and deformed geometry foreseen by the FEM is found     

Molecular methods for the rapid diagnosis of mycobacterial infections

With the aim to reduce the time for the diagnosis of mycobacterial infections, various molecular methods were developed recently which are particularly suitable for clinical mycobacteriology laboratories. These methods permit both the detection and rapid identification either directly from the pathological sample or culture. We hereby review the most recent advances in this field, particularly those involving hybridization with molecular probes, or nucleic amplification methods. We also review the strategy employed for current research developments for application in mycobacterial diagnosis as well as the future prospects.     

Transmitter-induced calcium responses differ in astrocytes acutely isolated from rat brain and in culture

Glial fibrillary acid protein (GFAP)-positive astrocytes isolated from the cerebral cortices of 3-10-day-old rats frequently showed increased intracellular Ca2+ concentration responses to L-glutamate and glutamate analogues. However, few of the acutely isolated cells responded to ATP, and no such cells responded to serotonin [5-hydroxytryptamine (5-HT)]. The same cell that failed to respond to ATP or 5-HT often responded to glutamate. Culturing acutely isolated cells in media containing horse serum decreased Ca2+ responses to glutamate but increased the responses to ATP and induced responses to 5-HT. In primary cultures prepared from the cerebral cortices of 1-day-old rats and cultured in horse serum, fewer of the cells responded to glutamate, but almost all cells responded to ATP and 5-HT. The lack of or limited response to, 5-HT or ATP in the acutely isolated cells seems unlikely to be due to selective damage to the respective receptors because acutely isolated GFAP-negative cells showed responses to ATP, several different proteases and mechanical dissociation yielded cells that also responded to glutamate but not to ATP, and exposure of primary cultures to papain did not abolish Ca2+ responses to several transmitters. The responses of the acutely isolated cells to glutamate but limited or lack of responses to ATP and 5-HT also correspond to what has been seen so far for astrocytes in situ. Thus, the present studies provide direct evidence that some of the receptors seen in primary astrocyte cultures may reflect a response to culture conditions and that, in the context of the relevant information so far available, acutely isolated astrocytes seem to reflect better the in vivo state.     

Preparation of Zn(In)Se films from alloyed precursors

Single-phase cubic Zn(In)Se thin film growth by Se vapor selenization of Zn(In) alloy precursors films is described. Depositing the Zn(In) precursor film at higher substrate temperatures changes the In/Zn composition and also yields highly crystalline Zn(In)Se films. The In/Zn ratio in the selenized film is higher in comparison to that of the precursor due to differential selenization kinetics and the complex In-Se and Zn-Se reaction chemistry. The resistivity of the Zn(In)Se film depends on the In/Zn ratio. Initially, the resistivity increases with increased indium incorporation due to increased defect concentration and then decreases at higher In/Zn ratios because of lower grain-boundary effects and reduced trap density owing to improvement in film crystallinity. Treatment with vapor-phase Zn compensates for Zn vacancies in the film, reduces electrically inactive defects, and increases doping efficiency, thereby lowering the resistivities to ∼1 Ω cm. Hot-probe and thermoelectric power measurements show that all low resistive ZnSe films are n-type.     

Current zinc intake and risk of diabetes and coronary artery disease and factors associated with insulin resistance in rural and urban populations of North India

OBJECTIVE: To determine the association between current zinc intake and prevalence of coronary artery disease (CAD) and diabetes as well as factors associated with insulin resistance. DESIGN, SUBJECTS AND METHODS: In this cross sectional survey, 3575 subjects, aged 25 to 64 years, including 1769 rural (894 men. 875 women) and 1806 urban (904 men, 902 women) subjects were studied. The survey methods included questionnaires for 7-day food intake record, physical examination, and electrocardiography using World Health Organization criteria. RESULTS: The prevalence of CAD, diabetes and glucose intolerance was significantly higher among subjects consuming lower intakes of dietary zinc. There was a higher prevalence of hypertension, hypertriglyceridemia and low high-density lipoprotein cholesterol levels which showed significant upward trend with lower zinc intakes. Serum lipoprotein (a) and 2-hour plasma insulin levels also were associated with low zinc intake. Multivariate logistic regression analysis after adjustment for age showed that zinc intake and CAD were inversely associated. Serum zinc (odds ratio:men 0.77, women 0.57), serum triglycerides (men 0.86, women 0.81), blood pressure (0.83 men, women 0.76), diabetes mellitus (men 0.90, women 0.85), central obesity (men 0.88, women 0.87), glucose intolerance (men 0.66, women 0.57) and low high-density lipoprotein cholesterol (men 0.72, women 0.70) were significant risk factors for CAD (explained by tertiles of zinc status) in urban subjects. These associations were not observed in rural subjects. CONCLUSION: Lower consumption of dietary zinc and low serum zinc levels were associated with an increased prevalence of CAD and diabetes and several of their associated risk factors including hypertension, hypertriglyceridemia and other factors suggestive of mild insulin resistance in urban subjects.     

Randomized, double-blind placebo-controlled trial of coenzyme Q10 in patients with acute myocardial infarction

Adherence with inhaled beta-agonists and corticosteroids in 24 asthmatic children was tracked over 3 months utilizing the metered-dose inhaler chronolog (MDIC). Patients seldom took all of their medications as prescribed, and failed to take any inhaled corticosteroid doses on a median of 41.8% of days or inhaled beta-agonists on 28.1% of days despite prescribed daily use. Medication nonadherence was correlated with lower levels of asthma knowledge (Asthma Knowledge Questionnaire) and family dysfunction (Family Assessment Device), but not child behavior disorder (Child Behavior Checklist). Patients tended to dramatically over-report medication use. Improved identification of the markers of nonadherence can directly facilitate more efficient targeting of behavioral interventions, resulting in improved adherence, better illness control, and less requirement of urgent medication intervention.     

Molecular typing of mycobacteria

Molecular typing is now widely used in mycobacteriology as a significant addition to conventional epidemiological studies. Several methods are available that differ with respect to their discriminating power and to the genetic marker used. Restriction fragment length polymorphism (RFLP) done using the IS6110 insertion sequence has been widely used for typing Mycobacterium tuberculosis. More recently, short repetitive DNA sequences have been used as RFLP markers, as well as for other typing methods involving gene amplification. One of these methods, the inter-direct repeat (DR)-based spoligotyping method, is rapid, directly applicable to clinical specimens, and highly discriminating.