Evidence for widespread cryptic sexual generations in apparently purely asexual Andricus gallwasps.
Oak gallwasps (Hymenoptera, Cynipidae, Cynipini) are one of seven major animal taxa that commonly reproduce by cyclical parthenogenesis (CP). A major question in research on CP taxa is the frequency with which lineages lose their sexual generations, and diversify as purely asexual radiations. Most oak gallwasp species are only known from an asexual generation, and secondary loss of sex has been conclusively demonstrated in several species, particularly members of the holarctic genus Andricus. This raises the possibility of widespread secondary loss of sex in the Cynipini, and of diversification within purely parthenogenetic lineages. We use two approaches based on analyses of allele frequency data to test for cryptic sexual generations in eight apparently asexual European species distributed through a major western palaearctic lineage of the gallwasp genus Andricus. All species showing adequate levels of polymorphism (7/8) showed signatures of sex compatible with cyclical parthenogenesis. We also use DNA sequence data to test the hypothesis that ignorance of these sexual generations (despite extensive study on this group) results from failure to discriminate among known but morphologically indistinguishable sexual generations. This hypothesis is supported: 35 sequences attributed by leading cynipid taxonomists to a single sexual adult morphospecies, Andricus burgundus, were found to represent the sexual generations of at least six Andricus species. We confirm cryptic sexual generations in a total of 11 Andricus species, suggesting that secondary loss of sex is rare in Andricus.


Imbalance of circulating lymphoid cells in Q fever endocarditis.
Q fever endocarditis is characterized by a defective cell-mediated immune response, which may be associated with the dysregulation of circulating subsets of immune cells. In this study, we found that naïve CD8(+) T lymphocytes and CD56dim natural killer cells were decreased patients whereas central memory CD8(+) T lymphocytes were increased. It is likely that these different subsets of immune cells play a role in the immunosuppression accompanying Q fever endocarditis.


Signature of recent historical events in the European Y-chromosomal STR haplotype distribution.
Previous studies of human Y-chromosomal single-nucleotide polymorphisms (Y-SNPs) established a link between the extant Y-SNP haplogroup distribution and the prehistoric demography of Europe. By contrast, our analysis of seven rapidly evolving Y-chromosomal short tandem repeat loci (Y-STRs) in over 12,700 samples from 91 different locations in Europe reveals a signature of more recent historic events, not previously detected by other genetic markers. Cluster analysis based upon molecular variance yields two clearly identifiable sub-clusters of Western and Eastern European Y-STR haplotypes, and a diverse transition zone in central Europe, where haplotype spectra change more rapidly with longitude than with latitude. This and other observed patterns of Y-STR similarity may plausibly be related to particular historical incidents, including, for example, the expansion of the Franconian and Ottoman Empires. We conclude that Y-STRs may be capable of resolving male genealogies to an unparalleled degree and could therefore provide a useful means to study local population structure and recent demographic history.


Hydrogen peroxide thermochemical oscillator as driver for primordial RNA replication.
This paper presents and tests a previously unrecognized mechanism for driving a replicating molecular system on the prebiotic earth. It is proposed that cell-free RNA replication in the primordial soup may have been driven by self-sustained oscillatory thermochemical reactions. To test this hypothesis, a well-characterized hydrogen peroxide oscillator was chosen as the driver and complementary RNA strands with known association and melting kinetics were used as the substrate. An open flow system model for the self-consistent, coupled evolution of the temperature and concentrations in a simple autocatalytic scheme is solved numerically, and it is shown that thermochemical cycling drives replication of the RNA strands. For the (justifiably realistic) values of parameters chosen for the simulated example system, the mean amount of replicant produced at steady state is 6.56 times the input amount, given a constant supply of substrate species. The spontaneous onset of sustained thermochemical oscillations via slowly drifting parameters is demonstrated, and a scheme is given for prebiotic production of complementary RNA strands on rock surfaces.


Alaska Mountain Wilderness Ski Classic: Alterations in Caloric Expenditure and Body Composition.
The Alaska Mountain Wilderness Ski Classic is a self-supported ultramarathon cross-country skiing event that traverses one of the mountain ranges of Alaska each winter. Unique aspects of this event challenge athletes with a significant amount of physical and mental stress while in the chronically cold conditions of the Arctic. Assessment of energy requirements or body composition has never been performed during this event. The objective of the study was to evaluate the influence of the 2016 Alaska Mountain Wilderness Ski Classic on caloric expenditure and body composition. Caloric expenditure was estimated using GT3x+ Actigraph accelerometers and ActiLife software. Lean tissue mass, total fat mass, visceral fat mass, and bone mineral density were measured using a General Electric iDXA before and after the event. Data are presented as mean±SD. Differences were analyzed using paired t tests with significance at P<0.05. Fifteen participants (age=32.7±6.6 years, body mass index=23.7±2.3 kg·m2) completed the study, 8 males and 7 females. Caloric expenditure was 6238±1390 kcal·day-1, and 37,163±8425 kcal for the entire event. Fat mass was reduced from pre-event (∆1.3±0.7 kg, P=0.00003). There was an increase in lean tissue mass (∆1.7±1.3 kg, P=0.0003) and relative skeletal muscle index (∆0.2±0.2 kg·m², P=0.001). There was no change in bone mineral density (∆0±0 g·cm3, P=0.4). Despite the metabolic demands, fat mass was reduced and lean tissue mass was preserved. Future studies of this event should be directed toward assessment of the nutritional provisions utilized and the mechanisms responsible for the preservation of lean tissue mass.


Should STEMI Patients Receive Opiate Analgesia? The Morphine Paradox.
The very significant benefit of P2Y12 receptor inhibitor administration in patients with ST-elevation myocardial infarction (STEMI), in reducing future ischaemic events and stent thrombosis, is undisputed. Morphine analgesia is very frequently co-administered to these patients for pain relief, along with antiplatelet therapy, at the time of presentation, and prior to reperfusion with primary percutaneous coronary intervention. Research and online content related to opiates use in STEMI was reviewed. Bibliographies of retrieved studies were searched manually for additional studies and reviews. There is sufficient data from pharmacokinetic and pharmacodynamic studies showing that the co-administration of morphine with oral P2Y12 receptor inhibitor results in delayed antiplatelet effects. However, whether this results in adverse outcomes remains unclear. Data from studies reporting the effect of morphine on clinical outcomes in STEMI are inconsistent, although they tend to be underpowered to show an effect on hard clinical outcomes, but some clearly show a relationship between morphine use and infarct size. Strategies to overcome the potentially significant negative impact of morphine on platelet reactivity in STEMI are discussed. Whilst clearly definitive, adequately powered, randomised controlled trials are lacking, we would recommend avoiding the combination of morphine with oral P2Y12 receptor inhibitors and recommend alternative strategies including intravenous platelet inhibitor strategies, in high risk patients.


Some aspects of multiple-beam interference techniques in digital data recording.
Some parameters related to the photographic recording of digital data by means of superposed diffraction gratings have been studied. The efficiencies of composite patterns formed by coherent multiple-beam interference depend strongly on the average exposure, beam balance ratios, spatial frequencies employed, and whether the exposures are made simultaneously or sequentially. Simultaneous exposures of N recording beams yield first-order outputs that are more intense by the factor N than the outputs of the same number of sequential exposures, other conditions being equal. The usable readout signal strength is limited primarily by the appearance of false images caused by nonlinear film transfer characteristics.


Postural stability of older female Scottish country dancers in comparison with physically active controls.
Physical activity assists older individuals' functional ability and postural stability. Recently, Scottish country dance (SCD) was reported as being a beneficial form of physical activity for functional ability in older females. This study aims to examine the effect of SCD on postural stability. Scottish country dancers (n = 20) were compared with physically active controls (n = 33) for static postural sway measured on a force platform. The Romberg and Tandem stances were used under 'eyes open' and 'eyes closed' conditions. Ninety-five percent ellipse area and sway velocity were calculated from the center of pressure displacement. Ninety-five percent ellipse area was the same for both groups in all tests. The control group had greater sway velocity for all tests (P < .01) except Tandem eyes closed. SCD participation resulted in similar postural sway as participation in other physical activities, however nondancers may need a greater amount of regulatory activity to maintain balance.


Elevated pulmonary artery systolic storage volume associated with improved ventilation-to-perfusion ratios in acute respiratory failure.
The possibility that an elevated pulmonary artery systolic storage volume (PASSV) correlates with improved overall ratios of ventilation-to-perfusion and hence benefits gas exchange in acute respiratory failure was examined. We examined this by assessing the correlation between PASSV and both the physiologic dead space to tidal volume ratio (VD/VT) and intrapulmonary shunt fraction (Qsp/Qt). The VD/VT and Qsp/Qt were used as an index of distribution of ventilation-to-perfusion as well as efficiency of pulmonary gas exchange. Twenty-eight patients suffering from acute respiratory failure were included. All required mechanical ventilation. PASSV was calculated from the pulmonary artery (PA) compliance and mean PA systolic distending pressure. Pulmonary arteriolar pressures were computed by Fourier analysis. PA compliance was derived from the PA time constant and the PA resistance. Storage volume fraction of stroke volume index (PASSV/SVI) was used to compare individual variations. There were inverse linear relationships between PASSV/SVI and VD/VT (r = 0.693, p less than 0.0001), and between PASSV/SVI and Qsp/Qt (r = 0.427, p = 0.012). Also, a direct correlation was found between VD/VT and PA time constant (r = 0.503, p = 0.002). The patients were divided into two groups based on PASSV/SVI to evaluate the effect of other hemodynamic data on PASSV. Comparison of the two groups revealed that VD/VT and Qsp/Qt were lower (p less than 0.0001, and p = 0.018, respectively), PA time constant was higher (p less than 0.001), and right ventricular stroke-work index was higher (p = 0.005) in the group with a high PASSV/SVI. There were no differences in other hemodynamic data between the two groups. These data suggest that an elevated PASSV may indeed benefit gas exchange in acute respiratory failure.


