Closed-reference metatranscriptomics enables in planta profiling of putative virulence activities in the grapevine trunk disease complex.
Grapevines, like other perennial crops, are affected by so-called 'trunk diseases', which damage the trunk and other woody tissues. Mature grapevines typically contract more than one trunk disease and often multiple grapevine trunk pathogens (GTPs) are recovered from infected tissues. The co-existence of different GTP species in complex and dynamic microbial communities complicates the study of the molecular mechanisms underlying disease development, especially under vineyard conditions. The objective of this study was to develop and optimize a community-level transcriptomics (i.e. metatranscriptomics) approach that could monitor simultaneously the virulence activities of multiple GTPs in planta. The availability of annotated genomes for the most relevant co-infecting GTPs in diseased grapevine wood provided the unprecedented opportunity to generate a multi-species reference for the mapping and quantification of DNA and RNA sequencing reads. We first evaluated popular sequence read mappers using permutations of multiple simulated datasets. Alignment parameters of the selected mapper were optimized to increase the specificity and sensitivity for its application to metagenomics and metatranscriptomics analyses. Initial testing on grapevine wood experimentally inoculated with individual GTPs confirmed the validity of the method. Using naturally infected field samples expressing a variety of trunk disease symptoms, we show that our approach provides quantitative assessments of species composition, as well as genome-wide transcriptional profiling of potential virulence factors, namely cell wall degradation, secondary metabolism and nutrient uptake for all co-infecting GTPs.


Solution structure of RING finger-like domain of retinoblastoma-binding protein-6 (RBBP6) suggests it functions as a U-box.
Retinoblastoma-binding protein-6 (RBBP6) plays a facilitating role, through its RING finger-like domain, in the ubiquitination of p53 by Hdm2 that is suggestive of E4-like activity. Although the presence of eight conserved cysteine residues makes it highly probable that the RING finger-like domain coordinates two zinc ions, analysis of the primary sequence suggests an alternative classification as a member of the U-box family, the members of which do not bind zinc ions. We show here that despite binding two zinc ions, the domain adopts a homodimeric structure highly similar to those of a number of U-boxes. Zinc ions could be replaced by cadmium ions without significantly disrupting the structure or the stability of the domain, although the rate of substitution was an order of magnitude slower than any previous measurement, suggesting that the structure is particularly stable, a conclusion supported by the high thermal stability of the domain. A hallmark of U-box-containing proteins is their association with chaperones, with which they cooperate in eliminating irretrievably unfolded proteins by tagging them for degradation by the proteasome. Using a yeast two-hybrid screen, we show that RBBP6 interacts with chaperones Hsp70 and Hsp40 through its N-terminal ubiquitin-like domain. Taken together with the structural similarities to U-box-containing proteins, our data suggest that RBBP6 plays a role in chaperone-mediated ubiquitination and possibly in protein quality control.


Intraclass correlation values for adolescent health outcomes in secondary schools in 21 European countries.
Cluster randomised controlled trials (CRCTs) are increasingly used to evaluate the effectiveness of interventions for improving health. A key feature of CRCTs is that individuals in clusters are often more alike than individuals in different clusters, irrespective of treatment. This similarity within clusters needs to be taken into account when planning CRCTs to obtain adequate sample sizes, and when analysing clustered data to obtain correct estimates. Nationally representative data from 15 to 16 year olds were analysed, from 21 of the 35 countries that participated in the 2007 European School Survey Project on Alcohol and Other Drugs. Within country school level intra-class correlation coefficients (ICCs) were calculated for substance use (self-reported alcohol use, regular alcohol use, binge drinking, any smoking, regular smoking, and illicit drug use) and psychosocial health (depressive mood and self-esteem). Unadjusted and adjusted ICCs are presented. ICCs are adjusted for student sex and socioeconomic status. ICCs ranged from 0.01 to 0.21, with the highest (0.21) reported for regular smoking. Within country school level ICCs varied substantially across health outcomes, and among countries for the same health outcomes. Estimated ICCs were consistently higher for substance use (range 0.01-0.21), than for psychosocial health (range 0.01-0.07). Within country ICCs for health outcomes varied by changes in the measurement of particular health outcomes, for example the ICCs for regular smoking (range 0.06-0.21) were higher than those for having smoked at all in the last month (range 0.03-0.17). For school level ICCs to be effectively utilised in informing sample size requirements for CRCTs and adjusting estimates from meta-analyses, the school level ICCs need to be both country and outcome specific.


Using a System Identification Approach to Investigate Subtask Control during Human Locomotion.
Here we apply a control theoretic view of movement to the behavior of human locomotion with the goal of using perturbations to learn about subtask control. Controlling one's speed and maintaining upright posture are two critical subtasks, or underlying functions, of human locomotion. How the nervous system simultaneously controls these two subtasks was investigated in this study. Continuous visual and mechanical perturbations were applied concurrently to subjects (n = 20) as probes to investigate these two subtasks during treadmill walking. Novel application of harmonic transfer function (HTF) analysis to human motor behavior was used, and these HTFs were converted to the time-domain based representation of phase-dependent impulse response functions (ϕIRFs). These ϕIRFs were used to identify the mapping from perturbation inputs to kinematic and electromyographic (EMG) outputs throughout the phases of the gait cycle. Mechanical perturbations caused an initial, passive change in trunk orientation and, at some phases of stimulus presentation, a corrective trunk EMG and orientation response. Visual perturbations elicited a trunk EMG response prior to a trunk orientation response, which was subsequently followed by an anterior-posterior displacement response. This finding supports the notion that there is a temporal hierarchy of functional subtasks during locomotion in which the control of upper-body posture precedes other subtasks. Moreover, the novel analysis we apply has the potential to probe a broad range of rhythmic behaviors to better understand their neural control.


Decoding of human hand actions to handle missing limbs in neuroprosthetics.
The only way we can interact with the world is through movements, and our primary interactions are via the hands, thus any loss of hand function has immediate impact on our quality of life. However, to date it has not been systematically assessed how coordination in the hand's joints affects every day actions. This is important for two fundamental reasons. Firstly, to understand the representations and computations underlying motor control "in-the-wild" situations, and secondly to develop smarter controllers for prosthetic hands that have the same functionality as natural limbs. In this work we exploit the correlation structure of our hand and finger movements in daily-life. The novelty of our idea is that instead of averaging variability out, we take the view that the structure of variability may contain valuable information about the task being performed. We asked seven subjects to interact in 17 daily-life situations, and quantified behavior in a principled manner using CyberGlove body sensor networks that, after accurate calibration, track all major joints of the hand. Our key findings are: (1) We confirmed that hand control in daily-life tasks is very low-dimensional, with four to five dimensions being sufficient to explain 80-90% of the variability in the natural movement data. (2) We established a universally applicable measure of manipulative complexity that allowed us to measure and compare limb movements across tasks. We used Bayesian latent variable models to model the low-dimensional structure of finger joint angles in natural actions. (3) This allowed us to build a naïve classifier that within the first 1000 ms of action initiation (from a flat hand start configuration) predicted which of the 17 actions was going to be executed-enabling us to reliably predict the action intention from very short-time-scale initial data, further revealing the foreseeable nature of hand movements for control of neuroprosthetics and tele operation purposes. (4) Using the Expectation-Maximization algorithm on our latent variable model permitted us to reconstruct with high accuracy (<5-6° MAE) the movement trajectory of missing fingers by simply tracking the remaining fingers. Overall, our results suggest the hypothesis that specific hand actions are orchestrated by the brain in such a way that in the natural tasks of daily-life there is sufficient redundancy and predictability to be directly exploitable for neuroprosthetics.


Mechanical stretch: physiological and pathological implications for human vascular endothelial cells.
Vascular endothelial cells are subjected to hemodynamic forces such as mechanical stretch due to the pulsatile nature of blood flow. Mechanical stretch of different intensities is detected by mechanoreceptors on the cell surface which enables the conversion of external mechanical stimuli to biochemical signals in the cell, activating downstream signaling pathways. This activation may vary depending on whether the cell is exposed to physiological or pathological stretch intensities. Substantial stretch associated with normal physiological functioning is important in maintaining vascular homeostasis as it is involved in the regulation of cell structure, vascular angiogenesis, proliferation and control of vascular tone. However, the elevated pressure that occurs with hypertension exposes cells to excessive mechanical load, and this may lead to pathological consequences through the formation of reactive oxygen species, inflammation and/or apoptosis. These processes are activated by downstream signaling through various pathways that determine the fate of cells. Identification of the proteins involved in these processes may help elucidate novel mechanisms involved in vascular disease associated with pathological mechanical stretch and could provide new insight into therapeutic strategies aimed at countering the mechanisms' negative effects.


Intrapartum care quality indicators: a systematic approach for achieving consensus.
To identify published maternity intrapartum quality indicators and rationalise them to a core set. Prospective qualitative consensus group exercise. A literature search identified sets of intrapartum quality indicators in the English language. These were rationalised to a shortlist using criteria adapted from 'The Good Clinical Indicator Guide'. An expert panel from key UK professions and organisations utilised a modified Delphi process to devise a final agreed set of intrapartum quality indicators that are specific, measurable, modifiable and relevant to clinical practice. Each indicator was rated in two rounds of web-based surveys, followed by a final face-to-face meeting of the panel. 290 clinical indicators were identified within 96 clinical categories with up to 18 different definitions. After rationalisation, 10 core and 24 additional quality indicators entered the Delphi process. A final panel of 12 indicators was defined. By using a simple Delphi process we have defined a set of broad clinical quality indicators that provide a comprehensive coverage of labour and delivery outcomes. We believe that this pragmatic portfolio will be useful for many wishing to develop performance monitoring and maternity dashboard systems.


