Additive multiple k-mer transcriptome of the keelworm Pomatoceros lamarckii (Annelida; Serpulidae) reveals annelid trochophore transcription factor cassette.
Recent advances in both next-generation sequencing and assembly programmes have made the low-cost construction of transcriptome datasets for non-model species feasible, capable of yielding a raft of information even from less well-transcribed genes. Here we present the results of assemblies performed on a 51-bp paired end Illumina dataset derived from a mixed larval sample of the annelid Pomatoceros lamarckii at 24, 48 and 72 h post-fertilization. We used Oases to assemble 36.5 million paired end reads with k-mer sizes from 21 to 29, followed by amalgamation of assemblies, redundancy removal with Vmatch and TGICL and removal of contigs less than 500 bp in length. This resulted in a final assembly of 50,151 contigs, with a mean length of 1,221 bp and covering 61.3 Mbp. A total of 34,846 (69.4 %) of these returned a BlastX hit above a cutoff of 1.0e (-3), and 17,967 (35.8 %) were assigned at least one GO annotation using Blast2GO. We used the assembly to identify genes belonging to the homeobox superclass and the Fox, Sox and Tbx classes, recovering 37, 16, four and three genes, respectively. This included orthologues of genes previously unidentified in lophotrochozoans and protostomes. Our study illustrates the utility of such transcriptomic assembly methods as a gene discovery tool and greatly expands our knowledge of transcription factor genes in annelids in general and in this species in particular.


One-dimensional ferromagnetically coupled bimetallic chains constructed with trans-[Ru(acac)2(CN)2]-: syntheses, structures, magnetic properties, and density functional theoretical study.
Four cyano-bridged 1D bimetallic polymers have been prepared by using the paramagnetic building block trans-[Ru(acac)(2)(CN)(2)](-) (Hacac=acetylacetone): {[{Ni(tren)}{Ru(acac)(2)(CN)(2)}][ClO(4)].CH(3)OH}(n) (1) (tren=tris(2-aminoethyl)amine), {[{Ni(cyclen)}{Ru(acac)(2)(CN)(2)}][ClO(4)].CH(3)OH}(n) (2) (cyclen=1,4,7,10-tetraazacyclododecane), {[{Fe(salen)}{Ru(acac)(2)(CN)(2)}]}(n) (3) (salen(2-)=N,N'-bis(salicylidene)-o-ethyldiamine dianion) and [{Mn(5,5'-Me(2)salen)}(2){Ru(acac)(2)(CN)(2)}][Ru(acac)(2)(CN)(2)].2CH(3)OH (4) (5,5'-Me(2)salen=N,N'-bis(5,5'-dimethylsalicylidene)-o-ethylenediimine). Compounds 1 and 2 are 1D, zigzagged NiRu chains that exhibit ferromagnetic coupling between Ni(II) and Ru(III) ions through cyano bridges with J=+1.92 cm(-1), zJ'=-1.37 cm(-1), g=2.20 for 1 and J=+0.85 cm(-1), zJ'=-0.16 cm(-1), g=2.24 for 2. Compound 3 has a 1D linear chain structure that exhibits intrachain ferromagnetic coupling (J=+0.62 cm(-1), zJ'=-0.09 cm(-1), g=2.08), but antiferromagnetic coupling occurs between FeRu chains, leading to metamagnetic behavior with T(N)=2.6 K. In compound 4, two Mn(III) ions are coordinated to trans-[Ru(acac)(2)(CN)(2)](-) to form trinuclear Mn(2)Ru units, which are linked together by pi-pi stacking and weak Mn...O* interactions to form a 1D chain. Compound 4 shows slow magnetic relaxation below 3.0 K with phi=0.25, characteristic of superparamagnetic behavior. The Mn(III)...Ru(III) coupling constant (through cyano bridges) and the Mn(III)...Mn(III) coupling constant (between the trimers) are +0.87 and +0.24 cm(-1), respectively. Compound 4 is a novel single-chain magnet built from Mn(2)Ru trimers through noncovalent interactions. Density functional theory (DFT) combined with the broken symmetry state method was used to calculate the molecular magnetic orbitals and the magnetic exchange interactions between Ru(III) and M (M=Ni(II), Fe(III), and Mn(III)) ions. To explain the somewhat unexpected ferromagnetic coupling between low-spin Ru(III) and high-spin Fe(III) and Mn(III) ions in compounds 3 and 4, respectively, it is proposed that apart from the relative symmetries, the relative energies of the magnetic orbitals may also be important in determining the overall magnetic coupling in these bimetallic assemblies.


Managing the social amplification of risk: a simulation of interacting actors.
A central problem in managing risk is dealing with social processes that either exaggerate or understate it. A longstanding approach to understanding such processes has been the social amplification of risk framework. But this implies that some true level of risk becomes distorted in social actors' perceptions. Many risk events are characterised by such uncertainties, disagreements and changes in scientific knowledge that it becomes unreasonable to speak of a true level of risk. The most we can often say in such cases is that different groups believe each other to be either amplifying or attenuating a risk. This inherent subjectivity raises the question as to whether risk managers can expect any particular kinds of outcome to emerge. This question is the basis for a case study of zoonotic disease outbreaks using systems dynamics as a modelling medium. The model shows that processes suggested in the social amplification of risk framework produce polarised risk responses among different actors, but that the subjectivity magnifies this polarisation considerably. As this subjectivity takes more complex forms it leaves problematic residues at the end of a disease outbreak, such as an indefinite drop in economic activity and an indefinite increase in anxiety.


Effect of acupuncture on patients with insomnia: study protocol for a randomized controlled trial.
Hypnotic drugs tend to be the dominant form of treatment of insomnia, but these come with a number of reported side effects. Acupuncture has been studied as an alternative, resulting in a rising need for methodological research towards verifying its efficacy as insomnia treatment. We describe a proposal for a single-center, patient-assessor-blinded, randomized controlled trial with two parallel arms. A total of 38 patients complete screening tests at the first visit, are registered into the clinical trial, and then randomly assigned to the experimental or sham control groups (19 patients for each group). All subjects are clinical insomnia patients who score a 6 or above on the Pittsburgh Sleep Quality Index (PSQI) and meet all inclusion criteria. All subjects are treated with acupuncture and intradermal acupuncture (IDA) three times during the first week. Five sham acupoints are used in the control group. In the experimental group, five real acupoints (PC6, SP6, HT7, KI6, and BL62) are used unilaterally in turn. Sham acupoints are over 1 cm away from each real acupoint.The primary outcomes are the scores on the Insomnia Severity Index (ISI) and PSQI. Secondary outcomes are the sleep log, the Beck Depression Inventory (BDI), the State-Trait Anxiety Inventory (STAI), the World Health Organization Quality of Life Abbreviated Version (WHOQOL-BREF), the Korean-Auditory Verbal Learning Test (K-AVLT), the Digit Span Test (DS), Event Related Potentials (ERPs) and heart rate variability (HRV) to assess emotional states, sleep quality, cognitive functioning, and electro-physiological changes.Subjects are assessed at three time points: baseline, post-treatment and follow-up. The duration of the clinical trial is 18 days. To study the enhancement of the effectiveness of acupuncture for insomnia, we test the intradermal acupuncture method, which is performed continuously on the subject's skin and stimulated at home by the subject every night. In the trial, objective measurements including ERPs and HRV are used to evaluate states of cognition and autonomic nervous system functioning and subjective self-report questionnaires assess insomnia symptoms.'Sham' acupuncture points provided by STRICTA are used for the control group. ClinicalTrials.gov: NCT01956760, registered 5 September 2013.


Comparison of two patient-based outcome measures for shoulder instability after nonoperative treatment.
This study compared the responsiveness and minimal clinically important differences of the Oxford Instability Shoulder Score (OISS) and Shoulder Rating Questionnaire (SRQ). Ninety-three patients referred to physiotherapy with shoulder instability (100 shoulders) completed the questionnaires at the initial appointment and at 1 and 9 months later. A subjective rating of change was also obtained. Both questionnaires were able to detect change (effect sizes 1.16 OISS and 0.99 SRQ). The OISS change scores correlated more consistently with patients' subjective responses. The minimal detectable change (MDC90) scores were 16% (7 points) for the OISS and 17% (8 unweighted points) for the SRQ. The minimal clinically important difference (MCID) was between 9%, 4.5 points ("slightly better" and "much better") and 13.5%, 6.5 points ("much better") for the OISS, dependent on the subjective descriptors used, and 11% (4 unweighted points) for SRQ. The MCID change scores varied in relation to the baseline level of function.


Introducing a Latin ballroom dance class to people with dementia living in care homes, benefits and concerns: a pilot study.
The objective of the study was to investigate the effect of introducing a dance-based psychomotor intervention using Danzón (Latin ballroom) for people with dementia in care homes. This was a grounded theory qualitative study. Thirteen of the 22 participants had dementia and were care-home residents. The remaining participants were care staff and facilitators of the dance sessions. Interviews were undertaken with seven people with dementia and nine staff, resulting in two separate sets of grounded theory methodologies. Two conceptual models, outlining positive outcomes and negative concerns of the use of Danzón were developed, depicting the experiences of people with dementia and care staff respectively. Danzón psychomotor intervention was found to enhance positive emotional states and general levels of satisfaction for both people with dementia and care staff. The details of these findings have been used to design a quantitative study.


A Linguistic Approach to Categorical Color Assignment for Data Visualization.
When data categories have strong color associations, it is useful to use these semantically meaningful concept-color associations in data visualizations. In this paper, we explore how linguistic information about the terms defining the data can be used to generate semantically meaningful colors. To do this effectively, we need first to establish that a term has a strong semantic color association, then discover which color or colors express it. Using co-occurrence measures of color name frequencies from Google n-grams, we define a measure for colorability that describes how strongly associated a given term is to any of a set of basic color terms. We then show how this colorability score can be used with additional semantic analysis to rank and retrieve a representative color from Google Images. Alternatively, we use symbolic relationships defined by WordNet to select identity colors for categories such as countries or brands. To create visually distinct color palettes, we use k-means clustering to create visually distinct sets, iteratively reassigning terms with multiple basic color associations as needed. This can be additionally constrained to use colors only in a predefined palette.