Clinician agreement on physical findings in child sexual abuse cases.
To measure agreement among experienced clinicians regarding the interpretation of physical findings in child sexual abuse cases and to determine whether knowledge of clinical history affects the interpretation of the physical findings. Experienced clinicians rated colposcopic photographs on a scale of 1 to 5 with 1 being normal and 5 being clear evidence of penetrating injury. To answer an additional study question of whether clinical history affected interpretation, 4 clinicians rated 69 cases in which they were blinded to the patients' histories and 70 cases in which the patients' histories were available. The other 3 clinicians then rated the same cases with the presence or absence of history reversed. All clinicians involved perform child sexual abuse examinations at tertiary care centers. A total of 139 girls with Tanner stage 1 or 2 genitalia who were referred to a general pediatric clinic at an academic medical center for examination of possible sexual abuse. Half of the photographs were interpreted as indicating little or no evidence of abuse. Of those photographic sets that both readers could interpret, 39% were in perfect agreement and 77% disagreed by 1 category or less. Perfect agreement across all possible pairs of readers was 34.5%. Agreement was better when the patient's clinical history was unknown (29.3% vs 38.9%, P = .005). The kappa, a measure of interrater reliability, indicated poor agreement among clinicians. The combined kappa for the first group of clinicians was 0.22 without knowledge of clinical history and 0.11 with knowledge of clinical history. For the second group of clinicians, the kappa was 0.31 without knowledge of clinical history and 0.15 with knowledge of clinical history. The overall kappa across all 7 clinicians disregarding clinical history was 0.20. Agreement was best for categories 1 (normal, kappa = 0.28) and 5 (clear evidence of a penetrating injury, kappa = 0.39). Clinicians educated and experienced in assessing child sexual abuse do not agree perfectly on the interpretation of photographs of genital findings in girls with Tanner stage 1 or 2 genitalia. Clinicians agree less when a patient's clinical history is available. Efforts should be directed at standardizing physical findings and avoiding overemphasis on physical findings in child sexual abuse cases.


Following tracks of hemichannels.
It has been suggested that plasma membrane-bound hemichannels perform physiological and pathophysiological functions per se. Such functions require the presence of hemichannels on the cell surface and their accessibility to the extracellular environment for at least some limited period of time. We have previously shown that hemichannels can be labeled by means of antibodies directed to an external loop domain of connexin (Cx) 43. We now provide evidence that trafficking of hemichannel vesicles can be visualized upon binding of a labeled homophilic peptide corresponding to a region of the first extracellular loop (EL1) of Cx43. In vivo imaging was performed after labeling hemichannels from the extracellular site with a mimetic peptide tagged with a fluorochrome (Alexa-546). Using a Cx43-CFP transfected HeLa cell line for incubation with the mimetic peptide, a significant number of double-labeled vesicles were found inside the cells. This double labeling indicates that a portion of Cx43 within the cell had accessed the cell surface as hemichannels where it bound to the peptide and was subsequently endocytosed. Pulse labeling with the peptide showed a decrease in the number of dual-labeled vesicles over time, indicating degradation and/or concurrent recycling of hemichannel vesicles.


Non-exponential resistive switching in Ag2S memristors: a key to nanometer-scale non-volatile memory devices.
The dynamics of resistive switchings in nanometer-scale metallic junctions formed between an inert metallic tip and an Ag film covered by a thin Ag2S layer are investigated. Our thorough experimental analysis and numerical simulations revealed that the resistance change upon a switching bias voltage pulse exhibits a strongly non-exponential behaviour yielding markedly different response times at different bias levels. Our results demonstrate the merits of Ag2S nanojunctions as nanometer-scale non-volatile memory cells with stable switching ratios, high endurance as well as fast response to write/erase, and an outstanding stability against read operations at technologically optimal bias and current levels.


Gaussian-beam profile shaping by acousto-optic Bragg diffraction.
We study how Gaussian laser-beam profiles can be modified into a desired form using acousto-optic Bragg diffraction. By exploiting the angular dependence of Bragg diffraction of plane waves by acoustic gratings, we demonstrate that the conversion from a Gaussian-profile beam into either a near-field or a far-field flattop profile is possible.


Zika virus infection disrupts neurovascular development and results in postnatal microcephaly with brain damage.
Zika virus (ZIKV) infection of pregnant women can result in fetal brain abnormalities. It has been established that ZIKV disrupts neural progenitor cells (NPCs) and leads to embryonic microcephaly. However, the fate of other cell types in the developing brain and their contributions to ZIKV-associated brain abnormalities remain largely unknown. Using intracerebral inoculation of embryonic mouse brains, we found that ZIKV infection leads to postnatal growth restriction including microcephaly. In addition to cell cycle arrest and apoptosis of NPCs, ZIKV infection causes massive neuronal death and axonal rarefaction, which phenocopy fetal brain abnormalities in humans. Importantly, ZIKV infection leads to abnormal vascular density and diameter in the developing brain, resulting in a leaky blood-brain barrier (BBB). Massive neuronal death and BBB leakage indicate brain damage, which is further supported by extensive microglial activation and astrogliosis in virally infected brains. Global gene analyses reveal dysregulation of genes associated with immune responses in virus-infected brains. Thus, our data suggest that ZIKV triggers a strong immune response and disrupts neurovascular development, resulting in postnatal microcephaly with extensive brain damage.


Attitudes towards the male contraceptive pill in men and women in casual and stable sexual relationships.
Men's and women's attitudes towards the male contraceptive pill and their trust in the effective use of the male pill were investigated, as well as the associated variables of reported health behaviours, perceived self-efficacy and type of sexual relationship, using a questionnaire survey. Although both sexes had a favourable attitude towards the male pill, females had a more positive attitude than men. Conversely, women had less trust that men would use the male pill effectively. Males in stable sexual relationships were more positive about the male pill than those in casual sexual relationships. Gender, relationship type and trust in the effective use of the male pill reliably predicted attitude towards the male pill. High perceived self-efficacy was related to engaging in more health behaviours, and in men a positive association between health behaviours and attitude towards the male pill has been found. A positive attitude towards the male pill does not automatically imply that the individual is confident about its effective use. Once the male pill is widely available, promotional campaigns could target not only men but also their female partners, as the latter tend to come into contact with health services more frequently. In order to increase confidence in effective implementation, a variety of presentations of the male pill should be made available in line with individual needs and lifestyles.


Structural and biochemical studies on procaspase-8: new insights on initiator caspase activation.
Caspases are proteases with an active-site cysteine and aspartate specificity in their substrates. They are involved in apoptotic cell death and inflammation, and dysfunction of these enzymes is directly linked to a variety of diseases. Caspase-8 initiates an apoptotic pathway triggered by external stimuli. It was previously characterized in its active inhibitor bound state by crystallography. Here we present the solution structure of the monomeric unprocessed catalytic domain of the caspase-8 zymogen, procaspase-8, showing for the first time the position of the linker and flexibility of the active site forming loops. Biophysical studies of carefully designed mutants allowed disentangling dimerization and processing, and we could demonstrate lack of activity of monomeric uncleaved procaspase-8 and of a processed but dimerization-incompetent mutant. The data provide experimental support in so-far unprecedented detail, and reveal why caspase-8 (and most likely other initiator caspases) needs the dimerization platform during activation.


Bone demarcation of the temporomandibular joint. Validity of clinical assessment of bone thickness by means of CT.
To study the CT depiction of bone demarcations in the temporomandibular joint, using conventional window level and window width; and to evaluate observer performance in estimating bone thickness in these images. Seven joint specimens were imaged by CT and then cryosectioned. The measurements of bone wall thickness in the images were compared to the true bone thickness at each cutting level. In addition, 4 experienced radiologists estimated the thickness of the bone walls in the images. The relative difference between the CT reproduction and the true bone thickness was small for bone walls thicker than 2 mm. This difference increased with the decrease in bone thickness and the increase in the inclination of the bone wall from the perpendicular to the image plane. Bone walls thinner than 1 mm were reproduced as considerably thicker than their true thickness. This resulted in a clinical overestimation of bone thickness. Both the CT representation and the interpretation of bone demarcation in the temporomandibular joint may constitute a problem. Partial volume averaging effects can result in an overestimation of bone dimensions amounting to 200% for thin bones. The central white zone in images of thin bone walls obtained with the parameters described here could serve as an indicator that could help to reduce the risk of overestimating bone thickness.


Treatment Outcomes for Patients with Failed Back Surgery.
Failed back surgery syndrome (FBSS) is a frequently encountered disease entity following lumbar spinal surgery. Although many plausible reasons have been investigated, the exact pathophysiology remains unknown. Various medications, reoperations, interventions such as spinal cord stimulation, epidural adhesiolysis or epidural injection, exercise therapy, and psychotherapy have been suggested treatment options. However, the evidence of the clinical outcome for each treatment has not been clearly determined. To evaluate the outcomes of each treatment modality and to present treatment guidelines for patients with FBSS. A systematic review of each treatment regimen in patients with FBSS. The available literature regarding each modality for the treatment of refractory back pain or radiating pain for FBSS was reviewed. The quality assessment and the level of evidence were analyzed using the "Methodology Checklist" of SIGN (Scottish Intercollegiate Guidelines Network). Data sources included relevant English language literature identified through searches of Pubmed, EMBASE, and Cochrane library from 1980 to Feb 2016. The primary outcome measure was pain relief of back pain or radiating pain for at least 3 months. Secondary outcome measures were improvement of the patient's functional status, health-related quality of life, return to work, and reduction of opioid use. Twenty-three articles were finally identified and reviewed. Based on our analysis, epidural adhesiolysis showed a short-term (6 to 24 months) effect (grade A) and spinal cord stimulation showed a mid-term (2 or 3 years) effect (grade B). Epidural injections showed a short-term (up to 2 years) effect (grade C). However, other treatments were recommended as grade D or inconclusive. The limitations of this systematic review included the rarity of relevant literature. Epidural adhesiolysis or spinal cord stimulation can be effective in order to control chronic back pain or leg pain due to FBSS, and its recommendation grades are A and B, respectively. Other treatments showed poor or inconclusive evidence.Key words: Failed back surgery syndrome, post spinal surgery syndrome, chronic low back pain, post lumbar surgery syndrome, epidural adhesiolysis, spinal cord stimulation, epidural injection, revision.


