Modeling ecological minimum requirements for distribution of greater sage-grouse leks: implications for population connectivity across their western range, U.S.A.
Greater sage-grouse Centrocercus urophasianus (Bonaparte) currently occupy approximately half of their historical distribution across western North America. Sage-grouse are a candidate for endangered species listing due to habitat and population fragmentation coupled with inadequate regulation to control development in critical areas. Conservation planning would benefit from accurate maps delineating required habitats and movement corridors. However, developing a species distribution model that incorporates the diversity of habitats used by sage-grouse across their widespread distribution has statistical and logistical challenges. We first identified the ecological minimums limiting sage-grouse, mapped similarity to the multivariate set of minimums, and delineated connectivity across a 920,000 km(2) region. We partitioned a Mahalanobis D (2) model of habitat use into k separate additive components each representing independent combinations of species-habitat relationships to identify the ecological minimums required by sage-grouse. We constructed the model from abiotic, land cover, and anthropogenic variables measured at leks (breeding) and surrounding areas within 5 km. We evaluated model partitions using a random subset of leks and historic locations and selected D (2) (k = 10) for mapping a habitat similarity index (HSI). Finally, we delineated connectivity by converting the mapped HSI to a resistance surface. Sage-grouse required sagebrush-dominated landscapes containing minimal levels of human land use. Sage-grouse used relatively arid regions characterized by shallow slopes, even terrain, and low amounts of forest, grassland, and agriculture in the surrounding landscape. Most populations were interconnected although several outlying populations were isolated because of distance or lack of habitat corridors for exchange. Land management agencies currently are revising land-use plans and designating critical habitat to conserve sage-grouse and avoid endangered species listing. Our results identifying attributes important for delineating habitats or modeling connectivity will facilitate conservation and management of landscapes important for supporting current and future sage-grouse populations.


The Role of the Anabolic Properties of Plant- versus Animal-Based Protein Sources in Supporting Muscle Mass Maintenance: A Critical Review.
Plant-sourced proteins offer environmental and health benefits, and research increasingly includes them in study formulas. However, plant-based proteins have less of an anabolic effect than animal proteins due to their lower digestibility, lower essential amino acid content (especially leucine), and deficiency in other essential amino acids, such as sulfur amino acids or lysine. Thus, plant amino acids are directed toward oxidation rather than used for muscle protein synthesis. In this review, we evaluate the ability of plant- versus animal-based proteins to help maintain skeletal muscle mass in healthy and especially older people and examine different nutritional strategies for improving the anabolic properties of plant-based proteins. Among these strategies, increasing protein intake has led to a positive acute postprandial muscle protein synthesis response and even positive long-term improvement in lean mass. Increasing the quality of protein intake by improving amino acid composition could also compensate for the lower anabolic potential of plant-based proteins. We evaluated and discussed four nutritional strategies for improving the amino acid composition of plant-based proteins: fortifying plant-based proteins with specific essential amino acids, selective breeding, blending several plant protein sources, and blending plant with animal-based protein sources. These nutritional approaches need to be profoundly examined in older individuals in order to optimize protein intake for this population who require a high-quality food protein intake to mitigate age-related muscle loss.


Eye protection in orthopaedic surgery. An in vitro study of various forms of eye protection and their effectiveness.
Conjunctival contamination from splashed debris during orthopaedic surgical procedures places surgeons at risk for communicable diseases such as human immunodeficiency virus (HIV) and hepatitis B and C. The purpose of this study was to compare the effectiveness of various types of protective eyewear in preventing conjunctival contamination. A simulation model was constructed with use of a mannequin head in a typical position of a surgeon's head during an operation. The head was placed at an appropriate distance from the surgical field, and a femoral osteotomy was performed on a cadaver thigh. Six experimental groups were tested to determine the ability of various types of eye protection to prevent contamination of the conjunctiva: (1) modern prescription glasses, (2) standard surgical telescopic loupes, (3) hard plastic contoured glasses, (4) disposable plastic glasses, (5) a combination facemask and eye shield, and (6) no protection (control). Thirty femoral osteotomies were performed, and contamination of both the protective devices and the simulated conjunctival surfaces were recorded. None of the tested devices were completely effective. The modern prescription glasses and the controls both were associated with conjunctival contamination rates of 83%. The other eye protective devices were associated with significantly lower rates of overall contamination, with a rate of 50% for the loupes, 30% for the facemask and eye shield, 17% for the hard plastic glasses, and 3% for the disposable plastic glasses. Modern prescription glasses provided no benefit over the control in our experimental model; therefore, we do not recommend that they be used as the sole eye protection, especially during surgical procedures in which there is a high rate of debris expulsion from the wound. Readily available and disposable plastic glasses were associated with the lowest rate of conjunctival contamination (3%) and are an effective means with which to protect the orthopaedic surgeon from communicable diseases by conjunctival contamination.


[Change in certain forms of aggressive behavior and the concentration of monoamines in the brain during selection of wild rats for taming].
Norway rats have been selected during 20 generations by the absence of aggressive reaction to man (tamed rats). From 7 up to 20th generations of selection, different forms of aggressive behaviour (reaction to glove, intermale, shock-induced aggression and predatory aggression) were studied, and the level of noradrenaline, serotonin and its metabolite 5-hydroxyindoleacetic acid was determined in the brain. In the absence of aggressive reaction to glove in tamed rats, the shock-induced aggression considerably decreased while the predatory aggressiveness (mouse-killing behaviour) and intermale aggressiveness did not change. Beginning from 15-16th generation of selection, a higher level of the 5-hydroxyindoleacetic acid in the hypothalamus was established, in the 20th generation an increased content of serotonin was revealed in the hypothalamus and the midbrain. In some generations of selection an increased level of noradrenaline in the hypothalamus in comparison to wild rats was observed. A conclusion is made that the selection of animals by taming unequally influences different kinds of aggressiveness and is accompanied by inherited consolidated reorganization of the monoamine brain systems.


Elevation of serum C-reactive protein levels is associated with obesity in boys.
This study aimed to reveal the relationships among C-reactive protein (CRP), obesity, blood pressure (BP), and serum lipids in children. Eighty-six obese and 58 non-obese boys aged an average of 11.2 years were examined. Serum CRP levels were measured by high sensitivity latex turbidimetric immunoassay and subjects with CRP levels below 0.3 mg/dl were adopted. Comparisons of serum CRP levels, BP, and serum lipids levels between age-matched obese and non-obese groups were performed. A comparison of serum CRP levels among the percentage of relative weight quartiles and the relationships among percentage of relative weight, BP, and serum lipids in CRP quartiles were analyzed. The relationships between CRP and other parameters were analyzed by simple and stepwise multiple regressions. Obese children had significantly higher high-sensitivity CRP (hs-CRP) levels than their non-obese counterparts. The mean hs-CRP level was 5.5-fold higher in the top quartile of the percentage of relative weight than in the bottom quartile. In the top quartile of CRP, the percentage of relative weight, systolic BP, diastolic BP, pulse pressure, and low density/high density lipoprotein-cholesterol (LDL-C/HDL-C) were significantly higher than in the bottom quartile. The percentage of relative weight, BP, LDL-C, and apolipoprotein B (ApoB) showed positive correlations and HDL-C showed a negative correlation with log CRP by simple regression. Stepwise multiple regression analysis indicated that only the percentage of relative weight was strongly related to CRP. In conclusion, this study revealed a significant relationship between CRP and obesity in children. Obese children tended to have high CRP levels, BP elevation, and slight dyslipidemia. These results support the findings that CRP is one of the useful indices of childhood obesity that would affect the progression to future atherosclerotic disease. We consider that a strategy of preventing obesity from childhood would contribute to a drop in the future incidence of metabolic syndromes.


Thermal cautery of the canine third eyelid for treatment of cartilage eversion.
To present a novel, minimally invasive technique for everted third eyelid cartilage correction in dogs that employs the use of low-energy cautery to remodel the cartilage. Twelve eyes of ten dogs had cautery performed under general anesthesia to correct everted third eyelid cartilage. The tip of a handheld cautery unit or an electrocautery handpiece was applied to the bulbar conjunctival surface of the third eyelid at the central location of cartilage convexity and treated to effect. This resulted in gradual conjunctival contraction and cartilage softening that remodeled the third eyelid to return to a more normal, physiologic position. When the tips of the cartilage bar were also curled, cautery was briefly applied to the convex surface to straighten the cartilage in a similar manner. Blanching of the conjunctiva at the site of treatment occurred. Char was sometimes present and was gently removed with a scalpel blade to improve postoperative patient comfort. Mild conjunctival hyperemia was noted in a few patients for 1-2 days after surgery, but there were no signs of discomfort or eyelid swelling. All dogs had good results in terms of cartilage correction with no recurrence; however, one of the Great Danes that had concurrent third eyelid gland prolapse required gland replacement surgery. Thermal cautery is a simple, inexpensive means of correcting third eyelid cartilage eversion in dogs with a high rate of success that preserves normal tissue while restoring function.


Altered Gene Response to Aflatoxin B1 in the Spleens of Susceptible and Resistant Turkeys.
Susceptibility and/or resistance to aflatoxin B1 (AFB1) is a threshold trait governed principally by glutathione S transferase (GST)-mediated detoxification. In poultry, domesticated turkeys are highly sensitive to AFB1, most likely due to dysfunction in hepatic GSTs. In contrast, wild turkeys are comparatively resistant to aflatoxicosis due to the presence of functional hepatic GSTAs and other possible physiological and immunological interactions. The underlying genetic basis for the disparate GST function in turkeys is unknown as are the broader molecular interactions that control the systemic response. This study quantifies the effects of dietary AFB1 on gene expression in the turkey spleen, specifically contrasting genetically distinct domesticated (DT, susceptible) and Eastern wild (EW, resistant) birds. Male turkey poults were subjected to a short-term AFB1 treatment protocol with feed supplemented with 320 ppb AFB1 beginning on day 15 of age and continuing for 14 days. Spleen tissues were harvested and subjected to deep RNA sequencing and transcriptome analysis. Analysis of differential gene expression found the effects of AFB1 treatment on the spleen transcriptomes considerably more prominent in the DT birds compared to EW. However, expression of the differentially expressed genes (DEGs) was directionally biased, with the majority showing higher expression in EW (i.e., down-regulation in DT). Significantly altered pathways included FXR/RXR and LXR/RXR activation, coagulation system, prothrombin activation, acute phase response, and atherosclerosis signaling. Differential extra-hepatic expression of acute phase protein genes was confirmed by quantitative real time PCR (qRT-PCR) in the original experiment and additional turkey lines. Results demonstrate that wild turkeys possess a capacity to more effectively respond to AFB1 exposure.


