Recent trends in molecular diagnostics of yeast infections : from PCR to NGS

The incidence of opportunistic yeast infections in humans has been increasing over recent years. These infections are difficult to treat and diagnose, in part due to the large number and broad diversity of species that can underlie the infection. In addition, resistance to one or several antifungal drugs in infecting strains is increasingly being reported, severely limiting therapeutic options and showcasing the need for rapid detection of the infecting agent and its drug susceptibility profile. Current methods for species and resistance identification lack satisfactory sensitivity and specificity, and often require prior culturing of the infecting agent, which delays diagnosis. Recently developed high-throughput technologies such as next generation sequencing or proteomics are opening completely new avenues for more sensitive, accurate and fast diagnosis of yeast pathogens. These approaches are the focus of intensive research, but translation into the clinics requires overcoming important challenges. In this review, we provide an overview of existing and recently emerged approaches that can be used in the identification of yeast pathogens and their drug resistance profiles. Throughout the text we highlight the advantages and disadvantages of each methodology and discuss the most promising developments in their path from bench to bedside

Global distribution of two fungal pathogens threatening endangered sea turtles

This work was supported by grants of Ministerio de Ciencia e Innovación, Spain (CGL2009-10032, CGL2012-32934). J.M.S.R was supported by PhD fellowship of the CSIC (JAEPre 0901804). The Natural Environment Research Council and the Biotechnology and Biological Sciences Research Council supported P.V.W. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Thanks Machalilla National Park in Ecuador, Pacuare Nature Reserve in Costa Rica, Foundations Natura 2000 in Cape Verde and Equilibrio Azul in Ecuador, Dr. Jesus Muñoz, Dr. Ian Bell, Dr. Juan Patiño for help and technical support during samplingPeer reviewedPublisher PD

Hsp21potentiates antifungal drug tolerance in Candida albicans

Peer reviewedPublisher PD

Small but crucial : the novel small heat shock protein Hsp21 mediates stress adaptation and virulence in Candida albicans

Peer reviewedPublisher PD

The 7.5 Magnitude Limit Sample of Bright Short-Period Binary Stars. I. How Many Contact Binaries Are There?

A sample of bright contact binary stars (W UMa-type or EW, and related: with beta Lyr light curves, EB, and ellipsoidal, ELL - in effect, all but the detached, EA), to the limit of Vmax = 7.5 magnitude is deemed to include all discoverable short-period (P<1 days) binaries with photometric variation larger than about 0.05 magnitude. Of the 32 systems in the final sample, 11 systems have been discovered by the Hipparcos satellite. The combined spatial density is evaluated at (1.02+/-0.24)x10^-5 pc^-3. The Relative Frequency of Occurrence (RFO), defined in relation to the Main Sequence stars, depends on the luminosity. An assumption of RFO~1/500 for MV>+1.5 is consistent with the data, although the number statistics is poor with the resulting uncertainty in the spatial density and the RFO by a factor of about two. The RFO rapidly decreases for brighter binaries to a level of 1/5,000 for MV<+1.5 and to 1/30,000 for MV<+0.5. The high RFO of 1/130, previously determined from the deep OGLE-I sample of Disk Population W UMa-type systems towards Baade's Window, is inconsistent with and unconfirmed by the new results. Possible reasons for the large discrepancy are discussed. They include several observational effects, but also a possibility of a genuine increase in the contact-binary density in the central parts of the Galaxy.Comment: AASTeX5, 11 figures, 3 tables. Table 1 is very wide; in case of problems send e-mail to [email protected] for a raw text versio

Maintenance of bone mineral density after implantation of a femoral neck hip prosthesis

<p>Abstract</p> <p>Background</p> <p>Stress shielding of the proximal femur has been observed in a number of conventional cementless implants used in total hip arthroplasty. Short femoral-neck implants are claiming less interference with the biomechanics of the proximal femur. The goal of this study was to investigate the changes of bone-mineral density in the proximal femur and the clinical outcome after implantation of a short femoral-neck prosthesis.</p> <p>Methods</p> <p>We prospectively assessed the clinical outcome and the changes of bone mineral density of the proximal femur up to one year after implantation of a short femoral neck prosthesis in 20 patients with a mean age of 47 years (range 17 to 65). Clinical outcome was assessed using the Harris Hip Score. The WOMAC was used as a patient-relevant outcome-measure. The bone mineral density was determined using dual energy x-ray absorptiometry, performed 10 days, three months and 12 months after surgery.</p> <p>Results</p> <p>The Harris Hip Score improved from an average preoperative score of 46 to a postoperative score at 12 months of 89 points, the global WOMAC index from 5,3 preoperatively to 0,8 at 12 months postoperatively. In contrast to conventional implants, the DEXA-scans overall revealed a slight increase of bone mineral density in the proximal femur in the 12 months following the implantation.</p> <p>Conclusion</p> <p>The short femoral neck stem lead to a distinct bone reaction. This was significantly different when compared to the changes in bone mineral density reported after implantation of conventional implants.</p

The EBLM Project I-Physical and orbital parameters, including spin-orbit angles, of two low-mass eclipsing binaries on opposite sides of the Brown Dwarf limit