Unusual life style of giant chlorella viruses.
Paramecium bursaria chlorella virus (PBCV-1) is the prototype of a family of large, icosahedral, plaque-forming, dsDNA viruses that replicate in certain unicellular, eukaryotic chlorella-like green algae. Its 330-kb genome contains approximately 373 protein-encoding genes and 11 tRNA genes. The predicted gene products of approximately 50% of these genes resemble proteins of known function, including many that are unexpected for a virus, e.g., ornithine decarboxylase, hyaluronan synthase, GDP-D-mannose 4,6 dehydratase, and a potassium ion channel protein. In addition to their large genome size, the chlorella viruses have other features that distinguish them from most viruses. These features include: (a) The viruses encode multiple DNA methyltransferases and DNA site-specific endonucleases. (b) The viruses encode at least some, if not all, of the enzymes required to glycosylate their proteins. (c) PBCV-1 has at least three types of introns, a self-splicing intron in a transcription factor-like gene, a spliceosomal processed intron in its DNA polymerase gene, and a small intron in one of its tRNA genes. (d) Many chlorella virus-encoded proteins are either the smallest or among the smallest proteins of their class. (e) Accumulating evidence indicates that the chlorella viruses have a very long evolutionary history.


Lessons learned in a European-Latin American collaboration for developing postgraduate education in public health.
New funding from the European Union (EU) allowed the collaborative development of postgraduate education in public health between five countries in South America and Europe. A planning meeting, two workshops, a final meeting and the use of e-mail and faxes for discussing curricula were employed. One new master of public health in Argentina has begun, plans for new masters in Chile have been developed. A new collaborative module using the Internet was started and is now in its third year, new European plans for distance learning are being developed. Possibilities for collaborative research have been identified and those involved developed a broader horizon and international contacts. The planned grant from the EU was 72,040 Euros. Staff time spent administering the project was much greater than time spent discussing curricula. This is the only publication. Although the aims were achieved, the opportunity costs were great, as methods of assessing European research do not value such work highly and the administrative burden of small EU grants is high. Once a network is established greater efforts should be put into electronic communication, but European programmes currently do not fund such 'remote' support.


Factor analytic mixed models for the provision of grower information from national crop variety testing programs.
Factor analytic mixed models for national crop variety testing programs have the potential to improve industry productivity through appropriate modelling and reporting to growers of variety by environment interaction. Crop variety testing programs are conducted in many countries world-wide. Within each program, data are combined across locations and seasons, and analysed in order to provide information to assist growers in choosing the best varieties for their conditions. Despite major advances in the statistical analysis of multi-environment trial data, such methodology has not been adopted within national variety testing programs. The most commonly used approach involves a variance component model that includes variety and environment main effects, and variety by environment (V × E) interaction effects. The variety predictions obtained from such an analysis, and subsequently reported to growers, are typically on a long-term regional basis. In Australia, the variance component model has been found to be inadequate in terms of modelling V × E interaction, and the reporting of information at a regional level often masks important local V × E interaction. In contrast, the factor analytic mixed model approach that is widely used in Australian plant breeding programs, has regularly been found to provide a parsimonious and informative model for V × E effects, and accurate predictions. In this paper we develop an approach for the analysis of crop variety evaluation data that is based on a factor analytic mixed model. The information obtained from such an analysis may well be superior, but will only enhance industry productivity if mechanisms exist for successful technology transfer. With this in mind, we offer a suggested reporting format that is user-friendly and contains far greater local information for individual growers than is currently the case.


Impaired NaV1.2 function and reduced cell surface expression in benign familial neonatal-infantile seizures.
Mutations in SCN2A, the gene encoding the brain voltage-gated sodium channel alpha-subunit Na(V)1.2, are associated with inherited epilepsies including benign familial neonatal-infantile seizures (BFNIS). Functional characterization of three BFNIS mutations was performed to identify defects in channel function that underlie this disease. We examined three BFNIS mutations (R1319Q, L1330F, and L1563V) using whole-cell patch-clamp recording of heterologously expressed human Na(V)1.2. Membrane biotinylation was employed to examine the cell surface protein expression of the four Na(V)1.2 alleles. R1319Q displayed mixed effects on activation and fast inactivation gating, consistent with a net loss of channel function. L1563V exhibited impaired fast inactivation predicting a net gain of channel function. The L1330F mutation significantly decreased overall channel availability during repetitive stimulation. Patch-clamp analysis also revealed that cells expressing BFNIS mutants exhibited lower levels of sodium current compared to wild type (WT) Na(V)1.2. Biochemical experiments demonstrated that all three BFNIS mutations exhibited a significant reduction in cell surface expression compared to WT. Our findings indicate that BFNIS is associated with a range of biophysical defects accompanied by reduced levels of channel protein at the plasma membrane.


Troglitazone increases the number of small adipocytes without the change of white adipose tissue mass in obese Zucker rats.
Troglitazone (CS-045) is one of the thiazolidinediones that activate the peroxisome proliferator-activated receptor gamma (PPARgamma), which is expressed primarily in adipose tissues. To elucidate the mechanism by which troglitazone relieves insulin resistance in vivo, we studied its effects on the white adipose tissues of an obese animal model (obese Zucker rat). Administration of troglitazone for 15 d normalized mild hyperglycemia and marked hyperinsulinemia in these rats. Plasma triglyceride level was decreased by troglitazone in both obese and lean rats. Troglitazone did not change the total weight of white adipose tissues but increased the number of small adipocytes (< 2,500 micron2) approximately fourfold in both retroperitoneal and subcutaneous adipose tissues of obese rats. It also decreased the number of large adipocytes (> 5,000 micron2) by approximately 50%. In fact, the percentage of apoptotic nuclei was approximately 2.5-fold higher in the troglitazone-treated retroperitoneal white adipose tissue than control. Concomitantly, troglitazone normalized the expression levels of TNF-alpha which were elevated by 2- and 1.4-fold in the retroperitoneal and mesenteric white adipose tissues of the obese rats, respectively. Troglitazone also caused a dramatic decrease in the expression levels of leptin, which were increased by 4-10-fold in the white adipose tissues of obese rats. These results suggest that the primary action of troglitazone may be to increase the number of small adipocytes in white adipose tissues, presumably via PPARgamma. The increased number of small adipocytes and the decreased number of large adipocytes in white adipose tissues of troglitazone-treated obese rats appear to be an important mechanism by which increased expression levels of TNF-alpha and higher levels of plasma lipids are normalized, leading to alleviation of insulin resistance.


Chiari malformation Type I surgery in pediatric patients. Part 1: validation of an ICD-9-CM code search algorithm.
OBJECTIVE Administrative billing data may facilitate large-scale assessments of treatment outcomes for pediatric Chiari malformation Type I (CM-I). Validated International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) code algorithms for identifying CM-I surgery are critical prerequisites for such studies but are currently only available for adults. The objective of this study was to validate two ICD-9-CM code algorithms using hospital billing data to identify pediatric patients undergoing CM-I decompression surgery. METHODS The authors retrospectively analyzed the validity of two ICD-9-CM code algorithms for identifying pediatric CM-I decompression surgery performed at 3 academic medical centers between 2001 and 2013. Algorithm 1 included any discharge diagnosis code of 348.4 (CM-I), as well as a procedure code of 01.24 (cranial decompression) or 03.09 (spinal decompression or laminectomy). Algorithm 2 restricted this group to the subset of patients with a primary discharge diagnosis of 348.4. The positive predictive value (PPV) and sensitivity of each algorithm were calculated. RESULTS Among 625 first-time admissions identified by Algorithm 1, the overall PPV for CM-I decompression was 92%. Among the 581 admissions identified by Algorithm 2, the PPV was 97%. The PPV for Algorithm 1 was lower in one center (84%) compared with the other centers (93%-94%), whereas the PPV of Algorithm 2 remained high (96%-98%) across all subgroups. The sensitivity of Algorithms 1 (91%) and 2 (89%) was very good and remained so across subgroups (82%-97%). CONCLUSIONS An ICD-9-CM algorithm requiring a primary diagnosis of CM-I has excellent PPV and very good sensitivity for identifying CM-I decompression surgery in pediatric patients. These results establish a basis for utilizing administrative billing data to assess pediatric CM-I treatment outcomes.


Awareness and use of electronic cigarettes and heat-not-burn tobacco products in Japan.
In addition to some electronic cigarettes (e-cigarettes), new heat-not-burn tobacco products Ploom and iQOS have recently begun to be sold by tobacco companies. These products are regulated differently in Japan, depending on whether the contents are liquid or tobacco leaf. Our objective was to estimate percentages of awareness and use of e-cigarettes and heat-not-burn tobacco products among the Japanese population, including minors. An internet survey (randomly sampled research agency panellists) with a propensity score adjustment for "being a respondent in an internet survey" using a nationally representative sample in Japan. A total of 8240 respondents aged 15-69 years in 2015 (4084 men and 4156 women). Adjusted percentages of awareness and use of e-cigarettes (nicotine or non-nicotine e-cigarettes) and heat-not-burn products among total participants; product types and percentages ever used among e-cigarettes ever users. Of respondents in Japan, 48% [95% confidence interval (CI) = 47-49] were aware of e-cigarettes and heat-not-burn tobacco products, 6.6% (95% CI = 6.1-7.1) had ever used, 1.3% (95% CI = 1.0-1.5) had used in the last 30 days and 1.3% (95% CI = 1.1-1.6) had experience of > 50 sessions. Seventy-two per cent (95% CI = 69-76) of ever users used non-nicotine e-cigarettes, while 33% (95% CI = 30-37) of them used nicotine e-cigarettes, which has the majority share world-wide; 7.8% (95% CI = 5.5-10.0) and 8.4% (95% CI = 6.1-10.7) of them used the new devices, Ploom and iQOS, respectively, with a relatively higher percentage among the younger population. Approximately half the respondents in a Japanese internet survey were aware of e-cigarettes and heat-not-burn tobacco products, 6.6% had ever used. More than 70% of ever users used non-nicotine e-cigarettes, the sale of which is not legally prohibited, even to minors, in Japan, and 33% of them used nicotine e-cigarettes; 3.5% of never smoking men and 1.3% of never smoking women had ever used e-cigarettes. Corresponding figures for use in the last 30 days were 0.6% and 0.3%, predominantly non-nicotine e-cigarettes.