Characterization of Brown Adipose-Like Tissue in Trauma-Induced Heterotopic Ossification in Humans.
Heterotopic ossification (HO), the abnormal formation of bone within soft tissues, is a major complication after severe trauma or amputation. Transient brown adipocytes have been shown to be a critical regulator of this process in a mouse model of HO. In this study, we evaluated the presence of brown fat within human HO lesions. Most of the excised tissue samples displayed histological characteristics of bone, fibroproliferative cells, blood vessels, and adipose tissue. Immunohistochemical analysis revealed extensive expression of uncoupling protein 1 (UCP1), a definitive marker of brown adipocytes, within HO-containing tissues but not normal tissues. As seen in the brown adipocytes observed during HO in the mouse, these UCP1+ cells also expressed the peroxisome proliferator-activated receptor γ coactivator 1α. However, further characterization showed these cells, like their mouse counterparts, did not express PR domain containing protein 16, a key factor present in brown adipocytes found in depots. Nor did they express factors present in beige adipocytes. These results identify a population of UCP1+ cells within human tissue undergoing HO that do not entirely resemble either classic brown or beige adipocytes, but rather a specialized form of brown adipocyte-like cells, which have a unique function. These cells may offer a new target to prevent this unwanted bone.


Normograde and retrograde pinning of the distal fragment in humeral fractures of the dog.
To determine if normograde and retrograde pinning of the distal humeral fragment, to maximize pin purchase in this fragment, would damage vital structures in and around the elbow and shoulder joints in dogs. Anatomic study. Cadaveric dogs (n = 12; 12 thoracic limb pairs). Twelve thoracic limb pairs were harvested from cross-breed dogs and 1 thoracic limb from each pair was allocated to 1 of 3 groups. A transverse osteotomy was created at the junction of the middle and distal thirds in the diaphyseal fracture group (n = 8) and proximal to the supracondylar foramen in the metaphyseal fracture group (n = 8). The humeri in the normograde group (n = 8) were left intact. In all specimens in the distal and supracondylar fracture groups, retrograde pinning of the distal fragment was performed with the elbow flexed. The pins were driven into the medial epicondyle until they exited the skin caudal to the elbow. Dissection of the soft tissues around the exit tract of the pin was performed. The fracture was then reduced and the pin was advanced until it exited the proximal humeral fragment. In the specimens in the normograde group, pinning was initiated on the distal aspect of the medial epicondyle. The pin was directed parallel to the caudal aspect of the medial epicondyle and driven proximally into the humerus until it exited the proximal humerus. Pins exiting the distal aspect of the medial epicondyle passed through muscle origins in 7 specimens in the diaphyseal osteotomy group and in all 8 specimens in the metaphyseal group. The ulnar nerve was entrapped in 1 specimen in the metaphyseal osteotomy group; however, it was not entrapped in any of the specimens in the diaphyseal osteotomy group. The articular cartilage of the trochlea was damaged in 2 specimens in each of the retrograde pinned groups. There was no damage to articular or periarticular structures by pins exiting the proximal humerus. Retrograde pinning of the distal fragment in humeral fractures in the dog may damage the articular cartilage and cannot be recommended.


Gene duplicability-connectivity-complexity across organisms and a neutral evolutionary explanation.
Gene duplication has long been acknowledged by biologists as a major evolutionary force shaping genomic architectures and characteristics across the Tree of Life. Major research has been conducting on elucidating the fate of duplicated genes in a variety of organisms, as well as factors that affect a gene's duplicability--that is, the tendency of certain genes to retain more duplicates than others. In particular, two studies have looked at the correlation between gene duplicability and its degree in a protein-protein interaction network in yeast, mouse, and human, and another has looked at the correlation between gene duplicability and its complexity (length, number of domains, etc.) in yeast. In this paper, we extend these studies to six species, and two trends emerge. There is an increase in the duplicability-connectivity correlation that agrees with the increase in the genome size as well as the phylogenetic relationship of the species. Further, the duplicability-complexity correlation seems to be constant across the species. We argue that the observed correlations can be explained by neutral evolutionary forces acting on the genomic regions containing the genes. For the duplicability-connectivity correlation, we show through simulations that an increasing trend can be obtained by adjusting parameters to approximate genomic characteristics of the respective species. Our results call for more research into factors, adaptive and non-adaptive alike, that determine a gene's duplicability.


Organic Bioelectronics: Bridging the Signaling Gap between Biology and Technology.
The electronics surrounding us in our daily lives rely almost exclusively on electrons as the dominant charge carrier. In stark contrast, biological systems rarely use electrons but rather use ions and molecules of varying size. Due to the unique combination of both electronic and ionic/molecular conductivity in conducting and semiconducting organic polymers and small molecules, these materials have emerged in recent decades as excellent tools for translating signals between these two realms and, therefore, providing a means to effectively interface biology with conventional electronics-thus, the field of organic bioelectronics. Today, organic bioelectronics defines a generic platform with unprecedented biological recording and regulation tools and is maturing toward applications ranging from life sciences to the clinic. In this Review, we introduce the field, from its early breakthroughs to its current results and future challenges.


Diagnosis of cardiovascular autonomic neuropathy in diabetes.
The utility of standard cardiovascular tests for diagnosis of cardiac autonomic neuropathy in diabetes has been well documented. Attention must be paid to standardizing the procedure with regard to time of day, metabolic status, distance from meal and insulin, coffee and smoking avoidance, and patient's collaboration. In the presence of cardiovascular disease or drugs affecting the cardiovascular or autonomic nervous system, some caution is needed in interpreting the results. More recent reflex tests, which evaluate mainly sympathetic or baroreflex activity, despite their ability to detect early autonomic involvement, lack sufficient standardization and still need to be proved as valid alternatives. Of the different methods of measuring heart rate variability, spectral analysis has a greater ability to differentiate vagal and sympathetic modulation of heart rate than do time-domain methods. However, since these latter methods are easier and more widely available, they can be used as a screening approach. Twenty-four-hour evaluation of heart rate variability provides data on the circadian rhythm of sympathovagal activity, which can be affected earlier than and differently from cardiovascular reflex tests. Information obtained could have prognostic implications in terms of cardiovascular morbidity and mortality and offer therapeutic opportunities. However, a wide consensus on many technical aspects of both time-domain and frequency-domain methods is needed. Furthermore, large prospective studies in the diabetic population to assess the prognostic value of 24-h heart rate variability parameters on cardiovascular morbidity and mortality are lacking. Recently, I123 meta-iodobenzylguanidine (MIBG) scintigraphy has documented abnormalities of sympathetic myocardial innervation also in newly diagnosed IDDM. The meaning of this finding, whether it is an expression of functional or structural defects, needs to be clarified. Preliminary data point to a possible pathogenetic meaning of the known association between autonomic neuropathy and other diabetic complications. This area of investigation could provide useful insights into the complex and multifactorial pathogenesis of diabetic complications.


How migratory thrushes conquered northern North America: a comparative phylogeography approach.
Five species of migratory thrushes (Turdidae) occupy a transcontinental distribution across northern North America. They have largely overlapping breeding ranges, relatively similar ecological niches, and mutualistic relationships with northern woodland communities as insectivores and seed-dispersing frugivores. As an assemblage of ecologically similar species, and given other vertebrate studies, we predicted a shared pattern of genetic divergence among these species between their eastern and western populations, and also that the timing of the coalescent events might be similar and coincident with historical glacial events. To determine how these five lineages effectively established transcontinental distributions, we used mitochondrial cytochrome b sequences to assess genetic structure and lineage coalescence from populations on each side of the continent. Two general patterns occur. Hermit and Swainson's thrushes (Catharus guttatus and C. ustulatus) have relatively deep divergences between eastern and western phylogroups, probably reflecting shared historic vicariance. The Veery (C. fuscescens), Gray-cheeked Thrush (C. minimus), and American Robin (Turdus migratorius) have relatively shallow divergences between eastern and western populations. However, coalescent and approximate Bayesian computational analyses indicated that among all species as many as five transcontinental divergence events occurred. Divergence within both Hermit and Swainson's thrushes resembled the divergence between Gray-cheeked Thrushes and Veeries and probably occurred during a similar time period. Despite these species' ecological similarities, the assemblage exhibits heterogeneity at the species level in how they came to occupy transcontinental northern North America but two general continental patterns at an among-species organizational level, likely related to lineage age.


4-hydroxynonenal induces a cellular redox status-related activation of the caspase cascade for apoptotic cell death.
4-Hydroxynonenal (HNE), a diffusible product of lipid peroxidation, has been suggested to be a key mediator of oxidative stress-induced cell death. In this study, we partially characterized the mechanism of HNE-mediated cytotoxicity. Incubation of human T lymphoma Jurkat cells with 20-50 microM HNE led to cell death accompanied by DNA fragmentation. Western blot analysis showed that HNE-treatment induced time- and dose-dependent activation of caspase-8, caspase-9 and caspase-3. HNE-induced caspase-3 processing was confirmed by a flow cytometric demonstration of increased catalytic activity on the substrate peptide. HNE treatment also led to remarkable cleavage of poly(ADP-ribose) polymerase (PARP), which was prevented by pretreatment of cells with DEVD-FMK as a caspase-3 inhibitor. The HNE-mediated activation of caspases, cleavage of PARP and DNA fragmentation were blocked by antioxidants cysteine, N-acety-L-cysteine and dithiothreitol, but not by two other HNE-reactive amino acids lysine and histidine, or by cystine, the oxidized form of cysteine. HNE rapidly decreased levels of intracellular reduced glutathione (GSH) and its oxidized form GSSG, and these were also attenuated by the reductants. Coincubation of Jurkat cells with a blocking anti-Fas antibody prevented Fas-induced but not HNE-induced activation of caspase-3. HNE also activated caspase-3 in K562 cells that do not express functional Fas. Our results thereby demonstrate that HNE triggers oxidative stress-linked apoptotic cell death through activation of the caspase cascade. The results also suggest a possible mechanism involving a direct scavenge of intracellular GSH by HNE.