HMGB1 genetic polymorphisms are biomarkers for the development and progression of breast cancer.
Breast cancer is a major cause of cancer mortality worldwide. High-mobility group box protein 1 (HMGB1) is a ubiquitous nuclear protein found in all mammal eukaryotic cells that participates in tumor progression, migration and metastasis. HMGB1 overexpression has been indicated in breast cancer patients. However, scant information is available regarding the association between HMGB1 single nucleotide polymorphisms (SNPs) and the risk or prognosis of breast cancer. We report on the association between 4 SNPs of the HMGB1 gene (rs1360485, rs1045411, rs2249825 and rs1412125) and breast cancer susceptibility as well as clinical outcomes in 313 patients with breast cancer and in 217 healthy controls. Patients with one G allele in the rs1360485 or rs2249825 domains are likely to progress to T2 tumor and lymph node metastasis. In addition, the presence of one G allele in SNPs rs1360485 or rs2249825 was associated with a higher risk of progressing to T2 tumor and distant metastasis amongst HER2-enriched and triple-negative breast cancer (TNBC) tumors compared with luminal A and luminal B tumors. Furthermore, having one C allele in the rs1412125 domain increased the risk of pathologic grade 3 disease in HER2-enriched and TNBC tumors. Our results indicate that genetic variations in the HMGB1 gene may serve as an important predictor of breast cancer progression and metastasis.


Why different countries manage death differently: a comparative analysis of modern urban societies.
The sociology of death, dying and bereavement tends to take as its implicit frame either the nation state or a homogenous modernity. Between-nation differences in the management of death and dying are either ignored or untheorized. This article seeks to identify the factors that can explain both similarities and differences in the management of death between different modern western nations. Structural factors which affect all modern nations include urbanization and the division of labour leading to the dominance of professionals, migration, rationality and bureaucracy, information technology and the risk society. How these sociologically familiar structural features are responded to, however, depends on national histories, institutions and cultures. Historically, key transitional periods to modernity, different in different nations, necessitated particular institutional responses in the management of dying and dead bodies. Culturally, key factors include individualism versus collectivism, religion, secularization, boundary regulation, and expressivism. Global flows of death practices depend significantly on subjugated nations' perceptions of colonialism, neo-colonialism and modernity, which can lead to a dominant power's death practices being either imitated or rejected.


Plasticity of empty major histocompatibility complex class I molecules determines peptide-selector function.
Major histocompatibility complex class I (MHC I) proteins provide protection from intracellular pathogens and cancer via each of a cell's MHC I molecules binding and presenting a peptide to cytotoxic T lymphocytes. MHC I genes are highly polymorphic and can have significant diversity, with polymorphisms predominantly localised in the peptide-binding groove where they can change peptide-binding specificity. However, polymorphic residues may also determine other functional properties, such as how dependent MHC I alleles are on the peptide-loading complex for optimal acquisition of peptide cargo. We describe how differences in the peptide-binding properties of two MHC I alleles correlates with altered conformational flexibility in the peptide-empty state. We hypothesise that plasticity is an intrinsic property encoded by the protein sequence, and that co-ordinated movements of the membrane-proximal and membrane-distal domains collectively determines how dependent MHC I are on the peptide-loading complex for efficient assembly with high affinity peptides.


Sedentary Behavior and Physical Activity Patterns in Older Adults After Hip Fracture: A Call to Action.
To characterize patterns of sedentary behavior and physical activity in older adults recovering from hip fracture and to determine characteristics associated with activity. Community-dwelling, Canadian adults (65 years+) who sustained hip fracture wore an accelerometer at the waist for seven days and provided information on quality of life, falls self-efficacy, cognitive functioning, and mobility. There were 53 older adults (mean age [SD] 79.5 [7.8] years) enrolled in the study; 49 had valid data and demonstrated high levels of sedentary time (median [p10, p90] 591.3 [482.2, 707.2] minutes/day), low levels of light activity (186.6 [72.6, 293.7]), and MVPA (2 [0.1, 27.6]), as well as few daily steps (2467.7 [617.1, 6820.4]). Regression analyses showed that age, gender, gait speed, and time since fracture were associated with outcomes. Older adults have long periods of sedentary time with minimal activity. Results are a call to action to encourage people to sit less and move more.


Mechanistic studies on norcoclaurine synthase of benzylisoquinoline alkaloid biosynthesis: an enzymatic Pictet-Spengler reaction.
Norcoclaurine synthase catalyzes an asymmetric Pictet-Spengler condensation of dopamine and 4-hydroxyphenylacetaldehyde to give (S)-norcoclaurine. This is the first committed step in the biosynthesis of the benzylisoquinoline alkaloids that include morphine and codeine. In this work, the gene encoding for the Thalictrum flavum norcoclaurine synthase is highly overexpressed in Escherichia coli and the resulting His-tagged recombinant enzyme is purified for the first time. A continuous assay based on circular dichroism spectroscopy is developed and used to monitor the kinetics of the enzymatic reaction. Dopamine analogues bearing a methoxy or hydrogen substituent in place of the C-1 phenolic group were readily accepted by the enzyme whereas those bearing the same substituents at C-2 were not. This supports a mechanism involving a two-step cyclization of the putative iminium ion intermediate that does not proceed via a spirocyclic intermediate. The reaction of [3,5,6-2H]dopamine was found to be slowed by a kinetic isotope effect of 1.7 +/- 0.1 on the value of kcat/KM. This is interpreted as showing that the deprotonation step causing rearomatization is partially rate determining in the overall reaction.


Febrile Illness Evaluation in a Broad Range of Endemicities (FIEBRE): protocol for a multisite prospective observational study of the causes of fever in Africa and Asia.
Fever commonly leads to healthcare seeking and hospital admission in sub-Saharan Africa and Asia. There is only limited guidance for clinicians managing non-malarial fevers, which often results in inappropriate treatment for patients. Furthermore, there is little evidence for estimates of disease burden, or to guide empirical therapy, control measures, resource allocation, prioritisation of clinical diagnostics or antimicrobial stewardship. The Febrile Illness Evaluation in a Broad Range of Endemicities (FIEBRE) study seeks to address these information gaps. FIEBRE investigates febrile illness in paediatric and adult outpatients and inpatients using standardised clinical, laboratory and social science protocols over a minimum 12-month period at five sites in sub-Saharan Africa and Southeastern and Southern Asia. Patients presenting with fever are enrolled and provide clinical data, pharyngeal swabs and a venous blood sample; selected participants also provide a urine sample. Laboratory assessments target infections that are treatable and/or preventable. Selected point-of-care tests, as well as blood and urine cultures and antimicrobial susceptibility testing, are performed on site. On day 28, patients provide a second venous blood sample for serology and information on clinical outcome. Further diagnostic assays are performed at international reference laboratories. Blood and pharyngeal samples from matched community controls enable calculation of AFs, and surveys of treatment seeking allow estimation of the incidence of common infections. Additional assays detect markers that may differentiate bacterial from non-bacterial causes of illness and/or prognosticate illness severity. Social science research on antimicrobial use will inform future recommendations for fever case management. Residual samples from participants are stored for future use. Ethics approval was obtained from all relevant institutional and national committees; written informed consent is obtained from all participants or parents/guardians. Final results will be shared with participating communities, and in open-access journals and other scientific fora. Study documents are available online (https://doi.org/10.17037/PUBS.04652739).


The relative magnitude of transgene-specific adaptive immune responses induced by human and chimpanzee adenovirus vectors differs between laboratory animals and a target species.
Adenovirus vaccine vectors generated from new viral serotypes are routinely screened in pre-clinical laboratory animal models to identify the most immunogenic and efficacious candidates for further evaluation in clinical human and veterinary settings. Here, we show that studies in a laboratory species do not necessarily predict the hierarchy of vector performance in other mammals. In mice, after intramuscular immunization, HAdV-5 (Human adenovirus C) based vectors elicited cellular and humoral adaptive responses of higher magnitudes compared to the chimpanzee adenovirus vectors ChAdOx1 and AdC68 from species Human adenovirus E. After HAdV-5 vaccination, transgene specific IFN-γ(+) CD8(+) T cell responses reached peak magnitude later than after ChAdOx1 and AdC68 vaccination, and exhibited a slower contraction to a memory phenotype. In cattle, cellular and humoral immune responses were at least equivalent, if not higher, in magnitude after ChAdOx1 vaccination compared to HAdV-5. Though we have not tested protective efficacy in a disease model, these findings have important implications for the selection of candidate vectors for further evaluation. We propose that vaccines based on ChAdOx1 or other Human adenovirus E serotypes could be at least as immunogenic as current licensed bovine vaccines based on HAdV-5.


The hazards of off-road motor sports: Are four wheels better than two?
Off-road motorsports are an increasing popular activity, yet the relative safety profile of all-terrain vehicles (ATV) to off-road motorcycles (ORMC) has not been compared. A retrospective review of the 2002-2006 US National Trauma Data Bank of ATV and ORMC crash victims. Patients were described according to demographic (age, sex, race and ethnicity, insurance status) and injury characteristics (Injury Severity Score, hypotension, motor component of the Glasgow Coma Score, presence of a severe head or extremity injury) known to affect trauma outcomes. Logistic regression evaluated the independent effect of an ATV vehicle on mortality, intensive care unit (ICU) admission, and placement on a ventilator relative to ORMC. The anatomic distribution of severe injuries was compared between survivors and decedents within each vehicle type. A total of 34,457 patients met inclusion criteria, of whom, 24,582 were ATV patients and 9875 were ORMC patients. ATV patients had 51% higher risk-adjusted odds of death (OR 1.51; 95% CI 1.03-2.20), 55% higher risk-adjusted odds of being admitted to an ICU (OR 1.55; 95% CI 1.42-1.70), and 42% higher risk-adjusted odds of being placed on a ventilator (OR 1.42, 95% CI 1.17-1.72) compared to ORMC crash victims. Decedents in both vehicle types were more likely to suffer severe head, thoracic, and abdominal injuries relative to their surviving counterparts. For injured riders, ATVs are associated with increased mortality and higher resource utilisation compared to ORMCs. Both groups suffer distinct anatomic injuries, suggesting the need for focused areas of injury prevention planning and research.


Sedentary Behaviors in Today's Youth: Approaches to the Prevention and Management of Childhood Obesity: A Scientific Statement From the American Heart Association.
This scientific statement is about sedentary behavior and its relationship to obesity and other cardiometabolic outcomes in youth. A deleterious effect of sedentary behavior on cardiometabolic health is most notable for screen-based behaviors and adiposity; however, this relation is less apparent for other cardiometabolic outcomes or when sedentary time is measured with objective movement counters or position monitors. Increasing trends of screen time are concerning; the portability of screen-based devices and abundant access to unlimited programming and online content may be leading to new patterns of consumption that are exposing youth to multiple pathways harmful to cardiometabolic health. This American Heart Association scientific statement provides an updated perspective on sedentary behaviors specific to modern youth and their impact on cardiometabolic health and obesity. As we reflect on implications for practice, research, and policy, what emerges is the importance of understanding the context in which sedentary behaviors occur. There is also a need to capture the nature of sedentary behavior more accurately, both quantitatively and qualitatively, especially with respect to recreational screen-based devices. Further evidence is required to better inform public health interventions and to establish detailed quantitative guidelines on specific sedentary behaviors in youth. In the meantime, we suggest that televisions and other recreational screen-based devices be removed from bedrooms and absent during meal times. Daily device-free social interactions and outdoor play should be encouraged. In addition, parents/guardians should be supported to devise and enforce appropriate screen time regulations and to model healthy screen-based behaviors.


