Emergence of Oxacillin Resistance in Stealth Methicillin-Resistant Staphylococcus aureus Due to mecA Sequence Instability.
Staphylococcus aureus strains that possess a mecA gene but are phenotypically susceptible to oxacillin and cefoxitin (OS-MRSA) have been recognized for over a decade and are a challenge for diagnostic laboratories. The mechanisms underlying the discrepancy vary from isolate to isolate. We characterized seven OS-MRSA clinical isolates of six different spa types from six different states by whole-genome sequencing to identify the nucleotide sequence changes leading to the OS-MRSA phenotype. The results demonstrated that oxacillin susceptibility was associated with mutations in regions of nucleotide repeats within mecA Subinhibitory antibiotic exposure selected for secondary mecA mutations that restored oxacillin resistance. Thus, strains of S. aureus that contain mecA but are phenotypically susceptible can become resistant after antibiotic exposure, which may result in treatment failure. OS-MRSA warrant follow-up susceptibility testing to ensure detection of resistant revertants.


No Association of Coronary Artery Disease with X-Chromosomal Variants in Comprehensive International Meta-Analysis.
In recent years, genome-wide association studies have identified 58 independent risk loci for coronary artery disease (CAD) on the autosome. However, due to the sex-specific data structure of the X chromosome, it has been excluded from most of these analyses. While females have 2 copies of chromosome X, males have only one. Also, one of the female X chromosomes may be inactivated. Therefore, special test statistics and quality control procedures are required. Thus, little is known about the role of X-chromosomal variants in CAD. To fill this gap, we conducted a comprehensive X-chromosome-wide meta-analysis including more than 43,000 CAD cases and 58,000 controls from 35 international study cohorts. For quality control, sex-specific filters were used to adequately take the special structure of X-chromosomal data into account. For single study analyses, several logistic regression models were calculated allowing for inactivation of one female X-chromosome, adjusting for sex and investigating interactions between sex and genetic variants. Then, meta-analyses including all 35 studies were conducted using random effects models. None of the investigated models revealed genome-wide significant associations for any variant. Although we analyzed the largest-to-date sample, currently available methods were not able to detect any associations of X-chromosomal variants with CAD.


Stabilization and anomalous hydration of collagen fibril under heating.
Type I collagen is the most common protein among higher vertebrates. It forms the basis of fibrous connective tissues (tendon, chord, skin, bones) and ensures mechanical stability and strength of these tissues. It is known, however, that separate triple-helical collagen macromolecules are unstable at physiological temperatures. We want to understand the mechanism of collagen stability at the intermolecular level. To this end, we study the collagen fibril, an intermediate level in the collagen hierarchy between triple-helical macromolecule and tendon. When heating a native fibril sample, its Young's modulus decreases in temperature range 20-58°C due to partial denaturation of triple-helices, but it is approximately constant at 58-75°C, because of stabilization by inter-molecular interactions. The stabilization temperature range 58-75°C has two further important features: here the fibril absorbs water under heating and the internal friction displays a peak. We relate these experimental findings to restructuring of collagen triple-helices in fibril. A theoretical description of the experimental results is provided via a generalization of the standard Zimm-Bragg model for the helix-coil transition. It takes into account intermolecular interactions of collagen triple-helices in fibril and describes water adsorption via the Langmuir mechanism. We uncovered an inter-molecular mechanism that stabilizes the fibril made of unstable collagen macromolecules. This mechanism can be relevant for explaining stability of collagen.


Cross-scanner and cross-protocol diffusion MRI data harmonisation: A benchmark database and evaluation of algorithms.
Diffusion MRI is being used increasingly in studies of the brain and other parts of the body for its ability to provide quantitative measures that are sensitive to changes in tissue microstructure. However, inter-scanner and inter-protocol differences are known to induce significant measurement variability, which in turn jeopardises the ability to obtain 'truly quantitative measures' and challenges the reliable combination of different datasets. Combining datasets from different scanners and/or acquired at different time points could dramatically increase the statistical power of clinical studies, and facilitate multi-centre research. Even though careful harmonisation of acquisition parameters can reduce variability, inter-protocol differences become almost inevitable with improvements in hardware and sequence design over time, even within a site. In this work, we present a benchmark diffusion MRI database of the same subjects acquired on three distinct scanners with different maximum gradient strength (40, 80, and 300 mT/m), and with 'standard' and 'state-of-the-art' protocols, where the latter have higher spatial and angular resolution. The dataset serves as a useful testbed for method development in cross-scanner/cross-protocol diffusion MRI harmonisation and quality enhancement. Using the database, we compare the performance of five different methods for estimating mappings between the scanners and protocols. The results show that cross-scanner harmonisation of single-shell diffusion data sets can reduce the variability between scanners, and highlight the promises and shortcomings of today's data harmonisation techniques.


Evaluating the Effect of Slow-Stroke Back Massage on the Anxiety of Candidates for Cataract Surgery.
The patients under cataract surgery often experience anxiety not only during the surgery, but also prior to the surgery. We sought to determine the effects of slow-stroke back massage on anxiety in patients undergoing cataract surgery. The study was conducted in the Amiral-momenin Hospital of Zabol city, south-east of Iran. A total of 60 candidates of cataract surgery participated in the study. The participants were randomly allocated to either control or intervention groups. The intervention group received slow-stroke back massages, while patients in control group received routine interventions. The slow-stroke back massage was performed on the patients assigned to the intervention group. The intervention was performed in the morning of the surgery day at 30 minutes before the surgery. The researcher performed each massage session in a sitting position. The duration of each massage session was 15 minutes. Anxiety was assessed in the both groups in the morning of the surgery, before and immediately after the intervention. Independent samples Student's t test, paired samples Student's t test, and chi-squared test were used to analyze the data. Anxiety was not significantly different between the two groups before and after the massage (p = .816). On the other hand, paired samples Student's t test showed a significant difference comparing the anxiety scores before (49.7±5.43) and after (45.16±3.89) the massage in the intervention group (p < .001). Based on our results, slow-stroke back massage, which is a low-cost and safe method, reduced anxiety in patients who were candidates for cataract surgery.


Circle of Willis variation in a complex stroke presentation: a case report.
The impact of circle of Willis anatomical variation upon the presentation of stroke is probably underrecognized. A 63-year-old right-handed woman developed a left hemiparesis and right leg weakness sequentially following a road traffic accident (RTA). Despite initial concern about the possibility of cervical spinal cord injury, the final diagnosis was bilateral artery-to-artery embolic cerebral infarction with dominant right internal carotid artery. The case illustrates the complex presentation of stroke as a pseudo-cervical cord lesion and the impact of circle of Willis anatomical variation upon the expression of large vessel cerebrovascular disease.


Evaluation of cytologic and biochemical variables in blood, plasma, and peritoneal fluid from calves before and after umbilical herniorrhaphy.
To establish reference intervals for cytologic and biochemical variables in peritoneal fluid, whole blood, and plasma in calves with congenital umbilical hernias (CUHs) before and after herniorrhaphy and to assess whether those variables in calves with CUHs were altered, compared with findings in clinically normal calves. 20 Holstein calves with or without a CUH. Procedures-10 calves with CUHs underwent herniorrhaphy. Blood and peritoneal fluid samples from all 20 calves were collected for cytologic and biochemical analyses on days 0 (before surgery), 1, 3, 5, 7, and 15. Data from the 2 groups were compared. Reference intervals for the variables of interest were established for each group. Before surgery, calves with CUHs had significantly greater plasma total protein concentration and creatine kinase (CK) and aspartate aminotransferase activities and peritoneal fluid specific gravity values, compared with values for calves without CUHs. At various time points after surgery, peritoneal fluid total protein concentration; fibrinogen concentration; nucleated cell, polymorphonuclear cell, and lymphocyte counts; specific gravity; and lactate dehydrogenase, aspartate aminotransferase, and CK activities in calves with CUHs were significantly different from values in calves without CUHs. Some plasma and blood variables (eg, total protein concentration, neutrophil count, and CK activity) were significantly different between the 2 groups. Values of certain cytologic and biochemical variables in peritoneal fluid, blood, and plasma were different between calves with and without CUHs. Thus, determination of reference intervals for these variables is important for interpreting diagnostic test results in calves with CUHs.


The DSF Family of Cell-Cell Signals: An Expanding Class of Bacterial Virulence Regulators.
Many pathogenic bacteria use cell-cell signaling systems involving the synthesis and perception of diffusible signal molecules to control virulence as a response to cell density or confinement to niches. Bacteria produce signals of diverse structural classes. Signal molecules of the diffusible signal factor (DSF) family are cis-2-unsaturated fatty acids. The paradigm is cis-11-methyl-2-dodecenoic acid from Xanthomonas campestris pv. campestris (Xcc), which controls virulence in this plant pathogen. Although DSF synthesis was thought to be restricted to the xanthomonads, it is now known that structurally related molecules are produced by the unrelated bacteria Burkholderia cenocepacia and Pseudomonas aeruginosa. Furthermore, signaling involving these DSF family members contributes to bacterial virulence, formation of biofilms and antibiotic tolerance in these important human pathogens. Here we review the recent advances in understanding DSF signaling and its regulatory role in different bacteria. These advances include the description of the pathway/mechanism of DSF biosynthesis, identification of novel DSF synthases and new members of the DSF family, the demonstration of a diversity of DSF sensors to include proteins with a Per-Arnt-Sim (PAS) domain and the description of some of the signal transduction mechanisms that impinge on virulence factor expression. In addition, we address the role of DSF family signals in interspecies signaling that modulates the behavior of other microorganisms. Finally, we consider a number of recently reported approaches for the control of bacterial virulence through the modulation of DSF signaling.