Evaluation of selenium status in pure bred duroc sows and their progeny.
The aim of this trial was to determine selenium status in pure bred duroc sows and their progeny and to compare it to Czech Large White x Landrace breed. The pregnant duroc sows (n=12) and pregnant Czech Large White x Landrace sows (n=12) were fed identical diets supplemented with sodium selenite. During lactation significantly higher serum Se concentrations (p<0.001) were found in duroc piglets. Also significantly higher serum GSH-Px activities (p<0.001) were found in duroc sows and piglets. No differences in concentrations of Se in colostrums and milk were found between the two breeds. Our findings support the hypothesis that there may be breed differences in indices of selenium status in pigs.


Efficient visible and near-infrared photoluminescent attapulgite-based lanthanide one-dimensional nanomaterials assembled by ion-pairing interactions.
Attapulgite, a one-dimensional fibrillar nanomaterial present in nature, with its extreme stability, is a promising material to act as a new carrier of luminescent lanthanide complexes for further applications. Herein, a series of lanthanide complexes Na[Ln(TTA)4] have been attached to attapulgite (Atta) via ion-pairing interactions, generating the first example of attapulgite-based visible and near-infrared (NIR) luminescent lanthanide one-dimensional nanomaterials, where TTA is 2-thenoyltrifluoroacetonate and Ln is Eu, Sm, Nd, Er or Yb. The hybrid materials were characterized by CHN elemental analysis, inductively coupled plasma-atomic emission spectroscopy (ICP), powder X-ray diffraction (PXRD), thermogravimetry (TG), transmission electron microscopy (TEM), and UV-vis absorption spectra. In order to investigate the photophysical behaviours of these materials, the visible and NIR luminescent spectra and the energy transfer process have been systematically investigated. Moreover, efforts have been made to produce Eu- and Sm-based plastic attapulgite materials by utilizing poly(methyl methacrylate) (PMMA) matrices, and the dispersibility of the lanthanide-doped hybrids in PMMA provides them with a high mechanical strength. The lanthanide-doped attapulgite appears to be an interesting material for photophysical applications. The results of this work would have potential significance for the design and assembly of luminescent lanthanide materials for light-emitting diodes (LED), sunlight-conversion films, optical amplifiers, solar concentrators, and lasers.


Processing of engulfed apoptotic bodies yields T cell epitopes.
Programmed death via apoptosis is the metazoan physiologic mode of cell death. Apoptotic cells are recognized by scavenger phagocytes via a number of membrane receptors and engulfed. Thereafter, little is known of their fate, or that of phagocytes. Here, we have traced apoptotic cells upon their engulfment by macrophages. After 3 h, apoptotic cells were contained in discrete well-defined vacuoles. Upon overnight chase, several small vesicles, possibly originating from the fragmentation of original vacuoles, were evident all over the macrophage body. Furthermore, Ags were diffused in the cytosol of some cells, which raises the possibility that epitopes from engulfed apoptotic cells may associate with macrophage MHC class I molecules and be recognized by T lymphocytes. Indeed, Ag-specific CTLs recognize and specifically lyse syngeneic macrophages upon phagocytosis of MHC class I-positive or -negative apoptotic cells, provided that they contain the relevant Ags. Synthesis and membrane expression of class I molecules by macrophages, together with functional transporters associated with Ag presentation, were necessary for recognition and lysis. The indirect presentation of epitopes from engulfed apoptotic cells by scavenger Ag-presenting phagocytes may, in the absence of "danger" signals, have implications for the establishment of central and peripheral self-tolerance.


Genetic testing for age-related macular degeneration: not indicated now.
Age-related macular degeneration is a very common condition that is caused by a complex interplay of genetic and environmental factors. It is likely that, in the future, genetic testing will allow physicians to achieve better clinical outcomes by administering specific treatments to patients based on their genotypes. However, improved outcomes for genotyped patients have not yet been demonstrated in a prospective clinical trial, and as a result, the costs and risks of routine genetic testing currently outweigh the benefits for patients with age-related macular degeneration.


Optimization of deposition uniformity for large-aperture National Ignition Facility substrates in a planetary rotation system.
Multilayer coatings on large substrates with increasingly complex spectral requirements are essential for a number of optical systems, placing stringent requirements on the error tolerances of individual layers. Each layer must be deposited quite uniformly over the entire substrate surface since any nonuniformity will add to the layer-thickness error level achieved. A deposition system containing a planetary rotation system with stationary uniformity masking is modeled, with refinements of the planetary gearing, source placement, and uniformity mask shape being utilized to achieve an optimal configuration. The impact of improper planetary gearing is demonstrated theoretically, as well as experimentally, providing more comprehensive requirements than simply avoiding repetition of previous paths through the vapor plume, until all possible combinations of gear teeth have been used. Deposition efficiency and the impact of changing vapor plume conditions on the uniformity achieved are used to validate improved source placement. Uniformity measurements performed on a mapping laser photometer demonstrate nonuniformities of less than 0.5% for 0.75 m optics in a 72 in. (1.8 m) coating chamber.


Fish utilisation of wetland nurseries with complex hydrological connectivity.
The physical and faunal characteristics of coastal wetlands are driven by dynamics of hydrological connectivity to adjacent habitats. Wetlands on estuary floodplains are particularly dynamic, driven by a complex interplay of tidal marine connections and seasonal freshwater flooding, often with unknown consequences for fish using these habitats. To understand the patterns and subsequent processes driving fish assemblage structure in such wetlands, we examined the nature and diversity of temporal utilisation patterns at a species or genus level over three annual cycles in a tropical Australian estuarine wetland system. Four general patterns of utilisation were apparent based on CPUE and size-structure dynamics: (i) classic nursery utlisation (use by recently settled recruits for their first year) (ii) interrupted peristence (iii) delayed recruitment (iv) facultative wetland residence. Despite the small self-recruiting 'facultative wetland resident' group, wetland occupancy seems largely driven by connectivity to the subtidal estuary channel. Variable connection regimes (i.e. frequency and timing of connections) within and between different wetland units (e.g. individual pools, lagoons, swamps) will therefore interact with the diversity of species recruitment schedules to generate variable wetland assemblages in time and space. In addition, the assemblage structure is heavily modified by freshwater flow, through simultaneously curtailing persistence of the 'interrupted persistence' group, establishing connectivity for freshwater spawned members of both the 'facultative wetland resident' and 'delayed recruitment group', and apparently mediating use of intermediate nursery habitats for marine-spawned members of the 'delayed recruitment' group. The diversity of utilisation pattern and the complexity of associated drivers means assemblage compositions, and therefore ecosystem functioning, is likely to vary among years depending on variations in hydrological connectivity. Consequently, there is a need to incorporate this diversity into understandings of habitat function, conservation and management.


The extended law of corresponding states when attractions meet repulsions.
Short-range attractive colloids show well-defined phase behaviour in the absence of repulsions, and highly intriguing equilibrium gelation in the presence of long-range repulsions. We present the state diagram of short-range attractive colloids with repulsions that range from fully screened to intermediately ranged, i.e. longer-ranged than the attractions, but shorter ranged than the colloid size. We demonstrate that although the macroscopic phase behaviour does not change perceptibly, there is a dramatic increase of inhomogeneities once the repulsions become longer-ranged than the attractions. The interaction potentials are characterized with small angle neutron scattering, and used to renormalize the state diagram with the minimum in the interaction potential, min[U(r)], and with the reduced second virial coefficient, B2*. We find that the extended law of corresponding states captures the onset of phase separation for shorter ranged repulsions, but fails for longer ranged repulsions. Instead, for a given model of U(r), the transition from visually homogeneous fluid to phase separation and/or gelation can be rescaled with min[U(r)] over the full range of repulsions. Finally, we suggest a generic state diagram to describe the effect of repulsions on short-range attractive systems.


Somatostatin receptor scintigraphy in metastatic breast cancer patients.
The purpose of this study was to evaluate the efficacy of somatostatin receptor scintigraphy (SRS) in imaging metastases in patients with advanced breast cancer (BC), and assess the relationship between exposure to chemotherapy and hormonotherapy with overexpression of somatostatin receptor (SS-R) on the breast cancer cell surface. Twelve patients with metastatic breast cancer were intravenously (i.v.) injected with In-111 pentatreotide (120 MBq). Early and later images were obtained with a double-head gamma camera equipped with medium-energy collimators. SPECT was performed when needed. Imaging results were compared with computed tomography and bone scan. Uptake levels were evaluated by site-specific visual analysis. Metastatic breast cancer can be visualized with SRS. Global sensitivity of imaging was 80% and specificity for correct prediction of tumor absence was 100%. Sensitivity was significantly higher for bone and lung metastases. SRS results related to the expression of SS-R on metastatic cell surfaces did not evidence a relationship with the biologic characteristics of the primary BC and drug exposure. In our series, SRS quantitative analysis demonstrated that tumor metastases differ greatly in uptake levels. Fifteen percent of metastatic sites in our series showed strong uptake. Our data support the important specificity of SRS in identifying BC metastases, mostly in cases of bone and lung disease, as well as the role of SRS in predicting responsiveness of metastatic BC cells to treatment with somatostatin analogues (SS), when SS-Rs are overexpressed on cell surfaces. If our results are confirmed in large scale studies, SRS shows the potential to treat selected patients with overexpressed SS-R on their tumoral cells with designed target therapies with SS analogue.