Genistein, a competitive PDE1-4 inhibitor, may bind on high-affinity rolipram binding sites of brain cell membranes and then induce gastrointestinal adverse effects.
The affinities of genistein on phosphodiesterase (PDE)1-4 and cause of gastrointestinal adverse effects of genistein remain unclear. Female BALB/c mice were actively sensitized by intraperitoneal injections of ovalbumin and challenged by aerosolized ovalbumin (1%). After secondary challenge, aerosolized methacholine (6.25-50mg/ml) induced increases of enhanced pause (P(enh)) values in conscious mice in a concentration-dependent manner. Genistein (30-100 micromol/kg, i.p.) markedly inhibited methacholine (12.5-50mg/ml)-induced increase of P(enh) value in the sensitized and challenged mice. In addition, genistein significantly reduced total inflammatory cells, macrophages, lymphocytes, neutrophils, and eosinophils in bronchoalveolar lavage fluid, with the exception that lymphocytes and neutrophils were not significantly inhibited by genistein at the lowest dose (10 micromol/kg). Genistein also markedly attenuated the release of cytokines, including interleukin (IL)-2, IL-4, IL-5, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha. Genistein competitively inhibited PDE1-4, with a K(i) value ranging from 4.3 to 13.7 microM. Genistein (3-300 microM) concentration-dependently displaced 2nM [(3)H]-rolipram bound on high-affinity rolipram binding sites of brain cell membranes. The therapeutic ratio of genistein was calculated to be 7.9. Genistein (100 micromol/kg, s.c.) significantly shortened xylazine/ketamine-induced anesthesia, suggesting that genistein administered at a higher dose may have gastrointestinal adverse effects. In conclusion, owing to the low therapeutic ratio of genistein, the gastrointestinal adverse effects may be induced via the binding of genistein on high-affinity rolipram binding sites of brain cell membranes, when it is used for a long term or at higher doses for treating allergies, asthma or chronic obstructive pulmonary disease.


Classification of Intervertebral Disc Disease.
Intervertebral disc disease (IVDD) has been recognized in dogs since the 1800s, when the first descriptions of extruded disc material within the vertebral canal were published. In the intervening time our understanding of intervertebral disc pathology in dogs and cats has increased dramatically, with many variations of IVDD described. Whilst the volume of literature and collective understanding of IVDD has expanded, there has also been scope for confusion as the definition of intervertebral disc disease, with its myriad different manifestations, becomes more complicated. A large volume of literature has aimed to combine the use of histopathology, diagnostic imaging and clinical findings to better understand the various ways in which IVDD can be classified. Much of this research has focused on the classification of mechanisms of intervertebral disc degeneration, centering around the differences between, and overlaps in, IVDD in chondrodystrophic and non-chondrodystrophic dog breeds. However, with the increasing availability of advanced imaging modalities allowing more accurate antemortem diagnosis, the concept of IVDD has expanded to include other clinical presentations that may not fit into traditional models of classification of IVDD. This review aims to provide an up to date overview of both historical and current systems of IVDD classification, highlighting the important findings and controversies underpinning them.


Alive and at home: Five-year outcomes in older adults following emergency general surgery.
While the short-term risks of emergency general surgery (EGS) admission among older adults have been studied, little is known about long-term functional outcomes in this population. Our objective was to evaluate the relationship between EGS admission and the probability of an older adult being alive and residing in their own home 5 years later. We also examined the extent to which specific EGS diagnoses, need for surgery, and frailty modified this relationship. We performed a population-based, retrospective cohort study of community-dwelling older adults (age, ≥65 years) admitted to hospital for one of eight EGS diagnoses (appendicitis, cholecystitis, diverticulitis, strangulated hernia, bowel obstruction, peptic ulcer disease, intestinal ischemia, or perforated viscus) between 2006 and 2018 in Ontario, Canada. Cases were matched to controls from the general population. Time spent alive and at home (measured as time to nursing home admission or death) was compared between cases and controls using Kaplan-Meier analysis and Cox models. A total of 90,245 older adults admitted with an EGS diagnosis were matched with controls. In the 5 years following an EGS admission, cases experienced significantly fewer months alive and at home compared with controls (mean time, 43 vs. 50 months; p < 0.001). Except for patients operated on for appendicitis and cholecystitis, all remaining patient subgroups experienced reduced time alive and at home compared with controls (p < 0.001). Cases remained at elevated risk of nursing home admission or death compared with controls for the entirety of the 5-year follow-up (hazard ratio, 1.17-5.11). Older adults who required hospitalization for an EGS diagnosis were at higher risk for death or admission to a nursing home for at least 5 years following admission compared with controls. However, most patients (57%) remained alive and living in their own home at the end of this 5-year period. Epidemiological, level III.


A study on reproducing silicone shade guide for maxillofacial prostheses matching Indian skin color.
The value and success of a well-fitting and anatomically correct prosthesis are compromised if the color does not match the adjoining tissues. The use of powder colors to help develop a simplified silicone shade guide to aid in the fabrication of silicone facial prostheses for Indian patients has been described here. Ten powder pigments were used to fabricate the silicone samples for three different subjects having light, medium and dark complexions who were separated into three groups depending on the value of their shades. Four-step wedge silicone samples with thickness variations of 1, 2, 4 and 6mm were fabricated. Visual assessment of the samples was done by four evaluators to check the agreement of color match. Data was statistically analyzed using kappa coefficients. The kappa values were found to be 0.09-0.44 for a light skin tone, -0.11 to 0.77 for medium skin and 0.44 to 0.85 for dark skin tones. This study showed that the samples of dark skin tone matched the skin tone well and showed a statistically good agreement. To further test the validity of these shade guides, facial silicone veneers were fabricated for three patients having light, medium and dark complexion. The color matching showed satisfactory results. The silicone veneers matched the skin color of all three patients. Hence, this shade guide will help clinicians to obtain a good intrinsic shade and minimize extrinsic coloration.


Ratio of motor nerve conduction velocity to F-wave conduction velocity in diabetic neuropathy.
To investigate the usefulness of a new parameter, the ratio of motor nerve conduction velocity to F-wave conduction velocity (M/F ratio), for the differential diagnosis of diabetic neuropathy. Nerve conduction studies were conducted in 95 patients with diabetic neuropathy, 44 nondiabetic patients with peripheral neuropathy, and 24 normal control subjects. Nondiabetic patients with neuropathy were grouped by clinical diagnosis as follows: segmental demyelination (n = 15), axonal neuropathy (n = 11), alcoholic polyneuropathy (n = 4), and other polyneuropathy (n = 14). Motor nerve conduction velocity (MCV) of post-tibial nerves, sensory nerve conduction velocity (SCV) of sural nerves, and F-wave conduction velocity (FWCV) of post-tibial nerves were measured by standardized techniques. The M/F ratio was calculated from these measurements. The MCV and SCV of diabetic patients were significantly slower and the M/F ratio was significantly lower than those of normal subjects: MCV, 43.7 +/- 5.4 vs. 47.1 +/- 2.9 m/s, P < 0.001; SCV, 44.7 +/- 11.1 vs. 48.3 +/- 5.7 m/s, P < 0.05; M/F ratio, 0.84 +/- 0.09 vs. 0.90 +/- 0.06, P < 0.001. The FWCV of nondiabetic patients with neuropathy was significantly slower (40.0 +/- 6.3 vs. 48.3 +/- 4.0 m/s, P < 0.001) and the M/F ratio was significantly higher (1.04 +/- 0.12, P < 0.001) than that of normal subjects, respectively. Although MCV, SCV, and FWCV were correlated with age in normal control subjects, the M/F ratio was independent of age in the diabetic as well as the nondiabetic patients with neuropathy. Results suggest that the M/F ratio, which is influenced by the neuronal damages in the distal segment of peripheral nerves, is useful in the differential diagnosis of diabetic neuropathy.


Receptor activator of NF-kB (RANK) expression in primary tumors associates with bone metastasis occurrence in breast cancer patients.
Receptor activator of NFkB (RANK), its ligand (RANKL) and the decoy receptor of RANKL (osteoprotegerin, OPG) play a pivotal role in bone remodeling by regulating osteoclasts formation and activity. RANKL stimulates migration of RANK-expressing tumor cells in vitro, conversely inhibited by OPG. We examined mRNA expression levels of RANKL/RANK/OPG in a publicly available microarray dataset of 295 primary breast cancer patients. We next analyzed RANK expression by immunohistochemistry in an independent series of 93 primary breast cancer specimens and investigated a possible association with clinicopathological parameters, bone recurrence and survival. Microarray analysis showed that lower RANK and high OPG mRNA levels correlate with longer overall survival (P = 0.0078 and 0.0335, respectively) and disease-free survival (P = 0.059 and 0.0402, respectively). Immunohistochemical analysis of RANK showed a positive correlation with the development of bone metastases (P = 0.023) and a shorter skeletal disease-free survival (SDFS, P = 0.037). Specifically, univariate analysis of survival showed that "RANK-negative" and "RANK-positive" patients had a SDFS of 105.7 months (95% CI: 73.9-124.4) and 58.9 months (95% CI: 34.7-68.5), respectively. RANK protein expression was also associated with accelerated bone metastasis formation in a multivariate analysis (P = 0.029). This is the first demonstration of the role of RANK expression in primary tumors as a predictive marker of bone metastasis occurrence and SDFS in a large population of breast cancer patients.


Suppression of developmental anomalies by maternal macrophages in mice.
We tested whether nonspecific tumoricidal immune cells can suppress congenital malformations by killing precursor cells destined to cause such defects. Pretreatment of pregnant ICR mice with synthetic (Pyran copolymer) and biological (Bacillus Calmette-Guérin) agents significantly suppressed radiation- and chemical-induced congenital malformations (cleft palate, digit anomalies, tail anomalies, etc.). Such suppressive effects were associated with the activation of maternal macrophages by these agents, but were lost either after the disruption of activated macrophages by supersonic waves or by inhibition of their lysosomal enzyme activity with trypan blue. These results indicate that a live activated macrophage with active lysosomal enzymes can be an effector cell to suppress maldevelopment. A similar reduction by activated macrophages was observed in strain CL/Fr, which has a high spontaneous frequency of cleft lips and palates. Furthermore, Pyran-activated maternal macrophages could pass through the placenta, and enhanced urethane-induced cell killing (but not somatic mutation) in the embryo. It is likely that a maternal immunosurveillance system eliminating preteratogenic cells allows for the replacement with normal totipotent blast cells during the pregnancy to protect abnormal development.