Cognitive and emotional variables predicting treatment outcome of cognitive behavior therapies for patients with medically unexplained symptoms: A meta-analysis.
Cognitive behavior therapy (CBT) is the best-evaluated psychological approach to treat patients with medically unexplained symptoms (MUS). We still need a better understanding of what characterizes patients with MUS who benefit more or less from CBT. This systematic review aimed to identify patients' cognitive-emotional characteristics predicting the outcome of CBT for MUS. A systematic literature search (PubMed, PsycINFO, Web of Science) revealed 37 eligible studies, 23 of these provided data for meta-analyses. Mean correlation coefficients between predictor variables and the outcomes (symptom intensity, physical or social-emotional functioning) were calculated using a random-effects model. Differences between syndromes of MUS were investigated with moderator analyses. Meta-analyses showed that patients with a comorbid mood disorder (r = 0.32, p < .01) or anxiety disorder (r = 0.18, p < .01), symptom catastrophizing and worries (r = 0.34, p < .01), tendencies of somatosensory amplification (r = 0.46, p = .04), and low symptom acceptance or self-efficacy (r = 0.25, p < .01) have a less favorable CBT outcome. Moderator analyses revealed that these associations between predictors and treatment outcome are pronounced in patients with chronic fatigue syndrome and irritable bowel syndrome. Our results show that pre-treatment differences in patients' cognitive-emotional characteristics predict patients' outcome in CBT. Patient-tailored CBT could be a promising approach to address MUS patients' widely varying needs more effectively. The protocol of this systematic review and meta-analysis was registered in the PROSPERO registry (CRD 42018098649).


Deductive biocomputing.
As biologists increasingly rely upon computational tools, it is imperative that they be able to appropriately apply these tools and clearly understand the methods the tools employ. Such tools must have access to all the relevant data and knowledge and, in some sense, "understand" biology so that they can serve biologists' goals appropriately and "explain" in biological terms how results are computed. We describe a deduction-based approach to biocomputation that semiautomatically combines knowledge, software, and data to satisfy goals expressed in a high-level biological language. The approach is implemented in an open source web-based biocomputing platform called BioDeducta, which combines SRI's SNARK theorem prover with the BioBike interactive integrated knowledge base. The biologist/user expresses a high-level conjecture, representing a biocomputational goal query, without indicating how this goal is to be achieved. A subject domain theory, represented in SNARK's logical language, transforms the terms in the conjecture into capabilities of the available resources and the background knowledge necessary to link them together. If the subject domain theory enables SNARK to prove the conjecture--that is, to find paths between the goal and BioBike resources--then the resulting proofs represent solutions to the conjecture/query. Such proofs provide provenance for each result, indicating in detail how they were computed. We demonstrate BioDeducta by showing how it can approximately replicate a previously published analysis of genes involved in the adaptation of cyanobacteria to different light niches. Through the use of automated deduction guided by a biological subject domain theory, this work is a step towards enabling biologists to conveniently and efficiently marshal integrated knowledge, data, and computational tools toward resolving complex biological queries.


Where and How Wolves (Canis lupus) Kill Beavers (Castor canadensis).
Beavers (Castor canadensis) can be a significant prey item for wolves (Canis lupus) in boreal ecosystems due to their abundance and vulnerability on land. How wolves hunt beavers in these systems is largely unknown, however, because observing predation is challenging. We inferred how wolves hunt beavers by identifying kill sites using clusters of locations from GPS-collared wolves in Voyageurs National Park, Minnesota. We identified 22 sites where wolves from 4 different packs killed beavers. We classified these kill sites into 8 categories based on the beaver-habitat type near which each kill occurred. Seasonal variation existed in types of kill sites as 7 of 12 (58%) kills in the spring occurred at sites below dams and on shorelines, and 8 of 10 (80%) kills in the fall occurred near feeding trails and canals. From these kill sites we deduced that the typical hunting strategy has 3 components: 1) waiting near areas of high beaver use (e.g., feeding trails) until a beaver comes near shore or ashore, 2) using vegetation, the dam, or other habitat features for concealment, and 3) immediately attacking the beaver, or ambushing the beaver by cutting off access to water. By identifying kill sites and inferring hunting behavior we have provided the most complete description available of how and where wolves hunt and kill beavers.


Intracranial extension of retrobulbar blastomycosis (Blastomyces dermatitidis) in a dog.
Blastomycosis (Blastomyces dermatitidis) is a fungal disease that is endemic in the southern United States. This case report illustrates the clinical, MRI and histopathologic findings in a dog with invasion of a retrobulbar blastomycotic lesion into the calvarium. A 5-year-old intact female Weimaraner was referred for a 2-month history of change in behavior and recent onset of visual deficits. Magnetic resonance imaging (MRI) examination revealed a large (5.8 × 2.0 × 2.5 cm) mass extending from the left orbit through a circular defect in the left cranioventral aspect of the calvarium caudally to the level of the pituitary fossa and interthalamic adhesion. The mass was heterogeneously iso- to hypointense on T2-W images, slightly hypointense on T1-W images, did not attenuate on fluid attenuated inversion recovery (FLAIR) images, and did not show evidence of susceptibility artifact on T2*-W gradient recalled echo (GRE) images. Vasogenic edema and associated mass effect were noted. The mass showed strong homogeneous contrast enhancement with well-defined margins and had thickening of the adjacent meninges (dural tail sign). Based on MRI findings a malignant neoplastic process was considered most likely and the patient was placed on oral prednisone to decrease peri-tumoral inflammation. The dog initially improved but was euthanized 3 weeks later for worsening clinical signs. Histopathologic assessment of the mass revealed marked pyogranulomatous optic neuritis with intralesional fungal yeasts consistent with blastomycosis (Blastomyces dermatitidis). To our knowledge this is the first report of invasion of a retrobulbar blastomycotic lesion into the calvarium in a dog.


Provision and awareness for isoniazid preventive therapy among PLHIV in Addis Ababa, Ethiopia.
The risk of acquiring tuberculosis by People living with HIV (PLHIV) could significantly be reduced through provision of isoniazid preventive therapy (IPT). In Ethiopia, it is neither practiced well nor researched in depth. Our objective was to assess IPT provision and awareness among PLHIV in Addis Ababa City Administration. Between February 2008 and May 2008, a cross sectional facility-based survey was conducted by exit interview of 406 PLHIV from six health facilities. The findings were analyzed and described in this report. The proportion of PLHIV ever had been provided with IPT were 74 of 231 TB free PLHIV (32.0%) and the proportion of having information about IPT among study participants was 29.8%. Females were about two times more informed about the provision of IPT in their health facilities than males [AOR (95%CI): 2.18 (1.31-3.61)]. We conclude that the practice of provision of IPT for PLHIV is high, but there is room for improvement. Provision of INH for TB free PLHIV has to be strengthened with better diagnostic facilities to certainly rule out active TB cases.


Distinct stage-specific transcriptional states of B cells derived from human tonsillar tissue.
B cells within secondary lymphoid tissues encompass a diversity of activation states and multiple maturation processes that reflect antigen recognition and transition through the germinal center (GC) reaction, in which mature B cells differentiate into memory and antibody-secreting cells (ASCs). Here, utilizing single-cell RNA-seq, we identify a range of distinct activation and maturation states of tonsil-derived B cells. In particular, we identify what we believe is a previously uncharacterized CCL4/CCL3 chemokine-expressing B cell population with an expression pattern consistent with B cell receptor/CD40 activation. Furthermore, we present a computational method that leverages regulatory network inference and pseudotemporal modeling to identify upstream transcription factor modulation along a GC-to-ASC axis of transcriptional maturation. Our data set provides valuable insight into diverse B cell functional profiles and will be a useful resource for further studies into the B cell immune compartment.


Conformational dynamics of the focal adhesion targeting domain control specific functions of focal adhesion kinase in cells.
Focal adhesion (FA) kinase (FAK) regulates cell survival and motility by transducing signals from membrane receptors. The C-terminal FA targeting (FAT) domain of FAK fulfils multiple functions, including recruitment to FAs through paxillin binding. Phosphorylation of FAT on Tyr(925) facilitates FA disassembly and connects to the MAPK pathway through Grb2 association, but requires dissociation of the first helix (H1) of the four-helix bundle of FAT. We investigated the importance of H1 opening in cells by comparing the properties of FAK molecules containing wild-type or mutated FAT with impaired or facilitated H1 openings. These mutations did not alter the activation of FAK, but selectively affected its cellular functions, including self-association, Tyr(925) phosphorylation, paxillin binding, and FA targeting and turnover. Phosphorylation of Tyr(861), located between the kinase and FAT domains, was also enhanced by the mutation that opened the FAT bundle. Similarly phosphorylation of Ser(910) by ERK in response to bombesin was increased by FAT opening. Although FAK molecules with the mutation favoring FAT opening were poorly recruited at FAs, they efficiently restored FA turnover and cell shape in FAK-deficient cells. In contrast, the mutation preventing H1 opening markedly impaired FAK function. Our data support the biological importance of conformational dynamics of the FAT domain and its functional interactions with other parts of the molecule.