Low-dose combination therapy with rosiglitazone and metformin to prevent type 2 diabetes mellitus (CANOE trial): a double-blind randomised controlled study.
The evolving epidemic of type 2 diabetes has challenged health-care providers to assess the safety and efficacy of various diabetes prevention strategies. The CANOE (CAnadian Normoglycemia Outcomes Evaluation) trial investigated whether low-dose combination therapy would affect development of type 2 diabetes. In this double-blind, randomised controlled trial undertaken in clinics in Canadian centres, 207 patients with impaired glucose tolerance were randomly assigned to receive combination rosiglitazone (2 mg) and metformin (500 mg) twice daily or matching placebo for a median of 3.9 years (IQR 3.0-4.6). Randomisation was computer-generated in blocks of four, with both participants and investigators masked to treatment allocation. The primary outcome was time to development of diabetes, measured by an oral glucose tolerance test or two fasting plasma glucose values of 7.0 mmol/L or greater. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00116932. 103 participants were assigned to rosiglitazone and metformin, and 104 to placebo; all were analysed. Vital status was obtained in 198 (96%) participants, and medication compliance (taking at least 80% of assigned medication) was 78% (n=77) in the metformin and rosiglitazone group and 81% (n=80) in the placebo group. Incident diabetes occurred in significantly fewer individuals in the active treatment group (n=14 [14%]) than in the placebo group (n=41 [39%]; p<0.0001). The relative risk reduction was 66% (95% CI 41-80) and the absolute risk reduction was 26% (14-37), yielding a number needed to treat of 4 (2.70-7.14). 70 (80%) patients in the treatment group regressed to normal glucose tolerance compared with 52 (53%) in the placebo group (p=0.0002). Insulin sensitivity decreased by study end in the placebo group (median -1.24, IQR -2.38 to -0.08) and remained unchanged with rosiglitazone and metformin treatment (-0.39, -1.30 to 0.84; p=0.0006 between groups). The change in beta-cell function, as measured by the insulin secretion-sensitivity index-2, did not differ between groups (placebo -252.3, -382.2 to -58.0 vs rosiglitazone and metformin -221.8, -330.4 to -87.8; p=0.28). We recorded an increase in diarrhoea in participants in the active treatment group compared with the placebo group (16 [16%] vs 6 [6%]; p=0.0253). Low-dose combination therapy with rosiglitazone and metformin was highly effective in prevention of type 2 diabetes in patients with impaired glucose tolerance, with little effect on the clinically relevant adverse events of these two drugs. GlaxoSmithKline.


Factors that influence the quality of paid support for adults with acquired neurological disability: scoping review and thematic synthesis.
Theoretically, individualised funding schemes empower people with disability (PWD) to choose high quality support services in line with their needs and preferences. Given the importance of support, the aim of this scoping review was to understand the factors that influence the quality of paid disability support for adults with acquired neurological disability. A comprehensive scoping review of the published literature from 2009-2019 was conducted on five databases: Medline, CINAHL, Embase, PsycINFO and Scopus. Of the 3391 records retrieved, 16 qualitative articles were eligible for review. Thematic synthesis of the findings revealed six key interrelated themes: (1) choice and control, (2) individualised support, (3) disability support worker (DSW) qualities, (4) DSW competence, (5) PWD - DSW relationship, and (6) accessing consistent support. The themes depict factors influencing the quality of paid disability support from the perspective of PWD, close others and DSWs. Although the evidence base is sparse, the factors identified were in line with international rights legislation and policy ideals. The findings can provide insights to PWD hiring and managing support, and facilitate the delivery of quality disability support. Further research is required to understand the interactions between the factors and how to optimise support in practice.Implications for rehabilitationThe quality of paid disability support is determined by a multitude of interrelated factors influenced by the disability support worker's qualities and competencies, the interaction between the person with disability and the disability support worker, as well as external contextual factors.Optimising choice and control for adults with acquired neurological disability and providing individualised support should be a significant focus for disability support workers.Training modules for disability support workers can be informed by five of the identified themes: (1) choice and control, (2) individualised support, (3) DSW qualities, (4) DSW competence and (5) the relationship between PWD and DSWs.


Leisure time physical activity and the risk of hip or knee replacement due to primary osteoarthritis: a population based cohort study (The HUNT Study).
The relationship between leisure time physical activity (LPA) and hip and knee OA and subsequent joint replacement has not yet been clearly defined. Some studies have found the risk of knee replacement (TKR) to increase with high levels of LPA, while others have found no overall relationship to either TKR or hip replacement (THR). The aim was to investigate the association between LPA and the risk of severe end-stage OA, defined as THR or TKR due to primary OA, in a large population-based cohort. Participants in the Nord-Trøndelag Health Study (HUNT) were followed prospectively to identify THR and TKR using the Norwegian Arthroplasty Register. Self-reported LPA was classified as inactive, low, moderate or high. The Cox proportional hazards model was used to calculate hazard ratios (HRs) according to levels of LPA with adjustments for confounding variables. Analyses were performed by age (<45, 45-59 and ≥60 years) and sex. A total of 66 964 participants (mean age 46.8 years (SD 16.3) were included in the analyses. We identified 1636 THRs and 1016 TKRs due to primary OA during 17.0 years (median) of follow-up. High LPA was significantly associated with THR for women <45 years (HR 1.78, 95 % CI 1.08-2.94) and men between 45-59 years (HR 1.53, 95 % CI 1.10-2.13) at baseline. A significant trend was found only among women < 45 years at baseline (p = 0.02). We found that LPA was significantly associated with TKR for women only (HR 1.45, 95 % CI 1.03-2.04). No measures of LPA were associated with TKR for men. In this population-based study, high level of LPA was associated with increased risk of THR where a significant trend of LPA was seen among women <45 years at baseline. For TKR, high LPA was associated with increased risk only in women. In contrast to previous studies, this study shows a possible association between high LPA and the risk of THR.


Biased predecisional processing of leading and nonleading alternatives.
When people obtain information about choice alternatives in a set one attribute at a time, they rapidly identify a leading alternative. Although previous research has established that people then distort incoming information, it is unclear whether distortion occurs through favoring of the leading alternative, disfavoring of the trailing alternative, or both. Prior examinations have not explored the predecisional treatment of the nonleading alternative (or alternatives) because they conceptualized distortion as a singular construct in binary choice and measured it using a relative item comparing the evaluation of both alternatives simultaneously. In this article, we introduce a measure of distortion at the level of the alternative, which allows for measuring whether predecisional distortion favors or disfavors every alternative being considered in choice sets of various sizes. We report that both proleader and antitrailer distortion occur and that the use of antitrailer processing differs between binary choices and multiple-options choices.


Absolute pK(a) determinations for substituted phenols.
The CBS-QB3 method was used to calculate the gas-phase free energy difference between 20 phenols and their respective anions, and the CPCM continuum solvation method was applied to calculate the free energy differences of solvation for the phenols and their anions. The CPCM solvation calculations were performed on both gas-phase and solvent-phase optimized structures. Absolute pK(a) calculations with solvated phase optimized structures for the CPCM calculations yielded standard deviations and root-mean-square errors of less than 0.4 pK(a) unit. This study is the most accurate absolute determination of the pK(a) values of phenols, and is among the most accurate of any such calculations for any group of compounds. The ability to make accurate predictions of pK(a) values using a coherent, well-defined approach, without external approximations or fitting to experimental data, is of general importance to the chemical community. The solvated phase optimized structures of the anions are absolutely critical to obtain this level of accuracy, and yield a more realistic charge separation between the negatively charged oxygen and the ring system of the phenoxide anions.


The Key Role of Side Chain Linkage in Structure Formation and Mixed Conduction of Ethylene Glycol Substituted Polythiophenes.
Functionalizing conjugated polymers with polar ethylene glycol side chains enables enhanced swelling and facilitates ion transport in addition to electronic transport in such systems. Here, we investigate three polythiophene homopolymers (P3MEET, P3MEEMT, and P3MEEET) having differently linked (without spacer and with methyl and ethyl spacer, respectively) diethylene glycol side chains. All the polymers were tested in organic electrochemical transistors (OECTs). They show drastic differences in the device performance. The highest μOECT C* product of 11.5 F/cm·V·s was obtained for ethyl-spaced P3MEEET. How the injection and transport of ions is influenced by the side-chain linkage was studied with electrochemical impedance spectroscopy, which shows a dramatic increase in volumetric capacitance from 80 ± 9 up to 242 ± 17 F/cm3 on going from P3MEET to P3MEEET. Thus, ethyl-spaced P3MEEET exhibits one of the highest reported volumetric capacitance values among p-type polymers. Moreover, P3MEEET exhibits in dry thin films an organic field-effect transistor (OFET) hole mobility of 0.005 cm2/V·s, highest among the three, which is one order of magnitude higher than that for P3MEEMT. The extracted hole mobility from OECT (oxidized swollen state) and the hole mobility in solid-state thin films (OFET) show contradictory trends for P3MEEMT and P3MEEET. In order to understand exactly the properties in the hydrated and dry states, the crystal structure of the polymers was investigated with wide-angle X-ray scattering (WAXS) and grazing incidence WAXS, and the water uptake under applied potential was monitored using electrochemical quartz crystal microbalance with dissipation monitoring (E-QCMD). These measurements reveal an amorphous state for P3MEET and a semicrystalline state for P3MEEMT and P3MEEEET. On the other hand, E-QCMD confirms that P3MEEET swells 10 times more than P3MEEMT in the oxidized state. Thus, the importance of the ethyl spacer toward crystallinity and mixed-conduction properties was clearly demonstrated, emphasizing the impact of side chain linkage of diethylene glycol. This detailed study offers a better understanding of how to design high-performance organic mixed conductors.


"Now What?" Perceived Factors Influencing Knowledge Exchange in School Health Research.
Increasing the uptake of school health research into practice is pivotal for improving adolescent health. COMPASS, a longitudinal study of Ontario and Alberta secondary students and schools (2012-2021), used a knowledge exchange process to enhance schools' use of research findings. Schools received annual summaries of their students' health behaviors and suggestions for action and were linked with a knowledge broker to support them in making changes to improve student health. The current research explored factors that influenced COMPASS knowledge exchange activities. Semistructured interviews were conducted with researchers (n = 13), school staff (n = 13), and public health stakeholders (n = 4). Interestingly, knowledge users focused more on factors that influenced their use of COMPASS findings than factors that influenced knowledge brokering. The factors identified by participants are similar to those that influence implementation of school health interventions (e.g., importance of school champions, competing priorities, inadequate resources). While knowledge exchange offers a way to reduce the gap between research and practice, schools that need the most support may not engage in knowledge exchange; hence, we must consider how to increase engagement of these schools to ultimately improve student health.