Evidence for association of GABA(B) receptors with Kir3 channels and regulators of G protein signalling (RGS4) proteins.
Many neurotransmitters and hormones signal by stimulating G protein-coupled neurotransmitter receptors (GPCRs), which activate G proteins and their downstream effectors. Whether these signalling proteins diffuse freely within the plasma membrane is not well understood. Recent studies have suggested that direct protein-protein interactions exist between GPCRs, G proteins and G protein-gated inwardly rectifying potassium (GIRK or Kir3) channels. Here, we used fluorescence resonance energy transfer (FRET) combined with total internal reflection fluorescence microscopy to investigate whether proteins within this signalling pathway move within 100 A of each other in the plasma membrane of living cells. GABA(B) R1 and R2 receptors, Kir3 channels, Galphao subunits and regulators of G protein signalling (RGS4) proteins were each fused to cyan fluorescent protein (CFP) or yellow fluorescent protein (YFP) and first assessed for functional expression in HEK293 cells. The presence of the fluorophore did not significantly alter the signalling properties of these proteins. Possible FRET was then investigated for different protein pair combinations. As a positive control, FRET was measured between tagged GABA(B) R1 and R2 subunits ( approximately 12% FRET), which are known to form heterodimers. We measured significant FRET between tagged RGS4 and GABA(B) R1 or R2 subunits ( approximately 13% FRET), and between Galphao and GABA(B) R1 or R2 subunits ( approximately 10% FRET). Surprisingly, FRET also occurred between tagged Kir3.2a/Kir3.4 channels and GABA(B) R1 or R2 subunits ( approximately 10% FRET). FRET was not detected between Kir3.2a and RGS4 nor between Kir3.2a and Galphao. These data are discussed in terms of a model in which GABA(B) receptors, G proteins, RGS4 proteins and Kir3 channels are closely associated in a signalling complex.


Antibody-induced internalisation of retroviral envelope glycoproteins is a signal initiation event.
As obligate parasites, viruses highjack, modify and repurpose the cellular machinery for their own replication. Viral proteins have, therefore, evolved biological functions, such as signalling potential, that alter host cell physiology in ways that are still incompletely understood. Retroviral envelope glycoproteins interact with several host proteins, extracellularly with their cellular receptor and anti-envelope antibodies, and intracellularly with proteins of the cytoskeleton or sorting, endocytosis and recirculation pathways. Here, we examined the impact of endogenous retroviral envelope glycoprotein expression and interaction with host proteins, particularly antibodies, on the cell, independently of retroviral infection. We found that in the commonly used C57BL/6 substrains of mice, where murine leukaemia virus (MLV) envelope glycoproteins are expressed by several endogenous MLV proviruses, the highest expressed MLV envelope glycoprotein is under the control of an immune-responsive cellular promoter, thus linking MLV envelope glycoprotein expression with immune activation. We further showed that antibody ligation induces extensive internalisation from the plasma membrane into endocytic compartments of MLV envelope glycoproteins, which are not normally subject to constitutive endocytosis. Importantly, antibody binding and internalisation of MLV envelope glycoproteins initiates signalling cascades in envelope-expressing murine lymphocytic cell lines, leading to cellular activation. Similar effects were observed by MLV envelope glycoprotein ligation by its cellular receptor mCAT-1, and by overexpression in human lymphocytic cells, where it required an intact tyrosine-based YXXΦ motif in the envelope glycoprotein cytoplasmic tail. Together, these results suggest that signalling potential is a general property of retroviral envelope glycoproteins and, therefore, a target for intervention.


Coordinate phosphorylation of insulin-receptor kinase and its 175,000-Mr endogenous substrate in rat hepatocytes.
To investigate the early events in insulin signal transmission in liver, isolated rat hepatocytes were labeled with 32P, and proteins phosphorylated in response to insulin were detected by immunoprecipitation with anti-phosphotyrosine and anti-receptor antibodies and analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and autoradiography. In these cells, insulin rapidly stimulated tyrosine phosphorylation of the 95,000-Mr beta-subunit of the insulin receptor and a 175,000-Mr phosphoprotein (pp175). Both proteins were precipitated by anti-phosphotyrosine antibody, whereas only the insulin receptor was recognized with anti-insulin-receptor antibody. In the insulin-stimulated state, both pp175 and the receptor beta-subunit were found to be phosphorylated on tyrosine and serine residues. Based on precipitation by the two antibodies, receptor phosphorylation was biphasic with an initial increase in tyrosine phosphorylation followed by a more gradual increase in serine phosphorylation over the first 30 min of stimulation. The time course of phosphorylation of pp175 was rapid and paralleled that of the beta-subunit of the insulin receptor. The pp175 was clearly distinguished from the insulin receptor, because it was detected only when boiling SDS was used to extract cellular phosphoproteins, whereas the insulin receptor was extracted with either Triton X-100 or SDS. In addition, the tryptic peptide maps of the two proteins were distinct. The dose-response curve for insulin stimulation was shifted slightly to the left of the insulin receptor, suggesting some signal amplification at this step. These data suggest that pp175 is a major endogenous substrate of the insulin receptor in liver and may be a cytoskeletal-associated protein.(ABSTRACT TRUNCATED AT 250 WORDS)


On-line preconcentration strategies for trace analysis of metabolites by capillary electrophoresis.
Analysis of low concentrations of metabolites is required for new fields of biological research, such as metabolomics. In this review, recent work in our laboratory aimed at developing improved strategies for on-line sample preconcentration of metabolites by capillary electrophoresis (CE) is presented. Dynamic pH junction, sweeping and dynamic pH junction-sweeping represent three complementary methods for electrokinetic focusing of large volumes of sample directly on-capillary. Focusing selectivity and focusing efficiency are two factors that can be used to assess the suitability of each method for different classes of metabolites. Buffer properties can be selected to enhance the focusing of specific types of metabolites based on knowledge of the analyte physicochemical properties. The application of on-line preconcentration CE for trace analysis of metabolites in real samples of interest, such as biological fluids and cellular extracts, is also demonstrated. Under optimum conditions, up to three orders of magnitude increase in concentration sensitivity can be realized for several classes of metabolites, including catecholamines, purines, nucleosides, nucleotides, amino acids, steroids and coenzymes. Recent work on hyphenating on-line preconcentration with multiplexed CE is highlighted as a promising platform for sensitive and high-throughput analyses of metabolites.


Resolution enhancement of lung 4D-CT via group-sparsity.
4D-CT typically delivers more accurate information about anatomical structures in the lung, over 3D-CT, due to its ability to capture visual information of the lung motion across different respiratory phases. This helps to better determine the dose during radiation therapy for lung cancer. However, a critical concern with 4D-CT that substantially compromises this advantage is the low superior-inferior resolution due to less number of acquired slices, in order to control the CT radiation dose. To address this limitation, the authors propose an approach to reconstruct missing intermediate slices, so as to improve the superior-inferior resolution. In this method the authors exploit the observation that sampling information across respiratory phases in 4D-CT can be complimentary due to lung motion. The authors' approach uses this locally complimentary information across phases in a patch-based sparse-representation framework. Moreover, unlike some recent approaches that treat local patches independently, the authors' approach employs the group-sparsity framework that imposes neighborhood and similarity constraints between patches. This helps in mitigating the trade-off between noise robustness and structure preservation, which is an important consideration in resolution enhancement. The authors discuss the regularizing ability of group-sparsity, which helps in reducing the effect of noise and enables better structural localization and enhancement. The authors perform extensive experiments on the publicly available DIR-Lab Lung 4D-CT dataset [R. Castillo, E. Castillo, R. Guerra, V. Johnson, T. McPhail, A. Garg, and T. Guerrero, "A framework for evaluation of deformable image registration spatial accuracy using large landmark point sets," Phys. Med. Biol. 54, 1849-1870 (2009)]. First, the authors carry out empirical parametric analysis of some important parameters in their approach. The authors then demonstrate, qualitatively as well as quantitatively, the ability of their approach to achieve more accurate and better localized results over bicubic interpolation as well as a related state-of-the-art approach. The authors also show results on some datasets with tumor, to further emphasize the clinical importance of their method. The authors have proposed to improve the superior-inferior resolution of 4D-CT by estimating intermediate slices. The authors' approach exploits neighboring constraints in the group-sparsity framework, toward the goal of achieving better localization and noise robustness. The authors' results are encouraging, and positively demonstrate the role of group-sparsity for 4D-CT resolution enhancement.


A Method for Isolation of Pasteuria penetrans Endospores for Bioassay and Genomic Studies.
A rapid method for collection of Pasteuria penetrans endospores was developed. Roots containing P. penetrans-infected root-knot nematode females were softened by pectinase digestion, mechanically processed, and filtered to collect large numbers of viable endospores. This method obviates laborious handpicking of Pasteuria-infected females and yields endospores competent to attach to and infect nematodes. Endospores are suitable for morphology studies and DNA preparations.


Aging mechanism of soman inhibited acetylcholinesterase.
Acetylcholinesterase (AChE) is a crucial enzyme in the cholinergic nervous system that hydrolyzes neurotransmitter acetylcholine (ACh) and terminates synaptic signals. The catalytic serine of AChE can be phosphonylated by soman, one of the most potent nerve agents, and subsequently undergo an aging reaction. This phosphonylation and aging process leads to irreversible AChE inhibition, results in accumulation of excess ACh at the synaptic clefts, and causes neuromuscular paralysis. By employing Born-Oppenheimer ab initio QM/MM molecular dynamics simulations with umbrella sampling, a state-of-the-art approach to simulate enzyme reactions, we have characterized the aging mechanism of soman phosphonylated AChE and determined its free energy profile. This aging reaction starts with the scission of the O2-Cα bond, which is followed by methyl migration, and results in a tertiary carbenium intermediate. At the transition state, the scissile O2-Cα bond is already cleaved with an average O-C distance of 3.2 ± 0.3 Å and the migrating methyl group is shared between Cα and Cβ carbons with C-C distances of 1.9 ± 0.1 and 1.8 ± 0.1 Å, respectively. The negatively charged phosphonate group is stabilized by a salt bridge with the imidazole ring of the catalytic histidine. A major product of aging, 2,3-dimethyl-2-butanol can be formed swiftly by the reaction of a water molecule. Our characterized mechanism and simulation results provide new detailed insights into this important biochemical process.