Surveillance for waterborne-disease outbreaks--United States, 1999-2000.
Since 1971, CDC, the U.S. Environmental Protection Agency (EPA), and the Council of State and Territorial Epidemiologists (CSTE) have maintained a collaborative surveillance system for the occurrences and causes of waterborne-disease outbreaks (WBDOs).This surveillance system is the primary source of data concerning the scope and effects of waterborne diseases on persons in the United States. This summary includes data regarding outbreaks occurring during January 1999-December 2000 and previously unreported outbreaks occurring in 1995 and 1997. The surveillance system includes data for outbreaks associated with drinking water and recreational water. State, territorial, and local public health departments are primarily responsible for detecting and investigating WBDOs and voluntarily reporting them to CDC on a standard form. The unit of analysis for the WBDO surveillance system is an outbreak, not an individual case of a waterborne disease. Two criteria must be met for an event to be defined as a WBDO. First, > or = 2 persons must have experienced a similar illness after either ingestion of drinking water or exposure to water encountered in recreational or occupational settings. This criterion is waived for single cases of laboratory-confirmed primary amebic meningoencephalitis and for single cases of chemical poisoning if water-quality data indicate contamination by the chemical. Second, epidemiologic evidence must implicate water as the probable source of the illness. During 1999-2000, a total of 39 outbreaks associated with drinking water was reported by 25 states. Included among these 39 outbreaks was one outbreak that spanned 10 states. These 39 outbreaks caused illness among an estimated 2,068 persons and were linked to two deaths. The microbe or chemical that caused the outbreak was identified for 22 (56.4%) of the 39 outbreaks; 20 of the 22 identified outbreaks were associated with pathogens, and two were associated with chemical poisoning. Of the 17 outbreaks involving acute gastroenteritis of unknown etiology, one was a suspected chemical poisoning, and the remaining 16 were suspected as having an infectious cause. Twenty-eight (71.8%) of 39 outbreaks were linked to groundwater sources; 18 (64.3%) of these 28 groundwater outbreaks were associated with private or noncommunity wells that were not regulated by EPA. Fifty-nine outbreaks from 23 states were attributed to recreational water exposure and affected an estimated 2,093 persons. Thirty-six (61.0%) of the 59 were outbreaks involving gastroenteritis. The etiologic agent was identified in 30 (83.3%) of 36 outbreaks involving gastroenteritis. Twenty-two (61.1%) of 36 gastroenteritis-related outbreaks were associated with pools or interactive fountains. Four (6.8%) of the 59 recreational water outbreaks were attributed to single cases of primary amebic meningoencephalitis (PAM) caused by Naegleria fowleri. All four cases were fatal. Fifteen (25.4%) of the 59 outbreaks were associated with dermatitis; 12 (80.0%) of 15 were associated with hot tubs or pools. In addition, recreational water outbreaks of leptospirosis, Pontiac fever, and chemical keratitis, as well as two outbreaks of leptospirosis and Pontiac fever associated with occupational exposure were also reported to CDC. The proportion of drinking water outbreaks associated with surface water increased from 11.8% during 1997-1998 to 17.9% in 1999-2000. The proportion of outbreaks (28) associated with groundwater sources increased 87% from the previous reporting period (15 outbreaks), and these outbreaks were primarily associated (60.7%) with consumption of untreated groundwater. Recreational water outbreaks involving gastroenteritis doubled (36 outbreaks) from the number of outbreaks reported in the previous reporting period (18 outbreaks). These outbreaks were most frequently associated with Cryptosporidium parvum (68.2%) in treated water venues (e.g., swimming pools or interactive fountains) and by Escherichia coli O157:H7 (21.4%) in freshwater venues. The increase in the number of outbreaks probably reflects improved surveillance and reporting at the local and state level as well as a true increase in the number of WBDOs. CDC and others have used surveillance data to identify the types of water systems, their deficiencies, and the etiologic agents associated with outbreaks and evaluated current technologies for providing safe drinking water and safe recreational water. Surveillance data are used also to establish research priorities, which can lead to improved water-quality regulations. Only the groundwater systems under the influence of surface water are required to disinfect their water supplies, but EPA is developing a groundwater rule that specifies when corrective action (including disinfection) is required. CDC and EPA are conducting epidemiologic studies to assess the level of waterborne illness attributable to municipal drinking water in nonoutbreak conditions. Rules under development by EPA--the Ground Water Rule (GWR), the Long Term 2 Enhanced Surface Water Treatment Rule (LT2ESWTR), and Stage 2 Disinfection Byproduct Rules (DBPR)--are expected to further protect the public from contaminants and disinfection byproducts in drinking water. Efforts by EPA under the Beaches Environmental Assessment, Closure, and Health (BEACH) program are aimed at reducing the risks for infection attributed to ambient recreational water by strengthening beach standards and testing; providing faster laboratory test methods; predicting pollution; investing in health and methods research; and improving public access to information regarding both the quality of the water at beaches and information concerning health risks associated with swimming in polluted water. EPA's Beach Watch (available at http://www.epa.gov/waterscience/beaches) provides online information regarding water quality at U.S. beaches, local protection programs, and other beach-related programs. CDC partnered with a consortium of local and national pool associations to develop a series of health communication materials for the general public who attend treated recreational water venues and to staff who work at those venues. CDC has also developed a recreational water outbreak investigation toolkit that can be used by public health professionals. All of the CDC materials are accessible at the CDC Healthy Swimming website (http://www.cdc.gov/healthyswimming).


Programmed cell death in the embryonic central nervous system of Drosophila melanogaster.
Although programmed cell death (PCD) plays a crucial role throughout Drosophila CNS development, its pattern and incidence remain largely uninvestigated. We provide here a detailed analysis of the occurrence of PCD in the embryonic ventral nerve cord (VNC). We traced the spatio-temporal pattern of PCD and compared the appearance of, and total cell numbers in, thoracic and abdominal neuromeres of wild-type and PCD-deficient H99 mutant embryos. Furthermore, we have examined the clonal origin and fate of superfluous cells in H99 mutants by DiI labeling almost all neuroblasts, with special attention to segment-specific differences within the individually identified neuroblast lineages. Our data reveal that although PCD-deficient mutants appear morphologically well-structured, there is significant hyperplasia in the VNC. The majority of neuroblast lineages comprise superfluous cells, and a specific set of these lineages shows segment-specific characteristics. The superfluous cells can be specified as neurons with extended wild-type-like or abnormal axonal projections, but not as glia. The lineage data also provide indications towards the identities of neuroblasts that normally die in the late embryo and of those that become postembryonic and resume proliferation in the larva. Using cell-specific markers we were able to precisely identify some of the progeny cells, including the GW neuron, the U motoneurons and one of the RP motoneurons, all of which undergo segment-specific cell death. The data obtained in this analysis form the basis for further investigations into the mechanisms involved in the regulation of PCD and its role in segmental patterning in the embryonic CNS.


Hydroxyethyl starches in the perioperative period. A review on the efficacy and safety of starch solutions.
Several randomized controlled trials have raised alarming concerns about the safety of hydroxyethyl starches (HES) for the hemodynamic stabilization of critically ill patients. It has been repeatedly demonstrated that the use of HES in patients treated in an intensive care unit was associated with an increased occurrence of serious adverse events, including a higher incidence of renal injury or failure, a higher need for renal replacement therapy (RRT), and (in one study) increased mortality. HES solutions are also widely used in the perioperative period, although high-level evidence on both the efficacy and safety of HES in patients undergoing surgery is sparse. It is unknown to which extent the concerning findings in critically ill patients can be extrapolated to the perioperative setting, where a variety of distinct patient populations is encountered. However, caution and an increased awareness for possible adverse effects of HES solutions in the perioperative setting are warranted. In 2013, the European Medicines Agency (EMA) concluded that the benefits of HES solutions no longer outweigh their risks, and therefore recommended that the marketing authorizations for these medicines be suspended (1). Upon request of some of the marketing-authorization holders (MAH), the EMA re-analyzed its recommendation. After a thorough review of the available data, the Pharmacovigilance Risk Assessment Committee (PRAC) concluded, on November 11th, that the increased risk of mortality and RRT or renal failure associated with the use of hydroxyethyl starch containing medicinal products outweighs their limited clinical benefits in the approved indications, and in any patient population. In June 2013, the U.S. Food and Drug Administration also communicated a serious warning with respect to the use of HES (2). The present (non-systematic) review summarizes the evidence upon which these remarkable recommendations are based. Moreover, current guidelines on the use of HES are quoted.


Outcome measures in the clinical evaluation of ambulatory Charcot-Marie-Tooth 1A subjects.
The outcome measures (OMs) in clinical trials for Charcot-Marie-Tooth disease (CMT) still represent an issue. A recent study highlighted that three additional clinical OMs, the 10-Meter Walk Test (10MWT), the 9-Hole Peg Test, and foot dorsal flexion dynamometry, further improve discrimination between severely and mildly affected patients. Another study has recently assessed the validity and reliability of the 6-Minute Walk Test (6MWT). The aim of this study was to identify the most useful scales in the clinical evaluation of CMT1A patients. Observational study of the baseline data collected in a multicenter, prospective, randomized, single blind, controlled study to evaluate the efficacy and safety of an innovative rehabilitation protocol based on treadmill training, stretching, respiratory, and proprioceptive exercises (TreSPE study) in CMT1A patients. The outpatient service of the four Italian centers involved, which are specialized in hereditary neuropathies. Fifty-three subjects with a clinical and genetically confirmed diagnosis of CMT1A. At baseline, in addition to the CMT Neuropathy Score, all subjects underwent walking evaluation (6MWT, 10MWT), balance assessment (Berg Balance Scale [BBS], Short Physical Performance Battery [SPPB]) and a subjective evaluation of quality of life (SF36) and walking ability (Walk12). Analyzing the baseline data, as expected, we found a strong correlation between walk and balance evaluation, proving the validity of these tests in investigating the functional impairment of CMT1A subjects. Particularly, we found that subjects with better balance control walk at higher speed and perceive less limitations in their physical activities or motor skills. This can be reconducted to the fact that ankle stability depends upon different factors such as anatomy integrity, muscle strength and proprioception. We identify the 6MWT, 10MWT, and SPPB as the most useful scales, in addition to the CMTNS, to evaluate the functional impairment of CMT1A patients who retain their walking capability and we suggest the use of SPPB because of its rapidity to assess balance and gait disorders in clinical settings. In the clinical practice it is important to evaluate patients comprehensively but rapidly. These outcome measures can help us to correctly assess balance and walking ability in CMT1A patients.


Factors Associated With Women's Reporting of Intimate Partner Violence in Spain.
In Spain, in 2013, the 20% of women who were murdered by their partner had reported him previously. We analyze the 2011 Spanish-Macrosurvey on Gender Violence to identify and analyze the prevalence of and the principal factors associated with reporting a situation of intimate partner violence (IPV) and the main reasons women cite for not filing such reports, or for subsequently deciding to withdraw their complaint. Overall, 72.8% of women exposed to IPV did not report their aggressor. The most frequent reasons for not reporting were not giving importance to the situation (33.9%), and fear and lack of trust in the reporting process (21.3%). The main reasons for withdrawing the complaint were cessation of the violence (20.0%), and fear and threats (18.2%). The probability of reporting increased among women with young children who were abused, prevalence ratio (95% confidence interval [CI]): 2.14 [1.54, 2.98], and those whose mother was abused, prevalence ratio (95% CI): 2.25 [1.42, 3.57]. Always focusing on the need to protect women who report abuse, it is necessary to promote the availability of and access to legal resources especially among women who use them less: women who do not have children and women who do not have previous family exposure to violence.