Presynaptic potentials and facilitation of transmitter release in the squid giant synapse.
Presynaptic potentials were studied during facilitation of transmitter release in the squid giant synapse. Changes in action potentials were found to cause some, but not all, of the facilitation during twin-pulse stimulation. During trains of action potentials, there were no progressive changes in presynaptic action potentials which could account for the growth of facilitation. Facilitation could still be detected in terminals which had undergone conditioning depolarization or hyperpolarization. Facilitation could be produced by small action potentials in low [Ca++]o and by small depolarizations in the presence of tetrodotoxin. Although the production of facilitation varied somewhat with presynaptic depolarization, nevertheless, approximately equal amounts of facilitation could be produced by depolarizations which caused the release of very different amounts of transmitter.


Elastic properties of the ascending aorta in patients with beta-thalassemia major.
Beta-thalassemia major (beta-TM) is a congenital hemolytic disorder characterized by impaired left ventricular and endothelial function. However, elastic properties of the aorta have not been sufficiently investigated in patients with beta-TM. We investigated whether beta-TM is related to impaired ascending aortic elastic properties. We studied 36 patients with beta-TM (age: 15.8 +/- 2.6 years) and 30 age- and sex-matched control subjects by echocardiography. Aortic elastic indexes, aortic strain (%), distensibility (cm(2) dyn(-1) 10(-3)), and stiffness index were calculated from the echocardiographically derived thoracic aortic diameters (mm/m(2)), and the measurement of pulse pressure obtained by cuff sphygmomanometry. Patients versus control subjects had increased aortic diameters (P < 0.001), lower mean aortic strain (9 +/- 3.6 vs. 14.9 +/- 3.2, P < 0.001) and distensibility (0.6 +/- 0.36 vs. 0.8 +/- 0.2, P < 0.012), and higher mean stiffness index (5.3 +/- 2.4 vs. 2.8 +/- 0.6, P < 0.001). Aortic elastic indexes were significantly associated with ferritin level, while stiffness index was significantly related to platelet count. Elastic properties of ascending aorta are impaired in patients with beta-TM. Impaired functions of aorta may lead to deterioration of left ventricular function.


Hypothalamic-pituitary-adrenal (HPA) axis and aging.
Human aging is associated with increasing frailty and morbidity which can result in significant disability. Dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis may contribute to aging-related diseases like depression, cognitive deficits, and Alzheimer's disease in some older individuals. In addition to neuro-cognitive dysfunction, it has also been associated with declining physical performance possibly due to sarcopenia. This article reviews the pathophysiology of HPA dysfunction with respect to increased basal adrenocorticotropic hormone (ACTH) and cortisol secretion, decreased glucocorticoid (GC) negative feedback at the level of the paraventricular nucleus (PVN) of the hypothalamus, hippocampus (HC), and prefrontal cortex (PFC), and flattening of diurnal pattern of cortisol release. It is possible that the increased cortisol secretion is secondary to peripheral conversion from cortisone. There is a decline in pregnolone secretion and C-19 steroids (DHEA) with aging. There is a small decrease in aldosterone with aging, but a subset of the older population have a genetic predisposition to develop hyperaldosteronism due to the increased ACTH stimulation. The understanding of the HPA axis and aging remains a complex area with conflicting studies leading to controversial interpretations.


Familial inclusion body myositis in a mother and son with different ancestral MHC haplotypes.
An Ashkenazi Jewish family in which the mother and a son both have inclusion body myositis (IBM) is reported. The condition developed at an earlier age and was more rapidly progressive and less responsive to treatment in the son than in the mother or other IBM patients in our clinic. Genetic analysis showed that the mother carried alleles of the 8.1 MHC ancestral haplotype (AH; HLA-B8, DRB1*0301), which is found in 85% of IBM patients in Western Australia. The son did not inherit this haplotype, but carried alleles characteristic of the 52.1AH (HLA-B5, DRB1*1502) of paternal origin. The findings indicate that in this family either the 8.1AH or 52.1AH may carry susceptibility for IBM and that the 52.1AH is associated with a more severe and treatment-resistant form of the disease.


Large-scale proteomic identification of S100 proteins in breast cancer tissues.
Attempts to reduce morbidity and mortality in breast cancer is based on efforts to identify novel biomarkers to support prognosis and therapeutic choices. The present study has focussed on S100 proteins as a potentially promising group of markers in cancer development and progression. One reason of interest in this family of proteins is because the majority of the S100 genes are clustered on a region of human chromosome 1q21 that is prone to genomic rearrangements. Moreover, there is increasing evidence that S100 proteins are often up-regulated in many cancers, including breast, and this is frequently associated with tumour progression. Samples of breast cancer tissues were obtained during surgical intervention, according to the bioethical recommendations, and cryo-preserved until used. Tissue extracts were submitted to proteomic preparations for 2D-IPG. Protein identification was performed by N-terminal sequencing and/or peptide mass finger printing. The majority of the detected S100 proteins were absent, or present at very low levels, in the non-tumoral tissues adjacent to the primary tumor. This finding strengthens the role of S100 proteins as putative biomarkers. The proteomic screening of 100 cryo-preserved breast cancer tissues showed that some proteins were ubiquitously expressed in almost all patients while others appeared more sporadic. Most, if not all, of the detected S100 members appeared reciprocally correlated. Finally, from the perspective of biomarkers establishment, a promising finding was the observation that patients which developed distant metastases after a three year follow-up showed a general tendency of higher S100 protein expression, compared to the disease-free group. This article reports for the first time the comparative proteomic screening of several S100 protein members among a large group of breast cancer patients. The results obtained strongly support the hypothesis that a significant deregulation of multiple S100 protein members is associated with breast cancer progression, and suggest that these proteins might act as potential prognostic factors for patient stratification. We propose that this may offer a significant contribution to the knowledge and clinical applications of the S100 protein family to breast cancer.


Pause time alters the preparation of two-component movements.
Targeted reciprocal aiming movements are pervasive in everyday life, but it is unclear how the timing parameters between task elements affect the preparation of these movements. This study used a loud (124 dB) startling acoustic stimulus (SAS) to probe how changes in the pause time between the outward and return components of a reciprocal aiming task affected the preparatory state of the motor system. Participants performed a visually guided wrist extension-flexion task to a target located at 20° from the start position and were instructed to pause the movement within the wrist extension target zone for either 50, 200, or 500 ms. A SAS was presented during 25 % of trials before either the onset of the wrist extension (out) or flexion (return) components of the task to determine how motor preparation was affected by task requirements. Results showed that the presentation of a SAS prior to the initial outward movement led to significantly earlier onsets of both the outward and return components (p < .05), indicating that the pause time in the planned action was pre-planned. For the longer (200, 500 ms) pause-time conditions, a SAS delivered prior to returning from the target region triggered the return portion of the movement early. These findings suggest that the shortest pause-time movement (50 ms) was preplanned as a single action, whereas for reciprocal movements with longer pause times at least the initial part of the movement and the timing of the pause were preplanned and integrated, while the return portion was more independent.


An Adjunct Indicator for the Diagnosis of Fracture-Related Infections: Platelet Count to Mean Platelet Volume Ratio.
Introduction: Fracture-related infection (FRI) is a common complication associated with orthopaedic fracture care. Diagnosing these complications in the preoperative setting is difficult. Platelets are a known acute phase reactant with indices that change in accordance with infection and inflammation. The purpose of our study was to assess the diagnostic utility of platelet indices at assessing FRI. Methods: A retrospective review performed for all patients who underwent revision surgery for fracture nonunion between 2013 and 2018. Radiographs were employed to define nonunion. Intraoperative cultures were used to define FRI. Receiver operator characteristic (ROC) curve analysis was used to assess the diagnostic ability of preoperative erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and the platelet count/mean platelet volume ratio (P/V) at recognizing FRI. Results: Of the 53 revision surgeries that were performed for fracture nonunion, 17 (32.1%) were identified as FRI. There were no significant demographic differences between the two cohorts. Patients with FRIs exhibited higher values for ESR (54.82 vs. 19.16, p<0.001), CRP (0.90 vs. 0.35, p=0.003), and P/V (37.4 vs. 22.8, p<0.001) as compared to those within the aseptic nonunion cohort. ROC curve analysis for P/V demonstrated that at an optimal ratio of 23, area under the curve (AUC) is 0.814, specificity is 55.6%, and sensitivity is 100.0%. There was no significant difference in the diagnostic performance of the serum biomarkers but only ESR and P/V had an AUC greater than 0.80. The negative predictive value (NPV) for P/V, ESR, and CRP was 100.0%, 84.6%, and 78.6%, respectively. Conclusion: The P/V ratio may serve as a reliable screening test for FRI.


Quantification of allospecific and nonspecific corneal endothelial cell damage after corneal transplantation.
To investigate the effect of host immunity (allospecific) and surgical manipulation (non-allospecific) on corneal endothelial cells (CECs) in corneal transplantation. Draining lymph nodes and grafted C57BL/6 corneas were harvested from syngeneic recipients, allograft acceptors, and allograft rejectors (BALB/c) 1, 3, and 8 weeks after transplantation. We analyzed CEC apoptosis using an ex vivo cornea-in-the-cup assay, and visualized cell-to-cell junctions using immunohistochemical staining (ZO-1). Automatic cell analysis using Confoscan software was used to measure CEC density as well as changes in CEC morphology by quantifying the coefficient of variation in cell size (polymegethism) and shape (pleomorphism). The cornea-in-the-cup assay showed that allogeneic acceptor T cells and to an even greater extent rejector T cells (but not syngeneic T cells) induced CEC apoptosis. CEC density after corneal transplantation was significantly reduced in allogeneic acceptors compared with syngeneic grafts (P<0.001), and CEC density was even further reduced in the allo-rejector group compared with the allo-acceptor group. Allogeneic grafts showed a greater increase in the coefficient of variation in cell size (polymegethism) when compared with syngeneic grafts 1 week after transplantation (P=P<0.001). However, pleomorphism was not significantly different between syngeneic and allo-acceptor grafts, indicating that polymegethism (but not pleomorphism or cell density) is a sensitive indicator of the effect of alloimmunity on CECs. Our data demonstrate that host alloimmunity rather than surgical manipulation alone is the major cause of CEC damage in corneal transplantation, and such morphologic changes of CECs can be detected before the clinically visible onset of allograft rejection.