Variability of Potassium Channel Blockers in Mesobuthus eupeus Scorpion Venom with Focus on Kv1.1: AN INTEGRATED TRANSCRIPTOMIC AND PROTEOMIC STUDY.
The lesser Asian scorpion Mesobuthus eupeus (Buthidae) is one of the most widely spread and dispersed species of the Mesobuthus genus, and its venom is actively studied. Nevertheless, a considerable amount of active compounds is still under-investigated due to the high complexity of this venom. Here, we report a comprehensive analysis of putative potassium channel toxins (KTxs) from the cDNA library of M. eupeus venom glands, and we compare the deduced KTx structures with peptides purified from the venom. For the transcriptome analysis, we used conventional tools as well as a search for structural motifs characteristic of scorpion venom components in the form of regular expressions. We found 59 candidate KTxs distributed in 30 subfamilies and presenting the cysteine-stabilized α/β and inhibitor cystine knot types of fold. M. eupeus venom was then separated to individual components by multistage chromatography. A facile fluorescent system based on the expression of the KcsA-Kv1.1 hybrid channels in Escherichia coli and utilization of a labeled scorpion toxin was elaborated and applied to follow Kv1.1 pore binding activity during venom separation. As a result, eight high affinity Kv1.1 channel blockers were identified, including five novel peptides, which extend the panel of potential pharmacologically important Kv1 ligands. Activity of the new peptides against rat Kv1.1 channel was confirmed (IC50 in the range of 1-780 nm) by the two-electrode voltage clamp technique using a standard Xenopus oocyte system. Our integrated approach is of general utility and efficiency to mine natural venoms for KTxs.


Vertically fused tracheal cartilage. An underrecognized anomaly.
Anterior vertical fusion of the tracheal cartilages with normal pars membranacea is a rarely described tracheal anomaly. We report four cases of this anomaly, three associated with craniosynostosis syndromes and one with Goldenhar's syndrome. The anomaly was documented at postmortem examination in one case, both endoscopically and at tracheotomy in two cases, and endoscopically in one case. Documentation from gross postmortem photographs, photomicrographs, and intraoperative endoscopic photographs is presented so that the anatomical aspects of this anomaly can be thoroughly understood and recognized. All four newborn patients developed recurrent lower respiratory tract infections, reactive airway disease, and chronically retained secretions attributable to this anomaly. Neither tracheal stenosis nor stridor was encountered. In the three surviving patients, the infections grew less frequent and less severe in later childhood. These observations suggest that this tracheobronchial anomaly may be associated with a variety of craniofacial syndromes and may alter airflow dynamics in a way that predisposes to retained secretions and chronic pulmonary disease.


Single subunits of the GABAA receptor form ion channels with properties of the native receptor.
The alpha and beta subunits of the gamma-aminobutyric acidA (GABAA) receptor were expressed individually in Xenopus oocytes by injection of RNA synthesized from their cloned DNAs. GABA-sensitive chloride channels were detected several days after injection with any one of three different alpha RNAs (alpha 1, alpha 2, and alpha 3) or with beta RNA. The channels induced by each of the alpha-subunit RNAs were indistinguishable, they had multiple conductance levels (10, 19, 28, and 42 picosiemens), and their activity was potentiated by pentobarbital and inhibited by picrotoxin. The beta channels usually expressed poorly but showed similar single channel conductance levels (10, 18, 27, and 40 picosiemens), potentiation by pentobarbital and inhibition by picrotoxin. The finding that both alpha and beta subunits, examined separately, form GABA-sensitive ion channels with permeation properties and regulatory sites characteristic of the native receptor suggests that the amino acid sequences that confer these properties are within the homologous domains shared by the subunits.


Peer-Led, Postanatomy Reflection Exercise in Dissection Teams: Curriculum and Training Materials.
The importance of emphasizing empathy, reflection, and professionalism during anatomy dissection has been well established. This small-group curriculum was developed to fulfill a need for structured reflection at the end of anatomy. In this 45-minute reflection session, each dissection team of first-year medical students in anatomy is led by one or two peer facilitators recruited from the second-year medical student class. The session is designed to include a time for sharing of personal reflections, a clinical observation activity about the cadaver's cause of death, and an appreciative inquiry approach to the dissection team experience. In addition to the reflection session curriculum, materials also include a 1-hour presession training module, containing a small-group facilitator skill-building and role-play and a 30-minute postsession debrief for peer facilitators. We found that the majority of anatomy students felt that the end-of-course reflection was a meaningful way to conclude the course and that the session had a positive impact on their relationship with their dissection team. Fifteen peer facilitators participated in focus groups, and common themes included the value of giving back, making meaning of past experiences, countering burnout by recognizing one's own growth, and continued learning through peer teaching. This anatomy reflection curriculum has been incorporated into our 7-week anatomy course and has been well received by both anatomy students and peer facilitators. We believe that peer-led small-group reflection sessions after intensive experiences in medical school can promote personal and professional growth among both junior and senior medical students.


Stridor in multiple system atrophy: Consensus statement on diagnosis, prognosis, and treatment.
Multiple system atrophy (MSA) is a neurodegenerative disorder characterized by a combination of autonomic failure, cerebellar ataxia, and parkinsonism. Laryngeal stridor is an additional feature for MSA diagnosis, showing a high diagnostic positive predictive value, and its early occurrence might contribute to shorten survival. A consensus definition of stridor in MSA is lacking, and disagreement persists about its diagnosis, prognosis, and treatment. An International Consensus Conference among experts with methodological support was convened in Bologna in 2017 to define stridor in MSA and to reach consensus statements for the diagnosis, prognosis, and treatment. Stridor was defined as a strained, high-pitched, harsh respiratory sound, mainly inspiratory, occurring only during sleep or during both sleep and wakefulness, and caused by laryngeal dysfunction leading to narrowing of the rima glottidis. According to the consensus, stridor may be recognized clinically by the physician if present at the time of examination, with the help of a witness, or by listening to an audio recording. Laryngoscopy is suggested to exclude mechanical lesions or functional vocal cord abnormalities related to different neurologic conditions. If the suspicion of stridor needs confirmation, drug-induced sleep endoscopy or video polysomnography may be useful. The impact of stridor on survival and quality of life remains uncertain. Continuous positive airway pressure and tracheostomy are both suggested as symptomatic treatment of stridor, but whether they improve survival is uncertain. Several research gaps emerged involving diagnosis, prognosis, and treatment. Unmet needs for research were identified.


MR imaging assessment of experimental hepatic dysfunction with Mn-DPDP.
Manganese (II) N,N'-dipyridoxylethylenediamine-N,N'-diacetate 5,5'-bis(phosphate) (DPDP) is a paramagnetic magnetic resonance (MR) contrast agent that enhances the liver and is predominantly excreted through the biliary tree. The authors evaluated its utility in diffuse liver disease by assessing liver and gallbladder enhancement in 24 rabbits. Total (n = 6) or segmental (n = 6) biliary occlusion or galactosamine-induced hepatitis (n = 6) was induced 3 days before imaging. Six rabbits served as normal controls. T1- and T2-weighted axial MR images were acquired at baseline, followed by T1-weighted images every 10 minutes for 1 hour after the intravenous administration of 20 mumol/kg Mn-DPDP. Except for the segmental occlusion group, the baseline study did not allow distinction between normal and abnormal livers. The temporal hepatic enhancement pattern was statistically different for each group. The normal, segmental occlusion, and hepatitis groups showed patterns similar to one another but markedly higher signal intensity than the total-occlusion group throughout the observation period. In contrast, the gallbladder showed a greater difference in both degree of enhancement and time to peak enhancement among the four groups. Mn-DPDP produces a temporal hepatic enhancement pattern that allows recognition of markedly impaired livers, and gallbladder enhancement patterns that allow distinction of more subtly impaired livers.


An event-related functional MRI study comparing interference effects in the Simon and Stroop tasks.
The Stroop and Simon tasks typify a class of interference effects in which the introduction of task-irrelevant stimulus characteristics robustly slows reaction times. Behavioral studies have not succeeded in determining whether the neural basis for the resolution of these interference effects during successful task performance is similar or different across tasks. Event-related functional magnetic resonance imaging (fMRI) studies were obtained in 10 healthy young adults during performance of the Stroop and Simon tasks. Activation during the Stroop task replicated findings from two earlier fMRI studies. These activations were remarkably similar to those observed during the Simon task, and included anterior cingulate, supplementary motor, visual association, inferior temporal, inferior parietal, inferior frontal, and dorsolateral prefrontal cortices, as well as the caudate nuclei. The time courses of activation were also similar across tasks. Resolution of interference effects in the Simon and Stroop tasks engage similar brain regions, and with a similar time course. Therefore, despite the widely differing stimulus characteristics employed by these tasks, the neural systems that subserve successful task performance are likely to be similar as well.


Early Childhood Obesity Risk Factors: Socioeconomic Adversity, Family Dysfunction, Offspring Distress, and Junk Food Self-Medication.
To explore the sequence and interaction of infancy and early childhood risk factors, particularly relating to disturbances in the social environment, and how the consequences of such exposures can promote weight gain and obesity. This review will argue that socioeconomic adversity is a key upstream catalyst that sets the stage for critical midstream risk factors such as family strain and dysfunction, offspring insecurity, stress, emotional turmoil, low self-esteem, and poor mental health. These midstream risk factors, particularly stress and emotional turmoil, create a more or less perfect foil for calorie-dense junk food self-medication and subtle addiction, to alleviate uncomfortable psychological and emotional states. Disturbances in the social environment during infancy and early childhood appear to play a critical role in weight gain and obesity, through such mechanisms as insecurity, stress, and emotional turmoil, eventually leading to junk food self-medication and subtle addiction.