Coupling of human immunodeficiency virus type 1 fusion to virion maturation: a novel role of the gp41 cytoplasmic tail.
Retrovirus particles are not infectious until they undergo proteolytic maturation to form a functional core. Here we report a link between human immunodeficiency virus type 1 (HIV-1) core maturation and the ability of the virus to fuse with target cells. Using a recently developed reporter assay of HIV-1 virus-cell fusion, we show that immature HIV-1 particles are 5- to 10-fold less active for fusion with target cells than are mature virions. The fusion of mature and immature virions was rendered equivalent by truncating the gp41 cytoplasmic domain or by pseudotyping viruses with the glycoprotein of vesicular stomatitis virus. An analysis of a panel of mutants containing mutated cleavage sites indicated that HIV-1 fusion competence is activated by the cleavage of Gag at any site between the MA and NC segments and not as an indirect consequence of an altered core structure. These results suggest a mechanism by which binding of the gp41 cytoplasmic tail to Gag within immature HIV-1 particles inhibits Env conformational changes on the surface of the virion that are required for membrane fusion. This "inside-out" regulation of HIV-1 fusion could play an important role in the virus life cycle by preventing the entry of immature, noninfectious particles.


Electrical energy storage for the grid: a battery of choices.
The increasing interest in energy storage for the grid can be attributed to multiple factors, including the capital costs of managing peak demands, the investments needed for grid reliability, and the integration of renewable energy sources. Although existing energy storage is dominated by pumped hydroelectric, there is the recognition that battery systems can offer a number of high-value opportunities, provided that lower costs can be obtained. The battery systems reviewed here include sodium-sulfur batteries that are commercially available for grid applications, redox-flow batteries that offer low cost, and lithium-ion batteries whose development for commercial electronics and electric vehicles is being applied to grid storage.


Visible foliar injury of red spruce seedlings subjected to simulated acid mist.
Two-year-old red spruce seedlings [Picea rubens Sarg. syn. P. rubra (Du Roi) Link] were subjected to 6 simulated acid mist treatments (pH 2.5, 2.7, 3.0, 3.5, 4.0, 5.0) in a replicated experiment using open-top chambers. Acid mist solutions containing equimolar (NH4 )2 SO4 , and HNO3 were applied twice weekly for 22 weeks, each application being equivalent to 2 mm of precipitation. Visible symptoms of foliar damage were observed on the 3 most acidic mist treatments (pH 2.5, 2.7, 3.0). The inputs of nitrogen, sulphur and acidity in the most acidic treatment were 55, 42, 1,3 kg ha-1 , respectively, over a 10 week period. The plants subjected to the pH 2.5 treatment were found to be most severely damaged with approximately 40% foliar necrosis after 10 weeks of treatment. On approximately 80% of seedlings, necrosis was confined to current year needles only. These damaged needles were initially light brown or light orange in colour turning a deeper red 3 to 5 weeks after initial necrosis. Percentage foliar damage was linearly related to concentration (of NH4 + , NO3 - , SO4 2- and H+ ) with 62% foliar damage in the pH 2.5 treatment after a 22-week period. Spray application stopped in December 1987. Observations during the following spring showed that the pH 2.5 and pH 3 treatments induced earlier Hushing, requiring 60 day °C less thermal time than the pH 5-0 treatment. In 1988, this decrease in thermal requirement was equivalent to flushing 11 days earlier. There was no evidence of acid mist treatments inducing bud mortality.


Prophylactic correction of the international normalized ratio in neurosurgery: a brief review of a brief literature.
Prophylactic fresh-frozen plasma (FFP) transfusion is often undertaken in hemodynamically stable patients with a minimally elevated international normalized ratio (INR) prior to invasive procedures, despite little evidence in support of this practice. The authors review the current literature in an attempt to clarify best clinical practice with regard to this issue. Although the activated partial thromboplastin time and prothrombin time-INR are useful laboratory tests to measure specific clotting factors in the coagulation cascade, in the absence of active bleeding or a preexisting coagulopathy, their utility as predictors of overall bleeding risk is limited. Several studies have shown an imperfect correlation between mild elevations in the INR and subsequent bleeding tendency. Furthermore, FFP transfusion is not always sufficient to achieve normal INR values in patients who have mild elevations (< 2) to begin with. Finally, there are risks associated with FFP transfusion, including potential transfusion-associated [disease] exposures as well as the time delay imposed by laboratory testing and transfusion administration prior to initiation of procedures. The authors propose that the current concept of a "normal" INR value warrants redefinition to make it a more meaningful clinical tool. Based on their review of the literature, the authors suggest that in a hemodynamically stable patient population there is a range of mildly prolonged INR values for which FFP transfusion is not beneficial, and is potentially harmful.


Randomised controlled trial of a Calcium Channel or Angiotensin Converting Enzyme Inhibitor/Angiotensin Receptor Blocker Regime to Reduce Blood Pressure Variability following Ischaemic Stroke (CAARBS): a protocol for a feasibility study.
Raised blood pressure (BP) is common after stroke and is associated with a poor prognosis, yet trials of BP lowering in the immediate poststroke period have not demonstrated a benefit. One possible explanation for this may be that BP variability (BPV) rather than absolute levels predicts outcome, as BPV is increased after stroke and is associated with poor outcomes. Furthermore, there is evidence of distinct antihypertensive class effects on BPV despite similar BP-lowering effects. However, whether BPV in the immediate poststroke period is a therapeutic target has not been prospectively investigated.The objectives of this trial are to assess the feasibility and safety of recruiting patients following an acute ischaemic stroke or transient ischaemic attack (TIA) to an interventional randomised controlled trial comparing the effects of two different antihypertensive drug classes on BPV. Secondary exploratory objectives are to assess if different therapeutic strategies have diverse effects on levels of BPV and if this has an impact on outcomes. 150 adult patients with first-ever ischaemic stroke or TIA who require antihypertensive therapy for secondary prevention will be recruited within 7 days of the event from stroke services across three sites. After baseline assessments they will be randomly assigned to treatment with a calcium channel blocker or ACE inhibitor/angiotensin receptor blocker-based regimen and followed up for a period of three months. Ethical and regulatory approvals have been granted. Dissemination is planned via publication in peer-reviewed medical journals and presentation at relevant conferences. ISRCTN10853487.


Sphingosine 1-phosphate protects primary human keratinocytes from apoptosis via nitric oxide formation through the receptor subtype S1P₃.
Although the lipid mediator sphingosine 1-phosphate (S1P) has been identified to induce cell growth arrest of human keratinocytes, the sphingolipid effectively protects these epidermal cells from apoptosis. The molecular mechanism of the anti-apoptotic action induced by S1P is less characterized. Apart from S1P, endogenously produced nitric oxide (NO•) has been recognized as a potent modulator of apoptosis in keratinocytes. Therefore, it was of great interest to elucidate whether S1P protects human keratinocytes via a NO•-dependent signalling pathway. Indeed, S1P induced an activation of endothelial nitric oxide synthase (eNOS) in human keratinocytes leading to an enhanced formation of NO•. Most interestingly, the cell protective effect of S1P was almost completely abolished in the presence of the eNOS inhibitor L-NAME as well as in eNOS-deficient keratinocytes indicating that the sphingolipid metabolite S1P protects human keratinocytes from apoptosis via eNOS activation and subsequent production of protective amounts of NO•. It is well established that most of the known actions of S1P are mediated by a family of five specific G protein-coupled receptors. Therefore, the involvement of S1P-receptor subtypes in S1P-mediated eNOS activation has been examined. Indeed, this study clearly shows that the S1P(3) is the exclusive receptor subtype in human keratinocytes which mediates eNOS activation and NO• formation in response to S1P. In congruence, when the S1P(3) receptor subtype is abrogated, S1P almost completely lost its ability to protect human keratinocytes from apoptosis.


Hepatitis B virus genotypes A and D in Uganda.
The prevalence of hepatitis B virus (HBV) infection in Uganda is 10%. Hepatitis B virus genotypes impact on treatment response, rate of spontaneous recovery and progression of chronic HBV infection and hepatocellular carcinoma. There is little information on the HBV genotypic distribution in Uganda. To determine HBV genotypes in Uganda. The MBN clinical laboratory performs HBV viral load and genotype testing in Uganda. It receives hepatitis B surface antigen (HBsAg)-positive samples from all over the country for additional HBV testing. Samples are stored for 6 months before being discarded. Our study used delinked stored samples. PCR-positive samples had DNA extracted and used as template for HBV genome amplification by nested PCR. Reverse hybridisation was performed and genotypes were determined by the line probe assay method (INNO-LiPA). One hundred stored HBsAg-positive plasma samples with detectable viral loads were analysed. Of these, 93 samples showed PCR amplification products and gave genotype-specific probe lines on the INNO-LiPA assay. Of the patients, where gender was recorded, 60.9% were female, and the overall median age (IQR) was 25 (2-60) years. There was a predominance of HBV genotype D (47 patients; 50.5%), followed by genotype A, (16 patients; 17.2%). One patient (1.1%) had genotype E. In 28% of the samples mixed infections were detected with genotypes A/E (9.7%) and A/D (6.5%) being most common. Genotypes B, C, E and H only occurred as part of mixed infections. Hepatitis B genotypes D and A were predominant in our study population.


A Nuclease Protection ELISA Assay for Colorimetric and Electrochemical Detection of Nucleic Acids.
Early and accurate diagnosis is crucial to monitor infection outcomes and provide timely interventions. However, gold standard polymerase chain reaction assays (PCR) are labor-intensive and require expensive reagents and instrumentation. Nuclease protection has been used for decades to detect and quantify nucleic acid but has not yet been investigated as a diagnostic tool for infectious disease. In this work, we describe a nuclease protection enzyme-linked immunosorbent assay (NP-ELISA) for accurate and sensitive detection of nucleic acid. Briefly, binding of a nucleic acid target to an oligo probe protects it from digestion of un-hybridized nucleic acid by S1 nuclease. Following the workflow of an ELISA, a horseradish peroxidase (HRP)-conjugated antibody binds the probe and oxidizes its substrate to generate signal. The assay was validated with three HRP substrates for absorbance, chemiluminescence, and electrochemical readouts, demonstrating great versatility. Electrochemical detection with 3,3',5,5'-Tetramethylbenzidine (TMB) gave the highest assay sensitivity with a limit of detection of 3.72×103 molecules mL-1. Furthermore, non-complementary targets did not generate a response, indicating a high degree of specificity. This proof of principle serves as a stepping stone towards developing miniaturized, multiplexed nuclease protection assays for point-of-care diagnosis.


Conceptual design of a neutron camera for MAST Upgrade.
This paper presents two different conceptual designs of neutron cameras for Mega Ampere Spherical Tokamak (MAST) Upgrade. The first one consists of two horizontal cameras, one equatorial and one vertically down-shifted by 65 cm. The second design, viewing the plasma in a poloidal section, also consists of two cameras, one radial and the other one with a diagonal view. Design parameters for the different cameras were selected on the basis of neutron transport calculations and on a set of target measurement requirements taking into account the predicted neutron emissivities in the different MAST Upgrade operating scenarios. Based on a comparison of the cameras' profile resolving power, the horizontal cameras are suggested as the best option.