Psychedelics.
Psychedelics (serotonergic hallucinogens) are powerful psychoactive substances that alter perception and mood and affect numerous cognitive processes. They are generally considered physiologically safe and do not lead to dependence or addiction. Their origin predates written history, and they were employed by early cultures in many sociocultural and ritual contexts. After the virtually contemporaneous discovery of (5R,8R)-(+)-lysergic acid-N,N-diethylamide (LSD)-25 and the identification of serotonin in the brain, early research focused intensively on the possibility that LSD and other psychedelics had a serotonergic basis for their action. Today there is a consensus that psychedelics are agonists or partial agonists at brain serotonin 5-hydroxytryptamine 2A receptors, with particular importance on those expressed on apical dendrites of neocortical pyramidal cells in layer V. Several useful rodent models have been developed over the years to help unravel the neurochemical correlates of serotonin 5-hydroxytryptamine 2A receptor activation in the brain, and a variety of imaging techniques have been employed to identify key brain areas that are directly affected by psychedelics. Recent and exciting developments in the field have occurred in clinical research, where several double-blind placebo-controlled phase 2 studies of psilocybin-assisted psychotherapy in patients with cancer-related psychosocial distress have demonstrated unprecedented positive relief of anxiety and depression. Two small pilot studies of psilocybin-assisted psychotherapy also have shown positive benefit in treating both alcohol and nicotine addiction. Recently, blood oxygen level-dependent functional magnetic resonance imaging and magnetoencephalography have been employed for in vivo brain imaging in humans after administration of a psychedelic, and results indicate that intravenously administered psilocybin and LSD produce decreases in oscillatory power in areas of the brain's default mode network.


Microtomography-based comparison of reciprocating single-file F2 ProTaper technique versus rotary full sequence.
A preparation technique with only 1 single instrument was proposed on the basis of the reciprocating movement of the F2 ProTaper instrument. The present study was designed to quantitatively assess canal preparation outcomes achieved by this technique. Twenty-five extracted human mandibular first molars with 2 separate mesial root canals were selected. Canals were randomly assigned to 1 of the 2 experimental groups: group 1, rotary conventional preparation by using ProTaper, and group 2, reciprocate instrumentation with 1 single ProTaper F2 instrument. Specimens were scanned initially and after root canal preparation with an isotropic resolution of 20 μm by using a micro-computed tomography system. The following parameters were assessed: changes in dentin volume, percentage of shaped canal walls, and degree of canal transportation. In addition, the time required to reach working length with the F2 instrument was recorded. Preoperatively, there were no differences regarding root canal curvature and volume between experimental groups. Overall, instrumentation led to enlarged canal shapes with no evidence of preparation errors. There were no statistical differences between the 2 preparation techniques in the anatomical parameters assessed (P > .01), except for a significantly higher canal transportation caused by the reciprocating file in the coronal canal third. On the other hand, preparation was faster by using the single-file technique (P < .01). Shaping outcomes with the single-file F2 ProTaper technique and conventional ProTaper full-sequence rotary approach were similar. However, the single-file F2 ProTaper technique was markedly faster in reaching working length.


A simple method for EEG guided transcranial electrical stimulation without models.
There is longstanding interest in using EEG measurements to inform transcranial Electrical Stimulation (tES) but adoption is lacking because users need a simple and adaptable recipe. The conventional approach is to use anatomical head-models for both source localization (the EEG inverse problem) and current flow modeling (the tES forward model), but this approach is computationally demanding, requires an anatomical MRI, and strict assumptions about the target brain regions. We evaluate techniques whereby tES dose is derived from EEG without the need for an anatomical head model, target assumptions, difficult case-by-case conjecture, or many stimulation electrodes. We developed a simple two-step approach to EEG-guided tES that based on the topography of the EEG: (1) selects locations to be used for stimulation; (2) determines current applied to each electrode. Each step is performed based solely on the EEG with no need for head models or source localization. Cortical dipoles represent idealized brain targets. EEG-guided tES strategies are verified using a finite element method simulation of the EEG generated by a dipole, oriented either tangential or radial to the scalp surface, and then simulating the tES-generated electric field produced by each model-free technique. These model-free approaches are compared to a 'gold standard' numerically optimized dose of tES that assumes perfect understanding of the dipole location and head anatomy. We vary the number of electrodes from a few to over three hundred, with focality or intensity as optimization criterion. Model-free approaches evaluated include (1) voltage-to-voltage, (2) voltage-to-current; (3) Laplacian; and two Ad-Hoc techniques (4) dipole sink-to-sink; and (5) sink to concentric. Our results demonstrate that simple ad hoc approaches can achieve reasonable targeting for the case of a cortical dipole, remarkably with only 2-8 electrodes and no need for a model of the head. Our approach is verified directly only for a theoretically localized source, but may be potentially applied to an arbitrary EEG topography. For its simplicity and linearity, our recipe for model-free EEG guided tES lends itself to broad adoption and can be applied to static (tDCS), time-variant (e.g., tACS, tRNS, tPCS), or closed-loop tES.


Radioactive labeling of protein carboxyl groups on factor VIII: use of carbodiimides for nuclear medicine.
Carbodiimides have been used to study macromolecular structure and to produce immunologically active antigens. We have used this method to label a labile coagulation protein, factor VIII, with 14C-glycine-ethyl-ester. No discernible chemical change, loss of biologic function in vitro, or alteration of the plasma disappearance of factor VIII resulted. The carbodiimide labeling method has potentially broad application because many biologic molecules contain carboxyl groups that are generally not critical to their chemical or immunologic character. This method can be used to incorporate short-lived positron emitters, such as 11C and 13N, into biologic compounds, or to attach ligands to useful antibodies for subsequent chelation to radioactive metals, such as 111In. Carbodiimides are especially useful for radionuclidic labeling of labile proteins because of the mild conditions, rapid reaction, and firmly attached label.


Pathologies of the digestive system caused by marine debris in Chelonia mydas.
The growth of human population and deficient pollution control measures pose significant challenge to the environment. Despite conservation efforts, all sea turtle species are at some risk of extinction. The present study investigated the effect of marine debris on the gastrointestinal tract of green turtles in southeastern Brazil. Of the 777 animals evaluated, 290 showed marine debris in one segment of the gastrointestinal tract. The presence of these materials in the gastrointestinal tract may be harmful, independent of the segment involved, and increases the risk of impaction. Marine debris has become a significant hazard to Chelonia mydas in the region surveyed, causing perforation, rupture, or fecal impaction that, when not treated, is potentially fatal, exposing the intestine to bacterial infection.


Preparing for pandemics: a systematic review of pandemic influenza clinical management guidelines.
The COVID-19 pandemic has highlighted the importance of evidence-based clinical decision-making. Clinical management guidelines (CMGs) may help reduce morbidity and mortality by improving the quality of clinical decisions. This systematic review aims to evaluate the availability, inclusivity, and quality of pandemic influenza CMGs, to identify gaps that can be addressed to strengthen pandemic preparedness in this area. Ovid Medline, Ovid Embase, TRIP (Turning Research Into Practice), and Guideline Central were searched systematically from January 2008 to 23rd June 2022, complemented by a grey literature search till 16th June 2022. Pandemic influenza CMGs including supportive care or empirical treatment recommendations were included. Two reviewers independently extracted data from the included studies and assessed their quality using AGREE II (Appraisal of Guidelines for Research & Evaluation). The findings are presented narratively. Forty-eight CMGs were included. They were produced in high- (42%, 20/48), upper-middle- (40%, 19/48), and lower-middle (8%, 4/48) income countries, or by international organisations (10%, 5/48). Most CMGs (81%, 39/48) were over 5 years old. Guidelines included treatment recommendations for children (75%, 36/48), pregnant women (54%, 26/48), people with immunosuppression (33%, 16/48), and older adults (29%, 14/48). Many CMGs were of low quality (median overall score: 3 out of 7 (range 1-7). All recommended oseltamivir; recommendations for other neuraminidase inhibitors and supportive care were limited and at times contradictory. Only 56% (27/48) and 27% (13/48) addressed oxygen and fluid therapy, respectively. Our data highlights the limited availability of up-to-date pandemic influenza CMGs globally. Of those identified, many were limited in scope and quality and several lacked recommendations for specific at-risk populations. Recommendations on supportive care, the mainstay of treatment, were limited and heterogeneous. The most recent guideline highlighted that the evidence-base to support antiviral treatment recommendations is still limited. There is an urgent need for trials into treatment and supportive care strategies including for different risk populations. New evidence should be incorporated into globally accessible guidelines, to benefit patient outcomes. A 'living guideline' framework is recommended and further research into guideline implementation in different resourced settings, particularly low- and middle-income countries.


Investigating the Origin of Enhanced C2+ Selectivity in Oxide-/Hydroxide-Derived Copper Electrodes during CO2 Electroreduction.
Oxide-/hydroxide-derived copper electrodes exhibit excellent selectivity toward C2+ products during the electrocatalytic CO2 reduction reaction (CO2RR). However, the origin of such enhanced selectivity remains controversial. Here, we prepared two Cu-based electrodes with mixed oxidation states, namely, HQ-Cu (containing Cu, Cu2O, CuO) and AN-Cu (containing Cu, Cu(OH)2). We extracted an ultrathin specimen from the electrodes using a focused ion beam to investigate the distribution and evolution of various Cu species by electron microscopy and electron energy loss spectroscopy. We found that at the steady stage of the CO2RR, the electrodes have all been reduced to Cu0, regardless of the initial states, suggesting that the high C2+ selectivities are not associated with specific oxidation states of Cu. We verified this conclusion by control experiments in which HQ-Cu and AN-Cu were pretreated to fully reduce oxides/hydroxides to Cu0, and the pretreated electrodes showed even higher C2+ selectivity compared with their unpretreated counterparts. We observed that the oxide/hydroxide crystals in HQ-Cu and AN-Cu were fragmented into nanosized irregular Cu grains under the applied negative potentials. Such a fragmentation process, which is the consequence of an oxidation-reduction cycle and does not occur in electropolished Cu, not only built an intricate network of grain boundaries but also exposed a variety of high-index facets. These two features greatly facilitated the C-C coupling, thus accounting for the enhanced C2+ selectivity. Our work demonstrates that the use of advanced characterization techniques enables investigating the structural and chemical states of electrodes in unprecedented detail to gain new insights into a widely studied system.