Two types of Alzheimer's beta-amyloid (1-40) peptide membrane interactions: aggregation preventing transmembrane anchoring versus accelerated surface fibril formation.
The 39-42 amino acid long, amphipathic amyloid-beta peptide (Abeta) is one of the key components involved in Alzheimer's disease (AD). In the neuropathology of AD, Abeta presumably exerts its neurotoxic action via interactions with neuronal membranes. In our studies a combination of 31P MAS NMR (magic angle spinning nuclear magnetic resonance) and CD (circular dichroism) spectroscopy suggest fundamental differences in the functional organization of supramolecular Abeta(1-40) membrane assemblies for two different scenarios with potential implication in AD: Abeta peptide can either be firmly anchored in a membrane upon proteolytic cleavage, thereby being prevented against release and aggregation, or it can have fundamentally adverse effects when bound to membrane surfaces by undergoing accelerated aggregation, causing neuronal apoptotic cell death. Acidic lipids can prevent release of membrane inserted Abeta(1-40) by stabilizing its hydrophobic transmembrane C-terminal part (residue 29-40) in an alpha-helical conformation via an electrostatic anchor between its basic Lys28 residue and the negatively charged membrane interface. However, if Abeta(1-40) is released as a soluble monomer, charged membranes act as two-dimensional aggregation-templates where an increasing amount of charged lipids (possible pathological degradation products) causes a dramatic accumulation of surface-associated Abeta(1-40) peptide followed by accelerated aggregation into toxic structures. These results suggest that two different molecular mechanisms of peptide-membrane assemblies are involved in Abeta's pathophysiology with the finely balanced type of Abeta-lipid interactions against release of Abeta from neuronal membranes being overcompensated by an Abeta-membrane assembly which causes toxic beta-structured aggregates in AD. Therefore, pathological interactions of Abeta peptide with neuronal membranes might not only depend on the oligomerization state of the peptide, but also the type and nature of the supramolecular Abeta-membrane assemblies inherited from Abeta's origin.


2022 ACC/AHA Guideline for the Diagnosis and Management of Aortic Disease: A Report of the American Heart Association/American College of Cardiology Joint Committee on Clinical Practice Guidelines.
The "2022 ACC/AHA Guideline for the Diagnosis and Management of Aortic Disease" provides recommendations to guide clinicians in the diagnosis, genetic evaluation and family screening, medical therapy, endovascular and surgical treatment, and long-term surveillance of patients with aortic disease across its multiple clinical presentation subsets (ie, asymptomatic, stable symptomatic, and acute aortic syndromes). A comprehensive literature search was conducted from January 2021 to April 2021, encompassing studies, reviews, and other evidence conducted on human subjects that were published in English from PubMed, EMBASE, the Cochrane Library, CINHL Complete, and other selected databases relevant to this guideline. Additional relevant studies, published through June 2022 during the guideline writing process, were also considered by the writing committee, where appropriate. Structure: Recommendations from previously published AHA/ACC guidelines on thoracic aortic disease, peripheral artery disease, and bicuspid aortic valve disease have been updated with new evidence to guide clinicians. In addition, new recommendations addressing comprehensive care for patients with aortic disease have been developed. There is added emphasis on the role of shared decision making, especially in the management of patients with aortic disease both before and during pregnancy. The is also an increased emphasis on the importance of institutional interventional volume and multidisciplinary aortic team expertise in the care of patients with aortic disease.


Dipyridamole, an Anti-platelet Agent, Reduces the Vascular Endothelial Cell Injury Induced by Active Oxygen Species.
The effects of the anti-platelet agent dipyridamole on vascular endothelial cells were assessed by measurement of the injury index induced by oxygen stress. Vascular endothelial cell injury was assayed by measuring (51)Cr release from labeled vascular endothelial cells. Leukocytes activated by phorbol 12-myristate 13-acetate (PMA) were used to induce the injury of endothelial cells. In this system, dipyridamole suppressed endothelial cell injury in a dose dependent manner (0.1-10 4mUM) while it had no effect on production of superoxide anion in PMA-activated leukocytes. Treatment of endothelial cells with hydrogen peroxide also induced endothelial cell injury in a dose dependent manner (50-150 μM). Dipyridamole also prevented the endothelial cell injury induced by hydrogen peroxide with a dose dependent fashion (1-10 μM). There were no significant changes in the activities of catalyzing enzymes such as catalase and glutathione peroxidase in the endothelial cells following dipyridamole treatment. In contrast, dipyridamole significantly increased the cyclic GMP content of endothelial cells in a dose dependent manner (1-10 μM). Addition of 8-bromo-cyclic GMP (1 mM) to the culture also protected endothelial cells from injury induced by hydrogen peroxide, but 8-bromo-cyclic AMP did not. These data suggest that the protective effect of dipyridamole against oxygen stress is correlated with the increase in the cyclic GMP content of the endothelial cells.


miRNA signature and Dicer requirement during human endometrial stromal decidualization in vitro.
Decidualization is a morphological and biochemical transformation of endometrial stromal fibroblast into differentiated decidual cells, which is critical for embryo implantation and pregnancy establishment. The complex regulatory networks have been elucidated at both the transcriptome and the proteome levels, however very little is known about the post-transcriptional regulation of this process. miRNAs regulate multiple physiological pathways and their de-regulation is associated with human disorders including gynaecological conditions such as endometriosis and preeclampsia. In this study we profile the miRNAs expression throughout human endometrial stromal (hESCs) decidualization and analyze the requirement of the miRNA biogenesis enzyme Dicer during this process. A total of 26 miRNAs were upregulated and 17 miRNAs downregulated in decidualized hESCs compared to non-decidualized hESCs. Three miRNAs families, miR-181, miR-183 and miR-200, are down-regulated during the decidualization process. Using miRNAs target prediction algorithms we have identified the potential targets and pathways regulated by these miRNAs. The knockdown of Dicer has a minor effect on hESCs during in vitro decidualization. We have analyzed a battery of decidualization markers such as cell morphology, Prolactin, IGFBP-1, MPIF-1 and TIMP-3 secretion as well as HOXA10, COX2, SP1, C/EBPß and FOXO1 expression in decidualized hESCs with decreased Dicer function. We found decreased levels of HOXA10 and altered intracellular organization of actin filaments in Dicer knockdown decidualized hESCs compared to control. Our results provide the miRNA signature of hESC during the decidualization process in vitro. We also provide the first functional characterization of Dicer during human endometrial decidualization although surprisingly we found that Dicer plays a minor role regulating this process suggesting that alternative biogenesis miRNAs pathways must be involved in human endometrial decidualization.


Sources of information about MDMA (3,4-methylenedioxymethamphetamine): perceived accuracy, importance, and implications for prevention among young adult users.
The goal of this cross-sectional study was to assess the perceived accuracy and the importance of various sources of information about MDMA/ecstasy among young adult users. A respondent driven sampling plan was used to recruit a community sample of recent ecstasy users (n = 304), aged 18-30, in Ohio, who responded to structured interviews. Friends, drug abuse treatment programs, and physicians were perceived to be the most accurate sources of information about ecstasy by 45.7, 37.2, and 30.3% of the sample, respectively. Friends were considered the most important source of information about ecstasy (40.2%), followed by web sites like DanceSafe (16.2%), and MTV/VH1 television specials (6.9%). About half the sample used the Internet to obtain information about ecstasy, with younger and more educated participants significantly more likely to do so. Educated users were also significantly more likely to consider the Internet to be an important source of information. Web sites like DanceSafe were visited by four times as many users as government-sponsored web sites. Findings support the development of peer-oriented, network strategies to reach ecstasy users with prevention messages. Efforts to make prevention web sites more attractive should be considered.


Adaptive Radiation in Socially Advanced Stem-Group Ants from the Cretaceous.
Across terrestrial ecosystems, modern ants are ubiquitous. As many as 94 out of every 100 individual arthropods in rainforests are ants, and they constitute up to 15% of animal biomass in the Amazon. Moreover, ants are pervasive agents of natural selection as over 10,000 arthropod species are specialized inquilines or myrmecomorphs living among ants or defending themselves through mimicry. Such impact is traditionally explained by sociality: ants are the first major group of ground-dwelling predatory insects to become eusocial, increasing efficiency of tasks and establishing competitive superiority over solitary species. A wealth of specimens from rich deposits of 99 million-year-old Burmese amber resolves ambiguity regarding sociality and diversity in the earliest ants. The stem-group genus Gerontoformica maintained distinct reproductive castes including morphotypes unknown in solitary aculeate (stinging) wasps, providing insight into early behavior. We present rare aggregations of workers, indicating group recruitment as well as an instance of interspecific combat; such aggression is a social feature of modern ants. Two species and an unusual new genus are described, further expanding the remarkable diversity of early ants. Stem-group ants are recovered as a paraphyletic assemblage at the base of modern lineages varying greatly in size, form, and mouthpart structure, interpreted here as an adaptive radiation. Though Cretaceous stem-group ants were eusocial and adaptively diverse, we hypothesize that their extinction resulted from the rise of competitively superior crown-group taxa that today form massive colonies, consistent with Wilson and Hölldobler's concept of "dynastic succession."


High seas fisheries play a negligible role in addressing global food security.
Recent international negotiations have highlighted the need to protect marine diversity on the high seas-the ocean area beyond national jurisdiction. However, restricting fishing access on the high seas raises many concerns, including how such restrictions would affect food security. We analyze high seas catches and trade data to determine the contribution of the high seas catch to global seafood production, the main species caught on the high seas, and the primary markets where these species are sold. By volume, the total catch from the high seas accounts for 4.2% of annual marine capture fisheries production and 2.4% of total seafood production, including freshwater fisheries and aquaculture. Thirty-nine fish and invertebrate species account for 99.5% of the high seas targeted catch, but only one species, Antarctic toothfish, is caught exclusively on the high seas. The remaining catch, which is caught both on the high seas and in national jurisdictions, is made up primarily of tunas, billfishes, small pelagic fishes, pelagic squids, toothfish, and krill. Most high seas species are destined for upscale food and supplement markets in developed, food-secure countries, such as Japan, the European Union, and the United States, suggesting that, in aggregate, high seas fisheries play a negligible role in ensuring global food security.