Exposure to Bordetella pertussis adenylate cyclase toxin affects integrin-mediated adhesion and mechanics in alveolar epithelial cells.
The adenylate cyclase (CyaA) toxin is a major virulent factor of Bordetella pertussis, the causative agent of whooping cough. CyaA toxin is able to invade eukaryotic cells where it produces high levels of cyclic adenosine monophosphate (cAMP) affecting cellular physiology. Whether CyaA toxin can modulate cell matrix adhesion and mechanics of infected cells remains largely unknown. In this study, we use a recently proposed multiple bond force spectroscopy (MFS) with an atomic force microscope to assess the early phase of cell adhesion (maximal detachment and local rupture forces) and cell rigidity (Young's modulus) in alveolar epithelial cells (A549) for toxin exposure <1 h. At 30 min of exposure, CyaA toxin has a minimal effect on cell viability (>95%) at CyaA concentration of 0.5 nM, but a significant effect (≈81%) at 10 nM. MFS performed on A549 for three different concentrations (0.5, 5 and 10 nM) demonstrates that CyaA toxin significantly affects both cell adhesion (detachment forces are decreased) and cell mechanics (Young's modulus is increased). CyaA toxin (at 0.5 nM) assessed at three indentation/retraction speeds (2, 5 and 10 μm/s) significantly affects global detachment forces, local rupture events and Young modulus compared with control conditions, while an enzymatically inactive variant CyaAE5 has no effect. These results reveal the loading rate dependence of the multiple bonds newly formed between the cell and integrin-specific coated probe as well as the individual bond kinetics which are only slightly affected by the patho-physiological dose of CyaA toxin. Finally, theory of multiple bond force rupture enables us to deduce the bond number N which is reduced by a factor of 2 upon CyaA exposure (N ≈ 6 versus N ≈ 12 in control conditions). MFS measurements demonstrate that adhesion and mechanical properties of A549 are deeply affected by exposure to the CyaA toxin but not to an enzymatically inactive variant. This indicates that the alteration of cell mechanics triggered by CyaA is a consequence of the increase in intracellular cAMP in these target cells. These results suggest that mechanical and adhesion properties of the cells appear as pertinent markers of cytotoxicity of CyaA toxin.


Review of combinations of experimental and computational techniques to identify and understand genes involved in innate immunity and effector-triggered defence.
The innate immune system includes a first layer of defence that recognises conserved pathogen-associated molecular patterns that are essential for microbial fitness. Resistance (R) gene-based recognition of pathogen effectors, which function in modulation or avoidance of host immunity, activates a second layer of plant defence. In this review, experimental and computational techniques are considered to improve understanding of the plant immune system. Biocomputation contributes to discovery of the molecular genetic basis of host resistance against pathogens. Sequenced genomes have been used to identify R genes in plants. Resistance gene enrichment sequencing based on conserved protein domains has increased the number of R genes with nucleotide-binding site and leucine-rich repeat domains. Network analysis will contribute to an improved understanding of the innate immune system and identify novel genes for partial disease resistance. Machine learning algorithms are expected to become important in defining aspects of the immune system that are less well characterised, including identification of R genes that lack conserved protein domains.


Adaptation of EPEC-EM Curriculum in a Residency with Asynchronous Learning.
The Education in Palliative and End-of-life Care for Emergency Medicine Project (EPEC™-EM) is a comprehensive curriculum in palliative and end-of-life care for emergency providers. We assessed the adaptation of this course to an EM residency program using synchronous and asynchronous learning. Curriculum adaptation followed Kern's standardized six-step curriculum design process. Post-graduate year (PGY) 1-4 residents were taught all EPEC™-EM cognitive domains, divided as seven synchronous and seven asynchronous modules. All synchronous modules featured large group didactic lectures and review of EPEC™-EM course materials. Asynchronous modules use only EPEC™-EM electronic course media for resident self-study. Targeted evaluation for EPEC™-EM knowledge objectives was conducted by a prospective case-control crossover study, with synchronous learning serving as the quasi-control, using validated exam tools. We compared de-identified test scores for effectiveness of learning method, using aggregate group performance means for each learning strategy. Of 45 eligible residents 55% participated in a pre-test for local needs analysis, and 78% completed a post-test to measure teaching method effect. Post-test scores improved across all EPEC™-EM domains, with a mean improvement for synchronous modules of +28% (SD=9) and a mean improvement for asynchronous modules of +30% (SD=18). The aggregate mean difference between learning methods was 1.9% (95% CI -15.3, +19.0). Mean test scores of the residents who completed the post-test were: synchronous modules 77% (SD=12); asynchronous modules 83% (SD=13); all modules 80% (SD=12). EPEC™-EM adapted materials can improve resident knowledge of palliative medicine domains, as assessed through validated testing of course objectives. Synchronous and asynchronous learning methods appear to result in similar knowledge transfer, feasibly allowing some course content to be effectively delivered outside of large group lectures.


Histamine poisoning (scombroid fish poisoning): an allergy-like intoxication.
Histamine poisoning results from the consumption of foods, typically certain types of fish and cheeses, that contain unusually high levels of histamine. Spoiled fish of the families, Scombridae and Scomberesocidae (e.g. tuna, mackerel, bonito), are commonly implicated in incidents of histamine poisoning, which leads to the common usage of the term, "scombroid fish poisoning", to describe this illness. However, certain non-scombroid fish, most notably mahi-mahi, bluefish, and sardines, when spoiled are also commonly implicated in histamine poisoning. Also, on rare occasions, cheeses especially Swiss cheese, can be implicated in histamine poisoning. The symptoms of histamine poisoning generally resemble the symptoms encountered with IgE-mediated food allergies. The symptoms include nausea, vomiting, diarrhea, an oral burning sensation or peppery taste, hives, itching, red rash, and hypotension. The onset of the symptoms usually occurs within a few minutes after ingestion of the implicated food, and the duration of symptoms ranges from a few hours to 24 h. Antihistamines can be used effectively to treat this intoxication. Histamine is formed in foods by certain bacteria that are able to decarboxylate the amino acid, histidine. However, foods containing unusually high levels of histamine may not appear to be outwardly spoiled. Foods with histamine concentrations exceeding 50 mg per 100 g of food are generally considered to be hazardous. Histamine formation in fish can be prevented by proper handling and refrigerated storage while the control of histamine formation in cheese seems dependent on insuring that histamine-producing bacteria are not present in significant numbers in the raw milk.


Rheumatoid arthritis and dietary interventions: systematic review of clinical trials.
The impact of various dietary interventions on rheumatoid arthritis (RA), characterized by immune-inflammatory response, has been subject to increased attention. A systematic review was conducted to update the current knowledge on the effects of nutritional, dietary supplement, and fasting interventions on RA outcomes. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, with prespecification of all methods, Medline and Embase were systematically searched for relevant articles. Data were extracted by 2 independent reviewers. A total of 70 human studies were identified. Administration of omega-3 polyunsaturated fatty acids at high doses resulted in a reduction in RA disease activity and a lower failure rate of pharmacotherapy. Vitamin D supplementation and dietary sodium restriction were beneficial on some RA outcomes. Fasting resulted in significant but transient subjective improvements. While the Mediterranean diet demonstrated improvements in some RA disease activity measures, outcomes from vegetarian, elimination, peptide, or elemental diets suggested that responses are very individualized. Some dietary approaches may improve RA symptoms and thus it is recommended that nutrition should be routinely addressed.


Ethnic and gender differences regarding the insulin-blood pressure relationship.
Ageing is associated with increased insulin and C-peptide levels. Due to a lack of data, our first aim was to establish whether this also holds true for Africans from South Africa. Our second aim was to determine whether an association between insulin/C-peptide levels and blood pressure exist within an African and Caucasian population with increasing age, as well as to establish gender differences. African men and women (N=260) and Caucasian men and women (N=369) were recruited and stratified into age groups (18-35 years, 36-45 years and >45 years). ANCOVAs and partial correlations were performed. Results showed opposing changes in insulin/C-peptide levels of African and Caucasian men with increasing age. Insulin/C-peptide tended to decrease in African men, whereas insulin tended to increase and C-peptide increased significantly (p=0.03) in Caucasian men. Despite similar obesity levels, the oldest African women had significantly lower insulin (p<0.01) and C-peptide (p<0.01) levels compared to their Caucasian counterparts. In conclusion, insulin/C-peptide levels tended to decrease in the African population with increasing age. Despite significantly lower levels of insulin, blood pressure levels of African men seems to be affected more detrimentally compared to their Caucasian counterparts, leaving them more vulnerable for the development of cardiovascular diseases.


Role of biomass adaptation in the removal of formic acid in sequencing batch reactors.
This study deals with formic acid removal in activated sludge processes, in particular in the processes carried out in sequencing batch reactors (SBRs). Formic acid removal has been investigated in a SBR fed with acetic and formic acids at equimolar concentrations. Biomass performance in the reactor has been investigated both by the analysis of the removal of the two substrates and by batch tests. Regarding SBR process, the obtained results show that a relevant difference occurred between formic and acetic acid profiles. Acetic acid was never found in the effluent and was always completely removed during the reaction phase. On the other hand, formic acid removal was determined by biomass acclimation, which is in turn determined by sludge age imposed to the system. Batch tests confirmed that formic acid removal occurs only if biomass is acclimated. It has been shown that the minimal sludge age to obtain complete formic acid removal is much higher than those predictable with the classical models of microbial growth in wastewater treatment processes. The advantages of SBRs over continuous-flow systems in the removal of formic acid have also been highlighted.