This paper introduces a series of papers aiming to study the dozens of low mass eclipsing binaries (EBLM), with F, G, K primaries, that have been discovered in the course of the WASP survey. Our objects are mostly single-line binaries whose eclipses have been detected by WASP and were initially followed up as potential planetary transit candidates. These have bright primaries, which facilitates spectroscopic observations during transit and allows the study of the spin-orbit distribution of F, G, K+M eclipsing binaries through the Rossiter-McLaughlin effect. Here we report on the spin-orbit angle of WASP-30b, a transiting brown dwarf, and improve its orbital parameters. We also present the mass, radius, spin-orbit angle and orbital parameters of a new eclipsing binary, J1219-39b (1SWAPJ121921.03-395125.6, TYC 7760-484-1), which, with a mass of 95 +/- 2 Mjup, is close to the limit between brown dwarfs and stars. We find that both objects orbit in planes that appear aligned with their primaries' equatorial planes. Neither primaries are synchronous. J1219-39b has a modestly eccentric orbit and is in agreement with the theoretical mass--radius relationship, whereas WASP-30b lies above it.Comment: 12 pages, 7 figures, data in appendices, submitted to A&A (taking in account 1st referee report

Candida albicans Possesses Sap7 as a Pepstatin A-Insensitive Secreted Aspartic Protease

BACKGROUND: Candida albicans, a commensal organism, is a part of the normal flora of healthy individuals. However, once the host immunity is compromised, C. albicans opportunistically causes recurrent superficial or fatal systemic candidiasis. Secreted aspartic proteases (Sap), encoded by 10 types of SAP genes, have been suggested to contribute to various virulence processes. Thus, it is important to elucidate their biochemical properties for better understanding of the molecular mechanisms that how Sap isozymes damage host tissues. METHODOLOGY/PRINCIPAL FINDINGS: The SAP7 gene was cloned from C. albicans SC5314 and heterogeneously produced by Pichia pastoris. Measurement of Sap7 proteolytic activity using the FRETS-25Ala library showed that Sap7 was a pepstatin A-insensitive protease. To understand why Sap7 was insensitive to pepstatin A, alanine substitution mutants of Sap7 were constructed. We found that M242A and T467A mutants had normal proteolytic activity and sensitivity to pepstatin A. M242 and T467 were located in close proximity to the entrance to an active site, and alanine substitution at these positions widened the entrance. Our results suggest that this alteration might allow increased accessibility of pepstatin A to the active site. This inference was supported by the observation that the T467A mutant has stronger proteolytic activity than the wild type. CONCLUSIONS/SIGNIFICANCE: We found that Sap7 was a pepstatin A-insensitive protease, and that M242 and T467 restricted the accessibility of pepstatin A to the active site. This finding will lead to the development of a novel protease inhibitor beyond pepstatin A. Such a novel inhibitor will be an important research tool as well as pharmaceutical agent for patients suffering from candidiasis

Global Research Alliance N2O chamber methodology guidelines: considerations for automated flux measurement

Nitrous oxide (N2O) emissions are highly episodic in response to nitrogen additions and changes in soil moisture. Automated gas sampling provides the necessary high temporal frequency to capture these emission events in real time, ensuring the development of accurate N2O inventories and effective mitigation strategies to reduce global warming. This paper outlines the design and operational considerations of automated chamber systems including chamber design and deployment, frequency of gas sampling, and options in terms of the analysis of gas samples. The basic hardware and software requirements for automated chambers are described, including the major challenges and obstacles in their implementation and operation in a wide range of environments. Detailed descriptions are provided of automated systems that have been deployed to assess the impacts of agronomy on the emissions of N2O and other significant greenhouse gases. This information will assist researchers across the world in the successful deployment and operation of automated N2O chamber systems

Interaction between Hydrogenase Maturation Factors HypA and HypB Is Required for [NiFe]-Hydrogenase Maturation

The active site of [NiFe]-hydrogenase contains nickel and iron coordinated by cysteine residues, cyanide and carbon monoxide. Metal chaperone proteins HypA and HypB are required for the nickel insertion step of [NiFe]-hydrogenase maturation. How HypA and HypB work together to deliver nickel to the catalytic core remains elusive. Here we demonstrated that HypA and HypB from Archaeoglobus fulgidus form 1∶1 heterodimer in solution and HypA does not interact with HypB dimer preloaded with GMPPNP and Ni. Based on the crystal structure of A. fulgidus HypB, mutants were designed to map the HypA binding site on HypB. Our results showed that two conserved residues, Tyr-4 and Leu-6, of A. fulgidus HypB are required for the interaction with HypA. Consistent with this observation, we demonstrated that the corresponding residues, Leu-78 and Val-80, located at the N-terminus of the GTPase domain of Escherichia coli HypB were required for HypA/HypB interaction. We further showed that L78A and V80A mutants of HypB failed to reactivate hydrogenase in an E. coli ΔhypB strain. Our results suggest that the formation of the HypA/HypB complex is essential to the maturation process of hydrogenase. The HypA binding site is in proximity to the metal binding site of HypB, suggesting that the HypA/HypB interaction may facilitate nickel transfer between the two proteins