The pharmacological regulation of cellular mitophagy

Small molecules are pharmacological tools of considerable value for dissecting complex biological processes and identifying potential therapeutic interventions. Recently, the cellular quality-control process of mitophagy has attracted considerable research interest; however, the limited availability of suitable chemical probes has restricted our understanding of the molecular mechanisms involved. Current approaches to initiate mitophagy include acute dissipation of the mitochondrial membrane potential (ΔΨm) by mitochondrial uncouplers (for example, FCCP/CCCP) and the use of antimycin A and oligomycin to impair respiration. Both approaches impair mitochondrial homeostasis and therefore limit the scope for dissection of subtle, bioenergy-related regulatory phenomena. Recently, novel mitophagy activators acting independently of the respiration collapse have been reported, offering new opportunities to understand the process and potential for therapeutic exploitation. We have summarized the current status of mitophagy modulators and analyzed the available chemical tools, commenting on their advantages, limitations and current applications

Development of electrochemical biosensors by e-beam lithography for medical diagnostics

In this work the fabrication and characterization of boron-doped diamond (BDD) nanoelectrode arrays are discussed. The use of boron-doped diamond electrodes is very attractive due to advantageous properties including high reproducibility, stability, and robustness under extreme conditions, where conventional electrode materials may undergo severe erosion. BDD electrodes have also proved to be very useful because they show an extremely wide potential window in aqueous solutions without oxidation of the electrode itself. This allows electrochemical detection, at tiny background currents, of a number of substances that oxidize at very positive potentials, where other electrodic materials are not suitable. BDD based NEAs were prepared using Si substrates coated with a layer of Boron doped diamond as macroelectrode. NEAs were obtained by creating an array of nanoholes by electron beam lithography (EBL) in a thin film of polycarbonate deposited on top of the macroelectrode. This approach leads to the formation of recessed nanoelectrodes. The parameters for using polycarbonate as a novel electron beam resist have been optimized and successfully used for fabrication of NEAs. The most interesting properties of this polymer for nanofabrication purposes are the high lithographic contrast, which allows the creation of structures of dimensions less than 100 nm; chemical stability, which guarantees a long-term use in electrochemical solutions and the possibility of functionalization with biological molecules (DNA and proteins). NEAs have been characterized with cyclic voltammetry and have provided voltammetric signals controlled by pure radial diffusion. The low background current of BDD added to the properties of NEAs indicate that this system can be applied for the development of sensors with high sensitivity. Polycarbonate surface of NEAs was successfully functionalized with small ss-DNA sequence, confirming the possibility of exploiting these systems as diagnostic biosensors

Obesity and post-operative cognitive dysfunction: a systematic review and meta-analysis

Background: Post-operative cognitive dysfunction (POCD) occurs frequently after surgery, and is related to dementia and premature death. Obesity increases the risk of late-life cognitive impairment, but little is known about its role in POCD. We conducted a systematic review and meta-analysis of studies on the association between obesity and risk of POCD. Methods: PubMed and the Cochrane Library were systematically searched. Studies were included if they had prospective designs, reported on human adults undergoing surgery, if cognitive function was measured pre- and post-surgery, if obesity, body mass index (BMI) and/or body weight were ascertained, and if associations with POCD were reported as relative risks or odds ratios. Underweight, weight loss, and post-operative delirium were not considered. Results: Inclusion criteria were met by six articles. Samples totaled 1432 older patients (mean age ≥62 years) who were followed up for 24 hours to twelve months after surgery. Analysis of studies with obesity defined as a categorical measure found a non-significantly higher risk of POCD among persons with BMI > 30 kg/m2 versus ≤30 kg/m2 (RR 1.27; 95% CI 0.95, 1.70; p = 0.10). No such associations were found for studies that analyzed BMI or body weight continuously as predictors of POCD (RR 0.98 per kg/m2; 95% CI 0.93, 1.03, p = 0.45; RR 0.99 per kg; 95% CI 0.89, 1.09; p = 0.83, respectively). Conclusions: Few studies have addressed the topic, and the results of these studies provide only limited support for an increased risk of POCD in patients who are obese. Further large-scale, prospective investigations are necessary for clarification

Characterization of coagulase-negative staphylococcal isolates from blood with reduced susceptibility to glycopeptides and therapeutic options