Analyzing variability and the rate of decline of migratory shorebirds in Moreton Bay, Australia.
Estimating the abundance of migratory species is difficult because sources of variability differ substantially among species and populations. Recently developed state-space models address this variability issue by directly modeling both environmental and measurement error, although their efficacy in detecting declines is relatively untested for empirical data. We applied state-space modeling, generalized least squares (with autoregression error structure), and standard linear regression to data on abundance of wetland birds (shorebirds and terns) at Moreton Bay in southeast Queensland, Australia. There are internationally significant numbers of 8 species of waterbirds in the bay, and it is a major terminus of the large East Asian-Australasian Flyway. In our analyses, we considered 22 migrant and 8 resident species. State-space models identified abundances of 7 species of migrants as significantly declining and abundance of one species as significantly increasing. Declines in migrant abundance over 15 years were 43-79%. Generalized least squares with an autoregressive error structure showed abundance changes in 11 species, and standard linear regression showed abundance changes in 15 species. The higher power of the regression models meant they detected more declines, but they also were associated with a higher rate of false detections. If the declines in Moreton Bay are consistent with trends from other sites across the flyway as a whole, then a large number of species are in significant decline.


Partnering with patients--a concept ready for action.
This paper is a report of a concept analysis of partnership within the context of a professional-patient relationship. The concept of partnership has been previously characterized as immature, with a need for further consensus and consistency. Critical attributes previously reported include relationship, power sharing and negotiation, with empowerment as the primary consequence. Rodgers' evolutionary method of analysis for concept development was used to re-examine the concept of partnership. Historical documents and previously published conceptual papers were reviewed for context. A search of multidisciplinary literature published between 2000 and 2004 was undertaken using the keywords of 'partnership' and 'partnering', combined with nurse/professional/physician-client relationship. Attributes, uses, antecedents and consequences were inductively derived from the citations analysed (n = 62). Previous authors affirmed that partnership involves a process and a consistent set of eight attributes. Relationship, shared power, shared decision-making and patient autonomy are attributes that distinguish partnership from other related concepts. Most of the literature, however, consisted of expert opinion or descriptive research. Little progress has been made in applying theory, developing tools to test the process, identifying when partnerships are needed and what specific outcomes occur when they are present. Partnerships between healthcare providers and patients develop over time. They are created to support patients in having a greater voice in their care and to empower them in self-management. A descriptive model of partnership is proposed to support researchers in generating or applying existing theory to the development of research designs and tools that could test how this process actually works.


[Tuberculosis of the knee joint: a case report].
Tuberculosis may develop in the skeletal system apart from its primary location, the lungs. A 22-year-old male patient presented with complaints of severe pain and swelling in the left knee and difficulty in walking. The symptoms first appeared 12 years before and he underwent open surgical debridement and arthroscopic debridement at two other centers, at which time aspiration cultures taken from the knee yielded no growth. Physical examination showed a painful left knee with swelling, increased temperature, effusion, and limited range of motion. Based on history, physical examination, radiological studies, and laboratory findings, knee osteoarthritis was considered secondary to nonspecific septic arthritis and knee arthrodesis was performed. At surgery, marked destruction of articular cartilage and synovial hypertrophy were observed. In addition, a cavitary lesion was detected, about 1 x 1 cm in size, in the lateral femoral condyle without articular involvement. Biopsy cultures taken from the lesion showed growth of Mycobacterium tuberculosis in the third week. This enabled a diagnosis of tuberculosis of the knee joint and antituberculous drug therapy was instituted. At six months postoperatively, the patient was on antituberculous treatment and had a painless knee.


Low Body Mass Index in Females May Portend Inferior Outcomes After Primary Hip Arthroscopy: A Propensity-Matched Analysis With Minimum 2-Year Follow-up.
There is a paucity of literature evaluating the effect of low body mass index (BMI) in female patients undergoing primary hip arthroscopy. (1) To report minimum 2-year patient-reported outcome scores for female patients with low BMI who underwent primary hip arthroscopy for femoroacetabular impingement syndrome and (2) to compare these results with a propensity-matched control group of female patients with normal BMI. Cohort study; Level of evidence, 3. Data were collected on all female patients who had low BMI (<18.5) and underwent primary hip arthroscopy between April 2008 and January 2019. Patients were excluded if they had a previous hip arthroscopy; had workers' compensation; were unwilling to participate in the study; or had Tönnis osteoarthritis grade >1, acetabular dysplasia, or a previous hip condition. Minimum 2-year patient-reported outcomes were collected for the modified Harris Hip Score (mHHS), Non-Arthritic Hip Score, Hip Outcome Score-Sport Specific Subscale (HOS-SSS), visual analog scale for pain, and satisfaction. The percentage of patients achieving the minimal clinically important difference (MCID), Patient Acceptable Symptom State (PASS), and maximum outcome improvement satisfaction threshold (MOIST) was also recorded. These patients were propensity matched in a 1:2 ratio to female patients with normal BMI for comparison. A total of 43 patients (49 hips) were included with a mean ± SD follow-up of 26.9 ± 8.3 months, age of 24.1 ± 12.6 years, and BMI of 17.7 ± 0.67. These patients demonstrated significant improvement from presurgery to minimum 2-year follow-up for the mHHS, Non-Arthritic Hip Score, HOS-SSS, and visual analog scale (P < .05). When outcomes were compared with 93 control patients (97 hips), female patients with low BMI demonstrated lower rates of achieving the PASS for the mHHS (61.2% vs 77.3%; P = .041) and HOS-SSS (39.5% vs 57.1%; P = .031). Rates of achieving the MCID and MOIST were similar between the groups (P > .05). Female patients with low BMI also had higher rates of revision when compared with the control group, but this did not reach statistical significance (14.2% vs 7.2%; P = .171). Female patients with low BMI undergoing primary hip arthroscopy for femoroacetabular impingement syndrome demonstrated significant improvement in patient-reported outcomes at minimum 2-year follow-up. When compared with a propensity-matched control group with normal BMI, these patients demonstrated lower rates of achieving the PASS for the mHHS and HOS-SSS. Low BMI in females undergoing primary hip arthroscopy for FAIS may adversely affect outcomes at short-term follow-up.


Applications of mouse airway epithelial cell culture for asthma research.
Primary airway epithelial cell culture provides a valuable tool for studying cell differentiation, cell-cell interactions, and the role of immune system factors in asthma pathogenesis. In this chapter, we discuss the application of mouse tracheal epithelial cell cultures for the study of asthma biology. A major advantage of this system is the ability to use airway epithelial cells from mice with defined genetic backgrounds. The in vitro proliferation and differentiation of mouse airway epithelial cells uses the air-liquid interface condition to generate well-differentiated epithelia with characteristics of native airways. Protocols are provided for manipulation of differentiation, induction of mucous cell metaplasia, genetic modification, and cell and pathogen coculture. Assays for the assessment of gene expression, responses of cells, and analysis of specific cell subpopulations within the airway epithelium are included.


Developmental expression of the putative transcription factor Egr-1 suggests that Egr-1 and c-fos are coregulated in some tissues.
We have investigated developmental expression of the gene Egr-1, which encodes a protein containing three zinc fingers. Egr-1 like c-fos is a serum inducible, early response gene, which is co-induced with c-fos in a variety of quite different situations. A single 3.7-kb RNA was detected throughout fetal mouse development, which increased in absolute levels in total fetal RNA from 9.5 to 12.5 days post coitum (p.c.). In situ hybridization to 14.5- and 17.5-day p.c. fetal tissues demonstrated Egr-1 accumulation at several specific sites. These included mesenchymal components of the developing tooth germs and salivary and nasal glands; an ectodermally derived component of the whisker pad and developing muscle, cartilage, and bone. Expression of Egr-1 in cartilage and bone showed a strikingly similar expression to previously published reports of c-fos in these tissues. High levels of Egr-1 RNA was observed at the perichondrial interface of opposing cartilaginous elements and in interstitial cells that lie in between. Bone expression was observed in membranous bone of the head, alveolar bone around the tooth germs, and at periosteal and endochondral ossification sites in the limb bones. Our data support the idea that Egr-1 and c-fos may be coregulated in vivo and together may regulate normal development of the skeleton.


Undocumented adult migrants in Sweden: mental health and associated factors.
Undocumented migrants (UMs) in Europe constitute a heterogeneous group. They are typically in a vulnerable and marginalised situation, since most of them have exhausted their options for gaining asylum and protection from war and persecution, many are traumatised and fear disclosure and deportation, and they typically lack basic social security. The present study investigates living conditions, access to human rights and mental health of UMs living in Sweden. A cross-sectional study with adult UMs was performed in the three largest cities in Sweden in 2014-2016. Sampling was done via informal networks. A socioeconomic questionnaire was constructed, and psychiatric symptoms were screened for using Beck's Depression Inventory II, Beck's Anxiety Inventory and the PTSD Checklist (PCL) for civilians. Trained field workers conducted the interviews. Descriptive statistics, chi-square tests and logistic regression models were used. A total number of 104 individuals participated. Preliminary findings show that 68% of respondents were suffering from either moderate or severe anxiety, 71% from either moderate or severe depression and 58% from PTSD. No statistically significant gender differences occurred, but age was statistically significant in relation to anxiety and depression. The majority feared returning to their country of origin, for political reasons, due to war in progress there and/or because they belonged to a minority and feared harassment. Almost all had an unstable housing situation and were often forced to move. Fifty-seven percent experienced food insecurity. The psychosocial situation among UMs in Sweden, in addition to insecure living conditions without a guarantee of basic needs being met is stressful, and many UMs live in constant fear of disclosure and deportation, all of which has a detrimental effect of the mental health. It is important to understand both associated risk factors for ill-health and coping strategies in this vulnerable population in order try to reduce ongoing stress.