Generalization of color by chickens: experimental observations and a Bayesian model.
Sensory generalization influences animals' responses to novel stimuli. Because color forms a perceptual continuum, it is a good subject for studying generalization. Moreover, because different causes of variation in spectral signals, such as pigmentation, gloss, and illumination, have differing behavioral significance, it may be beneficial to have adaptable generalization. We report on generalization by poultry chicks following differential training to rewarded (T(+)) and unrewarded (T(-)) colors, in particular on the phenomenon of peak shift, which leads to subjects preferring stimuli displaced away from T(-). The first three experiments test effects of learning either a fine or a coarse discrimination. In experiments 1 and 2, peak shift occurs, but contrary to some predictions, the shift is smaller after the animal learned a fine discrimination than after it learned a coarse discrimination. Experiment 3 finds a similar effect for generalization on a color axis orthogonal to that separating T(+) from T(-). Experiment 4 shows that generalization is rapidly modified by experience. These results imply that the scale of a "perceptual ruler" is set by experience. We show that the observations are consistent with generalization following principles of Bayesian inference, which forms a powerful framework for understanding this type of behavior.


High-Flow Nasal Cannula Versus Standard Oxygen Therapy After Extubation in Liver Transplantation: A Matched Controlled Study.
High-flow nasal cannula (HFNC) is a key component of oxygen therapy and has largely been used in patients with acute respiratory failure. We conducted a matched controlled study with the aim to compare the preventive use of oxygen therapy delivered by HFNC versus via air-entrainment mask (standard O2) after extubation in adult subjects with liver transplantation for reducing postextubation hypoxemia. Twenty-nine subjects with liver transplantation who received HFNC after extubation (HFNC group) were matched 1:1 with 29 controls (standard O2 group) chosen from an historical group of 90 subjects admitted to the ICU during the previous 36 months. The primary outcome of the study was the incidence of hypoxemia at 1 h and 24 h after extubation. Secondary outcomes were the rate of weaning failure, ICU length of stay, and 28-d mortality. The incidence of hypoxemia was not significantly different between the HFNC and standard O2 groups at 1 h and 24 h after extubation. In the HFNC group, there was a trend toward a lower rate of weaning failure compared with the standard O2 group. ICU length of stay and 28-d mortality were similar in both groups. Early application of HFNC in the subjects with liver transplantation did not reduce the incidence of hypoxemia after extubation compared with standard O2 and did not modify the incidence of weaning failure, ICU length of stay, and 28-d mortality in this high-risk population of subjects. (ClinicalTrials.gov registration NCT03441854.).


Systemic treatments for metastatic cutaneous melanoma.
The prognosis of people with metastatic cutaneous melanoma, a skin cancer, is generally poor. Recently, new classes of drugs (e.g. immune checkpoint inhibitors and small-molecule targeted drugs) have significantly improved patient prognosis, which has drastically changed the landscape of melanoma therapeutic management. This is an update of a Cochrane Review published in 2000. To assess the beneficial and harmful effects of systemic treatments for metastatic cutaneous melanoma. We searched the following databases up to October 2017: the Cochrane Skin Group Specialised Register, CENTRAL, MEDLINE, Embase and LILACS. We also searched five trials registers and the ASCO database in February 2017, and checked the reference lists of included studies for further references to relevant randomised controlled trials (RCTs). We considered RCTs of systemic therapies for people with unresectable lymph node metastasis and distant metastatic cutaneous melanoma compared to any other treatment. We checked the reference lists of selected articles to identify further references to relevant trials. Two review authors extracted data, and a third review author independently verified extracted data. We implemented a network meta-analysis approach to make indirect comparisons and rank treatments according to their effectiveness (as measured by the impact on survival) and harm (as measured by occurrence of high-grade toxicity). The same two review authors independently assessed the risk of bias of eligible studies according to Cochrane standards and assessed evidence quality based on the GRADE criteria. We included 122 RCTs (28,561 participants). Of these, 83 RCTs, encompassing 21 different comparisons, were included in meta-analyses. Included participants were men and women with a mean age of 57.5 years who were recruited from hospital settings. Twenty-nine studies included people whose cancer had spread to their brains. Interventions were categorised into five groups: conventional chemotherapy (including single agent and polychemotherapy), biochemotherapy (combining chemotherapy with cytokines such as interleukin-2 and interferon-alpha), immune checkpoint inhibitors (such as anti-CTLA4 and anti-PD1 monoclonal antibodies), small-molecule targeted drugs used for melanomas with specific gene changes (such as BRAF inhibitors and MEK inhibitors), and other agents (such as anti-angiogenic drugs). Most interventions were compared with chemotherapy. In many cases, trials were sponsored by pharmaceutical companies producing the tested drug: this was especially true for new classes of drugs, such as immune checkpoint inhibitors and small-molecule targeted drugs.When compared to single agent chemotherapy, the combination of multiple chemotherapeutic agents (polychemotherapy) did not translate into significantly better survival (overall survival: HR 0.99, 95% CI 0.85 to 1.16, 6 studies, 594 participants; high-quality evidence; progression-free survival: HR 1.07, 95% CI 0.91 to 1.25, 5 studies, 398 participants; high-quality evidence. Those who received combined treatment are probably burdened by higher toxicity rates (RR 1.97, 95% CI 1.44 to 2.71, 3 studies, 390 participants; moderate-quality evidence). (We defined toxicity as the occurrence of grade 3 (G3) or higher adverse events according to the World Health Organization scale.)Compared to chemotherapy, biochemotherapy (chemotherapy combined with both interferon-alpha and interleukin-2) improved progression-free survival (HR 0.90, 95% CI 0.83 to 0.99, 6 studies, 964 participants; high-quality evidence), but did not significantly improve overall survival (HR 0.94, 95% CI 0.84 to 1.06, 7 studies, 1317 participants; high-quality evidence). Biochemotherapy had higher toxicity rates (RR 1.35, 95% CI 1.14 to 1.61, 2 studies, 631 participants; high-quality evidence).With regard to immune checkpoint inhibitors, anti-CTLA4 monoclonal antibodies plus chemotherapy probably increased the chance of progression-free survival compared to chemotherapy alone (HR 0.76, 95% CI 0.63 to 0.92, 1 study, 502 participants; moderate-quality evidence), but may not significantly improve overall survival (HR 0.81, 95% CI 0.65 to 1.01, 2 studies, 1157 participants; low-quality evidence). Compared to chemotherapy alone, anti-CTLA4 monoclonal antibodies is likely to be associated with higher toxicity rates (RR 1.69, 95% CI 1.19 to 2.42, 2 studies, 1142 participants; moderate-quality evidence).Compared to chemotherapy, anti-PD1 monoclonal antibodies (immune checkpoint inhibitors) improved overall survival (HR 0.42, 95% CI 0.37 to 0.48, 1 study, 418 participants; high-quality evidence) and probably improved progression-free survival (HR 0.49, 95% CI 0.39 to 0.61, 2 studies, 957 participants; moderate-quality evidence). Anti-PD1 monoclonal antibodies may also result in less toxicity than chemotherapy (RR 0.55, 95% CI 0.31 to 0.97, 3 studies, 1360 participants; low-quality evidence).Anti-PD1 monoclonal antibodies performed better than anti-CTLA4 monoclonal antibodies in terms of overall survival (HR 0.63, 95% CI 0.60 to 0.66, 1 study, 764 participants; high-quality evidence) and progression-free survival (HR 0.54, 95% CI 0.50 to 0.60, 2 studies, 1465 participants; high-quality evidence). Anti-PD1 monoclonal antibodies may result in better toxicity outcomes than anti-CTLA4 monoclonal antibodies (RR 0.70, 95% CI 0.54 to 0.91, 2 studies, 1465 participants; low-quality evidence).Compared to anti-CTLA4 monoclonal antibodies alone, the combination of anti-CTLA4 plus anti-PD1 monoclonal antibodies was associated with better progression-free survival (HR 0.40, 95% CI 0.35 to 0.46, 2 studies, 738 participants; high-quality evidence). There may be no significant difference in toxicity outcomes (RR 1.57, 95% CI 0.85 to 2.92, 2 studies, 764 participants; low-quality evidence) (no data for overall survival were available).The class of small-molecule targeted drugs, BRAF inhibitors (which are active exclusively against BRAF-mutated melanoma), performed better than chemotherapy in terms of overall survival (HR 0.40, 95% CI 0.28 to 0.57, 2 studies, 925 participants; high-quality evidence) and progression-free survival (HR 0.27, 95% CI 0.21 to 0.34, 2 studies, 925 participants; high-quality evidence), and there may be no significant difference in toxicity (RR 1.27, 95% CI 0.48 to 3.33, 2 studies, 408 participants; low-quality evidence).Compared to chemotherapy, MEK inhibitors (which are active exclusively against BRAF-mutated melanoma) may not significantly improve overall survival (HR 0.85, 95% CI 0.58 to 1.25, 3 studies, 496 participants; low-quality evidence), but they probably lead to better progression-free survival (HR 0.58, 95% CI 0.42 to 0.80, 3 studies, 496 participants; moderate-quality evidence). However, MEK inhibitors probably have higher toxicity rates (RR 1.61, 95% CI 1.08 to 2.41, 1 study, 91 participants; moderate-quality evidence).Compared to BRAF inhibitors, the combination of BRAF plus MEK inhibitors was associated with better overall survival (HR 0.70, 95% CI 0.59 to 0.82, 4 studies, 1784 participants; high-quality evidence). BRAF plus MEK inhibitors was also probably better in terms of progression-free survival (HR 0.56, 95% CI 0.44 to 0.71, 4 studies, 1784 participants; moderate-quality evidence), and there appears likely to be no significant difference in toxicity (RR 1.01, 95% CI 0.85 to 1.20, 4 studies, 1774 participants; moderate-quality evidence).Compared to chemotherapy, the combination of chemotherapy plus anti-angiogenic drugs was probably associated with better overall survival (HR 0.60, 95% CI 0.45 to 0.81; moderate-quality evidence) and progression-free survival (HR 0.69, 95% CI 0.52 to 0.92; moderate-quality evidence). There may be no difference in terms of toxicity (RR 0.68, 95% CI 0.09 to 5.32; low-quality evidence). All results for this comparison were based on 324 participants from 2 studies.Network meta-analysis focused on chemotherapy as the common comparator and currently approved treatments for which high- to moderate-quality evidence of efficacy (as represented by treatment effect on progression-free survival) was available (based on the above results) for: biochemotherapy (with both interferon-alpha and interleukin-2); anti-CTLA4 monoclonal antibodies; anti-PD1 monoclonal antibodies; anti-CTLA4 plus anti-PD1 monoclonal antibodies; BRAF inhibitors; MEK inhibitors, and BRAF plus MEK inhibitors. Analysis (which included 19 RCTs and 7632 participants) generated 21 indirect comparisons.The best evidence (moderate-quality evidence) for progression-free survival was found for the following indirect comparisons:• both combinations of immune checkpoint inhibitors (HR 0.30, 95% CI 0.17 to 0.51) and small-molecule targeted drugs (HR 0.17, 95% CI 0.11 to 0.26) probably improved progression-free survival compared to chemotherapy;• both BRAF inhibitors (HR 0.40, 95% CI 0.23 to 0.68) and combinations of small-molecule targeted drugs (HR 0.22, 95% CI 0.12 to 0.39) were probably associated with better progression-free survival compared to anti-CTLA4 monoclonal antibodies;• biochemotherapy (HR 2.81, 95% CI 1.76 to 4.51) probably lead to worse progression-free survival compared to BRAF inhibitors;• the combination of small-molecule targeted drugs probably improved progression-free survival (HR 0.38, 95% CI 0.21 to 0.68) compared to anti-PD1 monoclonal antibodies;• both biochemotherapy (HR 5.05, 95% CI 3.01 to 8.45) and MEK inhibitors (HR 3.16, 95% CI 1.77 to 5.65) were probably associated with worse progression-free survival compared to the combination of small-molecule targeted drugs; and• biochemotherapy was probably associated with worse progression-free survival (HR 2.81, 95% CI 1.54 to 5.11) compared to the combination of immune checkpoint inhibitors.The best evidence (moderate-quality evidence) for toxicity was found for the following indirect comparisons:• combination of immune checkpoint inhibitors (RR 3.49, 95% CI 2.12 to 5.77) probably increased toxicity compared to chemotherapy;• combination of immune checkpoint inhibitors probably increased toxicity (RR 2.50, 95% CI 1.20 to 5.20) compared to BRAF inhibitors;• the combination of immune checkpoint inhibitors probably increased toxicity (RR 3.83, 95% CI 2.59 to 5.68) compared to anti-PD1 monoclonal antibodies; and• biochemotherapy was probably associated with lower toxicity (RR 0.41, 95% CI 0.24 to 0.71) compared to the combination of immune checkpoint inhibitors.Network meta-analysis-based ranking suggested that the combination of BRAF plus MEK inhibitors is the most effective strategy in terms of progression-free survival, whereas anti-PD1 monoclonal antibodies are associated with the lowest toxicity.Overall, the risk of bias of the included trials can be considered as limited. When considering the 122 trials included in this review and the seven types of bias we assessed, we performed 854 evaluations only seven of which (< 1%) assigned high risk to six trials. We found high-quality evidence that many treatments offer better efficacy than chemotherapy, especially recently implemented treatments, such as small-molecule targeted drugs, which are used to treat melanoma with specific gene mutations. Compared with chemotherapy, biochemotherapy (in this case, chemotherapy combined with both interferon-alpha and interleukin-2) and BRAF inhibitors improved progression-free survival; BRAF inhibitors (for BRAF-mutated melanoma) and anti-PD1 monoclonal antibodies improved overall survival. However, there was no difference between polychemotherapy and monochemotherapy in terms of achieving progression-free survival and overall survival. Biochemotherapy did not significantly improve overall survival and has higher toxicity rates compared with chemotherapy.There was some evidence that combined treatments worked better than single treatments: anti-PD1 monoclonal antibodies, alone or with anti-CTLA4, improved progression-free survival compared with anti-CTLA4 monoclonal antibodies alone. Anti-PD1 monoclonal antibodies performed better than anti-CTLA4 monoclonal antibodies in terms of overall survival, and a combination of BRAF plus MEK inhibitors was associated with better overall survival for BRAF-mutated melanoma, compared to BRAF inhibitors alone.The combination of BRAF plus MEK inhibitors (which can only be administered to people with BRAF-mutated melanoma) appeared to be the most effective treatment (based on results for progression-free survival), whereas anti-PD1 monoclonal antibodies appeared to be the least toxic, and most acceptable, treatment.Evidence quality was reduced due to imprecision, between-study heterogeneity, and substandard reporting of trials. Future research should ensure that those diminishing influences are addressed. Clinical areas of future investigation should include the longer-term effect of new therapeutic agents (i.e. immune checkpoint inhibitors and targeted therapies) on overall survival, as well as the combination of drugs used in melanoma treatment; research should also investigate the potential influence of biomarkers.


