Tbx5a lineage tracing shows cardiomyocyte plasticity during zebrafish heart regeneration.
During development, mesodermal progenitors from the first heart field (FHF) form a primitive cardiac tube, to which progenitors from the second heart field (SHF) are added. The contribution of FHF and SHF progenitors to the adult zebrafish heart has not been studied to date. Here we find, using genetic tbx5a lineage tracing tools, that the ventricular myocardium in the adult zebrafish is mainly derived from tbx5a+ cells, with a small contribution from tbx5a- SHF progenitors. Notably, ablation of ventricular tbx5a+-derived cardiomyocytes in the embryo is compensated by expansion of SHF-derived cells. In the adult, tbx5a expression is restricted to the trabeculae and excluded from the outer cortical layer. tbx5a-lineage tracing revealed that trabecular cardiomyocytes can switch their fate and differentiate into cortical myocardium during adult heart regeneration. We conclude that a high degree of cardiomyocyte cell fate plasticity contributes to efficient regeneration.


The SUDEP Risk Inventory: Association with postictal generalized EEG suppression.
To help identify patients at greatest risk for sudden unexpected death in epilepsy (SUDEP), screening inventories like the SUDEP-7 Inventory can be useful. In this study, we examined the strength of association between this inventory's risk factors and postictal generalized EEG suppression (PGES), a biomarker of SUDEP risk. We reanalyzed data from an epilepsy monitoring unit study of 37 children. We performed a 2 by 2 contingency table analysis to determine the association between "yes" responses on the inventory questions and PGES following >=1 seizure. Having a history of >3 generalized tonic-clonic seizures (GTCS) in the past year had the strongest association with PGES (Pearson chi-square p<0.001, Cramer's V=0.75). Having >=1 GTCS in the past year was also strongly associated with PGES (Pearson chi-square p<0.001, Cramer's V=0.636). Histories of >50 seizures of any type/month (Pearson chi-square p=0.14, Cramer's V=0.241) and intellectual disability (Pearson chi-square p=0.04, Cramer's V=0.337) were not as robustly associated with PGES. Current use of >=3 AEDs had the weakest association with PGES (Pearson chi-square p=0.66, Cramer's V=0.072). Given that all study patients had >=1 seizure per year and epilepsy durations <30 years, the strength of association with these questions and PGES could not be analyzed.


Optimal configuration of the spiral linking technique for tendon repair. Biomechanical evaluation.
A previous study described a new spiral linking technique for tendon repairs and demonstrated that it was strong enough to be used in clinical practice as an alternative to the Pulvertaft tendon weave repair. However the repairs were less stiff, needed slightly more tendon length for the same repair and were a little bulkier. In this study two variables have been changed with a view to improving the spiral technique. At first the number of spirals was reduced consecutively, keeping the same number of standard mattress sutures. Once the optimal number of spirals had been identified, repairs with different numbers of sutures were tested using an alternative cross-stitch technique. The spiral repair technique using two spirals linked with six sutures was at least as strong and stiff as a four-weave Pulvertaft technique and was also easier to do.


Down-regulation of protein kinase C alpha/ezrin signals in light-induced phagocytic crisis of retinal pigment epithelium cells.
To investigate the roles of PKC-α/ezrin signals in phagocytosis crisis of retinal pigment epithelium (RPE) cells in light damage model. Light induced mice RPE injury model was established by continuously irradiating cool white light at different exposure time (0, 4, 8h light intensity: 4.18×10-6 J/cm2). In vitro, human ARPE-19 cells treated with the doses and intensity (1.57×10-6 J/cm2) of laser irradiation. Histology analysis was evaluated by hematoxylin and eosin (HE) staining. In vivo RPE phagocytosis was quantified by measuring the accumulation of photoreceptor outer segments in the sub-retinal space. In vitro RPE phagocytosis was assessed by calculating the relative fluorescence intensity of FITC-labeled microspheres in ARPE-19 cells. To further investigate the molecular mechanism, the activation of PKC-α/ezrin signal was evaluated by Western blot in vivo and in vitro. HE staining revealed that the thickness of outer nuclear layer decreased significantly after 4 and 8h light exposure. By immunostaining with rhodopsin, a significant greater accumulation of photoreceptor outer segment was noticed after light injury. In vitro, light injured RPE cells had less phagocytic activity in a dose dependent manner than that of the normal control (P<0.01). Western blot suggested the activation of PKC-α/ezrin signaling was down-regulated in a dose-dependent manner after light exposure. Our data suggest that light induced phagocytic crisis of RPE cells may result from the down-regulation of PKC-α/ezrin signaling.