Drinking water source and human Toxoplasma gondii infection in the United States: a cross-sectional analysis of NHANES data.
Toxoplasma gondii imparts a considerable burden to public health. Human toxoplasmosis can be life-threatening in immunocompromised individuals, has been associated with psychiatric disorders, and can cause severe congenital pathologies, spontaneous abortion, or stillbirth. Environmental modes of transmission contributing to the incidence of human toxoplasmosis are poorly understood. We sought to examine National Health and Nutrition Examination Survey (NHANES) data for risk factors associated with T. gondii seroprevalence. T. gondii serology results reported for Continuous NHANES survey years 1999-2004 and 2009-10 were examined. To explore associations with toxoplasmosis seropositivity, covariates of interest were selected a priori, including source and home treatment of tap water. Associations between potential risk factors and evidence of IgG antibodies against T. gondii were assessed using multivariable logistic regression. Among 23,030 participants with available T. gondii serology across 8 years of continuous NHANES survey data (1999-2004; 2009-2010), persons born outside the United States were significantly more likely to be seropositive, and seropositivity was inversely associated with years spent in the United States. Among US-born participants, participants with homes on well water (both those who used at-home water treatment devices and those who did not), as well as participants with public/private company-provided tap water who did not use at-home water treatment devices, were significantly more likely to be seropositive compared to participants who used home treatment devices on tap water provided by a private or public water company. A comparative subpopulation analysis revealed age-adjusted seroprevalence among US-born persons 12-49 yrs old significantly declined to 6.6% (95% CI, 5.2-8.0) (P <0.0001) in 2009-10, compared to previously published reports for NHANES data from 1988-1994 (14.1%) and 1999-2004 (9.0%). Data suggests that T. gondii infections continue to decline in the United States, but the overall infection rate remains substantial at nearly 7%. Despite the limitations in the Continuous NHANES cross-sectional survey, the association between well water use and T. gondii infection warrants further research.


(Z)3,4,5,4'-trans-tetramethoxystilbene, a new analogue of resveratrol, inhibits gefitinb-resistant non-small cell lung cancer via selectively elevating intracellular calcium level.
Calcium is a second messenger which is required for regulation of many cellular processes. However, excessive elevation or prolonged activation of calcium signaling would lead to cell death. As such, selectively regulating calcium signaling could be an alternative approach for anti-cancer therapy. Recently, we have identified an effective analogue of resveratrol, (Z)3,4,5,4'-trans-tetramethoxystilbene (TMS) which selectively elevated the intracellular calcium level in gefitinib-resistant (G-R) non-small-cell lung cancer (NSCLC) cells. TMS exhibited significant inhibitory effect on G-R NSCLC cells, but not other NSCLC cells and normal lung epithelial cells. The phosphorylation and activation of EGFR were inhibited by TMS in G-R cells. TMS induced caspase-independent apoptosis and autophagy by directly binding to SERCA and causing endoplasmic reticulum (ER) stress and AMPK activation. Proteomics analysis also further confirmed that mTOR pathway, which is the downstream of AMPK, was significantly suppressed by TMS. JNK, the cross-linker of ER stress and mTOR pathway was significantly activated by TMS. In addition, the inhibition of JNK activation can partially block the effect of TMS. Taken together, TMS showed promising anti-cancer activity by mediating calcium signaling pathway and inducing apoptosis as well as autophagy in G-R NSCLC cells, providing strategy in designing multi-targeting drug for treating G-R patients.


Mortality from occupational exposure to relatively pure chrysotile: a 39-year study.
Asbestos exposure is related to serious adverse health effects. However, there is disagreement about the relationship between chrysotile exposure and mesothelioma or lung cancer. Our aim was to investigate the mortality rate among workers exposed to relatively pure chrysotile in an asbestos cement factory. In an asbestos cement plant opened in 1968, we prospectively studied all 317 workers. A quantity of 2,000 tons of chrysotile, with minimal amphibole contamination, was used annually until 1 January 2005. Asbestos fiber concentration was measured regularly. Date and cause of death were recorded among active and retired workers. Asbestos fiber concentration was always below permissible levels. Fifty-two workers died during the study. The cause was cancer in 28 subjects; lung cancer was diagnosed in 16 of them. No case of mesothelioma was reported. Death was attributed to cardiovascular diseases in 23 subjects and to liver cirrhosis in 1. Overall mortality rate was significantly lower than that of the Greek general population, standardized mortality ratio (SMR) was 0.71 (95% CI 0.53-0.93). Mortality due to cancer was increased (SMR 1.15, 95% CI 0.77-1.67), mainly due to lung cancer mortality (SMR 1.71, 95% CI 0.98-2.78), but not significantly. Occupational exposure to relatively pure chrysotile within permissible levels was not associated with a significant increase in lung cancer or with mesothelioma. Decreased overall mortality of workers indicates a healthy worker effect, which--together with the relatively small cohort size--could have prevented small risks to be detected.


Brain aging in major depressive disorder: results from the ENIGMA major depressive disorder working group.
Major depressive disorder (MDD) is associated with an increased risk of brain atrophy, aging-related diseases, and mortality. We examined potential advanced brain aging in adult MDD patients, and whether this process is associated with clinical characteristics in a large multicenter international dataset. We performed a mega-analysis by pooling brain measures derived from T1-weighted MRI scans from 19 samples worldwide. Healthy brain aging was estimated by predicting chronological age (18-75 years) from 7 subcortical volumes, 34 cortical thickness and 34 surface area, lateral ventricles and total intracranial volume measures separately in 952 male and 1236 female controls from the ENIGMA MDD working group. The learned model coefficients were applied to 927 male controls and 986 depressed males, and 1199 female controls and 1689 depressed females to obtain independent unbiased brain-based age predictions. The difference between predicted "brain age" and chronological age was calculated to indicate brain-predicted age difference (brain-PAD). On average, MDD patients showed a higher brain-PAD of +1.08 (SE 0.22) years (Cohen's d = 0.14, 95% CI: 0.08-0.20) compared with controls. However, this difference did not seem to be driven by specific clinical characteristics (recurrent status, remission status, antidepressant medication use, age of onset, or symptom severity). This highly powered collaborative effort showed subtle patterns of age-related structural brain abnormalities in MDD. Substantial within-group variance and overlap between groups were observed. Longitudinal studies of MDD and somatic health outcomes are needed to further assess the clinical value of these brain-PAD estimates.


Electrostatics, proton sensor, and networks governing the gating transition in GLIC, a proton-gated pentameric ion channel.
The pentameric ligand-gated ion channel (pLGIC) from Gloeobacter violaceus (GLIC) has provided insightful structure-function views on the permeation process and the allosteric regulation of the pLGICs family. However, GLIC is activated by pH instead of a neurotransmitter and a clear picture for the gating transition driven by protons is still lacking. We used an electrostatics-based (finite difference Poisson-Boltzmann/Debye-Hückel) method to predict the acidities of all aspartic and glutamic residues in GLIC, both in its active and closed-channel states. Those residues with a predicted pKa close to the experimental pH50 were individually replaced by alanine and the resulting variant receptors were titrated by ATR/FTIR spectroscopy. E35, located in front of loop F far away from the orthosteric site, appears as the key proton sensor with a measured individual pKa at 5.8. In the GLIC open conformation, E35 is connected through a water-mediated hydrogen-bond network first to the highly conserved electrostatic triad R192-D122-D32 and then to Y197-Y119-K248, both located at the extracellular domain-transmembrane domain interface. The second triad controls a cluster of hydrophobic side chains from the M2-M3 loop that is remodeled during the gating transition. We solved 12 crystal structures of GLIC mutants, 6 of them being trapped in an agonist-bound but nonconductive conformation. Combined with previous data, this reveals two branches of a continuous network originating from E35 that reach, independently, the middle transmembrane region of two adjacent subunits. We conclude that GLIC's gating proceeds by making use of loop F, already known as an allosteric site in other pLGICs, instead of the classic orthosteric site.


Well-being in older Mexican American spouses.
There is a strong connection between marriage and well-being, with evidence suggesting that the well-being of one spouse is closely correlated with that of the other. However, among older Mexican Americans, there is little information about this phenomenon. To address this, we explore two research questions: Does one spouse's well-being predict the other spouse's well-being? Are there gender differences in these effects? We assess information from 553 couples who participated in Wave 1 (1993-1994) of the Hispanic Established Populations for the Epidemiologic Studies of the Elderly. Using structural equation models, we examined three aspects of well-being among older Mexican American couples: depressive symptoms, life satisfaction, and self-rated health. The findings revealed evidence of an association between the well-being of one spouse and that of the other. Specifically, the self-rated health of husbands and wives predicts that of their partners. However, there is evidence that husbands' depressive symptoms and life satisfaction influence wives' well-being, but not the reverse. The findings from this study are important because they add to the literature on the connection of well-being among spouses, point to important gender differences, focus on an understudied minority group with unique cultural characteristics, and have implications for the examination of well-being within a marriage framework.


Computation of likelihood ratios in fingerprint identification for configurations of any number of minutiae.
Recent court challenges have highlighted the need for statistical research on fingerprint identification. This paper proposes a model for computing likelihood ratios (LRs) to assess the evidential value of comparisons with any number of minutiae. The model considers minutiae type, direction and relative spatial relationships. It expands on previous work on three minutiae by adopting a spatial modeling using radial triangulation and a probabilistic distortion model for assessing the numerator of the LR. The model has been tested on a sample of 686 ulnar loops and 204 arches. Features vectors used for statistical analysis have been obtained following a preprocessing step based on Gabor filtering and image processing to extract minutiae data. The metric used to assess similarity between two feature vectors is based on an Euclidean distance measure. Tippett plots and rates of misleading evidence have been used as performance indicators of the model. The model has shown encouraging behavior with low rates of misleading evidence and a LR power of the model increasing significantly with the number of minutiae. The LRs that it provides are highly indicative of identity of source on a significant proportion of cases, even when considering configurations with few minutiae. In contrast with previous research, the model, in addition to minutia type and direction, incorporates spatial relationships of minutiae without introducing probabilistic independence assumptions. The model also accounts for finger distortion.