<p>Abstract</p> <p>Background</p> <p>Coagulase-negative staphylococci (CoNS) are a major cause of nosocomial blood stream infection, especially in critically ill and haematology patients. CoNS are usually multidrug-resistant and glycopeptide antibiotics have been to date considered the drugs of choice for treatment. The aim of this study was to characterize CoNS with reduced susceptibility to glycopeptides causing blood stream infection (BSI) in critically ill and haematology patients at the University Hospital Tor Vergata, Rome, Italy, in 2007.</p> <p>Methods</p> <p>Hospital microbiology records for transplant haematology and ICU were reviewed to identify CoNS with elevated MICs for glycopeptides, and isolates were matched to clinical records to determine whether the isolates caused a BSI. The isolates were tested for susceptibility to new drugs daptomicin and tigecycline and the genetic relationship was assessed using f-AFLP.</p> <p>Results</p> <p>Of a total of 17,418 blood cultures, 1,609 were positive for CoNS and of these, 87 (5.4%) displayed reduced susceptibility to glycopeptides. Clinical review revealed that in 13 cases (7 in haematology and 6 in ICU), CoNS with reduced susceptibility to glycopeptides were responsible for a BSI. <it>Staphylococcus epidermidis </it>was the causative organism in 11 instances and <it>Staphylococcus haemolyticus </it>in 2. The incidence of oxacillin resistance was high (77%), although all isolates remained susceptible to linezolid, daptomycin and tigecycline. Fingerprinting of CoNS identified one clonal relationship between two isolates.</p> <p>Conclusion</p> <p>Multi-resistant CoNS with reduced susceptibility to glycopeptides, although still relatively infrequent in our hospital, are emerging pathogens of clinical concern. Surveillance by antibiotyping with attention to multi-resistant profile, and warning to clinicians, is necessary.</p

Phylogenetic Analysis of Seven WRKY Genes across the Palm Subtribe Attaleinae (Arecaceae) Identifies Syagrus as Sister Group of the Coconut

BACKGROUND:The Cocoseae is one of 13 tribes of Arecaceae subfam. Arecoideae, and contains a number of palms with significant economic importance, including the monotypic and pantropical Cocos nucifera L., the coconut, the origins of which have been one of the "abominable mysteries" of palm systematics for decades. Previous studies with predominantly plastid genes weakly supported American ancestry for the coconut but ambiguous sister relationships. In this paper, we use multiple single copy nuclear loci to address the phylogeny of the Cocoseae subtribe Attaleinae, and resolve the closest extant relative of the coconut. METHODOLOGY/PRINCIPAL FINDINGS:We present the results of combined analysis of DNA sequences of seven WRKY transcription factor loci across 72 samples of Arecaceae tribe Cocoseae subtribe Attaleinae, representing all genera classified within the subtribe, and three outgroup taxa with maximum parsimony, maximum likelihood, and Bayesian approaches, producing highly congruent and well-resolved trees that robustly identify the genus Syagrus as sister to Cocos and resolve novel and well-supported relationships among the other genera of the Attaleinae. We also address incongruence among the gene trees with gene tree reconciliation analysis, and assign estimated ages to the nodes of our tree. CONCLUSIONS/SIGNIFICANCE:This study represents the as yet most extensive phylogenetic analyses of Cocoseae subtribe Attaleinae. We present a well-resolved and supported phylogeny of the subtribe that robustly indicates a sister relationship between Cocos and Syagrus. This is not only of biogeographic interest, but will also open fruitful avenues of inquiry regarding evolution of functional genes useful for crop improvement. Establishment of two major clades of American Attaleinae occurred in the Oligocene (ca. 37 MYBP) in Eastern Brazil. The divergence of Cocos from Syagrus is estimated at 35 MYBP. The biogeographic and morphological congruence that we see for clades resolved in the Attaleinae suggests that WRKY loci are informative markers for investigating the phylogenetic relationships of the palm family

Patterns of Reproductive Isolation in Toads

Understanding the general features of speciation is an important goal in evolutionary biology, and despite significant progress, several unresolved questions remain. We analyzed an extensive comparative dataset consisting of more than 1900 crosses between 92 species of toads to infer patterns of reproductive isolation. This unique dataset provides an opportunity to examine the strength of reproductive isolation, the development and sex ratios of hybrid offspring, patterns of fertility and infertility, and polyploidization in hybrids all in the context of genetic divergence between parental species. We found that the strength of intrinsic postzygotic isolation increases with genetic divergence, but relatively high levels of divergence are necessary before reproductive isolation is complete in toads. Fertilization rates were not correlated to genetic divergence, but hatching success, the number of larvae produced, and the percentage of tadpoles reaching metamorphosis were all inversely related with genetic divergence. Hybrids between species with lower levels of divergence developed to metamorphosis, while hybrids with higher levels of divergence stopped developing in gastrula and larval stages. Sex ratios of hybrid offspring were biased towards males in 70% of crosses and biased towards females in 30% of crosses. Hybrid females from crosses between closely related species were completely fertile, while approximately half (53%) of hybrid males were sterile, with sterility predicted by genetic divergence. The degree of abnormal ploidy in hybrids was positively related to genetic divergence between parental species, but surprisingly, polyploidization had no effect on patterns of asymmetrical inviability. We discuss explanations for these patterns, including the role of Haldane's rule in toads and anurans in general, and suggest mechanisms generating patterns of reproductive isolation in anurans