The mirror like expression of genes involved in the FOXO signaling pathway could be effective in the pathogenesis of human lymphotropic virus type 1 (HTLV-1) through disruption of the downstream pathways.
Human lymphotropic virus type 1 (HTLV-1) is the cause of two major diseases, ATLL and HAM/TSP in a percentage of carriers. Despite progress in understanding the pathogenesis of these two diseases, the exact pathogenesis mechanism is still not well understood. High-throughput technologies have revolutionized medical research. This study aims to investigate the mechanism of pathogenesis of these two diseases using the results of high-throughput analysis of microarray datasets. A total of 100 differentially expressed genes were found between ATLL and HAM/TSP. After constructing protein-protein network and further analyzing, proteins including ATM, CD8, CXCR4, PIK3R1 and CD2 were found as the hub ones between ATLL and HAM/TSP. Finding the modules of the subnetwork revealed the enrichment of two common pathways including FOXO signaling pathway and Cell cycle with two common genes including ATM and CDKN2D. Unlike ATLL, ATM gene had higher expressions in HAM/TSP patients. The expression of CDKN2D was increased in ATLL patients. The results of this study could be helpful for understanding the pathogenic mechanism of these two diseases in the same signaling pathways.


Phosphosites of the yeast centrosome component Spc110 contribute to cell cycle progression and mitotic exit.
Spc110 is an essential component of the spindle pole body (SPB), the yeast equivalent of the centrosome, that recruits the γ-tubulin complex to the nuclear side of the SPB to produce the microtubules that form the mitotic spindle. Here, we identified phosphosites S11 and S36 in maternally originated Spc110 and explored their functions in vivo. Yeast expressing non-phosphorylatable Spc110S11A had a distinct spindle phenotype characterised by higher levels of α-tubulin, which was frequently asymmetrically distributed between the two SPBs. Furthermore, expression of the double mutant Spc110S11AS36A had a delayed cell cycle progression. Specifically, the final steps of mitosis were delayed in Spc110S11AS36A cells, including expression and degradation of the mitotic cyclin Clb2, disassembling the mitotic spindle and re-localizing Cdc14 to the nucleoli, resulting in late mitotic exit and entry in G1. Thus, we propose that Spc110 phosphorylation at S11 and S36 is required to regulate timely cell cycle progression in budding yeast. This article has an associated First Person interview with the first author of the paper.


A locus on mouse chromosome 6 that determines resistance to herpes simplex virus also influences reactivation, while an unlinked locus augments resistance of female mice.
During studies to determine a role for tumor necrosis factor (TNF) in herpes simplex virus type 1 (HSV-1) infection using TNF receptor null mutant mice, we discovered a genetic locus, closely linked to the TNF p55 receptor (Tnfrsf1a) gene on mouse chromosome 6 (c6), that determines resistance or susceptibility to HSV-1. We named this locus the herpes resistance locus, Hrl, and showed that it also mediates resistance to HSV-2. Hrl has at least two alleles, Hrl(r), expressed by resistant strains like C57BL/6 (B6), and Hrl(s), expressed by susceptible strains like 129S6 (129) and BALB/c. Although Hrl is inherited as an autosomal dominant gene, resistance to HSV-1 is strongly sex biased such that female mice are significantly more resistant than male mice. Analysis of backcrosses between resistant B6 and susceptible 129 mice revealed that a second locus, tentatively named the sex modifier locus, Sml, functions to augment resistance of female mice. Besides determining resistance, Hrl is one of several genes involved in the control of HSV-1 replication in the eye and ganglion. Remarkably, Hrl also affects reactivation of HSV-1, possibly by interaction with some unknown gene(s). We showed that Hrl is distinct from Cmv1, the gene that determines resistance to murine cytomegalovirus, which is encoded in the major NK cell complex just distal of p55 on c6. Hrl has been mapped to a roughly 5-centimorgan interval on c6, and current efforts are focused on obtaining a high-resolution map for Hrl.