Mortality, asthma, smoking and acute chest syndrome in young adults with sickle cell disease.
Sickle cell disease (SCD) patients with asthma have an increased risk of death. Acute chest syndrome (ACS) is a major cause of mortality in patients with SCD, and ACS may be more common in SCD patients who smoke. The purpose of this study was to test the hypothesis that mortality in young adults with SCD would be greater than that of controls during a 10-year period and to determine whether asthma, reduced lung function, ACS episodes, and/or smoking predicted mortality during the follow-up period. The outcomes during a 10-year period were ascertained of SCD patients and race-matched controls who had taken part in a pulmonary function study when they were between age 19 and 27 years. Smoking and asthma status and whether they had had ACS episodes were determined, and lung function was measured at the initial assessment. Seventy-five subjects with SCD were followed for 683 patient years. There were 11 deaths with a mortality rate of 1.6 deaths per 100 patient years, which was higher than that of the controls; one death in 47 controls was observed for 469 patient years with a mortality rate of 0.2 per 100 patient years (p = 0.03). There were no significant associations of body mass index, recurrent episodes of acute chest, steady state haemoglobin, or gender with mortality. Adjusting for baseline lung function in SCD patients, "current" asthma [hazard ratio (HR) 11.2; 95 % confidence interval (CI) 2.5-50.6; p = 0.002] and smoking [HR 2.7; (95 % CI 1.3-5.5); p = 0.006] were significantly associated with mortality during the 10-year period. Our results indicate that young adults with SCD should be discouraged from smoking and their asthma aggressively treated.


7000 years of Emiliania huxleyi viruses in the Black Sea.
A 7000-year record of Coccolithovirus and its host, the calcifying haptophyte Emiliania huxleyi, was reconstructed on the basis of genetic signatures preserved in sediments underlying the Black Sea. The data show that the same virus and host populations can persist for centuries. Major changes in virus and host populations occurred during early sapropel deposition, ~5600 years ago, and throughout the formation of the coccolith-bearing sediments of Unit I during the past 2500 years, when the Black Sea experienced dramatic changes in hydrologic and nutrient regimes. Unit I saw a reoccurrence of the same host genotype thousands of years later in the presence of a different subset of viruses. Historical plankton virus populations can thus be included in paleoecological and paleoenvironmental studies.


Arylamine N-acetyltransferases. Expression in Escherichia coli, purification, and substrate specificities of recombinant hamster monomorphic and polymorphic isozymes.
Two isozymes of arylamine N-acetyltransferases (NATs) catalyze the biotransformation of arylamines to arylamides, and the bioactivation of carcinogenic arylhydroxylamines and arylhydroxamic acids to reactive electrophiles capable of forming a variety of DNA and protein adducts. As part of a project directed toward delineation of the molecular factors responsible for the pronounced differences in the substrate specificity of the isozymes, we have recently reported the expression in Escherichia coli and purification of hamster NAT1 (NAT1 8) as a fusion protein to an antibody-reactive amino terminus FLAG peptide capable of being removed by digestion with enterokinase. Unfortunately, the conditions necessary for the removal of the peptide by enterokinase resulted in incomplete protease digestion and substantial loss of NAT1 activity. Consequently, we have constructed the plasmid pPH8 in which an 11 amino acid thrombin proteolysis site has been inserted between the FLAG peptide and the amino terminus of NAT1. In addition, a plasmid that expresses hamster NAT2 (NAT2 15) was constructed by exchanging the gene sequence for NAT1 with the cloned sequence for NAT2. Both NAT fusion proteins were expressed in JM105 cells. Analysis of catalytically active cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that approximately 30-fold more soluble NAT2 was expressed than NAT1 in the bacterial cytosol. The fusion proteins were purified to homogeneity by immunoaffinity chromatography, followed by gel filtration to remove high molecular weight contaminants. The FLAG peptide was subsequently removed by treatment with human thrombin, followed by an additional pass over the immunoaffinity column. Unlike the results obtained from proteolysis by enterokinase, the activities of the rNAT1 and rNAT2 were shown to be unaffected by treatment with thrombin. Moreover, the substrate specificities for the recombinant NATs closely matched those observed for NAT1 and NAT2 isolated from hamster liver.


Explaining equity gains in child survival in Bangladesh: scale, speed, and selectivity in health and development.
By disaggregating gains in child health in Bangladesh over the past several decades, significant improvements in gender and socioeconomic inequities have been revealed. With the use of a social determinants of health approach, key features of the country's development experience can be identified that help explain its unexpected health trajectory. The systematic equity orientation of health and socioeconomic development in Bangladesh, and the implementation attributes of scale, speed, and selectivity, have been important drivers of health improvement. Despite this impressive pro-equity trajectory, there remain significant residual inequities in survival of girls and lower wealth quintiles as well as a host of new health and development challenges such as urbanisation, chronic disease, and climate change. Further progress in sustaining and enhancing equity-oriented achievements in health hinges on stronger governance and longer-term systems thinking regarding how to effectively promote inclusive and equitable development within and beyond the health system.


An evaluation of the information gained from the use of intraoperative nerve recording in the management of suspected brachial plexus root avulsion.
The authors examine the information gained from the use of intraoperative nerve recording in the management of suspected brachial plexus root avulsion. A retrospective chart review examined 25 patients who had undergone intraoperative nerve recording for a brachial plexus injury. Thirty-seven preganglionic root avulsions were demonstrated by somatosensory evoked potentials, C4 to T1, during intraoperative nerve recording. To measure the information gain derived from intraoperative nerve recording, the authors compared the number of roots diagnosed as preganglionic root avulsions preoperatively to those diagnosed by intraoperative nerve recording. From this, the authors can demonstrate the positive and negative predictive values of their preoperative multimodality assessment for brachial plexus root avulsion and compare this to the result of intraoperative nerve recording to derive the gain of information. In addition, the authors examined the validity of the intraoperative nerve recording somatosensory evoked potentials when this produced a diagnosis of an intact root in this cohort by performing a clinical outcome analysis for those roots used for reconstruction. Twenty-five patients underwent intraoperative nerve recording for unilateral brachial plexus injury; 15 patients were diagnosed with 55 preganglionic root avulsions from C4 to T1 preoperatively by multimodality assessment. Fourteen of 55 roots thought to be avulsed preoperatively were found to be intact with intraoperative nerve recording, representing a gain of information of 25 percent derived from intraoperative nerve recording for roots thought to be avulsed preoperatively. Intraoperative nerve recording adds useful information during exploratory brachial plexus surgery.


Integration of the physical and genetic linkage map for human chromosome 13.
We have used a panel of somatic cell hybrids containing different rearrangements of human chromosome 13 to integrate genetic and physical maps of this chromosome. The positions of 17 translocation/deletion breakpoints on human chromosome 13 have been determined relative to the microsatellite markers on the genetic linkage map compiled by Généthon. Because markers on maps from several other Consortium groups have also been analyzed using many of the same hybrids, it was possible to relate these with the Généthon map. The position of all of the chromosome breakpoints have been placed, wherever possible, between two adjacent markers on the genetic linkage maps using PCR analysis for the presence/absence of the markers in the somatic cell hybrids. The positions of the breakpoints have already been determined cytogenetically, and some of these breakpoints are located at landmark positions on the chromosome. The relative density of markers along the chromosome differs between independently derived maps, and, based on the known locations of certain breakpoints in the physical map, inconsistencies in the genetic maps have been identified.


Design considerations to minimize the impact of drug absorption in polymer-based organ-on-a-chip platforms.
Biocompatible polymers, such as polydimethylsiloxane (PDMS), are the materials of choice for creating organ-on-a-chip microfluidic platforms. Desirable qualities include ease of fabrication, optical clarity, and hydrophobicity, the latter of which facilitates oxygen transport to encased cells. An emerging and important application of organ-on-a-chip technology is drug discovery; however, a potential issue for polymer-based microfluidic devices has been highlighted by recent studies with PDMS, which have demonstrated absorption (and thus loss) of hydrophobic drugs into PDMS under certain experimental conditions. Absorption of drug in the polymer can also lead to undesirable transfer of drug between adjacent microfluidic lines. Given the benefits of polymers, it is essential to develop a comprehensive understanding of drug absorption. In this study, we considered convection, dissolution, and diffusion of a drug within a polymer-based microfluidic device to characterize the dynamics of drug loss in a quantitative manner. We solved Fick's 2nd law of diffusion (unsteady diffusion-convection) by finite element analysis in COMSOL®, and experimentally validated the numerical model for loss of three hydrophobic molecules (rhodamine B, cyanine NHS ester, and paclitaxel) in PDMS. Drug loss, as well as the unintended mixing of drugs by adjacent microfluidic channels, depends strongly on platform design parameters, experimental conditions, and the physico-chemical properties of the drug, and can be captured in a simple quantitate relationship that employs four scalable dimensionless numbers. This simple quantitative framework can be used in the design of a wide range of polymer-based microfluidic devices to minimize the impact of drug absorption.


Population-based register of acute myocardial infarction: manual of operations.
Cardiovascular disease is the leading cause of death and hospitalization in both sexes in nearly all countries of Europe. The main forms of cardiovascular disease are ischaemic heart disease and stroke. The magnitude of the problem contrasts with the shortage, weak quality and comparability of data available in most European countries. Innovations in medical, invasive and biological treatments have substantially contributed to the escalating costs of health services. It is therefore urgent to obtain reliable information on the magnitude and distribution of the disease for both adequate health planning (including preventive strategies) and clinical decision making with correct cost-benefit assessments.A stepwise surveillance procedure based on standardized data collection, appropriate record linkage and validation methods was set up by the EUROCISS Project (EUROpean Cardiovascular Indicators Surveillance Set) to build up comparable and reliable indicators (attack rate and case fatality) for the surveillance of acute myocardial infarction/acute coronary syndrome at population level. This manual of operations is intended for health professionals and policy makers and provides a standardized and simple model for the implementation of a population-based register. It recommends to start from a minimum data set and then follow a stepwise procedure. Before implementing a population-based register, it is important to identify the target population under surveillance which should preferably cover a well-defined geographical and administrative area or region representative of the whole country for which population data and vital statistics (mortality and hospital discharge records at minimum) are routinely collected and easily available each year. All cases among residents should be recorded even if the case occurs outside the area. Validation of a sample of fatal and nonfatal events is mandatory.