Establishing the Canadian HIV Women's Sexual and Reproductive Health Cohort Study (CHIWOS): Operationalizing Community-based Research in a Large National Quantitative Study.
Community-based research has gained increasing recognition in health research over the last two decades. Such participatory research approaches are lauded for their ability to anchor research in lived experiences, ensuring cultural appropriateness, accessing local knowledge, reaching marginalized communities, building capacity, and facilitating research-to-action. While having these positive attributes, the community-based health research literature is predominantly composed of small projects, using qualitative methods, and set within geographically limited communities. Its use in larger health studies, including clinical trials and cohorts, is limited. We present the Canadian HIV Women's Sexual and Reproductive Health Cohort Study (CHIWOS), a large-scale, multi-site, national, longitudinal quantitative study that has operationalized community-based research in all steps of the research process. Successes, challenges and further considerations are offered. Through the integration of community-based research principles, we have been successful in: facilitating a two-year long formative phase for this study; developing a novel survey instrument with national involvement; training 39 Peer Research Associates (PRAs); offering ongoing comprehensive support to PRAs; and engaging in an ongoing iterative community-based research process. Our community-based research approach within CHIWOS demanded that we be cognizant of challenges managing a large national team, inherent power imbalances and challenges with communication, compensation and volunteering considerations, and extensive delays in institutional processes. It is important to consider the iterative nature of community-based research and to work through tensions that emerge given the diverse perspectives of numerous team members. Community-based research, as an approach to large-scale quantitative health research projects, is an increasingly viable methodological option. Community-based research has several advantages that go hand-in-hand with its obstacles. We offer guidance on implementing this approach, such that the process can be better planned and result in success.


Long noncoding RNA LISPR1 is required for S1P signaling and endothelial cell function.
Sphingosine-1-Phosphate (S1P) is a potent signaling lipid. The effects of S1P are mediated by the five S1P receptors (S1PR). In the endothelium S1PR1 is the predominant receptor and thus S1PR1 abundance limits S1P signaling. Recently, lncRNAs were identified as a novel class of molecules regulating gene expression. Interestingly, the lncRNA NONHSAT004848 (LISPR1, Long intergenic noncoding RNA antisense to S1PR1), is closely positioned to the S1P1 receptors gene and in part shares its promoter region. We hypothesize that LISPR1 controls endothelial S1PR1 expression and thus S1P-induced signaling in endothelial cells. In vitro transcription and translation as well as coding potential assessment showed that LISPR1 is indeed noncoding. LISPR1 was localized in both cytoplasm and nucleus and harbored a PolyA tail at the 3'end. In human umbilical vein endothelial cells, as well as human lung tissue, qRT-PCR and RNA-Seq revealed high expression of LISPR1. S1PR1 and LISPR1 were downregulated in human pulmonary diseases such as COPD. LISPR1 but also S1PR1 were induced by inflammation, shear stress and statins. Knockdown of LISPR1 attenuated endothelial S1P-induced migration and spheroid outgrowth of endothelial cells. LISPR1 knockdown decreased S1PR1 expression, which was paralleled by an increase of the binding of the transcriptional repressor ZNF354C to the S1PR1 promoter and a reduction of the recruitment of RNA Polymerase II to the S1PR1 5'end. This resulted in attenuated S1PR1 expression and attenuated S1P downstream signaling. Collectively, the disease relevant lncRNA LISPR1 acts as a novel regulatory unit important for S1PR1 expression and endothelial cell function.


The combined roles of ADAMTS13 and VWF in murine models of TTP, endotoxemia, and thrombosis.
Ultralarge von Willebrand factor (UL-VWF) multimers are thought to play a central role in pathogenesis of the disease thrombotic thrombocytopenic purpura (TTP); however, experimental evidence in support of this hypothesis has been difficult to establish. Therefore, to examine directly the requirement for VWF in TTP pathogenesis, we generated ADAMTS13-deficient mice on a TTP-susceptible genetic background that were also either haploinsufficient (Vwf+/-) or completely deficient (Vwf-/-) in VWF. Absence of VWF resulted in complete protection from shigatoxin (Stx)-induced thrombocytopenia, demonstrating an absolute requirement for VWF in this model (Stx has been shown previously to trigger TTP in ADAMTS13-deficient mice). We next investigated the requirements for ADAMTS13 and VWF in a murine model of endotoxemia. Unlike Stx-induced TTP findings, LPS-induced thrombocytopenia and mortality were not affected by either VWF or ADAMTS13 deficiency, suggesting divergent mechanisms of thrombocytopenia between these 2 disorders. Finally, we show that VWF deficiency abrogates the ADAMTS13-deficient prothrombotic state, suggesting VWF as the only relevant ADAMTS13 substrate under these conditions. Together, these findings shed new light on the potential roles played by ADAMTS13 and VWF in TTP, endotoxemia, and normal hemostasis.


Comparison between smartphone pedometer applications and traditional pedometers for improving physical activity and body mass index in community-dwelling older adults.
[Purpose] The effectiveness of a smartphone pedometer application was compared with that of a traditional pedometer for improving the physical activity and weight status of community-dwelling older adults. [Subjects and Methods] This study had a nonequivalent pretest-posttest control group design. Ninety-seven older adults (mean age ± SD, 60.1 ± 5.5 years) joined the smartphone pedometer group and underwent a 2-week walking intervention based on a smartphone pedometer application. Fifty-four older adults (mean age ± SD, 65.3 ± 8.7 years) joined the traditional pedometer group and underwent a 2-week walking intervention based on a traditional pedometer. The participants' physical activity was evaluated using the International Physical Activity Questionnaire-Short Form, and their weight status was quantified by calculating the body mass index. The daily pedometer count was also documented. [Results] No significant time, group, or time-by-group interaction effects were found for any of the outcome variables. However, trends of improvement in physical activity and body mass index were seen only in the smartphone pedometer group. [Conclusion] A smartphone pedometer application might be more favorable than a traditional pedometer in improving physical activity and body mass index in community-dwelling older adults. However, further experimental studies are necessary to confirm the results.


Rodent Models of Spondyloarthritis Have Decreased White and Bone Marrow Adipose Tissue Depots.
Bone marrow adipose tissue (BMAT) has recently been recognized as a distinct fat depot with endocrine functions. However, if and how it is regulated by chronic inflammation remains unknown. Here, we investigate the amount of white fat and BMAT in HLA-B27 transgenic rats and curdlan-challenged SKG mice, two well-established models of chronic inflammatory spondyloarthritis (SpA). Subcutaneous and gonadal white adipose tissue and BMAT was reduced by 65-70% and by up to 90% in both experimental models. Consistently, B27 rats had a 2-3-fold decrease in the serum concentrations of the adipocyte-derived cytokines adiponectin and leptin as well as a 2-fold lower concentration of triglycerides. The bone marrow of B27 rats was further characterized by higher numbers of neutrophils, lower numbers of erythroblast precursors, and higher numbers of IL-17 producing CD4+ T cells. IL-17 concentration was also increased in the serum of B27 rats. Using a cell culture model, we show that high levels of IL-17 in the serum of B27 rats negatively impacted adipogenesis (-76%), an effect that was reversed in the presence of neutralizing anti-IL-17 antibody. In summary, these findings show BMAT is severely reduced in two experimental models of chronic inflammatory SpA and suggest that IL-17 is involved in this process.


Chinese immigrant men smokers' sources of cigarettes in Canada: A qualitative study.
Immigrants often experience economic hardship in their host country and tend to belong to economically disadvantaged groups. Individuals of lower socioeconomic status tend to be more sensitive to cigarette price changes. This study explores the cigarette purchasing patterns among Chinese Canadian male immigrants. Semi-structured in-depth interviews were conducted with 22 Chinese Canadian immigrants who were smoking or had quit smoking in the last five years. Because of financial pressures experienced by participants, the high price of Canadian cigarettes posed a significant challenge to their continued smoking. While some immigrants bought fully-taxed cigarettes from licensed retailers, more often they sought low-cost cigarettes from a variety of sources. The two most important sources were cigarettes imported during travels to China and online purchases of Chinese cigarettes. The cigarettes obtained through online transactions were imported by smoking or non-smoking Chinese immigrants and visitors, suggesting the Chinese community were involved or complicit in sustaining this form of purchasing behavior. Other less common sources included Canada-USA cross border purchasing, roll your-own pouch tobacco, and buying cigarettes available on First Nations reserves. Chinese Canadian immigrant men used various means to obtain cheap cigarettes. Future research studies could explore more detailed features of access to expose gaps in policy and improve tobacco regulatory frameworks.


Polyhydroxyalkanoic acids from structurally-unrelated carbon sources in Escherichia coli.
Polyhydroxyalkanoates (PHAs) that contain varied monomers with different chain lengths/structures were normally synthesized when a structurally-related precursor was present. The biosynthesis of PHAs from unrelated carbon sources in microorganisms including Escherichia coli met many challenges in the past. Recently, with the development of metabolic engineering and synthetic biology, the production of PHAs from unrelated carbon sources obtained a breakthrough. Polyesters containing 2-hydroxypropionate, 3-hydroxypropionate, 4-hydroxybutyrate, 3-hydroxyvalarate, and medium-chain-length 3-hydroxyalkanoate monomers can all be synthesized in E. coli by integrating exogenous or endogenous pathways and/or genes. This review will summarize the progresses in this area. In addition, the strategies that lead to the production of PHAs with varied monomers and high polymer content in the cell are discussed.


Mathematical modelling of hepatitis C treatment for injecting drug users.
Hepatitis C virus (HCV) is a blood-borne infection that can lead to progressive liver failure, cirrhosis, hepatocellular carcinoma and death. In developed countries, the majority of HCV infections are transmitted via injecting drug users (IDUs). Despite effective antiviral treatment for HCV, very few active IDUs are treated. Reluctance to treat is partially due to the risk of reinfection. We develop a mathematical model of HCV transmission amongst active IDUs, and examine the potential effect of antiviral treatment. As most mathematical models of interventions utilise a treatment function proportional to the infected population, but many policy implementations set fixed yearly targets for specific numbers treated, we study the effects of using two different treatment terms: annually treating a proportion of infecteds or a fixed number of infecteds. We examine the behaviour of the two treatment models and find different bifurcation behaviours in each case. We calculate analytical solutions for the treatment level needed for disease clearance or control, and observe that achievable levels of treatment can result in control or eradication across a wide range of prevalence levels. Finally, we calculate the sensitivity of the critical treatment threshold to the model parameters, and find that for a given observed prevalence, the injecting duration and infection risk play the most important role in determining the treatment level needed. By contrast, the sensitivity analysis indicates the presence (or absence) of immunity does not alter the treatment threshold. We conclude by discussing the public health implications of this work, and comment on the importance and feasibility of utilising treatment as prevention for HCV spread amongst IDUs.