Specific allergen immunotherapy for the treatment of allergic asthma: a review of current evidence.
Asthma is frequently associated with atopy, characterized by the production of specific immunoglobulin E in response to environmental allergens. Currently, two types of allergen immunotherapy (AIT) are used in clinical practice: subcutaneous and sublingual immunotherapy, both accepted as key components of the therapeutic repertoire for allergic rhinitis and conjunctivitis. However, their role in asthma remains controversial. The present document is aimed at providing the clinicians with a review of the evidence on the use of AIT in asthma, focusing on the most relevant aspects of its mechanism of action, its efficacy, and existing data on safety, tolerability, and cost-effectivity, both in pediatric and adult populations. A systematic search of MEDLINE, Cochrane, and Clinical Trials databases from 2000 to April of 2016 was carried out by a panel of experts from the Spanish Allergy and Clinical Immunology Scientific Society. Relevant studies prior to the year 2000 included in ulterior systematic reviews were also considered. More than 4000 articles were identified during the search and 241 were selected to retrieve available evidence on AIT, which was graded according to the Oxford classification. All the group members reviewed the resulting text until the final version reached the consensual agreement. A summary of recommendations on the more relevant topics are proposed. The role of AIT as a valuable therapeutic strategy for prevention of exacerbation and progressive decline in lung function is highlighted. Future research should include specific tools for asthma evaluation when assessing AIT effectiveness in asthmatic patients.


Gastric cancer surgery in cirrhotic patients: result of gastrectomy with D2 lymph node dissection.
To explore the feasibility of performing gastrectomy with D2 lymphadenectomy in gastric cancer patients with liver cirrhosis. A total of 7,178 patients were admitted with a diagnosis of liver cirrhosis from January 1993 to December 2003. We reviewed the records of 142 patients who were diagnosed with liver cirrhosis and gastric adenocarcinoma during the same period. Gastrectomy with D2 lymph node dissection for carcinoma of the stomach was performed in 94 patients with histologically proven hepatic cirrhosis. All but 12 patients were classified as Child's class A. Only 35 patients (37.2%) were diagnosed with cirrhosis before operation. Seventy-three patients underwent a subtotal gastrectomy (77.7%) and 21 patients (22.3%) underwent a total gastrectomy, each with D2 or more lymph node dissection. Two patients (3.8%) who had prophylactic intra-operative drain placement, died of postoperative complications from hepatorenal failure with intractable ascites. Thirty-seven patients (39.4%) experienced postoperative complications. The extent of gastric resection did not influence the morbidity whereas serum aspartate aminotransferase level (P = 0.011) and transfusion did (P = 0.008). The most common postoperative complication was ascites (13.9%) followed by wound infection (10.6%). We concluded that the presence of compensated cirrhosis, i.e. Child class A, is not a contraindication against gastrectomy with D2 or more lymph node dissection, when curative resection for gastric cancer is possible. Hepatic reserve and meticulous hemostasis are the likely determinants of operative prognosis.


Narrow band 3 × 3 Mueller polarimetric endoscopy.
Mueller matrix polarimetric imaging has shown potential in tissue diagnosis but is challenging to implement endoscopically. In this work, a narrow band 3 × 3 Mueller matrix polarimetric endoscope was designed by rotating the endoscope to generate 0°, 45° and 90° linearly polarized illumination and positioning a rotating filter wheel in front of the camera containing three polarisers to permit polarization state analysis for backscattered light. The system was validated with a rotating linear polarizer and a diffuse reflection target. Initial measurements of 3 × 3 Mueller matrices on a rat are demonstrated, followed by matrix decomposition into the depolarization and retardance matrices for further analysis. Our work shows the feasibility of implementing polarimetric imaging in a rigid endoscope conveniently and economically in order to reveal diagnostic information.


Identification of undiagnosed diabetes and quality of diabetes care in the United States: cross-sectional study of 11.5 million primary care electronic records.
Electronic diabetes registers promote structured care and enable identification of undiagnosed diabetes, but they require consistent coding of the diagnosis in electronic medical records. We investigated the potential of electronic medical records to identify undiagnosed diabetes and to support diabetes management in a large primary care population in the United States. We conducted a cross-sectional study and retrospective observational cohort analysis of primary care electronic medical records from a nationally representative US database (GE Centricity). We tested the feasibility of identifying patients with undiagnosed diabetes by applying simple algorithms to the electronic medical record data. We compared the quality of care provided to patients in the United States who had diabetes (coded and uncoded) for at least 15 months with the quality of care provided in England using a set of 16 indicators. We included 11 540 454 electronic medical records from more than 9000 primary care clinics across the United States. Of the 1 110 398 records indicating diagnosed diabetes, only 61.9% contained a diagnostic code. Of the 10 430 056 records for nondiabetic patients, 0.4% (n = 40 359) had at least 2 abnormal fasting or random blood glucose values, and 0.2% (n = 23 261) of the remaining records had at least 1 documented glycated hemoglobin (HbA1c) value of 6.5% or higher. Among the 622 260 patients for whom information on quality-of-care indicators was available, those with a coded diagnosis of diabetes had a significantly higher level of quality of care than those with uncoded diabetes (p < 0.01); however, the quality of care was generally lower than that indicated in England. We were able to identify a substantial number of patients with uncoded diabetes and probable undiagnosed diabetes using simple algorithms applied to the primary care electronic records. Electronic coding of the diagnosis was associated with improved quality of care. Electronic diabetes registers are underused in US primary care and provide opportunities to facilitate the systematic, structured approach that is established in England.


Current management of adrenal tumors.
Adrenal tumors evoke considerable interest and diagnostic challenges. This rare group of tumors includes functional tumors with a gamut of clinical presentations, as well as adrenocortical carcinoma, with its advanced disease at presentation and dismal prognosis posing additional challenge. Increasing detection of incidentalomas adds further interest with the concomitant diagnostic and management dilemmas. Significant advances have been made in diagnostic imaging modalities for identifying malignancy risk in adrenal incidentalomas. Considerable progress has occurred in understanding adrenocortical carcinoma pathogenesis from the study of genetics at the germline level in familial carcinomas, as well as at the somatic level by analyzing molecular alterations in sporadic tumors; this research supplies opportunities to develop novel therapeutic agents against a tumor with poor prognosis. Laparoscopic adrenalectomy has emerged as standard of care in the treatment of functional benign adenomas and nonfunctional tumors larger than 4 cm when adrenocortical carcinoma is not suspected. Open adrenalectomy with en-bloc excision has been the mainstay for primary and recurrent adrenocortical carcinoma due to the lack of effective adjuvant therapy. International consensus conferences have attempted to standardize diagnostic and treatment approaches in the management of adrenal tumors; further research is necessary.