Behavioural, ecological and evolutionary responses to extreme climatic events: challenges and directions.
More extreme climatic events (ECEs) are among the most prominent consequences of climate change. Despite a long-standing recognition of the importance of ECEs by paleo-ecologists and macro-evolutionary biologists, ECEs have only recently received a strong interest in the wider ecological and evolutionary community. However, as with many rapidly expanding fields, it lacks structure and cohesiveness, which strongly limits scientific progress. Furthermore, due to the descriptive and anecdotal nature of many ECE studies it is still unclear what the most relevant questions and long-term consequences are of ECEs. To improve synthesis, we first discuss ways to define ECEs that facilitate comparison among studies. We then argue that biologists should adhere to more rigorous attribution and mechanistic methods to assess ECE impacts. Subsequently, we discuss conceptual and methodological links with climatology and disturbance-, tipping point- and paleo-ecology. These research fields have close linkages with ECE research, but differ in the identity and/or the relative severity of environmental factors. By summarizing the contributions to this theme issue we draw parallels between behavioural, ecological and evolutionary ECE studies, and suggest that an overarching challenge is that most empirical and theoretical evidence points towards responses being highly idiosyncratic, and thus predictability being low. Finally, we suggest a roadmap based on the proposition that an increased focus on the mechanisms behind the biological response function will be crucial for increased understanding and predictability of the impacts of ECE.This article is part of the themed issue 'Behavioural, ecological and evolutionary responses to extreme climatic events'.


Characteristics of thyroidectomy in Taiwan.
Thyroidectomy is a common treatment for thyroid disorders in Taiwan, and constitutes a significant percentage of medical expenses. This study investigated the characteristics of thyroidectomy in Taiwan. A total of 21 senior general surgeons from 16 medical centers and 5 regional hospitals participated. One surgeon from each hospital reviewed the hospital's medical records of thyroid operations performed in the year 2001. Medical records for 3846 thyroidectomies were retrospectively analyzed, including surgical indications and modalities, complications, pathology reports, and the use of antibiotics. Most of the patients were women (85%). Indications for surgery included proven malignancy (9%), suspicious malignancy (30%), evident compression symptoms (20%), hyperthyroidism (20%), and cosmetic reasons (12%). The majority of patients (78%) underwent a surgical procedure with lobectomy or bilateral thyroidectomy; 13% had unilateral partial thyroidectomy. Prophylactic antibiotics were administered in 46% of procedures, and continued postoperatively in 12%. Postoperative complications occurred in 12% of patients. Hypocalcemia was the most common complication (8%), and its incidence was significantly correlated with the frequency of total thyroidectomy (p < 0.01). Complete pathology reports for the thyroidectomized specimens were available in only 65% of the cases. The frequency of cosmetic reasons for surgery, unilateral subtotal resection, routine antibiotic administration, and incomplete pathology reports were significantly higher in regional hospitals than in medical centers. Reduction in the high rates of cosmetic reasons for surgery, unilateral partial thyroidectomy, incomplete pathology reports, and use of antibiotic prophylaxis are needed to improve the quality of thyroidectomy in Taiwan.


Methane Emission in a Specific Riparian-Zone Sediment Decreased with Bioelectrochemical Manipulation and Corresponded to the Microbial Community Dynamics.
Dissimilatory metal-reducing bacteria are widespread in terrestrial ecosystems, especially in anaerobic soils and sediments. Thermodynamically, dissimilatory metal reduction is more favorable than sulfate reduction and methanogenesis but less favorable than denitrification and aerobic respiration. It is critical to understand the complex relationships, including the absence or presence of terminal electron acceptors, that govern microbial competition and coexistence in anaerobic soils and sediments, because subsurface microbial processes can effect greenhouse gas emissions from soils, possibly resulting in impacts at the global scale. Here, we elucidated the effect of an inexhaustible, ferrous-iron and humic-substance mimicking terminal electron acceptor by deploying potentiostatically poised electrodes in the sediment of a very specific stream riparian zone in Upstate New York state. At two sites within the same stream riparian zone during the course of 6 weeks in the spring of 2013, we measured CH4 and N2/N2O emissions from soil chambers containing either poised or unpoised electrodes, and we harvested biofilms from the electrodes to quantify microbial community dynamics. At the upstream site, which had a lower vegetation cover and highest soil temperatures, the poised electrodes inhibited CH4 emissions by ∼45% (when normalized to remove temporal effects). CH4 emissions were not significantly impacted at the downstream site. N2/N2O emissions were generally low at both sites and were not impacted by poised electrodes. We did not find a direct link between bioelectrochemical treatment and microbial community membership; however, we did find a correspondence between environment/function and microbial community dynamics.


Comparing the DNA hypermethylome with gene mutations in human colorectal cancer.
We have developed a transcriptome-wide approach to identify genes affected by promoter CpG island DNA hypermethylation and transcriptional silencing in colorectal cancer. By screening cell lines and validating tumor-specific hypermethylation in a panel of primary human colorectal cancer samples, we estimate that nearly 5% or more of all known genes may be promoter methylated in an individual tumor. When directly compared to gene mutations, we find larger numbers of genes hypermethylated in individual tumors, and a higher frequency of hypermethylation within individual genes harboring either genetic or epigenetic changes. Thus, to enumerate the full spectrum of alterations in the human cancer genome, and to facilitate the most efficacious grouping of tumors to identify cancer biomarkers and tailor therapeutic approaches, both genetic and epigenetic screens should be undertaken.


Genetic control of diabetes and insulitis in the nonobese diabetic (NOD) mouse.
Genetic analysis of the development of diabetes and insulitis has been performed in the nonobese diabetic (NOD) mouse strain, a model of insulin-dependent (type I) diabetes mellitus. (NOD X C57BL/10)F1, F2, and (F1 X NOD) first-, second-, and third-backcross generations were studied. The data obtained were consistent with the hypothesis that diabetes is controlled by at least three functionally recessive diabetogenic genes, or gene complexes, one of which is linked to the MHC of the NOD. In contrast, pancreatic inflammation leading to insulitis was found to be controlled by a single incompletely dominant gene. One of the two diabetogenic loci that is not linked to the MHC appears to be essential for the development of severe insulitis. This diabetogenic gene may be identical to the gene that controls the initiation of the autoimmune response that progresses to insulitis. Although this gene appears to be functionally recessive in its control of diabetes, it is incompletely dominant in its control of insulitis. The MHC-linked diabetogenic gene, although not required for the development of insulitis, apparently influences the progression of the autoimmune response since NOD MHC homozygotes in the backcross generations displayed the highest incidence and most severe cases of insulitis. Interestingly, we have found two MHC heterozygous backcross females that have become diabetic, suggesting that either the MHC-linked diabetogenic gene is not strictly recessive or that a recombination event has occurred between the diabetogenic gene and the K or I-A regions of the MHC. The third diabetogenic locus appears to influence the progression of severe insulitis to overt diabetes. In animals homozygous at this locus, diabetes may result from a decreased ability to develop a protective suppressor response to the autoimmune process.


An Iliopsoas Impingement Lesion in the Absence of Painful Internal Snapping May Not Require Iliopsoas Fractional Lengthening.
Iliopsoas impingement (IPI) has been associated with a distinct lesion on the anterior labrum. Iliopsoas fractional lengthening (IFL) can treat IPI in instances of painful internal snapping (PIS) and mechanical groin pain. To report minimum 2-year outcomes of patients without PIS who had an IPI lesion diagnosed intraoperatively that did not undergo IFL (+IPI -PIS -IFL) as compared with a matched group of patients with PIS and an IPI lesion that was treated with IFL (+IPI +PIS +IFL). Cohort study; Level of evidence, 3. Data on all patients who underwent primary hip arthroscopy between May 2009 and June 2017 were retrospectively reviewed. Patients were included if they underwent hip arthroscopy for femoroacetabular impingement-related pathology, an IPI lesion was diagnosed intraoperatively, and they had minimum 2-year postoperative scores for the following: modified Harris Hip Score (mHHS), Nonarthritic Hip Score (NAHS), Hip Outcome Score-Sports Specific Subscale (HOS-SSS), iHOT-12 (International Hip Outcome Tool-12), patient satisfaction, and visual analog score (VAS) for pain. Patients were propensity score matched based on the following criteria: age, body mass index, follow-up time, sex, labral treatment, femoroplasty, and acetabuloplasty. A total of 412 hips were eligible for the current study, of which 336 (81.6%) had 2-year follow-up. The matching process established 37 hips in the +IPI -PIS -IFL group and 87 hips in the +IPI +PIS +IFL group. Both groups experienced significant improvements from presurgery to latest follow-up for all recorded patient-reported outcomes (PROs). The +IPI -PIS -IFL group compared favorably with the +IPI +PIS +IFL group for mHHS (86.0 vs 86.1; P = .53), NAHS (83.0 vs 84.7; P = .40), and HOS-SSS (78.1 vs 76.5; P = .87). Additionally, iHOT-12, VAS, patient satisfaction, and rates of achieving the minimal clinically important difference for mHHS, NAHS, and HOS-SSS were similar between groups at the latest follow-up. Patients without PIS who were diagnosed with an IPI lesion intraoperatively and did not undergo IFL had similar and favorable improvements in PROs, VAS, and satisfaction to a matched cohort with PIS who had IFL performed. Thus, an IPI lesion in the absence of PIS may not require IFL.


TGF-beta receptor II in epithelia versus mesenchyme plays distinct roles in the developing lung.
Transforming growth factor (TGF)-beta signalling plays important roles in regulating lung development. However, the specific regulatory functions of TGF-beta signalling in developing lung epithelial versus mesenchymal cells are still unknown. By immunostaining, the expression pattern of the TGF-beta type II receptor (TbetaRII) was first determined in the developing mouse lung. The functions of TbetaRII in developing lung were then determined by conditionally knocking out TbetaRII in the lung epithelium of floxed-TbetaRII/surfactant protein C-reverse tetracycline transactivator/TetO-Cre mice versus mesenchyme of floxed-TbetaRII/Dermo1-Cre mice. TbetaRII was expressed only in distal airway epithelium at early gestation (embryonic day (E)11.5), but in both airway epithelium and mesenchyme from mid-gestation (E14.5) to post-natal day 14. Abrogation of TbetaRII in mouse lung epithelium resulted in retardation of post-natal lung alveolarisation, with markedly decreased type I alveolar epithelial cells, while no abnormality in prenatal lung development was observed. In contrast, blockade of TbetaRII in mesoderm-derived tissues, including lung mesenchyme, resulted in mildly abnormal lung branching and reduced cell proliferation after mid-gestation, accompanied by multiple defects in other organs, including diaphragmatic hernia. The primary lung branching defect was verified in embryonic lung explant culture. The novel findings of the present study suggest that transforming growth factor-beta type II receptor-mediated transforming growth factor-beta signalling plays distinct roles in lung epithelium versus mesenchyme to differentially control specific stages of lung development.