Outcomes After Listing for Liver Transplant in Patients With Acute-on-Chronic Liver Failure: The Multicenter North American Consortium for the Study of End-Stage Liver Disease Experience.
Acute-on-chronic liver failure (ACLF) characterized with ≥2 extrahepatic organ failures in cirrhosis carries a high mortality. Outcomes of patients listed for liver transplantation (LT) after ACLF and after LT are largely unknown. The North American Consortium for the Study of End-Stage Liver Disease prospectively enrolled 2793 nonelectively hospitalized patients with cirrhosis; 768 were listed for LT. Within 3 months, 265 (35%) received a LT, 395 remained alive without LT, and 108 died/delisted. Compared with nonlisted patients, those listed were younger and more often had ACLF, acute kidney injury, and a higher admission Model for End-Stage Liver Disease (MELD) score. ACLF was most common in patients who died/delisted, followed by those alive with and without LT respectively, (30%, 22%, and 7%, respectively; P < 0.001). At LT, median MELD was 27.9% and 70% were inpatients; median time from hospitalization to LT was 26 days. Post-LT survival at 6 months was unchanged between those with and without ACLF (93% each at 6 months). There was no difference in 3- and 6-month mean post-LT creatinine in those with and without ACLF, despite those with ACLF having a higher mean pre-LT creatinine and a higher rate of perioperative dialysis (61%). In conclusion, patients with and without ACLF had similar survival after transplant with excellent renal recovery in both groups.


Acute stent thrombosis and/or atrial fibrillation occurring after bee sting.
Acute stent thrombosis may result from many causes. Bee sting is a very rare cause of this situation. As bee venom activates some inflammatory and vascular events resulting in occlusion in vessels or diseases in electrical system of heart, it may cause myocardial infarction or atrial fibrillation. This problem may be temporary or fatal. Because of its pathologic effects, to be able to catch on and treat them shortly is very important.


Ascorbate and Cu(II)-induced oxidative degradation of high-molar-mass hyaluronan. Pro- and antioxidative effects of some thiols.
This study presents the results of antioxidative and pro-oxidative efficacy of cysteamine and D-penicillamine (D-pen) in comparison to L-glutathione (L-GSH) on high-molar-mass hyaluronan (HA) degradation by cupric ions plus ascorbic acid. The substance tested was applied in the degradative system cupric ions plus ascorbate: (i) before the reaction onset or also (ii) 1 h after the reaction started. The results obtained were compared with that one recorded by using the degradative system in the absence of the substance tested. To monitor HA degradation kinetics, rotational viscometry was applied. Moreover, the standard ABTS and DPPH assays were used. By using the method of rotational viscometry, D-pen showed dual effect: initial inhibitory effect on •OH radicals was changed to a pro-oxidative one in the dose and time dependent manner. Both L-GSH and cysteamine were recorded to be more effective scavengers of •OH radicals than D-pen. Cysteamine demonstrated to be an excellent scavenger also of alkoxyl- and peroxyl- type radicals. Based on IC50 values, gained by ABTS assay, it is evident that D-pen showed higher radical scavenging capacity compared to cysteamine. Similar results were observed also in DPPH assay, although in this assay less effective radical scavenging capacities of both substances tested were recorded. On the basis of the results obtained, it can be stated that D-pen can produce hydrogen peroxide or •OH radicals and can inhibit the production of these oxidants. Our results showed that both L-GSH and cysteamine are similarly effective in inhibiting of HA degradation. Moreover, cysteamine demonstrated to be a significant inhibitor of alkoxyl- and peroxyl- type radicals generated from C-type macroradical of HA.


Risk Factors for Local Recurrence after Emergency Resection for Colon Cancer: Scenario in Sweden.
Patients undergoing emergency resection for colon cancer have a worse outcome both in terms of short- and long-term survival than those having elective surgery. The aim of this population-based study was to determine factors associated with increased risk for local recurrence following emergency resection. The Stockholm-Gotland Healthcare Region Colon Cancer Register was used to identify all colon cancer patients who had undergone emergency colon resection with curative intent in that region 1997-2007. Patient records were scrutinised to obtain any missing information. The influence of the following factors was assessed: indication for emergency resection; time between admission and surgery; surgery daytime or at night; American Association of Anesthesiologists score; volume of blood lost; and T- and N-stage. Our endpoint was loco-regional recurrence. Apart from stage, perforation as indication for emergency surgery was the only factor that influenced the risk for local recurrence (hazard ratio 1.96; 95% CI 1.12-3.43). In this study, the only factor associated with local recurrence after emergency resection for colon cancer was preoperative perforation. This implies that changes in our current management algorithm would be unlikely to lead to improvement. Efforts should therefore concentrate on reducing the proportion of patients operated on an emergency basis.


Nitrogen supply differentially affects litter decomposition rates and nitrogen dynamics of sub-arctic bog species.
High-latitude peatlands are important soil carbon sinks. In these ecosystems, the mineralization of carbon and nitrogen are constrained by low temperatures and low nutrient concentrations in plant litter and soil organic matter. Global warming is predicted to increase soil N availability for plants at high-latitude sites. We applied N fertilizer as an experimental analogue for this increase. In a three-year field experiment we studied N fertilization effects on leaf litter decomposition and N dynamics of the four dominant plant species (comprising >75% of total aboveground biomass) in a sub-arctic bog in northern Sweden. The species were Empetrum nigrum (evergreen shrub), Eriophorum vaginatum (graminoid), Betula nana (deciduous shrub) and Rubus chamaemorus (perennial forb). In the controls, litter mass loss rates increased in the order: Empetrum<Eriophorum<Betula<Rubus. Increased N availability had variable, species-specific effects: litter mass loss rates (expressed per unit litter mass) increased in Empetrum, did not change in Eriophorum and Betula and decreased in Rubus. In the leaf litter from the controls, we measured no or only slight net N mineralization even after three years. In the N-fertilized treatments we found strong net N immobilization, especially in Eriophorum and Betula. This suggests that, probably owing to substantial chemical and/or microbial immobilization, additional N supply does not increase the rate of N cycling for at least the first three years.


A comparative study of the elastic fibre system within the mouse and human cornea.
The cornea relies on its organised extracellular matrix for maintaining transparency and biomechanical strength. Studies have identified an elastic fibre system within the human posterior cornea, thought to allow for slight deformations in response to internal pressure fluctuations within the eye. However, the type of elastic fibres that exist within the cornea and their roles remain elusive. The aim of this study was to compare the distribution and organisation of the elastic fibres within the posterior peripheral mouse and human cornea, and elucidate how these fibres integrate with the trabecular meshwork, whilst characterising the distribution of their main likely components (fibrillin-1, elastin and type VI collagen) in different parts of the cornea and adjacent sclera. We identified key differences in the elastic fibre system between the human and mouse cornea. True elastic fibres (containing elastin) were identified within the human posterior peripheral cornea. Elastic fibres appeared to present as an extensive network throughout the mouse corneal stroma, but as fibrillin-rich microfibril bundles rather than true elastic fibres. However, tropoelastin staining indicated the possibility that true elastic fibres had yet to develop in the young mice studied. Differences were also apparent within the anatomy of the trabecular meshwork. The human trabecular meshwork appeared to insert between the corneal stroma and Descemet's membrane, with elastic fibres continuing into the stroma from the trabecular meshwork anterior to Descemet's membrane. Within the mouse cornea, no clear insertion point of the trabecular meshwork was seen, instead the elastic fibres within the trabecular meshwork continued into Descemet's membrane, with the trabecular meshwork joining posterior to Descemet's membrane.


Mapping QTL for white striping in relation to breast muscle yield and meat quality traits in broiler chickens.
White striping (WS) is an emerging muscular defect occurring on breast and thigh muscles of broiler chickens. It is characterized by the presence of white striations parallel to the muscle fibers and has significant consequences for meat quality. The etiology of WS remains poorly understood, even if previous studies demonstrated that the defect prevalence is related to broiler growth and muscle development. Moreover, recent studies showed moderate to high heritability values of WS, which emphasized the role of genetics in the expression of the muscle defect. The aim of this study was to identify the first quantitative trait loci (QTLs) for WS as well as breast muscle yield (BMY) and meat quality traits using a genome-wide association study (GWAS). We took advantage of two divergent lines of chickens selected for meat quality through Pectoralis major ultimate pH (pHu) and which exhibit the muscular defect. An expression QTL (eQTL) detection was further performed for some candidate genes, either suggested by GWAS analysis or based on their biological function. Forty-two single nucleotide polymorphisms (SNPs) associated with WS and other meat quality traits were identified. They defined 18 QTL regions located on 13 chromosomes. These results supported a polygenic inheritance of the studied traits and highlighted a few pleiotropic regions. A set of 16 positional and/or functional candidate genes was designed for further eQTL detection. A total of 132 SNPs were associated with molecular phenotypes and defined 21 eQTL regions located on 16 chromosomes. Interestingly, several co-localizations between QTL and eQTL regions were observed which could suggest causative genes and gene networks involved in the variability of meat quality traits and BMY. The QTL mapping carried out in the current study for WS did not support the existence of a major gene, but rather suggested a polygenic inheritance of the defect and of other studied meat quality traits. We identified several candidate genes involved in muscle metabolism and structure and in muscular dystrophies. The eQTL analyses showed that they were part of molecular networks associated with WS and meat quality phenotypes and suggested a few putative causative genes.


Two-wavelength wrinkling patterns in helicoidal plywood surfaces: imprinting energy landscapes onto geometric landscapes.
We present a model to investigate the formation of two-length scale surface patterns in biological and synthetic anisotropic soft matter materials through the high order interaction of anisotropic interfacial tension and capillarity at their free surfaces. The unique pattern-formation mechanism emerging from the presented model is based on the interaction between lower and higher order anchoring modes. Analytical and numerical solutions are used to shed light on why and how simple anisotropic anchoring generates two-lengthscale wrinkles whose amplitudes are given in terms of anchoring coefficients. The novel finding is that the surface energy landscape with its maxima and minima can be imprinted onto the surface geometric landscape. Symmetry relations and scaling laws are used to provide the explicit relations between the anchoring constants and surface profile of the two length scale wrinkles. These new findings establish a new paradigm for characterizing surface wrinkling in biological liquid crystals, and inspire the design of novel functional surface structures.