Stem cell research: paths to cancer therapies and regenerative medicine.
Most tissues in complex metazoans contain a rare subset of cells that, at the single-cell level, can self-renew and also give rise to mature daughter cells. Such stem cells likely in development build tissues and are retained in adult life to regenerate them. Cancers and leukemias are apparently not an exception: rare leukemia stem cells and cancer stem cells have been isolated that contain all of the tumorigenicity of the whole tumor, and it is their properties that will guide future therapies. None of this was apparent just 20 years ago, yet this kind of stem cell thinking already provides new perspectives in medical science and could usher in new therapies. Today, political, religious, and ethical issues surround embryonic stem cell and patient-specific pluripotent stem cell research and are center stage in the attempts by governments to ban these fields for discovery and potential therapies. These interventions require physicians and physician-scientists to determine for themselves whether patient welfare or personal ethics will dominate in their practices, and whether all aspects of stem cell research can be pursued in a safe and regulated fashion.


Sentinel lymph node biopsy in melanoma: The Oxford ten year clinical experience.
Sentinel lymph node biopsy (SLNB) has become an established investigation for assessing microscopic nodal metastasis in melanoma. The American Joint Committee on Cancer (AJCC) incorporates the sentinel node status in its staging criteria for melanoma. We present our clinical evaluation of performing SLNB in a single UK centre between 1998 and 2008. There were 697 patients with a mean age 53 years (range 13-92). We were able to surgically harvest at least one sentinel node in 694 patients of which 532 (76%) were negative. Of the 162 positive patients, 129 underwent further completion lymphadenectomy with 29 showing further pathologically positive nodes. At median follow up of 46 months, mortality from melanoma for SLN positive and negative patients was 32% and 4%, respectively. Disease recurrence was noted in 10% of the SLN negative group. Survival curves showed significant difference (p<0.001) in outcomes for patients grouped by Breslow thickness. Postoperative complications were noted in 6% of patients. No life-threatening complications were noted. Our results are comparable to other national and international studies. We await the outcomes of ongoing trials to assess the therapeutic value of SLNB for melanoma.


How do marital status, work effort, and wage rates interact?
How marital status interacts with men's earnings is an important analytic and policy issue, especially in the context of debates in the United States over programs that encourage healthy marriage. This paper generates new findings about the earnings-marriage relationship by estimating the linkages among flows into and out of marriage, work effort, and wage rates. The estimates are based on National Longitudinal Survey of Youth panel data, covering 23 years of marital and labor market outcomes, and control for unobserved heterogeneity. We estimate marriage effects on hours worked (our proxy for work effort) and on wage rates for all men and for black and low-skilled men separately. The estimates reveal that entering marriage raises hours worked quickly and substantially but that marriage's effect on wage rates takes place more slowly while men continue in marriage. Together; the stimulus to hours worked and wage rates generates an 18%-19% increase in earnings, with about one-third to one-half of the marriage earnings premium attributable to higher work effort. At the same time, higher wage rates and hours worked encourage men to marry and to stay married. Thus, being married and having high earnings reinforce each other over time.


Differential effects of corticosterone on the slow afterhyperpolarization in the basolateral amygdala and CA1 region: possible role of calcium channel subunits.
The stress hormone corticosterone increases the amplitude of the slow afterhyperpolarization (sAHP) in CA1 pyramidal neurons, without affecting resting membrane potential, input resistance, or action potential characteristics. We here examined how corticosterone affects these properties in the basolateral amygdala (BLA). In the amygdala, corticosterone does not change the AHP amplitude, nor any of the passive and active membrane properties studied. The lack of effect on the AHP is surprising since in both areas corticosterone increases high-voltage-activated sustained calcium currents, which supposedly regulate the sAHP. We wondered whether corticosterone targets different calcium channel subunits in the two areas because currents through only one of the subunits (Cav1.3) are thought to alter the AHP amplitude. In situ hybridization studies revealed that CA1 cells express Cav1.2 and Cav1.3 subunits; corticosterone does not transcriptionally regulate Cav1.2 but increases Cav1.3 expression compared with vehicle treatment. In the BLA, Cav1.3 expression was not detectable, both after control and corticosterone treatment. Cav1.2 is moderately expressed. In a modeling study, we examined putative consequences of changes in specific calcium channel subunit expression and calcium extrusion by corticosterone for the AHP in CA1 and amygdala neurons. A differential distribution and transcriptional regulation of Cav1.2 and Cav1.3 in the CA1 area versus BLA partly explain the observed differences in AHP amplitude. The functional implication of these findings could be that stress-induced arousal of activity in the BLA is more prolonged than that in the CA1 hippocampal area, so that information with an emotional component is more effectively encoded.


Synthetic and Mechanistic Implications of Chlorine Photoelimination in Nickel/Photoredox C(sp3)-H Cross-Coupling.
In recent years, the development of light-driven reactions has contributed numerous advances in synthetic organic chemistry. A particularly active research area combines photoredox catalysis with nickel catalysis to accomplish otherwise inaccessible cross-coupling reactions. In these reactions, the photoredox catalyst absorbs light to generate an electronically excited charge-transfer state that can engage in electron or energy transfer with a substrate and the nickel catalyst. Our group questioned whether photoinduced activation of the nickel catalyst itself could also contribute new approaches to cross-coupling. Over the past 5 years, we have sought to advance this hypothesis for the development of a suite of mild and site-selective C(sp3)-H cross-coupling reactions with chloride-containing coupling partners via photoelimination of a Ni-Cl bond.On the basis of a report from the Nocera laboratory, we reasoned that photolysis of a Ni(III) aryl chloride species, generated by single-electron oxidation of a typical Ni(II) intermediate in cross-coupling, might allow for the catalytic generation of chlorine atoms. Combining this with the ability of Ni(II) to accept alkyl radicals, we hypothesized that photocatalytically generated chlorine atoms could mediate hydrogen atom transfer (HAT) with C(sp3)-H bonds to generate a substrate-derived alkyl radical that is captured by the Ni center in cross-coupling. A photoredox catalyst was envisioned to promote the necessary single-electron oxidation and reduction of the Ni catalyst to facilitate an overall redox-neutral process. Overall, this strategy would offer a visible-light-driven mechanism for chlorine radical formation enabled by the sequential capture of two photons.As an initial demonstration, we developed a Ni/photoredox-catalyzed α-oxy C(sp3)-H arylation of cyclic and acyclic ethers. This method was extended to a mild formylation of abundant and complex aryl chlorides through selective 2-functionalization of 1,3-dioxolane. Seeking to develop a suite of reactions that introduce carbon at all different oxidation states, we explored C(sp3)-H cross-coupling with trimethyl orthoformate, a common laboratory solvent. We found that trimethyl orthoformate serves as a source of methyl radical for a methylation reaction via β-scission from a tertiary radical generated upon chlorine-mediated HAT. Since chlorine radical is capable of abstracting unactivated C(sp3)-H bonds, our efforts have also been directed at cross-coupling with a range of feedstock chemicals, such as alkanes and toluenes, along with late-stage intermediates, using chloroformates as coupling partners. Overall, this platform enables access to valuable synthetic transformations with (hetero)aryl chlorides, which despite being the most ubiquitous and inexpensive aryl halide coupling partners, are rarely reactive in Ni/photoredox catalysis.Little is known about the photophysics and photochemistry of organometallic Ni complexes relevant to cross-coupling. We have conducted mechanistic investigations, including computational, spectroscopic, emission quenching, and stoichiometric oxidation studies, of Ni(II) aryl halide complexes common to Ni/photoredox reactions. These studies indicate that chlorine radical generation from excited Ni(III) is operative in the described C(sp3)-H functionalization methods. More generally, the studies illustrate that the photochemistry of cross-coupling catalysts cannot be ignored in metallaphotoredox reactions. We anticipate that further mechanistic understanding should facilitate new catalyst design and lead to the development of new synthetic methods.


Primaquine to reduce transmission of Plasmodium falciparum malaria in Mali: a single-blind, dose-ranging, adaptive randomised phase 2 trial.
Single low doses of primaquine, when added to artemisinin-based combination therapy, might prevent transmission of Plasmodium falciparum malaria to mosquitoes. We aimed to establish the activity and safety of four low doses of primaquine combined with dihydroartemisinin-piperaquine in male patients in Mali. In this phase 2, single-blind, dose-ranging, adaptive randomised trial, we enrolled boys and men with uncomplicated P falciparum malaria at the Malaria Research and Training Centre (MRTC) field site in Ouelessebougou, Mali. All participants were confirmed positive carriers of gametocytes through microscopy and had normal function of glucose-6-phosphate dehydrogenase (G6PD) on colorimetric quantification. In the first phase, participants were randomly assigned (1:1:1) to one of three primaquine doses: 0 mg/kg (control), 0·125 mg/kg, and 0·5 mg/kg. Randomisation was done with a computer-generated randomisation list (in block sizes of six) and concealed with sealed, opaque envelopes. In the second phase, different participants were sequentially assigned (1:1) to 0·25 mg/kg primaquine or 0·0625 mg/kg primaquine. Primaquine tablets were dissolved into a solution and administered orally in a single dose. Participants were also given a 3 day course of dihydroartemisinin-piperaquine, administered by weight (320 mg dihydroartemisinin and 40 mg piperaquine per tablet). Outcome assessors were masked to treatment allocation, but participants were permitted to find out group assignment. Infectivity was assessed through membrane-feeding assays, which were optimised through the beginning part of phase one. The primary efficacy endpoint was the mean within-person percentage change in mosquito infectivity 2 days after primaquine treatment in participants who completed the study after optimisation of the infectivity assay, had both a pre-treatment infectivity measurement and at least one follow-up infectivity measurement, and who were given the correct primaquine dose. The safety endpoint was the mean within-person change in haemoglobin concentration during 28 days of study follow-up in participants with at least one follow-up visit. This study is registered with ClinicalTrials.gov, number NCT01743820. Between Jan 2, 2013, and Nov 27, 2014, we enrolled 81 participants. In the primary analysis sample (n=71), participants in the 0·25 mg/kg primaquine dose group (n=15) and 0·5 mg/kg primaquine dose group (n=14) had significantly lower mean within-person reductions in infectivity at day 2-92·6% (95% CI 78·3-100; p=0·0014) for the 0·25 mg/kg group; and 75·0% (45·7-100; p=0·014) for the 0·5 mg/kg primaquine group-compared with those in the control group (n=14; 11·3% [-27·4 to 50·0]). Reductions were not significantly different from control for participants assigned to the 0·0625 mg/kg dose group (n=16; 41·9% [1·4-82·5]; p=0·16) and the 0·125 mg/kg dose group (n=12; 54·9% [13·4-96·3]; p=0·096). No clinically meaningful or statistically significant drops in haemoglobin were recorded in any individual in the haemoglobin analysis (n=70) during follow-up. No serious adverse events were reported and adverse events did not differ between treatment groups. A single dose of 0·25 mg/kg primaquine, given alongside dihydroartemisinin-piperaquine, was safe and efficacious for the prevention of P falciparum malaria transmission in boys and men who are not deficient in G6PD. Future studies should assess the safety of single-dose primaquine in G6PD-deficient individuals to define the therapeutic range of primaquine to enable the safe roll-out of community interventions with primaquine. Bill & Melinda Gates Foundation.