Identification of marrow-derived and thymus-derived small lymphocytes in the lymphoid tissue and thoracic duct lymph of normal rats.
These experiments show that small lymphocytes from the thoracic duct of rats are normally a mixture of thymus-derived and marrow-derived cells, and define the traffic areas in lymphoid tissues through which the two populations recirculate. Thoracic duct lymphocytes were labeled in vitro with uridine-(3)H and their histological distribution in the lymphoid tissues of normal recipients was demonstrated by radioautography. Labeled lymphocytes occupied two adjacent areas distinguished by a marked difference in the intensity of labeling; heavily labeled cells were found in thymus-dependent traffic areas of lymphocyte recirculation, while lightly labeled cells localized in the thymus-independent follicular areas around germinal centers. A corresponding heterogeneity of uridine uptake among small lymphocytes from normal donors was demonstrated by sedimentation at 1 g; slowly sedimenting cells incorporated little uridine and localized in follicular areas after transfusion while rapidly sedimenting cells incorporated more uridine and localized in thymus-dependent areas after transfusion. Experimentally prepared marrow-derived small lymphocytes behaved in sedimentation studies and after transfusion like a pure population of the lightly labeled small lymphocytes in normal lymph. Artificially reconstituted mixtures of marrow-derived and thymus-derived lymphocytes were qualitatively indistinguishable from normal lymphocyte populations.


Alveolar macrophages are essential for protection from respiratory failure and associated morbidity following influenza virus infection.
Alveolar macrophages (AM) are critical for defense against bacterial and fungal infections. However, a definitive role of AM in viral infections remains unclear. We here report that AM play a key role in survival to influenza and vaccinia virus infection by maintaining lung function and thereby protecting from asphyxiation. Absence of AM in GM-CSF-deficient (Csf2-/-) mice or selective AM depletion in wild-type mice resulted in impaired gas exchange and fatal hypoxia associated with severe morbidity to influenza virus infection, while viral clearance was affected moderately. Virus-induced morbidity was far more severe in Csf2-/- mice lacking AM, as compared to Batf3-deficient mice lacking CD8α+ and CD103+ DCs. Csf2-/- mice showed intact anti-viral CD8+ T cell responses despite slightly impaired CD103+ DC development. Importantly, selective reconstitution of AM development in Csf2rb-/- mice by neonatal transfer of wild-type AM progenitors prevented severe morbidity and mortality, demonstrating that absence of AM alone is responsible for disease severity in mice lacking GM-CSF or its receptor. In addition, CD11c-Cre/Ppargfl/fl mice with a defect in AM but normal adaptive immunity showed increased morbidity and lung failure to influenza virus. Taken together, our results suggest a superior role of AM compared to CD103+ DCs in protection from acute influenza and vaccinia virus infection-induced morbidity and mortality.


The efficacy of pediatric advanced life support training in emergency medical service providers.
Pediatric advanced life support (PALS) teaches skills unique to pediatric resuscitation. The purpose of this study was to assess the effect of PALS training among emergency medical service (EMS) providers in out-of-hospital trauma and medical resuscitations. A physician panel evaluated all EMS run sheets of pediatric traumas and medical resuscitations brought to a tertiary children's hospital/regional trauma center over a 3-year period. In 183 responses, EMS personnel were the sole providers of medical stabilization. Evaluation included the ability to secure an airway, establish vascular access, shock recognition, and appropriate cardiac rhythm assessment and resuscitation. The panel was blinded to the PALS training status of the responding EMS squad until completion of the review. Pediatric advanced life support-trained EMS personnel responded to 36% of the resuscitations reviewed. A significant difference in successful intubations was noted in PALS-trained squads compared with squads with no PALS training (85% vs 48%; P < 0.001). A significant difference was also noted in the ability to obtain vascular access in shock/arrest cases (100% vs 70%; P < 0.001). Similarly, PALS-trained squads were more successful in intraosseous line placement than non-PALS-trained squads (100% vs 55%; P < 0.01). However, despite better procedural skills, there was no difference in mortality rates between the groups (37% PALS vs 32% non-PALS). We conclude that PALS training improves procedural skills among EMS personnel and should be strongly considered as part of EMS training.


Divergent access to N-hydroxypyrroles and isoxazoles via the gold(i)- or Brønsted acid-catalysed regioselective cyclization of N-(2-trifluoromethyl-3-alkynyl) oximes.
The divergent regioselective cyclization of N-(2-trifluoromethyl-3-alkynyl) oximes by a suitable choice of a gold(i) or a Brønsted acid catalyst, leading to 4-trifluoromethyl-N-hydroxypyrroles or 4-trifluoromethyl-5-alkylisoxazoles was developed. In order to avoid the tedious separation of unstable N-(2-trifluoromethyl-3-alkynyl) oximes, an easy two-step, one-pot synthesis of 4-trifluoromethyl-5-alkylisoxazoles was realized via the sequential oxidation of the corresponding hydroxylamines and subsequent treatment with the reductant, sodium thiosulfate (Na2S2O3). This two-step, one-pot procedure is a complementary method for the synthesis of 4-trifluoromethyl-5-alkylisoxazole from those unstable N-(2-trifluoromethyl-3-alkynyl) oximes.


Effects of management, feeding, and treatment on clinical and biochemical variables in cattle with displaced abomasum.
To determine effects of management, feeding, and treatment on clinical and biochemical variables in cows with displaced abomasum (DA). 374 cows that received 470 treatments for DA. Blood and milk samples were obtained from 139 affected cows for analysis; for all cows, clinical data, management, feeding, and treatments were evaluated. Multiparous cows were more predisposed to DA than primiparous cows were, and Swedish Friesians were more predisposed than Swedish Red and Whites were. Eighty percent of cows had left-sided DA, and 20% had right-sided DA. In > 50% of affected cows, clinical signs appeared just before calving to 2 weeks after calving. Incidence of twin calves and periparturient diseases was significantly higher in affected cows than in the overall Swedish cow population. Content of neutral detergent fiber in the silage was low in herds with DA. Feeding a total mixed ration was a risk factor for DA. Treatment by surgical methods gave a significantly higher recovery rate than nonsurgical methods. Displaced abomasum is a periparturient nutritional disease. Feeding roughage with low neutral detergent content is a more important causative factor than the amount of concentrates fed at the time of calving. The basic principle for prevention of DA is to maintain good ruminal filling before and at calving. The amount of high-quality roughage fed before and at calving should be kept to a minimum. By changing routines for periparturient feeding, it should be possible to reduce the incidence of DA.


The temporal effect of platelet-rich plasma on pain and physical function in the treatment of knee osteoarthritis: systematic review and meta-analysis of randomized controlled trials.
Quite a few randomized controlled trials (RCTs) investigating the efficacy of platelet-rich plasma (PRP) for treatment of knee osteoarthritis (OA) have been recently published. Therefore, an updated systematic review was performed to evaluate the temporal effect of PRP on knee pain and physical function. Pubmed, Embase, Cochrane library, and Scopus were searched for human RCTs comparing the efficacy and/or safety of PRP infiltration with other intra-articular injections. A descriptive summary and quality assessment were performed for all the studies finally included for analysis. For studies reporting outcomes concerning Western Ontario and McMaster Universities Arthritis Index (WOMAC) or adverse events, a random-effects model was used for data synthesis. Fourteen RCTs comprising 1423 participants were included. The control included saline placebo, HA, ozone, and corticosteroids. The follow-up ranged from 12 weeks to 12 months. Risk of bias assessment showed that 4 studies were considered as moderate risk of bias and 10 as high risk of bias. Compared with control, PRP injections significantly reduced WOMAC pain subscores at 3, 6, and 12 months follow-up (p = 0.02, 0.004, <0.001, respectively); PRP significantly improved WOMAC physical function subscores at 3, 6, and 12 months (p = 0.002, 0.01, <0.001, respectively); PRP also significantly improved total WOMAC scores at 3, 6 and 12 months (all p < 0.001); nonetheless, PRP did not significantly increased the risk of post-injection adverse events (RR, 1.40 [95% CI, 0.80 to 2.45], I 2 = 59%, p = 0.24). Intra-articular PRP injections probably are more efficacious in the treatment of knee OA in terms of pain relief and self-reported function improvement at 3, 6 and 12 months follow-up, compared with other injections, including saline placebo, HA, ozone, and corticosteroids. PROSPERO CRD42016045410 . Registered 8 August 2016.


Meta-analysis of combined therapy with angiotensin receptor antagonists versus ACE inhibitors alone in patients with heart failure.
There is insufficient evidence whether the benefit of adding angiotensin II receptor blockers (ARBs) to angiotensin-converting enzyme (ACE) inhibitors outweighs the increased risk of adverse effects in patients with heart failure. Two independent reviewers searched and abstracted randomized controlled trials of ARBs and ACE inhibitors compared to ACE inhibitor therapy alone in patients with heart failure reporting mortality and hospitalizations having a follow-up of at least 6 months identified by a systematic literature search. Eight trials including a total of 18,061 patients fulfilled our inclusion criteria. There was no difference between patients treated with combination therapy and ACE inhibitor therapy alone for overall mortality, hospitalization for any reason, fatal or nonfatal MI. Combination therapy was, however, associated with fewer hospital admissions for heart failure (RR 0.81, 95%CI 0.72-0.91), although there was significant heterogeneity across trials (p-value for heterogeneity = 0.04; I(2) = 57% [95%CI 0-83%]). Patients treated with combination therapy had a higher risk of worsening renal function and symptomatic hypotension, and their trial medications were more often permanently discontinued. Lack of individual patient data precluded the analysis of time-to-event data and identification of subgroups which potentially benefit more from combination therapy such as younger patients with preserved renal function and thus at lower risk to experience worsening renal function or hyperkalemia. Combination therapy with ARBs and ACE inhibitors reduces admissions for heart failure in patients with congestive heart failure when compared to ACE inhibitor therapy alone, but does not reduce overall mortality or all-cause hospitalization and is associated with more adverse events. Thus, based on current evidence, combination therapy with ARBs and ACE inhibitors may be reserved for patients who remain symptomatic on therapy with ACE inhibitors under strict monitoring for any signs of worsening renal function and/or symptomatic hypotension.


You won't know if you don't ask: discrepancy and ambivalence in attitudes toward behavior change.
Behavior change is challenging following an acute cardiac event, and commonly, individuals are ambivalent. The objective of this study was to describe the experience of behavior change of survivors of an acute cardiac event. Semistructured interviews were undertaken with 25 participants attending 3 cardiac rehabilitation programs. An inductive process of qualitative thematic analysis was used to analyze the transcripts. Analysis revealed ambivalence to change, misconceptions, and confusion about terminology. Discrepancies between what participants felt they should be doing and what they actually were doing reflected their ambivalence. Further inconsistencies were reflected in participants' misunderstandings and confusion regarding disease processes and management of heart disease. These findings reflect the misconception and ambivalence regarding behavior change that individuals experience. Clinicians may require greater skills in detecting conflicting or ambivalent discourse to support patients through sustainable health behavior change.