Examining thiosulfate-driven autotrophic denitrification through respirometry.
Anoxic respirometry was applied to characterize a sulfide-oxidizing nitrate-reducing (SO-NR) culture obtained from an anoxic biogas desulfurizing biotrickling filter treating high loads of H2S. Immobilized biomass extracted from the biotrickling filter was grown in a suspended culture with thiosulfate as electron donor to obtain the biomass growth yield and the S2O3(2)(-)/NO3(-) consumed ratio. Afterward, respirometry was applied to describe thiosulfate oxidation under anoxic conditions. A pure culture of Thiobacillus denitrificans was also used as a control culture in order to validate the procedure proposed in this work to characterize the SO-NR biomass. Respirometric profiles obtained with this microbial culture showed that nitrite was formed as intermediate during nitrate reduction and revealed that no competitive inhibition appeared when both electron acceptors were present in the medium. Although final bioreaction products depended on the initial S2O3(2)(-)/NO3(-) ratio, such ratio did not affect thiosulfate oxidation or denitrification rates. Moreover, respirometric profiles showed that the specific nitrite uptake rate depended on the biomass characteristics being that of a SO-NR mixed culture (39.8mgNg(-1) VSSh(-1)) higher than that obtained from a pure culture of T. denitrificans (19.7mgNg(-1) VSSh(-1)). For the first time, the stoichiometry of the two-step denitrification mechanism with thiosulfate oxidation and biomass growth associated was solved for both reactions.


Functional diversity of human protein kinase splice variants marks significant expansion of human kinome.
Protein kinases are involved in diverse spectrum of cellular processes. Availability of draft version of the human genomic data in the year 2001 enabled recognition of repertoire of protein kinases. However, over the years the human genomic data is being refined and the current release of human genomic data has helped us to recognize a larger repertoire of over 900 human protein kinases represented mainly by splice variants. Many of these identified protein kinases are alternatively spliced products. Interestingly, some of the human kinase splice variants appear to be significantly diverged in terms of their functional properties as represented by incorporation or absence of one or more domains. Many sets of protein kinase splice variants have substantially different domain organization and in a few sets of splice variants kinase domains belong to different subfamilies of kinases suggesting potential participation in different signal transduction pathways. Addition or deletion of a domain between splice variants of multi-domain kinases appears to be a means of generating differences in the functional features of otherwise similar kinases. It is intriguing that marked sequence diversity within the catalytic regions of some of the splice variant kinases result in kinases belonging to different subfamilies. These human kinase splice variants with different functions might contribute to diversity of eukaryotic cellular signaling.


First Report of Alfalfa mosaic virus Occurrence in Hydrangea in the United States.
In spring of 2012, a previously unrecorded virus-like disease characterized by conspicuous yellow leaf blotching (calico symptoms) was observed in plants of Hydrangea macrophylla in a single location in Southampton, NY. Bacilliform and spherical particles resembling those of Alfalfa mosaic virus (AMV) were observed by transmission electron microscopy (TEM) in partially purified extracts from symptomatic leaf tissue. The identity of the virus was confirmed by immunosorbent electron microscopy (ISEM) (4) using antiserum to AMV (ATCC PVAS 92) that both trapped and decorated the virions. Three primer pairs designed from available AMV RNA 1, RNA 2, and RNA 3 genomic sequences were used to generate amplicons from the hydrangea AMV isolate. Reverse-transcription (RT)-PCR was done using total RNA extracted from symptomatic hydrangea leaf tissue with a Qiagen RNeasy kit, and Ready-to-Go RT-PCR beads (GE Healthcare). Amplicons of 1,049, 1,013, and 658 bp were obtained using the primer pairs AMV1F (5'-ATCCACCGATGCCAGCCTTA)/AMV1R (5'-TTCCGCCTCACTGCTGTCTG), AMV2F (5'-GATCGCCGGAAGTGATCCAG)/AMV2R (5'-TCACCGGAAGCAACAACGAA), and AMV3F (5'-GCCGGTTCTCCAAAGGGTCT)/AMV3R (5'-CGCGTCGAAGTCCAGACAGA), respectively. The PCR products were cloned using a TOPO TA cloning kit (Invitrogen) and three clones of each were sequenced. The sequences obtained from the hydrangea AMV RNA 1 (JX154090), RNA 2 (JX154091), and RNA 3 (JX154092) had 95 to 98% nucleotide sequence identity to homologous genomic sequences of known AMV isolates. To our knowledge, this is the first report of AMV occurrence in H. macrophylla in the United States. This virus has been reported to occur in H. macrophylla in British Columbia (3), but in a previous survey its presence was not detected in hydrangeas in the United States (1). A report of possible AMV infection in H. macrophylla in Italy (2) was based solely on symptomatology and cross-protection tests and therefore cannot be verified. The AMV-infected hydrangea plants were found by ISEM to also contain low concentrations of Hydrangea ringspot virus (HRSV) and Hydrangea chlorotic mottle virus (HdCMV). However, based on previous evidence of single and mixed infections (3), it is unlikely that the calico symptoms observed were influenced by the presence of HRSV and HdCMV. This report is of interest both because AMV, unlike HRSV and HdCMV, causes foliar symptoms that would render hydrangea plant unmarketable, and because the disease can be spread by a number of common aphid species that transmit AMV. It will also serve to alert growers and diagnosticians to the potential threat posed by AMV infection. References: (1) T. C. Allen et al. Acta Hortic. 164:85, 1985. (2) G. Belli. Phytopathol. Mediterr. 7:70, 1968. (3) A. W. Chiko and S. E. Godkin. Plant Dis. 70:541, 1986. (4) B. E. L. Lockhart et al. Phytopathology 82:691, 1992.


Correlation of sex steroids with IGF-1 and IGFBP-3 during different pubertal stages.
Insulin-like growth factor 1 (IGF-1) is the major factor that affects linear bone growth. Also, androgens and estrogens are necessary for increasing longitudinal bone growth during sexual maturation. The aim of this study was to investigate the relationships among IGF-1 axis and sex steroids during pubertal development in healthy adolescents. In this cross-sectional study, IGF-1, IGF binding protein-3 (IGFBP-3) and sex steroid levels (estradiol in girls, testosterone in boys) of 205 healthy adolescents (101 female, 104 male) aged 9-17 years were measured. All subjects were apparently healthy, with no growth retardation and with skeletal ages appropriate for chronological ages, and none were taking medications known to influence calcium homeostasis. Greulich and Pyle's Radiographic Atlas of Skeletal Development of the Hand and Wrist was used for determination of skeletal ages. Tanner's classification was used to determine the pubertal developmental stage. Fasting blood samples were obtained from subjects between 09:00-10:00 h. Serum IGF-1 and IGFBP-3 levels differed significantly between pubertal developmental stages. Serum IGF-1 levels and IGF-1/IGFBP-3 ratios increased with proceeding stages and maximum mean values were found at stages III-IV in girls and at stage IV in boys. Estradiol levels of girls and testosterone levels of boys differed significantly between stages, and in both sexes, serum IGF-1 levels and IGF-1/IGFBP-3 ratios were significantly correlated with sex steroid levels. Increase in growth hormone secretion increases IGF-1 levels. Furthermore, increasing sex steroids with pubertal development increase the IGF-1 levels and IGF-1/IGFBP-3 ratios that affect bone growth.


The receptor-dependent actions of 1,25-dihydroxyvitamin D are required for normal growth plate maturation in NPt2a knockout mice.
Rickets is a growth plate abnormality observed in growing animals and humans. Rachitic expansion of the hypertrophic chondrocyte layer of the growth plate, in the setting of hypophosphatemia, is due to impaired apoptosis of these cells. Rickets is observed in humans and mice with X-linked hypophosphatemia that is associated with renal phosphate wasting secondary to elevated levels of fibroblast growth factor-23. Rickets is also seen in settings of impaired vitamin D action, due to elevated PTH levels that increase renal phosphate excretion. However, mice with hypophosphatemia secondary to ablation of the renal sodium-dependent phosphate transport protein 2a (Npt2a), have not been reported to develop rickets. Because activation of the mitochondrial apoptotic pathway by phosphate is required for hypertrophic chondrocyte apoptosis in vivo, investigations were undertaken to address this paradox. Analyses of the Npt2a null growth plate demonstrate expansion of the hypertrophic chondrocyte layer at 2 wk of age, with resolution of this abnormality by 5 wk of age. This is temporally associated with an increase in circulating levels of 1,25-dihydroxyvitamin D. To address whether the receptor-dependent actions of this steroid hormone are required for normalization of the growth plate phenotype, the Npt2a null mice were mated with mice lacking the vitamin D receptor or were rendered vitamin D deficient. These studies demonstrate that the receptor-dependent actions of 1,25-dihydroxyvitamin D are required for maintenance of a normal growth plate phenotype in the Npt2a null mice.


First Report of Hop stunt viroid in Greenhouse Cucumber in Finland.
In July 2009, the occurrence of pale yellow, bottle-shaped greenhouse cucumber (Cucumis sativus L.) fruits was reported by a horticultural adviser from Kannus in western Finland. The grower had observed the first symptoms in greenhouse cucumber cv. Rapides in May. The most distinctive symptoms were found in fruits, but also flowers were crumpled. Symptoms had spread along plant rows. Estimated yield loss by the grower was 2 to 3%. Fruit and flower symptoms were typical of cucumber pale fruit disease (3) caused by a strain of Hop stunt viroid (HSVd) (2). The original samples were collected by a phytosanitary inspector and the farmer from approximately 10 symptomatic plants growing in the same greenhouse. Testing two samples, one a cucumber leaf and the other a cucumber fruit, by return-polyacrylamide gel electrophoresis gave clear electrophoretic bands at the same position. However, the position of the bands differed slightly from the positive control (Potato spindle tuber viroid) and two other pospiviroids tested on the same gel, indicating the presence of a different viroid with a shorter length than those from the genus Pospiviroid. This observation of size combined with symptoms on cucumber gave a strong indication of the presence of a viroid, likely to be HSVd. RNA was extracted from two subsamples of fruit and leaf samples with a RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Extracted RNA was examined with primer pair HSVdF1/R1 designed to detect HSVd from citrus (1). The PCR reactions were performed using a reverse transcription-PCR protocol developed for Pospiviroids (4). Both fruit and leaf samples gave a PCR amplicon of the same size. Sequencing of the amplicon of approximately 300 bp revealed 99% similarity with 11 GenBank citrus isolates of HSVd and 98% similarity with two cucumber isolates of HSVd (Accession Nos. X07405 and X00524) On the basis of these results, the viroid of these cucumber plants was identified as HSVd. To our knowledge, this is the first finding of this viroid in Finland. Although we could determine the causal agent, we could not find the origin of the infection. The seedling plants had been grown in the same greenhouse where the infection was detected. Even though HSVd is not known to be seed transmitted (3) the other production places that had used the same seed lot were inspected and found to be free of the viroid. Very strict phytosanitary measures were taken to eradicate the infection. Since the viroid is easily sap transmissible, there is a certain risk of spreading of HSVd via human action, e.g., visitors, staff, and the use of common packing facilities. References: (1) L. Bernard and N. Duran-Vila. Mol. Cell. Probes 20:105, 2006. (2) T. Sano et al. Nucleic Acids Res. 12:3427, 1984. (3) H. J. M. van Dorst and D. Peters. Neth. J. Plant Pathol. 80:85, 1974. (4) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004.