Pest categorisation of Ripersiella hibisci.
The EFSA Panel on Plant Health performed a pest categorisation of Ripersiella hibisci (Hemiptera: Rhizoecidae) for the EU. R. hibisci occurs in Japan, China and Taiwan and has spread to the USA: Florida, Hawaii and the territory of Puerto Rico. R. hibisci is a polyphagous mealybug recorded feeding on roots of monocotyledonous and dicotyledonous plants. Root damage reduces nutrient and water uptake, retards plant growth and may cause leaves to wilt or discolour, heavily infested plants can die. Literature most commonly refers to damage to greenhouse grown potted ornamentals such as Cuphea, Hibiscus, Pelargonium and Phoenix. All life stages occur in the soil and host plants for planting with growing media provide a pathway for eggs, nymphs and adults. Multiple overlapping generations occur in greenhouses each year. R. hibisci is listed in Annex IIA of EU Regulation 2016/2031, appearing with the synonym Rhizoecus hibisci. All plants for planting from third countries are regulated. The import of soil or growing medium attached to plants for planting from third countries (other than Switzerland) is prohibited and therefore reduces the likelihood, but does not prevent entry of R. hibisci, as individuals may remain attached to the roots. There have been interceptions of R. hibisci in the EU, usually on artificially dwarfed plants, i.e. bonsai/penjing. Findings in EU greenhouses have been eradicated. R. hibisci would be able to establish in the EU, greenhouse potted plant production would be most affected. Phytosanitary measures are available to lower the likelihood of introduction. R. hibisci satisfies the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest. R. hibisci does not meet the criterion of occurring in the EU for it to be regarded as a potential Union regulated non-quarantine pest.


The impact of climate change on food security in South Africa: Current realities and challenges ahead.
This article aims to examine the impact of climate change on food security in South Africa. For this purpose, the article adopted a desktop study approach. Previous studies, reports, surveys and policies on climate change and food (in)security. From this paper's analysis, climate change presents a high risk to food security in sub-Saharan countries from crop production to food distribution and consumption. In light of this, it is found that climate change, particularly global warming, affects food security through food availability, accessibility, utilisation and affordability. To mitigate these risks, there is a need for an integrated policy approach to protect the arable land against global warming. The argument advanced in this article is that South Africa's ability to adapt and protect its food items depends on the understanding of risks and the vulnerability of various food items to climate change. However, this poses a challenge in developing countries, including South Africa, because such countries have weak institutions and limited access to technology. Another concern is a wide gap between the cost of adapting and the necessary financial support from the government. There is also a need to invest in technologies that will resist risks on food systems.


Novel anti-obesity effects of beer hops compound xanthohumol: role of AMPK signaling pathway.
Obesity alters adipose tissue metabolic and endocrine functioning, leading to an increased adiposity and release of pro-inflammatory cytokines. Various phytochemicals have been reported to contribute to the beiging of white adipose tissue in order to ameliorate obesity by increasing thermogenesis. Here, we show that the prenylated chalcone, xanthohumol (XN), induces beiging of white adipocytes, stimulates lipolysis, and inhibits adipogenesis of murine 3T3-L1 adipocytes and primary human subcutaneous preadipocytes and these effects are partly mediated by the activation of the AMP-activated protein kinase (AMPK) signaling pathway. 3T3-L1 adipocytes and primary human subcutaneous preadipocytes were differentiated using a standard protocol and were treated with various concentrations of XN, dorsomorphin, an AMPK inhibitor, or AICAR, an AMPK activator, to investigate the effects on adipogenesis, beiging and lipolysis. XN induced beiging of white adipocytes as witnessed by the increased expression of beige markers CIDE-A and TBX-1. XN increased mitochondrial biogenesis, as evidenced by increased mitochondrial content, enhanced expression of PGC-1α, and the thermogenic protein UCP1. Following 24 h of treatment, XN also increased oxygen consumption rate. XN stimulated lipolysis of mature 3T3-L1 and primary human subcutaneous adipocytes and inhibited adipogenesis of maturing adipocytes. XN activated AMPK and in turn, XN-induced upregulation of UCP1, p-ACC, HSL, and ATGL was downregulated in the presence of dorsomorphin. Likewise, an XN-induced decrease in adipogenesis was reversed in the presence of dorsomorphin. Taken together, XN demonstrates anti-obesity effects by not only inducing beiging but also decreasing adipogenesis and inducing lipolysis. The anti-obesity effects of XN are partly mediated by AMPK signaling pathway suggesting that XN may have potential therapeutic implications for obesity.


Genome sequence of E. coli O104:H4 leads to rapid development of a targeted antimicrobial agent against this emerging pathogen.
A recent widespread outbreak of Escherichia coli O104:H4 in Germany demonstrates the dynamic nature of emerging and re-emerging food-borne pathogens, particularly STECs and related pathogenic E. coli. Rapid genome sequencing and public availability of these data from the German outbreak strain allowed us to identify an O-antigen-specific bacteriophage tail spike protein encoded in the genome. We synthesized this gene and fused it to the tail fiber gene of an R-type pyocin, a phage tail-like bacteriocin, and expressed the novel bacteriocin such that the tail fiber fusion was incorporated into the bacteriocin structure. The resulting particles have bactericidal activity specifically against E. coli strains that produce the O104 lipopolysaccharide antigen, including the outbreak strain. This O-antigen tailspike-R-type pyocin strategy provides a platform to respond rapidly to emerging pathogens upon the availability of the pathogen's genome sequence.


Genome-wide identification of PAX3-FKHR binding sites in rhabdomyosarcoma reveals candidate target genes important for development and cancer.
The PAX3-FKHR fusion protein is present in a majority of alveolar rhabdomyosarcomas associated with increased aggressiveness and poor prognosis. To better understand the molecular pathogenesis of PAX3-FKHR, we carried out the first, unbiased genome-wide identification of PAX3-FKHR binding sites and associated target genes in alveolar rhabdomyosarcoma. The data shows that PAX3-FKHR binds to the same sites as PAX3 at both MYF5 and MYOD enhancers. The genome-wide analysis reveals that the PAX3-FKHR sites are (a) mostly distal to transcription start sites, (b) conserved, (c) enriched for PAX3 motifs, and (d) strongly associated with genes overexpressed in PAX3-FKHR-positive rhabdomyosarcoma cells and tumors. There is little evidence in our data set for PAX3-FKHR binding at the promoter sequences. The genome-wide analysis further illustrates a strong association between PAX3 and E-box motifs in these binding sites, suggestive of a common coregulation for many target genes. We also provide the first direct evidence that FGFR4 and IGF1R are the targets for PAX3-FKHR. The map of PAX3-FKHR binding sites provides a framework for understanding the pathogenic roles of PAX3-FKHR, as well as its molecular targets to allow a systematic evaluation of agents against this aggressive rhabdomyosarcoma.


Construction, characterization, and complementation of a Puf- mutant of Rhodobacter sphaeroides.
A Puf- strain of Rhodobacter sphaeroides (PUFB1) was constructed by deleting a portion of the proximal region of the puf operon and inserting a kanamycin resistance gene cartridge. Southern blot analysis demonstrated that in PUFB1, the defective copy of the puf operon had replaced, through homologous recombination, the normal chromosomal copy. The Puf- phenotype was characterized by the inability of PUFB1 to grow photoheterotrophically (PS-), the lack of detectable puf-specific transcripts, the absence of the light-harvesting I complex and, by inference, the reaction center spectral complex, and greatly reduced levels of the light-harvesting II complex. The PS+ phenotype was restored to PUFB1 when a 13-kilobase BamHI restriction endonuclease fragment containing the entire puf operon and flanking regions was cloned into the broad-host-range plasmid vector RK2 derivative pRK404 and introduced by conjugation into PUFB1. In these complemented strains, there was an increased number of copies of the puf operon (four to six copies per defective chromosomal copy) as the result of plasmid copy number. However, there was no concomitant increase in either the specific bacteriochlorophyll content or the level of puf-specific transcripts when these strains were grown photoheterotrophically.


Enzymes of uracil catabolism in normal and neoplastic human tissues.
Enzymes of the pyrimidine base catabolism, dihydrouracil dehydrogenase (EC 22.214.171.124), dihydropyrimidinase (EC 220.127.116.11), and beta-ureidopropionase (EC 220.127.116.11) were compared in the cytosolic extract of several normal and neoplastic human tissues. The activity was measured by following the catabolism of [6-14C]-uracil to dihydrouracil, carbamyl-beta-alanine, and beta-alanine. Substrate inhibition, hysteresis, allosterism, and the lack of dihydropyrimidinase are pointed out as special problems in assaying enzymes of pyrimidine degradation. The activity of dihydrouracil dehydrogenase has been demonstrated in several human extrahepatic tissues and tumors. The enzyme is rate limiting in extrahepatic solid tumors but not in their normal counterparts. Some of these solid tumors contain greater amounts of activity than do their normal equivalents, which encourages the use of inhibitors of this enzyme in conjunction with treatment of these tumors by 5-fluorouracil. Because of the lack of a pattern in dihydrouracil dehydrogenase activity between tumors and normal tissues, the enzyme is not a good marker for tumorigenicity. Dihydropyrimidinase, on the other hand, is highly active in all solid tumors studied but not in their normal counterparts; therefore, we suggest that dihydropyrimidinase can serve as a good marker of tumorigenicity as well as a target for cancer chemotherapy of human solid tumors.