Role of Microalbuminuria in Predicting Cardiovascular Mortality in Individuals With Subclinical Hypothyroidism.
Studies suggest that subclinical hypothyroidism (SCH) is related to cardiovascular mortality (CVM). We explored the role of microalbuminuria (MIA) as a predictor of long-term CVM in population with and without SCH with normal kidney function. We examined the National Health and Nutrition Education Survey - III database (n = 6,812). Individuals younger than 40 years, thyroid-stimulating hormone levels ≥20 and ≤0.35mIU/L, estimated glomerular filtration rate <60mL/minute/1.73m2 and urine albumin-to-creatinine ratio of >250mg/g in men and >355mg/g in women were excluded. SCH was defined as thyroid-stimulating hormone levels between 5 and 19.99mIU/L and serum T4 levels between 5 and 12µg/dL. MIA was defined as urine albumin-to-creatinine ratio of 17-250mg/g in men and 25-355mg/g in women. Patients were categorized into the following 4 groups: (1) no SCH or MIA, (2) MIA, but no SCH, (3) SCH, but no MIA and (4) both SCH and MIA. Prevalence of MIA in the subclinical hypothyroid cohort was 21% compared to 16.4% in those without SCH (P = 0.03). SCH was a significant independent predictor of MIA (n = 6,812), after adjusting for traditional risk factors (unadjusted odds ratio = 1.75; 95% CI: 1.24-2.48; P = 0.002 and adjusted odds ratio = 1.83; 95% CI: 1.2-2.79; P = 0.006). MIA was a significant independent predictor of long-term all-cause (adjusted hazard ratio = 1.7, 95% CI: 1.24-2.33) and CVM (adjusted hazard ratio = 1.72, 95% CI: 1.07-2.76) in subclinical hypothyroid individuals. In a cohort of subclinical hypothyroid individuals, the presence of MIA predicts increased risk of CVM as compared to nonmicroalbuminurics with SCH. Further randomized trials are needed to assess the benefits of treating microalbuminuric subclinical hypothyroid individuals and impact on CVM.


Kinetics of contraction-induced GLUT4 translocation in skeletal muscle fibers from living mice.
Exercise is an important strategy for the treatment of type 2 diabetes. This is due in part to an increase in glucose transport that occurs in the working skeletal muscles. Glucose transport is regulated by GLUT4 translocation in muscle, but the molecular machinery mediating this process is poorly understood. The purpose of this study was to 1) use a novel imaging system to elucidate the kinetics of contraction-induced GLUT4 translocation in skeletal muscle and 2) determine the function of AMP-activated protein kinase alpha2 (AMPKalpha2) in this process. Confocal imaging was used to visualize GLUT4-enhanced green fluorescent protein (EGFP) in transfected quadriceps muscle fibers in living mice subjected to contractions or the AMPK-activator AICAR. Contraction increased GLUT4-EGFP translocation from intracellular vesicle depots to both the sarcolemma and t-tubules with similar kinetics, although translocation was greater with contractions elicited by higher voltage. Re-internalization of GLUT4 did not begin until 10 min after contractions ceased and was not complete until 130 min after contractions. AICAR increased GLUT4-EGFP translocation to both sarcolemma and t-tubules with similar kinetics. Ablation of AMPKalpha2 activity in AMPKalpha2 inactive transgenic mice did not change GLUT4-EGFP's basal localization, contraction-stimulated intracellular GLUT4-EGFP vesicle depletion, translocation, or re-internalization, but diminished AICAR-induced translocation. We have developed a novel imaging system to study contraction-stimulated GLUT4 translocation in living mice. Contractions increase GLUT4 translocation to the sarcolemma and t-tubules with similar kinetics and do not require AMPKalpha2 activity.


A method for assessing the potential for confounding applied to ionic strength in central Appalachian streams.
Causal relationships derived from field data are potentially confounded by variables that are correlated with both the cause and its effect. The present study presents a method for assessing the potential for confounding and applies it to the relationship between ionic strength and impairment of benthic invertebrate assemblages in central Appalachian streams. The method weighs all available evidence for and against confounding by each potential confounder. It identifies 10 types of evidence for confounding, presents a qualitative scoring system, and provides rules for applying the scores. Twelve potential confounders were evaluated: habitat, organic enrichment, nutrients, deposited sediments, pH, selenium, temperature, lack of headwaters, catchment area, settling ponds, dissolved oxygen, and metals. One potential confounder, low pH, was found to be biologically significant and eliminated by removing sites with pH < 6. Other potential confounders were eliminated based on the weight of evidence. This method was found to be useful and defensible. It could be applied to other environmental assessments that use field data to develop causal relationships, including contaminated site remediation or management of natural resources.


Inhibitory Effect of a French Maritime Pine Bark Extract-Based Nutritional Supplement on TNF-α-Induced Inflammation and Oxidative Stress in Human Coronary Artery Endothelial Cells.
Oxidative stress and inflammation, leading to endothelial dysfunction, contribute to the pathogenesis of atherosclerosis. The popularity of natural product supplements has increased in recent years, especially those with purported anti-inflammatory and/or antioxidant effects. The efficacy and mechanism of many of these products are not yet well understood. In this study, we tested the antioxidant and anti-inflammatory effects of a supplement, HIPER Health Supplement (HIPER), on cytokine-induced inflammation and oxidative stress in human coronary artery endothelial cells (HCAECs). HIPER is a mixture of French maritime pine bark extract (PBE), honey, aloe vera, and papaya extract. Treatment for 24 hours with HIPER reduced TNF-α-induced reactive oxygen species (ROS) generation that was associated with decreased NADPH oxidase 4 and increased superoxide dismutase-1 expression. HIPER inhibited TNF-α induced monocyte adhesion to HCAECs that was in keeping with decreased expression of vascular cell adhesion molecule-1 and intercellular cell adhesion molecule-1 and decreased nuclear factor-kappa B (NF-κB) activation. Further investigation of mechanism showed HIPER reduced TNF-α induced IκBα and p38 and MEK1/2 MAP kinases phosphorylation. Our findings show that HIPER has potent inhibitory effects on HCAECs inflammatory and oxidative stress responses that may protect against endothelial dysfunction that underlies early atherosclerotic lesion formation.


Global assessment of seasonal potential distribution of Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae).
The Mediterranean fruit fly (Medfly) is one of the world's most economically damaging pests. It displays highly seasonal population dynamics, and the environmental conditions suitable for its abundance are not constant throughout the year in most places. An extensive literature search was performed to obtain the most comprehensive data on the historical and contemporary spatio-temporal occurrence of the pest globally. The database constructed contained 2328 unique geo-located entries on Medfly detection sites from 43 countries and nearly 500 unique localities, as well as information on hosts, life stages and capture method. Of these, 125 localities had information on the month when Medfly was recorded and these data were complemented by additional material found in comprehensive databases available online. Records from 1980 until present were used for medfly environmental niche modeling. Maximum Entropy Algorithm (MaxEnt) and a set of seasonally varying environmental covariates were used to predict the fundamental niche of the Medfly on a global scale. Three seasonal maps were also produced: January-April, May-August and September-December. Models performed significantly better than random achieving high accuracy scores, indicating a good discrimination of suitable versus unsuitable areas for the presence of the species.


Choline administration: activation of tyrosine hydroxylase in dopaminergic neurons of rat brain.
The administration of choline in doses previously shown to elevate brain acetylcholine concentrations also increases the activity of tyrosine hydroxylase in rat caudate nuclei. This response can be blocked by atropine, a muscarinic antagonist. These findings indicate that choline-induced increases in acetylcholine concentrations may be associated with parallel changes in the amount of the neurotransmitter released into synapses.


How attention can create synaptic tags for the learning of working memories in sequential tasks.
Intelligence is our ability to learn appropriate responses to new stimuli and situations. Neurons in association cortex are thought to be essential for this ability. During learning these neurons become tuned to relevant features and start to represent them with persistent activity during memory delays. This learning process is not well understood. Here we develop a biologically plausible learning scheme that explains how trial-and-error learning induces neuronal selectivity and working memory representations for task-relevant information. We propose that the response selection stage sends attentional feedback signals to earlier processing levels, forming synaptic tags at those connections responsible for the stimulus-response mapping. Globally released neuromodulators then interact with tagged synapses to determine their plasticity. The resulting learning rule endows neural networks with the capacity to create new working memory representations of task relevant information as persistent activity. It is remarkably generic: it explains how association neurons learn to store task-relevant information for linear as well as non-linear stimulus-response mappings, how they become tuned to category boundaries or analog variables, depending on the task demands, and how they learn to integrate probabilistic evidence for perceptual decisions.


Nanoscale surface engineered living cells with extended substrate spectrum.
We report on cell surface engineering of living microorganisms by using Layer-by-Layer (LbL) technology to extend the substrate spectrum. The yeast Arxula adeninivorans LS3 (Arxula) was employed as a model organism and biological template. By using LbL technology, Arxula cells were encapsulated by polyelectrolyte and enzyme layers. The biological activity of the Arxula was retained after the encapsulation process. The polymeric capsule surrounding the Arxula provides a stable interface for surface engineering of living cells. LbL of polyelectrolytes followed by an enzyme layer of lactate oxidase were assembled. The outer enzyme layer provides an additional biological function for Arxula to convert the unfavourable substrate lactate into the favourable substrate pyruvate, thus extending the substrate spectrum of the organism. Moreover, capsule stability and enzyme conjugate stability of the surface engineered Arxula were studied.


A murine DC-SIGN homologue contributes to early host defense against Mycobacterium tuberculosis.
The C-type lectin dendritic cell-specific intercellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN) mediates the innate immune recognition of microbial carbohydrates. We investigated the function of this molecule in the host response to pathogens in vivo, by generating mouse lines lacking the DC-SIGN homologues SIGNR1, SIGNR3, and SIGNR5. Resistance to Mycobacterium tuberculosis was impaired only in SIGNR3-deficient animals. SIGNR3 was expressed in lung phagocytes during infection, and interacted with M. tuberculosis bacilli and mycobacterial surface glycoconjugates to induce secretion of critical host defense inflammatory cytokines, including tumor necrosis factor (TNF). SIGNR3 signaling was dependent on an intracellular tyrosine-based motif and the tyrosine kinase Syk. Thus, the mouse DC-SIGN homologue SIGNR3 makes a unique contribution to protection of the host against a pulmonary bacterial pathogen.