A Novel Murine Model of Radiation Keratopathy.
Radiation therapy results in severe chronic keratopathy and dry eye disease. We developed a novel mouse model for radiation keratopathy to allow future mechanistic studies. Six to 8-week-old BALB/c mice underwent sublethal irradiation to the head only from a Cesium-137 irradiator, 2 × 550 rad, 3-hours apart. Irradiated mice were clinically evaluated by corneal fluorescein staining (CFS) at 1, 2, and 3 months, after which corneas were excised and immunofluorescence histochemistry performed with anti-CD45, anti-MHC class II, and anti-β-tubulin antibodies. The survival rate after irradiation was 100%. Mice demonstrated significant CFS and hair loss around the eyes. Corneal nerve density decreased in the central and peripheral corneas (P < 0.01) at 2 and 3 months, respectively. CD45+ immune cell densities increased in the central and peripheral corneas (P < 0.005, P < 0.001) at 2 and 3 months, respectively. MHC class II, a sign of antigen presenting cell activation, significantly increased after irradiation in the central and peripheral corneas at 2 and 3 months (P = 0.02). A strong inverse correlation was noted between decreased corneal nerves and increase in CD45+ cells in the central cornea at 2 (P = 0.04, r = -0.89) and 3 months (P = 0.03, r = -0.91) after irradiation. We present a model of radiation keratopathy and demonstrate significant nerve loss and increase in immune cell influx and activation within months. This model will enable future investigations to understand the effects of radiation therapy on the eye, and to study mechanisms of neuro-immune crosstalk in the cornea.


Protocol for a systematic review of venous coupler devices versus hand-sewn anastomosis for microsurgical free flap reconstruction.
A patent microvascular anastomosis is of paramount importance in free tissue transfer. Anastomotic coupler devices provide an alternative to technically demanding hand-sewn venous anastomosis. Various advantages of these devices have been discussed but previous systematic reviews had methodological flaws or did not perform a meta-analysis. This review aims to evaluate the quality of the evidence and quantify the efficacy and safety of venous couplers compared to hand-sewn anastomosis. A PRISMA-compliant systematic review and meta-analysis will be performed. A comprehensive search strategy has been developed and will be applied to the databases MEDLINE and Embase from inception to October 2018. All clinical studies using anastomotic coupler devices for venous anastomoses in free tissue transfer will be eligible for inclusion. Screening of studies and data extraction will be performed independently by two authors. Our primary outcome is anastomotic venous thrombosis. Secondary outcomes will include time to complete the venous anastomosis, tearing of veins, anastomotic leakage, flap loss/failure and fiscal outcomes. The risk of bias for included studies will be assessed by using the ROBINS-I tool, and recommendations based on the evidence will be made using the GRADE approach. Descriptive statistical analyses will be used and if two or more studies report the same outcome, data will be pooled for comparative analysis. A direct comparison meta-analysis will be performed if possible. There has been no comparison of coupled and hand-sewn venous anastomoses using a robust and validated methodology preceded by a protocol and performing meta-analysis. Included studies are expected to be mainly observational and prone to bias; however, there is value in summarising the evidence, assessing its risk of bias and performing meta-analysis to guide clinicians. By using a broad approach including all types of flaps, we foresee inherent differences regarding the unit of analysis and different anatomic sites. This will limit the validity of our conclusions but is unavoidable. We will seek unpublished data from authors and perform subgroup analysis where appropriate. Limitations and areas of uncertainty will be discussed to guide future research. PROSPERO CRD42018110111.


Understanding the cellular function of TRPV2 channel through generation of specific monoclonal antibodies.
Transient receptor potential vanilloid 2 (TRPV2) is a Ca(2+)-permeable nonselective cation channel proposed to play a critical role in a wide array of cellular processes. Although TRPV2 surface expression was originally determined to be sensitive to growth factor signaling, regulated trafficking of TRPV2 has remained controversial. TRPV2 has proven difficult to study due to the lack of specific pharmacological tools to modulate channel activity; therefore, most studies of the cellular function of TRPV2 rely on immuno-detection techniques. Polyclonal antibodies against TRPV2 have not been properly validated and characterized, which may contribute to conflicting results regarding its function in the cell. Here, we developed monoclonal antibodies using full-length TRPV2 as an antigen. Extensive characterization of these antibodies and comparison to commonly used commercially available TRPV2 antibodies revealed that while monoclonal antibodies generated in our laboratory were suitable for detection of endogenous TRPV2 by western blot, immunoprecipitation and immunocytochemistry, the commercially available polyclonal antibodies we tested were not able to recognize endogenous TRPV2. We used our newly generated and validated TRPV2 antibodies to determine the effects of insulin-like growth factor 1 (IGF-1) on TRPV2 surface expression in heterologous and endogenous expression systems. We found that IGF-1 had little to no effect on trafficking and plasma membrane expression of TRPV2. Overall, these new TRPV2 monoclonal antibodies served to dispel the controversy of the effects of IGF-1 on TRPV2 plasma membrane expression and will clarify the role TRPV2 plays in cellular function. Furthermore, our strategy of using full-length tetrameric TRP channels may allow for the generation of antibodies against other TRP channels of unclear function.


Transmission of electronic substituent effects across the 1,12-dicarba-closo-dodecaborane cage: a computational study based on structural variation, atomic charges, and ¹³C NMR chemical shifts.
The ability of the 1,12-dicarba-closo-dodecaborane cage to transmit long-range substituent effects has been investigated by analyzing the structural variation of a phenyl probe bonded to C1, as caused by a remote substituent X at C12. The geometries of 41 Ph-CB10H10C-X molecules, including 11 charged species, have been determined by MO calculations at the B3LYP/6-311++G** level of theory. The structural variation of the phenyl probe is best represented by a linear combination of the internal ring angles, termed SF(CARB). Multiple regression analysis of SF(CARB), using appropriate explanatory variables, reveals the presence of resonance effects, superimposed onto the field effect of the remote substituent. The ability of the para-carborane cage to transmit resonance effects is, on average, about one-half of that of the para-phenylene frame in coplanar para-substituted biphenyls. Analysis of the π-charge variation of the phenyl probe confirms that the para-carborane frame is less capable than the coplanar para-phenylene frame of transmitting π-electrons from the remote substituent to the phenyl probe, or vice versa. The para-carborane cage is a better π-acceptor than π-donor; this makes π-donor substituents less effective than π-acceptors in exchanging π-electrons with the phenyl probe across the cage. When the remote substituent is an uncharged group, the para-carborane cage acts as a very weak π-acceptor toward the phenyl probe. The structural variation of the para-carborane cage has also been investigated. It consists primarily of a change of the C1···C12 nonbonded separation, coupled with a change of the five B-C-B angles at C12. This concerted geometrical change is controlled by the electronegativity of the substituent and the resonance interactions occurring between substituent and cage. These, however, appear to be important only when π-donor substituents are involved. The (13)C NMR chemical shifts of the para-carbon of the phenyl probe correlate nicely with SF(CARB), pointing to the reliability of these quantities as measures of long-range substituent effects. On the contrary, the (11)B and (13)C chemical shifts of the cage atoms do not convey information on electronic substituent effects.


Molecular evolution in large genetic networks: does connectivity equal constraint?
Genetic networks show a broad-tailed distribution of the number of interaction partners per protein, which is consistent with a power-law. It has been proposed that such broad-tailed distributions are observed because they confer robustness against mutations to the network. We evaluate this hypothesis for two genetic networks, that of the E. coli core intermediary metabolism and that of the yeast protein-interaction network. Specifically, we test the hypothesis through one of its key predictions: highly connected proteins should be more important to the cell and, thus, subject to more severe selective and evolutionary constraints. We find, however, that no correlation between highly connected proteins and evolutionary rate exists in the E. coli metabolic network and that there is only a weak correlation in the yeast protein-interaction network. Furthermore, we show that the observed correlation is function-specific within the protein-interaction network: only genes involved in the cell cycle and transcription show significant correlations. Our work sheds light on conflicting results by previous researchers by comparing data from multiple types of protein-interaction datasets and by using a closely related species as a reference taxon. The finding that highly connected proteins can tolerate just as many amino acid substitutions as other proteins leads us to conclude that power-laws in cellular networks do not reflect selection for mutational robustness.


Three-dimensional (13)C-spectroscopic imaging in the isolated infarcted rat heart.
Acquisition weighted (13)C-spectroscopic imaging with three spatial dimensions is demonstrated in the isolated, perfused rat heart. Experiments were performed at 11.75 T with a home-built double resonant (13)C-(1)H probehead. Three-dimensional chemical shift imaging was used to obtain (1)H-decoupled (13)C-spectra in 96-microl voxels in about 58 min. Acquisition weighting significantly reduced signal contamination and improved image quality, with no penalty in sensitivity. As a first application, infarcted hearts were studied during perfusion with [2-(13)C]-sodium acetate. The extent of the incorporation of the (13)C-label into glutamate allows us to distinguish intact and infarcted myocardium. Chemical shift images show a homogeneous glutamate distribution in intact tissue, but a negligible amount in the infarction scar.