Distinct processing of tone offset in two primary auditory cortices.
In the rodent auditory system, the primary cortex is subdivided into two regions, both receiving direct inputs from the auditory thalamus: the primary auditory cortex (A1) and the anterior auditory field (AAF). Although neurons in the two regions display different response properties, like response latency, firing threshold or tuning bandwidth, it is still not clear whether they process sound in a distinct way. Using in vivo electrophysiological recordings in the mouse auditory cortex, we found that AAF neurons have significantly stronger responses to tone offset than A1 neurons. AAF neurons also display faster and more transient responses than A1 neurons. Additionally, offset responses in AAF - unlike in A1, increase with sound duration. Local field potential (LFP) and laminar analyses suggest that the differences in sound responses between these two primary cortices are both of subcortical and intracortical origin. These results emphasize the potentially critical role of AAF for temporal processing. Our study reveals a distinct role of two primary auditory cortices in tone processing and highlights the complexity of sound encoding at the cortical level.


Epidemiology of fears and specific phobia in adolescence: results from the Mexican Adolescent Mental Health Survey.
Specific phobia is among the most common disorders in the general adolescent population, but also among the least studied. The objectives of this study were to estimate the prevalence of specific fears among adolescents, the proportion of those with fears who meet criteria for specific phobia, and the proportion who recognize their fears as excessive and to identify comorbidity with other disorders and factors associated with severity and treatment. 3,005 youth aged 12 to 17 years participated in the Mexican Adolescent Mental Health Survey, a stratified multistage probability sample representative of adolescents living in Mexico City in 2005. Fears, specific phobia, and 20 other DSM-IV psychiatric disorders were evaluated with the adolescent computerized version of the World Mental Health Composite International Diagnostic Interview, administered by trained lay interviewers in the participants' homes. Descriptive, logistic regression, and discrete-time survival analyses were employed. Most adolescents reported at least 1 fear (76.5%); 36.5% of those met lifetime criteria for specific phobia, and 27.3% met criteria in the prior 12 months. Blood-injection-injury and animal fears were the most common types. Females were more likely to report any fear, to have more fear types, and to meet diagnostic criteria. Increased numbers of fears were associated with increased odds of meeting specific phobia criteria and with increased impairment. Among teens with specific phobia, 71.3% recognized their fear as excessive, 9.7% to 12.3% reported severe impairment, and only 6.5% had received treatment. Severity of impairment and comorbid disorders were associated with treatment seeking. The large percentage of adolescents meeting diagnostic criteria coupled with the low proportion with serious impairment suggests that current diagnostic thresholds may be too low or not developmentally sensitive. Future research should address the nature of impairment for adolescents with specific phobia. Early detection and timely treatment are important given that specific phobia is persistent and highly comorbid and that few individuals seek treatment.


Parma international protocol: pilot study of DHAP followed by involved-field radiotherapy and BEAC with autologous bone marrow transplantation.
Fifty patients with intermediate- or high-grade non-Hodgkin's lymphoma (NHL) who had relapsed after a complete remission induced by an Adriamycin-containing chemotherapy regimen participated in this prospective pilot study. The patients ranged in age from 16 to 60 years (median 42 years). All patients received dexamethasone, high-dose cytarabine, and cisplatin (DHAP) for two courses at 3- to 4-week intervals. Patients achieving a partial or complete response were scheduled to receive involved-field radiotherapy and high-dose carmustine, etoposide, cytarabine, and cyclophosphamide (BEAC), followed by autologous bone marrow transplantation (ABMT). Among 48 evaluable patients (ie, 1 was lost to follow-up and 1 had no measurable disease) 7 patients obtained a complete response (CR) and another 21 patients achieved partial response (PR), whereas the remaining 20 patients failed. One responder died of treatment-related toxicity, and six others declined ABMT. The patient with no measurable disease did not progress on DHAP and was submitted to ABMT. Twenty-two patients underwent ABMT [20 with BEAC and 2 with cyclophosphamide plus total body irradiation (TBI)] of whom 2 (9%) died of toxicity and 10 relapsed. One patient was a suicide at 28 months post-ABMT in CCR and 9 are alive disease-free 24 months to 32 months (median 30 months) post-ABMT. The actuarial 2-year event-free survival for patients undergoing transplantation is 40%. This prospective multicenter trial documents the ability of DHAP followed by ABMT to produce durable complete remission in a significant proportion of patients with relapsed aggressive NHL. Forty-four percent of all patients with relapsed lymphoma who entered the study actually underwent ABMT and 20% of the total group are projected to be long-term disease-free survivors.


Registration of 3D freehand ultrasound to a bone model for orthopedic procedures of the forearm.
For guidance of orthopedic surgery, the registration of preoperative images and corresponding surgical plans with the surgical setting can be of great value. Ultrasound (US) is an ideal modality for surgical guidance, as it is non-ionizing, real time, easy to use, and requires minimal (magnetic/radiation) safety limitations. By extracting bone surfaces from 3D freehand US and registering these to preoperative bone models, complementary information from these modalities can be fused and presented in the surgical realm. A partial bone surface is extracted from US using phase symmetry and a factor graph-based approach. This is registered to the detailed 3D bone model, conventionally generated for preoperative planning, based on a proposed multi-initialization and surface-based scheme robust to partial surfaces. 36 forearm US volumes acquired using a tracked US probe were independently registered to a 3D model of the radius, manually extracted from MRI. Given intraoperative time restrictions, a computationally efficient algorithm was determined based on a comparison of different approaches. For all 36 registrations, a mean (± SD) point-to-point surface distance of [Formula: see text] was obtained from manual gold standard US bone annotations (not used during the registration) to the 3D bone model. A registration framework based on the bone surface extraction from 3D freehand US and a subsequent fast, automatic surface alignment robust to single-sided view and large false-positive rates from US was shown to achieve registration accuracy feasible for practical orthopedic scenarios and a qualitative outcome indicating good visual image alignment.


Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers.
Two-dimensional (2D) alignment and crystallization of membrane proteins (MPs) is increasingly important in characterizing their three-dimensional (3D) structure, in designing pharmacological agents, and in leveraging MPs for biomimetic devices. Large, highly ordered MP 2D crystals in block copolymer (BCP) matrices are challenging to fabricate, but a facile and scalable technique for aligning and crystallizing MPs in thin-film geometries would rapidly translate into applications. This work introduces a novel method to grow larger and potentially better ordered 2D crystals by performing the crystallization process in the presence of a strong magnetic field. We demonstrate the efficacy of this approach using a β-barrel MP, outer membrane protein F (OmpF), in short-chain polybutadiene-poly(ethylene oxide) (PB-PEO) membranes. Crystals grown in a magnetic field were up to 5 times larger than conventionally grown crystals, and a signal-to-noise (SNR) analysis of diffraction peaks in Fourier transforms of specimens imaged by negative-stain electron microscopy (EM) and cryo-EM showed twice as many high-SNR diffraction peaks, indicating that the magnetic field also improves crystal order.


The relationship between safety net activities and hospital financial performance.
During the 1990's hospitals in the U.S were faced with cost containment charges, which may have disproportionately impacted hospitals that serve poor patients. The purposes of this paper are to study the impact of safety net activities on total profit margins and operating expenditures, and to trace these relationships over the 1990s for all U.S urban hospitals, controlling for hospital and market characteristics. The primary data source used for this analysis is the Annual Survey of Hospitals from the American Hospital Association and Medicare Hospital Cost Reports for years 1990-1999. Ordinary least square, hospital fixed effects, and two-stage least square analyses were performed for years 1990-1999. Logged total profit margin and operating expenditure were the dependent variables. The safety net activities are the socioeconomic status of the population in the hospital serving area, and Medicaid intensity. In some specifications, we also included uncompensated care burden. We found little evidence of negative effects of safety net activities on total margin. However, hospitals serving a low socioeconomic population had lower expenditure raising concerns for the quality of the services provided. Despite potentially negative policy and market changes during the 1990s, safety net activities do not appear to have imperiled the survival of hospitals. There may, however, be concerns about the long-term quality of the services for hospitals serving low socioeconomic population.


Maternal serum adiponectin multimers in patients with a small-for-gestational-age newborn.
Several mechanisms of disease have been implicated in the pathophysiology of small-for-gestational-age (SGA) including an anti-angiogenic state, and an exaggerated intravascular pro-inflammatory response. Adiponectin plays a role in a wide range of biological activities including those that have been implicated in the pathophysiology SGA. Thus, the aim of this study was to determine if third trimester adiponectin concentrations differed between women with normal weight infants and those with an SGA neonate. This cross-sectional study included women with: 1) a normal pregnancy (n=234); and 2) an SGA neonate (n=78). SGA was defined as a birth weight below the 10th percentile for gestational age at birth. The study population was further stratified by first trimester body mass index (BMI) (normal weight <25 kg/m(2) vs. overweight/obese > or =25 kg/m(2)). Maternal serum adiponectin multimers [total, high-molecular-weight (HMW), medium-molecular-weight (MMW) and low-molecular-weight (LMW)] concentrations were determined by ELISA. Non-parametric statistics were used for analyses. 1) The median maternal serum concentrations of total, HMW and MMW adiponectin were significantly lower in patients with an SGA neonate than in those with normal pregnancies; 2) patients with an SGA neonate had a significantly lower median HMW/total adiponectin ratio and higher median MMW/total adiponectin and LMW/total adiponectin ratios than those with a normal pregnancy; 3) among patients with an SGA neonate, neither maternal serum concentrations of adiponectin multimers, nor their relative distribution differ between normal weight and overweight/obese patients. 1) Pregnancies complicated by an SGA neonate are characterized by a alterations in the maternal serum adiponectin multimers concentrations and their relative abundance; 2) the findings reported herein suggest that maternal adipose tissue may play a role, in the pathogenesis of SGA.