Vehicle Systems and Excipients Used in Minipig Drug Development Studies.
Minipigs have been used for dermal drug development studies for decades, and they are currently more frequently considered as the second nonrodent species for pivotal nonclinical studies, in lieu of the dog or nonhuman primate, for compounds delivered via standard systemic routes of administration. Little is known about the tolerability of different excipients in minipigs; sharing knowledge of excipient tolerability and compositions previously used in nonclinical studies may avoid testing of inadequate formulations, thereby contributing to reduced animal usage. This article reviews vehicles employed in the Göttingen(®)minipig based on the combined experience from a number of pharmaceutical companies and contract research organizations. The review includes vehicles tolerated for single or multiple dosing by the Göttingen minipig, some of which are not appropriate for administration to other common nonrodent species (e.g., dogs). By presenting these data for dermal, oral, subcutaneous, and intravenous routes of administration, studies to qualify these vehicles in minipigs can be minimized or avoided. Additionally, investigators may more frequently consider using the minipig in place of higher species if the tolerability of a vehicle in the minipig is known.


Whole Body Vibration Exposures and Health Status among Professional Truck Drivers: A Cross-sectional Analysis.
Many professional truck drivers suffer from low back pain (LBP) which is thought to be associated with exposure to whole-body vibration (WBV). The objectives of this study were to: (i) characterize general health, regional body pain and WBV exposures, (ii) evaluate the associations between different WBV parameters and health outcomes, and (iii) determine whether there were factors which affect a truck driver's WBV exposures. This study analyzed WBV exposures from 96 long-haul truck drivers over their regular work shift (6-15h) per International Standards Organization (ISO) 2631-1 and 2631-5 WBV standards. This study also evaluated regional body pain (10-point scale), low back disability (the Oswestry Disability Index), and physical and mental health (the Short Form 12-item Health Survey). The results demonstrated that the daily vector sum WBV exposures [A(8), VDV(8) and Sed(8)] were above action limits while the predominant z-axis exposures were below action limits. Among all the musculoskeletal outcomes, LBP was the most prevalent (72.5%) with average LBP score of 2.9 (SD: 2.0). The SF-12 health scores demonstrated that truck drivers in general had lower physical health status than the general US population (P's < 0.04) and that physical health status decreased as WBV exposures increased (P = 0.03). In addition, the correlations between the WBV measures and health outcomes indicated that A(8) exposure measures had a stronger link to musculoskeletal (LBP) and other health outcomes than the VDV(8) and Sed(8) measures. Finally, seat manufacturer and seat age were two factors which had a strong influence on WBV exposures.


Prevalence and Clinical Features of Inflammatory Bowel Diseases Associated With Monogenic Variants, Identified by Whole-Exome Sequencing in 1000 Children at a Single Center.
A proportion of infants and young children with inflammatory bowel diseases (IBDs) have subtypes associated with a single gene variant (monogenic IBD). We aimed to determine the prevalence of monogenic disease in a cohort of pediatric patients with IBD. We performed whole-exome sequencing analyses of blood samples from an unselected cohort of 1005 children with IBD, aged 0-18 years (median age at diagnosis, 11.96 years) at a single center in Canada and their family members (2305 samples total). Variants believed to cause IBD were validated using Sanger sequencing. Biopsies from patients were analyzed by immunofluorescence and histochemical analyses. We identified 40 rare variants associated with 21 monogenic genes among 31 of the 1005 children with IBD (including 5 variants in XIAP, 3 in DOCK8, and 2 each in FOXP3, GUCY2C, and LRBA). These variants occurred in 7.8% of children younger than 6 years and 2.3% of children aged 6-18 years. Of the 17 patients with monogenic Crohn's disease, 35% had abdominal pain, 24% had nonbloody loose stool, 18% had vomiting, 18% had weight loss, and 5% had intermittent bloody loose stool. The 14 patients with monogenic ulcerative colitis or IBD-unclassified received their diagnosis at a younger age, and their most predominant feature was bloody loose stool (78%). Features associated with monogenic IBD, compared to cases of IBD not associated with a single variant, were age of onset younger than 2 years (odds ratio [OR], 6.30; P = .020), family history of autoimmune disease (OR, 5.12; P = .002), extra-intestinal manifestations (OR, 15.36; P < .0001), and surgery (OR, 3.42; P = .042). Seventeen patients had variants in genes that could be corrected with allogeneic hematopoietic stem cell transplantation. In whole-exome sequencing analyses of more than 1000 children with IBD at a single center, we found that 3% had rare variants in genes previously associated with pediatric IBD. These were associated with different IBD phenotypes, and 1% of the patients had variants that could be potentially corrected with allogeneic hematopoietic stem cell transplantation. Monogenic IBD is rare, but should be considered in analysis of all patients with pediatric onset of IBD.


Development and validation of a general health literacy test in Singapore.
Due to the concern of equating correct pronunciation with comprehension and the differences in health care systems, existing health literacy (HL) instruments may not be appropriate for or applicable to English-speaking countries other than the USA. The purpose of this study was to develop and validate the Health Literacy Test for Singapore (HLTS), which is an adapted version of the Short-Test of Functional Health Literacy in Adults. Three hundred and two patients were interviewed and administered the HLTS, the Newest Vital Sign (NVS), a demographic questionnaire, and a knowledge test of chronic diseases. The convergent validity of HLTS was determined by examining the association between HLTS and NVS HL levels, whereas predictive validity was tested by examining the difference in knowledge of chronic conditions between the two HLTS HL (i.e. adequate and inadequate HL) groups. Bivariate correlation of HLTS HL levels with age and education was assessed to test a priori hypotheses that patients with inadequate HL were older and less educated. The results showed that HLTS displayed good internal reliability (Cronbach's alpha = 0.87). The correlation between HLTS and NVS was moderate (γ = 0.55; P = 0.005) and individuals with inadequate HL were older (P = 0.002) and less educated (P = 0.007). In addition, patients with adequate HL had a higher mean score on the chronic disease knowledge test (P = 0.036). In conclusion, the HLTS is a valid and reliable measure for assessing Singaporeans ability to read and comprehend health-related materials written in English.


Does the mast cell still have a key role in asthma?
In recent years, the emerging concept of bronchial inflammation as a prominent histopathologic characteristic of asthma has profoundly modified the view of the role of the mast cell, which was traditionally thought to be linked to the release of soluble chemical mediators substantially involved in the genesis of acute, immediate bronchospasm. The finding that the production of proinflammatory cytokines by mast cells in asthmatic airways is comparable, in some circumstances, to that of T-cell origin, has led to the hypothesis that mast cells, along with T lymphocytes and eosinophils, may also contribute to the genesis of chronic, persistent asthma. This hypothesis is further supported by the finding that mast cells are able to functionally interact with B cells (promoting IgE synthesis) and T lymphocytes (acting as antigen presenting cells), thus taking part in the immune network. Moreover, mast cells produce an exclusive family of proteases (tryptases and chymases) that exert many biological actions relevant to airways inflammation and remodeling. Future studies will better explain the role of mast cells in asthma and, more specifically, the links with bone marrow-where mast cell progenitors originate-and the airways, where mast cells develop, differentiate, and assume the functions of mature cells. This article reviews recent data available on these topics.


Magnetic resonance imaging structured reporting in infertility.
Our objective was to define and propose a standardized magnetic resonance (MR) imaging structured report in patients with infertility to have clinical completeness on possible diagnosis and severity. Patients should be studied preferable on 3T equipment with a surface coil. Standard MR protocol should include high-resolution fast spin-echo T2-weighted, diffusion-weighted images and gradient-echo T1-weighted fat suppression images. The report should include ovaries (polycystic, endometrioma, tumor), oviduct (hydrosalpinx, hematosalpinx, pyosalpinx, peritubal anomalies), uterus (agenesia, hypoplasia, unicornuate, uterus didelphys, bicornuate, septate uterus), myometrium (leiomyomas, adenomyosis), endometrium (polyps, synechia, atrophy, neoplasia), cervix and vagina (isthmoceles, mucosal-parietal irregularity, stenosis, neoplasia), peritoneum (deep endometriosis), and urinary system-associated abnormalities. To be clinically useful, radiology reports must be structured, use standardized terminology, and convey actionable information. The structured report must comprise complete, comprehensive, and accurate information, allowing radiologists to continuously interact with patients and referring physicians to confirm that the information is used properly to affect the decision making process.