Promoting Psychological Well-Being Through an Evidence-Based Mindfulness Training Program.
Psychological well-being is a core feature of mental health, and may be defined as including hedonic (enjoyment, pleasure) and eudaimonic (meaning, fulfillment) happiness, as well as resilience (coping, emotion regulation, healthy problem solving). To promote psychological well-being, it is helpful to understand the underlying mechanisms associated with this construct and then develop targeted and effective training programs. In this perspective article, we discuss key components and potential brain-body mechanisms related to psychological well-being and propose mindfulness training as a promising way to improve it. Based on a series of randomized controlled trial (RCT) studies of one form of mindfulness training in adolescents and adults, the integrative body-mind training (IBMT), we use IBMT as an exemplar to provide research evidence of the positive effects of mindfulness training on psychological well-being. We focus on one of the mechanisms by which IBMT enhances psychological well-being-the interaction between mind (mindfulness) and body (bodifulness)-which involves both the central nervous system (CNS) and the autonomic nervous system (ANS). We also highlight the role of brain self-control networks, including the anterior cingulate cortex/prefrontal cortex (ACC/PFC), in improving psychological well-being. We suggest that mindfulness training may be a promising program that promotes the synergistic engagement of mind and body to achieve the goals of enhancing psychological well-being.


Clinical Significance of Gender and Body Mass Index in Asian Patients with Colorectal Cancer.
Objective: Colorectal cancer is a disease closely associated with anthropometric values. This study aimed to evaluate the clinical relevance of gender and body mass index (BMI) with colorectal cancer using a Korean nationwide cohort. Methods: Data of colorectal cancer cohorts between 2012 and 2013 were acquired from the Health Insurance Review and Assessment Service. All patients underwent surgery due to colorectal cancers. Stage IV patients were excluded due to possible clinical heterogeneity. BMI was classified with the World Health Organization criteria. Results: A total of 31,756 patients were analyzed. The underweight group had 33% higher risk of stage III disease (p<0.001). The overweight and obese groups had 20% and 19% lower risk of stage III (p<0.001 and p=0.002, respectively). The underweight and obese groups had higher risk of longest hospitalization period quartile (≥19 days), with odds ratio of 2.26 (p<0.001) and 1.33 (p<0.001), respectively. The overweight group had a 22% lower risk of the longest hospitalization period quartile (p=0.002). Females had 12% lower risk of distal cancer than males (p<0.001). There was no significant relationship between cancer stage and gender. The proportions of patients who were <50 years and ≥70 years old were higher in the females, and the proportions of patients in their 50s and 60s were higher in the males. Conclusions: Cancer stages and hospitalization period varied depending on BMI. Disease location and the age distribution were affected by gender.


A systematic review of nonpharmacologic interventions for treatment-related symptoms in women with ovarian cancer.
Women with ovarian cancer have a continued high symptom burden in comparison to other cancer survivors secondary to ongoing chemotherapy treatment. Prolonged or ineffective management of treatment-related symptoms can contribute to treatment noncompliance, worsening of symptoms, and reduced health-related quality of life. This review of the literature was conducted to describe experimental and quasi-experimental research addressing nonpharmacologic interventions for the treatment-related symptoms of sleep disturbance, pain, anxiety, depression, and low energy or fatigue in women with ovarian cancer and to critique the quality of interventions. A systematic search of the literature was conducted in PubMed and yielded 136 articles. Eight articles met the inclusion criteria and were evaluated. Nonpharmacologic interventions for treatment-related symptoms were complex, with an average of 4.4 components. Intervention delivery, setting, and exposure varied widely across studies. Only three studies contained details sufficient to replicate the intervention. Lack of clarity in intervention reporting may explain perceptions of clinically inefficacious symptom management in this context. Greater attention to reporting would facilitate better translation of interventions into practice and when addressing complex cancer symptom clusters.


Myosin VI: cellular functions and motor properties.
Myosin VI has been localized in membrane ruffles at the leading edge of cells, at the trans-Golgi network compartment of the Golgi complex and in clathrin-coated pits or vesicles, indicating that it functions in a wide variety of intracellular processes. Myosin VI moves along actin filaments towards their minus end, which is the opposite direction to all of the other myosins so far studied (to our knowledge), and is therefore thought to have unique properties and functions. To investigate the cellular roles of myosin VI, we identified various myosin VI binding partners and are currently characterizing their interactions within the cell. As an alternative approach, we have expressed and purified full-length myosin VI and studied its in vitro properties. Previous studies assumed that myosin VI was a dimer, but our biochemical, biophysical and electron microscopic studies reveal that myosin VI can exist as a stable monomer. We observed, using an optical tweezers force transducer, that monomeric myosin VI is a non-processive motor which, despite a relatively short lever arm, generates a large working stroke of 18 nm. Whether monomer and/or dimer forms of myosin VI exist in cells and their possible functions will be discussed.


Elucidation of the Mode of Action of a New Antibacterial Compound Active against Staphylococcus aureus and Pseudomonas aeruginosa.
Nosocomial and community-acquired infections caused by multidrug resistant bacteria represent a major human health problem. Thus, there is an urgent need for the development of antibiotics with new modes of action. In this study, we investigated the antibacterial characteristics and mode of action of a new antimicrobial compound, SPI031 (N-alkylated 3, 6-dihalogenocarbazol 1-(sec-butylamino)-3-(3,6-dichloro-9H-carbazol-9-yl)propan-2-ol), which was previously identified in our group. This compound exhibits broad-spectrum antibacterial activity, including activity against the human pathogens Staphylococcus aureus and Pseudomonas aeruginosa. We found that SPI031 has rapid bactericidal activity (7-log reduction within 30 min at 4x MIC) and that the frequency of resistance development against SPI031 is low. To elucidate the mode of action of SPI031, we performed a macromolecular synthesis assay, which showed that SPI031 causes non-specific inhibition of macromolecular biosynthesis pathways. Liposome leakage and membrane permeability studies revealed that SPI031 rapidly exerts membrane damage, which is likely the primary cause of its antibacterial activity. These findings were supported by a mutational analysis of SPI031-resistant mutants, a transcriptome analysis and the identification of transposon mutants with altered sensitivity to the compound. In conclusion, our results show that SPI031 exerts its antimicrobial activity by causing membrane damage, making it an interesting starting point for the development of new antibacterial therapies.


Crosstalk Between Oxidative Stress and Endoplasmic Reticulum (ER) Stress in Endothelial Dysfunction and Aberrant Angiogenesis Associated With Diabetes: A Focus on the Protective Roles of Heme Oxygenase (HO)-1.
Type-2 diabetes prevalence is continuing to rise worldwide due to physical inactivity and obesity epidemic. Diabetes and fluctuations of blood sugar are related to multiple micro- and macrovascular complications, that are attributed to oxidative stress, endoplasmic reticulum (ER) activation and inflammatory processes, which lead to endothelial dysfunction characterized, among other features, by reduced availability of nitric oxide (NO) and aberrant angiogenic capacity. Several enzymatic anti-oxidant and anti-inflammatory agents have been found to play protective roles against oxidative stress and its downstream signaling pathways. Of particular interest, heme oxygenase (HO) isoforms, specifically HO-1, have attracted much attention as major cytoprotective players in conditions associated with inflammation and oxidative stress. HO operates as a key rate-limiting enzyme in the process of degradation of the iron-containing molecule, heme, yielding the following byproducts: carbon monoxide (CO), iron, and biliverdin. Because HO-1 induction was linked to pro-oxidant states, it has been regarded as a marker of oxidative stress; however, accumulating evidence has established multiple cytoprotective roles of the enzyme in metabolic and cardiovascular disorders. The cytoprotective effects of HO-1 depend on several cellular mechanisms including the generation of bilirubin, an anti-oxidant molecule, from the degradation of heme; the induction of ferritin, a strong chelator of free iron; and the release of CO, that displays multiple anti-inflammatory and anti-apoptotic actions. The current review article describes the major molecular mechanisms contributing to endothelial dysfunction and altered angiogenesis in diabetes with a special focus on the interplay between oxidative stress and ER stress response. The review summarizes the key cytoprotective roles of HO-1 against hyperglycemia-induced endothelial dysfunction and aberrant angiogenesis and discusses the major underlying cellular mechanisms associated with its protective effects.


Getting "SMART" on Shock Treatment: An Evidence-Based Mnemonic Acronym for the Initial Management of Hemorrhage in Trauma.
Treating hemorrhagic shock is challenging, the pathology is complex, and time is critical. Treatment requires resources in mental bandwidth (i.e., focused attention), drugs and blood products, equipment, and personnel. Providers must focus on treatment options in order of priority while also maintaining a dynamic assessment of the patient's response to treatment and considering potential differential diagnoses. In this process, the cognitive load is substantial. To avoid errors of clinical reasoning and practical errors of commission, omission, or becoming fixated, it is necessary to use evidence-based treatment recommendations that are concise, in priority order, and easily recalled. This is particularly the case in the austere, remote, or tactical environment. A simple mnemonic acronym, SMART, is presented in this article. It is a clinical heuristic that can be used as an aide-mémoire during the initial phases of resuscitation of the trauma patient with hemorrhagic shock: Start the clock and Stop the bleeding; Maintain perfusion; Administer antifibrinolytics; Retain heat; Titrate blood products and calcium; Think of alternative causes of shock.


Predators and nutrient availability favor protozoa-resisting bacteria in aquatic systems.
The long co-existence of bacteria and protozoa has led to the development of bacterial protozoa resistance strategies, which are suggested to serve as drivers for the evolution of pathogenic bacteria. However, the ecological mechanisms underpinning selection for protozoa-resistance in aquatic bacteria are poorly known. To assess the role of nutrient availability and predation-pressure on selection for protozoa-resisting bacteria (PRB), an enrichment-dilution experiment was designed using laboratory microcosms containing natural lake water. PRB was monitored by screening 16S rRNA amplicon sequence data for reads assigned to bacteria that previously has been shown to resist degradation by amoebae. To estimate the effects of the microbial food web dynamics (microscopy of; heterotrophic bacteria, phytoplankton, protozoa and rotifers) and physicochemical variables on the PRB abundance in the study system, a joint species distribution modelling approach was used. The predation-pressure (ratio between predator and bacterial biomass) had a positive effect on the abundance of the PRB genus Mycobacterium, while perturbation (enrichment and dilution) favored the PRB genus Pseudomonas that dominated the bacterial community in the disturbed systems. Our results show that PRB with different ecological strategies can be expected in water of high and intermediate nutrient levels and after major disturbances of an aquatic system.