Eating habits of preschool children with high migrant status in Switzerland according to a new food frequency questionnaire.
Assessment of eating habits in young children from multicultural backgrounds has seldom been conducted. Our objectives were to study the reproducibility and the results of a food frequency questionnaire (FFQ) developed to assess changes in eating habits of preschool children with a high migrant population, in the context of a multidisciplinary multilevel lifestyle intervention. Three kindergarten classes (53% from migrant backgrounds) in French-speaking Switzerland were randomly selected and included 16 girls and 28 boys (mean age +/- SD, 5.4 +/- 0.7 years). The FFQ was filled out twice within a 4-week interval by the parents. Spearman rank correlations between the first and the second FFQ for the 39 items of the food questions were as follows: low (r < 0.50) for 8 (7 P < .05 and 1 nonsignificant), moderate (0.50 <or= r < 0.70) for 22 (all P < .01), and high (r >or= 0.70) for 9 (all P < .01). In addition, 28 of 39 intraclass correlation coefficients were high (>0.50, all P < .01). Eighty-six percent of the children ate breakfast at home daily, but only 67% had lunch at home. The percentages of children eating at least once a week in front of the TV were as follows: 50% for breakfast, 33% for lunch, 38% for dinner, and 48% for snacks. Forty percent of children asked their parents to buy food previously seen in advertisements and ate fast food between once a week and once a month. Children generally consumed foods with a high-energy content. The FFQ yielded good test-retest reproducibility for most items of the food questions and gave relevant findings about the eating habits of preschool children in areas with a high migrant population.


Diagnosing acute myocardial infarction in patients with left bundle branch block.
We compared the clinical features, laboratory and coronary angiographic findings, treatments, and outcomes among patients with ST-segment elevation myocardial infarction (MI) with and without left bundle branch block (LBBB). We examined 5,742 patients with ST-segment elevation MI with and without LBBB treated with primary percutaneous coronary intervention in the Assessment of Pexelizumab in Acute Myocardial Infarction trial. The main outcome measures were obstructive coronary disease, MI, positive cardiac biomarkers, angiographic Thrombolysis In Myocardial Infarction flow, and death, MI, or congestive heart failure at 90 days. LBBB was present in 98 patients (1.7%). According to the protocol, patients with LBBB were eligible only if they had ≥1 mm concordant ST-segment elevation. Obstructive coronary artery disease was present in >87% of the patients with LBBB. Documented MI (elevated biomarkers) with an initially occluded infarct artery was more common in patients with LBBB with concordant ST-segment elevation (71.4%) than in patients without (44.1%; p = 0.027). The use of ST-segment elevation concordance criteria in the presence of LBBB was more often associated with documented MI with an identifiable culprit vessel with an initially occluded infarct-related artery. In conclusion, because a substantial proportion of patients with LBBB have acute MI with a culprit lesion and positive biomarkers, these data support immediate catheterization with the intent for primary percutaneous coronary intervention for all patients presenting with suspected ST-segment elevation myocardial infarction, ischemic symptoms, and presumed new LBBB, particularly if concordant ST-segment elevation is present.


Sperm-egg binding mediated by sperm alpha-L-fucosidase in the ascidian, Halocynthia roretzi.
Spermatozoa bind to the vitelline coat in the ascidians and many other animals. The binding of sperm in Halocynthia roretzi is mediated by a sperm alpha-L-fucosidase and complementary-L-fucosyl residues of glycoproteins in the vitelline coat. cDNA clones for alpha-L-fucosidase were isolated from growing testis mRNA. It contained a 1398 bp full-length cDNA insert (HrFuc'ase) that encoded the 466 amino acid residues of H. roretzi sperm alpha-L-fucosidase. A putative signal peptide of 21 amino acid residues proceeded the sequence for the mature protein (M.W. 52.4 kDa). The coding sequence for HrFuc'ase showed 47.7% sequence identity to the human liver fucosidase sequence. The polyclonal antibody was prepared against a lacZ-HrFuc'ase fusion protein expressed in E. coli. The antibody crossed to a 54 kDa protein in sperm on western blotting and inhibited fertilization in a dose dependent manner. These data suggest that sperm-egg binding is mediated by the sperm alpha-L-fucosidase, HrFuc'ase in the ascidian, H. roretzi.


Long-term effects on cardiovascular risk of a structured multidisciplinary lifestyle program in clinical practice.
Cardiovascular disease is still the leading cause of premature death world-wide with factors like abdominal obesity, hypertension and dyslipidemia being central risk factors in the etiology. The aim of the present study was to investigate the effects on cardiovascular risk factors and cardiovascular risk after 6 months and 1 year, in individuals with increased cardiovascular risk enrolled in a lifestyle multidisciplinary program in a clinical setting. Individuals with increased cardiovascular risk were referred from primary health care and hospitals to a program at an outpatient clinic at a department of cardiology. The program consisted of three individual visits including a health check-up with a physical examination and blood sampling, and a person-centered dialogue for support in behavioural change of unhealthy lifestyle habits (at baseline, 6 months and 1 year). Furthermore, five educational group sessions were given at baseline. Cardiovascular risk was assessed according to Framingham cardiovascular risk predicting model. One hundred individuals (mean age 59 years, 64% women) enrolled between 2008 and 2014 were included in the study. Waist circumference, systolic and diastolic blood pressure and total cholesterol decreased significantly over 1 year. In parallel, cardiovascular risk according to the cardiovascular risk profile based on Framingham 10-year risk prediction model, decreased with 15%. The risk reduction was seen in both men and women, and in participants with or without previous cardiovascular disease. Participating in a structured lifestyle program over a year was associated with significant improvement in multiple cardiovascular risk factors and decreased overall cardiovascular risk. The study is registered at www.clinicaltrials.gov (ClinicalTrial.gov ID: NCT02744157 ).


Detection of rickettsioses and Q fever in Sri Lanka.
Current serological evidence suggests the presence of scrub typhus and spotted fever group (SFG) rickettsiosis in Sri Lanka. Our objective was to identify rickettsial agents/Q fever as aetiological causes for patients who were presumed having rickettsioses by the presence of an eschar or a rash. Sera from patients with unknown origin fever from Matara were tested by immunofluorescence for SFG rickettsial antigens, typhus group rickettsiae, Orientia tsutsugamushi, and Coxiella burnetii antigens. Thirteen (7.3%) of the patients presented with a rash, 11 (6.1%) had an inoculation eschar, and 16 patients recalled a tick or flea bite. We found that 25 (14%) patients had scrub typhus, 6 (3%) SFG rickettsioses, 3 (1.6%) acute Q fever, 3 (1.6%) murine typhus, and 3 (1.6%) were infected by Rickettsia felis. In addition to already described scrub and murine typhus, we found that R. felis and C. burnetii infections should be considered in Sri Lanka.


Pathophysiology of functional mutations of the thiazide-sensitive Na-Cl cotransporter in Gitelman disease.
Most of the missense mutations that have been described in the human SLC12A3 gene encoding the thiazide-sensitive Na(+)-Cl(-) cotransporter (TSC, NCC, or NCCT), as the cause of Gitelman disease, block TSC function by interfering with normal protein processing and glycosylation. However, some mutations exhibit considerable activity. To investigate the pathogenesis of Gitelman disease mediated by such mutations and to gain insights into structure-function relationships on the cotransporter, five functional disease mutations were introduced into mouse TSC cDNA, and their expression was determined in Xenopus laevis oocytes. Western blot analysis revealed immunoreactive bands in all mutant TSCs that were undistinguishable from wild-type TSC. The activity profile was: wild-type TSC (100%) > G627V (66%) > R935Q (36%) = V995M (32%) > G610S (12%) > A585V (6%). Ion transport kinetics in all mutant clones were similar to wild-type TSC, except in G627V, in which a small but significant increase in affinity for extracellular Cl(-) was observed. In addition, G627V and G610S exhibited a small increase in metolazone affinity. The surface expression of wild-type and mutant TSCs was performed by laser-scanning confocal microscopy. All mutants exhibited a significant reduction in surface expression compared with wild-type TSC, with a profile similar to that observed in functional expression analysis. Our data show that biochemical and functional properties of the mutant TSCs are similar to wild-type TSC but that the surface expression is reduced, suggesting that these mutations impair the insertion of a functional protein into the plasma membrane. The small increase in Cl(-) and thiazide affinity in G610S and G627V suggests that the beginning of the COOH-terminal domain could be implicated in defining kinetic properties.


The secretory granule matrix: a fast-acting smart polymer.
The secretory granule matrix is a miniature biopolymer that consists of a charged polymer network that traps peptides and transmitters when it condenses and releases them on exocytotic decondensation. Models of exocytotic fusion have treated this matrix as a short circuit and have neglected its electrical contributions. This matrix responded to negative voltages by swelling, which was accompanied by a large increase in conductance, and to positive voltages by condensing. Thus, the matrix resembled a diode. The swollen matrix exerted large pressures on the order of 12 bar. The responses took place within milliseconds of the application of the electric field. These findings suggest that matrix decondensation, and therefore product release, is controlled by potential gradients.


An Update of the Virion Proteome of Kaposi Sarcoma-Associated Herpesvirus.
The virion proteins of Kaposi sarcoma-associated herpesvirus (KSHV) were initially characterized in 2005 in two separate studies that combined the detection of 24 viral proteins and a few cellular components via LC-MS/MS or MALDI-TOF. Despite considerable advances in the sensitivity and specificity of mass spectrometry instrumentation in recent years, leading to significantly higher yields in detections, the KSHV virion proteome has not been revisited. In this study, we have re-examined the protein composition of purified KSHV virions via ultra-high resolution Qq time-of-flight mass spectrometry (UHR-QqTOF). Our results confirm the detection of all previously reported virion proteins, in addition to 17 other viral proteins, some of which have been characterized as virion-associated using other methods, and 10 novel proteins identified as virion-associated for the first time in this study. These results add KSHV ORF9, ORF23, ORF35, ORF48, ORF58, ORF72/vCyclin, K3, K9/vIRF1, K10/vIRF4, and K10.5/vIRF3 to the list of KSHV proteins that can be incorporated into virions. The addition of these proteins to the KSHV virion proteome provides novel and important insight into early events in KSHV infection mediated by virion-associated proteins. Data are available via ProteomeXchange with identifier PXD022626.