Revealing the mechanisms underlying embolic stroke using computational modelling.
Computational forecasting of arterial blockages in a virtual patient has the potential to provide the next generation of advanced clinical monitoring aids for stroke prevention. As a first step towards a physiologically realistic virtual patient, we have created a computer model investigating the effects of emboli (particles or gas bubbles) as they become lodged in the brain. Our model provides a framework for predicting the severity of microvascular obstruction by simulating fundamental interactions between emboli and the fractal geometry of the arterial tree through which they travel. The model vasculature consisted of a bifurcating fractal tree comprising over a million branches ranging between 1 mm and 12 microm in diameter. Motion of emboli through the tree was investigated using a Monte Carlo simulation to evaluate the effects of the embolus size, clearance time and embolization rate on the number and persistence of blocked arterioles. Our simulations reveal with striking clarity that the relationship between embolus properties and vascular obstruction is nonlinear. We observe a rapid change between free-flowing and severely blocked arteries at specific combinations of the embolus size and embolization rate. The model predicts distinct patterns of cerebral injury for solid and gaseous emboli which are consistent with clinical observations. Solid emboli are predicted to be responsible for focal persistent injuries, while fast-clearing gas emboli produce diffuse transient blockages similar to global hypoperfusion. The impact of solid emboli was found to be dramatically reduced by embolus fragmentation. Computer simulations of embolization provide a novel means of investigating the role of emboli in producing neurological injury and assessing effective strategies for stroke prevention.


Levels of expressed emotion and relapse in depressed patients.
The relationship between spouses' levels of expressed emotion (EE) and relapse was examined in a sample of 39 depressed psychiatric hospital patients. Over a nine-month follow-up, 59% of patients with high-EE spouses relapsed, although no patients living with low-EE spouses did so. The results replicate the association found by Vaughn & Leff between EE and depression, and suggest that the EE construct has predictive validity in both schizophrenic and depressed populations.


A systematic review of mindfulness intervention for individuals with developmental disabilities: long-term practice and long lasting effects.
Can individuals with developmental disabilities learn mindfulness? If so, with what result? A systematic literature review identified 12 studies that taught mindfulness practice to individuals with mild to severe developmental disabilities, demonstrating that mindfulness intervention could significantly reduce the behavioural and/or psychological problems of this population. The majority of these mindfulness intervention studies were longitudinal, featuring long intervention periods and long lasting intervention effects. This paper analyses the characteristics and objectives of mindfulness interventions, along with their effects, focusing on the adjustments made to intervention content and instruction strategies to meet the specific requirements of individuals with developmental disabilities. The potential for improving mindfulness interventions for people with developmental disabilities is also discussed.


A meta-analysis of the impacts of internal migration on child health outcomes in China.
According to China's 2010 population census, 38.81 million children migrated from rural to urban areas in Mainland China, a phenomenon that has attracted much scholarly attention. Due to the lack of quantitative synthesis of migrant children's developmental outcomes, we undertook a meta-analysis to compare their developmental outcomes with those of their urban counterparts. We searched Applied Social Sciences Index and Abstracts (ASSIA), Australian Education Index, British Education Index, ERIC, ProQuest Education Journals, PsycINFO, Social Services Abstracts, Family & Society Studies Worldwide, Medline, Women's Studies International databases and the Chinese CNKI database to identify relevant studies. Studies reporting physical and mental health outcomes of migrant children as well as potential protective and risk factors of child developmental outcomes were included. We assessed study quality using a quality assessment checklist. We selected 25 studies from a total of 1592. Our results reveal that migrant children in public schools present significantly greater mental health problems and lower well-being than their urban counterparts, while migrant children in migrant schools do not present significantly different outcomes. In addition, migrant children were found to be more likely to be exposed to physical health risks due to limited utilization of health services. The disadvantageous health outcomes of migrant children were found to be related to a series of individual and social factors, including academic performance, social relationships, and discrimination. Migrant children are disadvantaged by the sociocultural circumstances in urban areas. Government should target them and provide appropriate support in order to improve their developmental status, which will have a positive impact on the stability and development of society.


Neurophysiology of Skin Thermal Sensations.
Undoubtedly, adjusting our thermoregulatory behavior represents the most effective mechanism to maintain thermal homeostasis and ensure survival in the diverse thermal environments that we face on this planet. Remarkably, our thermal behavior is entirely dependent on the ability to detect variations in our internal (i.e., body) and external environment, via sensing changes in skin temperature and wetness. In the past 30 years, we have seen a significant expansion of our understanding of the molecular, neuroanatomical, and neurophysiological mechanisms that allow humans to sense temperature and humidity. The discovery of temperature-activated ion channels which gate the generation of action potentials in thermosensitive neurons, along with the characterization of the spino-thalamo-cortical thermosensory pathway, and the development of neural models for the perception of skin wetness, are only some of the recent advances which have provided incredible insights on how biophysical changes in skin temperature and wetness are transduced into those neural signals which constitute the physiological substrate of skin thermal and wetness sensations. Understanding how afferent thermal inputs are integrated and how these contribute to behavioral and autonomic thermoregulatory responses under normal brain function is critical to determine how these mechanisms are disrupted in those neurological conditions, which see the concurrent presence of afferent thermosensory abnormalities and efferent thermoregulatory dysfunctions. Furthermore, advancing the knowledge on skin thermal and wetness sensations is crucial to support the development of neuroprosthetics. In light of the aforementioned text, this review will focus on the peripheral and central neurophysiological mechanisms underpinning skin thermal and wetness sensations in humans. © 2016 American Physiological Society. Compr Physiol 6:1279-1294, 2016.2016.


Spillover effects of a community-managed marine reserve.
The value of no-take marine reserves as fisheries-management tools is controversial, particularly in high-poverty areas where human populations depend heavily on fish as a source of protein. Spillover, the net export of adult fish, is one mechanism by which no-take marine reserves may have a positive influence on adjacent fisheries. Spillover can contribute to poverty alleviation, although its effect is modulated by the number of fishermen and fishing intensity. In this study, we quantify the effects of a community-managed marine reserve in a high poverty area of Northern Mozambique. For this purpose, underwater visual censuses of reef fish were undertaken at three different times: 3 years before (2003), at the time of establishment (2006) and 6 years after the marine reserve establishment (2012). The survey locations were chosen inside, outside and on the border of the marine reserve. Benthic cover composition was quantified at the same sites in 2006 and 2012. After the reserve establishment, fish sizes were also estimated. Regression tree models show that the distance from the border and the time after reserve establishment were the variables with the strongest effect on fish abundance. The extent and direction of the spillover depends on trophic group and fish size. Poisson Generalized Linear Models show that, prior to the reserve establishment, the survey sites did not differ but, after 6 years, the abundance of all fish inside the reserve has increased and caused spillover of herbivorous fish. Spillover was detected 1 km beyond the limit of the reserve for small herbivorous fishes. Six years after the establishment of a community-managed reserve, the fish assemblages have changed dramatically inside the reserve, and spillover is benefitting fish assemblages outside the reserve.


A novel rat orthologue and homologue for the Drosophila crooked neck gene in neural stem cells and their immediate descendants.
The crooked neck (crn) gene of Drosophila melanogaster encodes a scaffold protein carrying multiple tetratricopeptide repeat (TPR) motifs, and its mutation results in a reduction in the number of neuroblasts and lethality during larval stages. Here, we isolated two structurally related genes from a rat embryonic brain cDNA library. One gene is the rat orthologue of crn, which encodes 690 amino acids including 16 copies of TPR. The other gene, ATH55, encodes an 855 amino acid protein including 21 TPR motifs, which presumably represents a rat crn homologue and an orthologue of human XAB2. Both genes are highly expressed in embryonic brain but their expressions decrease during development. ATH55-like immunoreactivity is present in the ventricular zone and newly formed cortical plate, while CRN-like immunoreactivity is more abundant in a younger ventricular zone. In agreement, both proteins were found to be enriched in cultured neural stem cells and to decrease in response to cell differentiation signals. As indicated for the yeast CRN-like protein, ATH55 and CRN immunoreactivities were both recovered in the nuclear fraction and detected in the splicing complex carrying pre-mRNA. These findings suggest that both TPR-motif-containing proteins are involved in RNA processing of mammalian neural stem cells and their immediate descendants.


Reaction of single-stranded DNA with hydroxyl radical generated by iron(II)-ethylenediaminetetraacetic acid.
This study demonstrates that the reaction of Fe(II)-EDTA and hydrogen peroxide with the single-stranded nucleic acids d(pT)70 and a 29-base sequence containing a mixture of bases results in substantial damage which is not directly detected by gel electrophoresis. Cleavage of the DNA sugar backbone is enhanced significantly after the samples are incubated at 90 degrees C in the presence of piperidine. The latter reaction is used in traditional Maxam-Gilbert DNA sequencing to detect base damage, and the current results are consistent with reaction of the hydroxyl radical with the bases in single-stranded DNA (although reaction with sugar may also produce adducts that are uncleaved but labile to cleavage by piperidine). We propose that hydroxyl radicals may react preferentially with the nucleic acid bases in ssDNA and that reaction of the sugars in dsDNA is dominant because the bases are sequestered within the double helix. These results have implications both for the study of single-stranded DNA binding protein binding sites and for the interpretation of experiments using the hydroxyl radical to probe DNA structure or to footprint double-stranded DNA binding protein binding sites.


Molecular characterisation of the faecal microbiota in patients with type II diabetes.
The investigation provides molecular analyses of the faecal microbiota in type 2 diabetic patients. In order to characterise the gut microbiota in diabetic patients and to assess whether there are changes in the diversity and similarity of gut microbiota in diabetic patients when compared with healthy individuals, bacterial DNAs from 16 type 2 diabetic patients and 12 healthy individuals were extracted from faecal samples and characterised by PCR-denaturing gradient gel electrophoresis (DGGE) with primers specifically targeting V3 region of the 16S rRNA gene, as well as been sequenced for excised gel bands. The counts of Bacteroides vulgatus, Clostridium leptum subgroup and Bifidobacterium genus were assessed using quantitative PCR. By comparing species diversity profiles of two groups, we observed that there were no significant differences between diabetic and healthy group, although a few diabetic individuals (D6, D8) exhibited a remarkable decrease in species profiles. As for the similarity index, it was lower in inter-group than that in intra-group, which showed that the composition of gut microbiota in diabetic group might be changed due to diabetes status. Sequencing results also revealed that bacterial composition of diabetic group was different from that of the healthy group. B. vulgatus and Bifidobacterium genus were low represented in the microbiota of diabetic group, and the significant decrease was observed for Bifidobacterium by real-time PCR. Taken together, in this work we observed the characterisation of gut microbiota in diabetic patients, which suggests that the gut microbiota of diabetes patients have some changes associated with occurrence and development of diabetes.