Role of endothelin and endothelin A-type receptor in adaptation of the carotid body to chronic hypoxia.
Chronic exposure in a low-PO(2) environment (i.e., chronic hypoxia, CH) elicits an elevated hypoxic ventilatory response and increased hypoxic chemosensitivity in arterial chemoreceptors in the carotid body. In the present study, we examine the hypothesis that changes in chemosensitivity are mediated by endothelin (ET), a 21-amino-acid peptide, and ET(A) receptors, both of which are normally expressed by O(2)-sensitive type I cells. Immunocytochemical staining showed incremental increases in ET and ET(A) expression in type I cells after 3, 7, and 14 days of CH (380 Torr). Peptide and receptor upregulation was confirmed in quantitative RT-PCR assays conducted after 14 days of CH. In vitro recordings of carotid sinus nerve activity after in vivo exposure to CH for 1-16 days demonstrated a time-dependent increase in chemoreceptor activity evoked by acute hypoxia. In normal carotid body, the specific ET(A) antagonist BQ-123 (5 microM) inhibited 11% of the nerve discharge elicited by hypoxia, and after 3 days of CH the drug diminished the hypoxia-evoked discharge by 20% (P < 0.01). This inhibitory effect progressed to 45% at day 9 of CH and to nearly 50% after 12, 14, and 16 days of CH. Furthermore, in the presence of BQ-123, the magnitude of the activity evoked by hypoxia did not differ in normal vs. CH preparations, indicating that the increased activity was the result of endogenous ET acting on an increasing number of ET(A). Collectively, our data suggest that ET and ET(A) autoreceptors on O(2)-sensitive type I cells play a critical role in CH-induced increased chemosensitivity in the rat carotid body.


Scintigraphic detection of colon carcinomas with iodinated monoclonal antibodies.
The aim of this study was to evaluate the clinical reliability of the immunoscintigraphy with iodinated monoclonal antibodies for the detection of metastases and recurrences of colon carcinomas. A total of 45 patients with colon carcinoma was investigated with gamma camera, after intravenous application of iodinated monoclonal antibodies. The sensitivity of the method was 90%, specificity 86%, positive predictive value 93%, negative predictive value 80% and accuracy 87%. There was statistically significant relationship between immunoscintigraphic and ultrasonographic (US) findings (p=0.005). Also, there was significant relationship between immunoscintigraphy and Dukes stage (p=0.019). Tumor marker levels were not significantly correlated with immunoscintigraphic findings (p<0.05). Significant difference was noted in patients with positive findings for malignancy on US and immunoscintigraphic findings (p=0.006), i.e. patients with positive findings for malignancy had more frequently immunoscintigraphic findings of malignancy. Correlation with other diagnostic procedures (rectoscopy, colonoscopy, CT) did not show significant correlations. We conclude that immunoscintigraphy can be helpful in the detection of metastases and recurrences of colon carcinomas.


Xeromammographic screening: is it helping?
We have reviewed our experience with xeromammography over a five-year period in a setting where there is no protocol for mammographic screening. Only 8.9% of breast carcinomas were found by xeromammography alone, a ratio substantially smaller than that found in large screening programs. While 80% of the occult carcinomas were localized, only 47% of women with palpable disease had localized tumors. It appears that screening mammography is underused, and that more localized tumors would be detected if more mammograms were ordered.


Finding covert fluid: methods for detecting volume overload in children on dialysis.
Lung ultrasound is a novel technique for detecting generalized fluid overload in children and adults with end-stage renal disease (ESRD). Echocardiography and bioimpedance spectroscopy are established methods, albeit variably adopted in clinical practice. We compared the practicality and accuracy of lung ultrasound with current objective techniques for detecting fluid overload in children with ESRD. A prospective observational study was performed to compare lung ultrasound B-lines, echocardiographic measurement of inferior vena cava parameters and bioimpedance spectroscopy in the assessment of fluid overload in children with ESRD on dialysis. The utility of each technique in predicting fluid overload, based on short-term weight gain, was assessed. Multiple linear regression models to predict fluid overload by weight were explored. A total of 22 fluid assessments were performed in 13 children (8 on peritoneal dialysis, 5 on haemodialysis) with a median age of 4.0 (range 0.8-14.0) years. A significant linear correlation was observed between the number of B-lines detected by lung ultrasound and fluid overload by weight (r = 0.57, p = 0.005). A non-significant positive linear correlation was observed between fluid overload by weight and bioimpedance spectroscopy (r = 0.43, p = 0.2), systolic blood pressure (r = 0.19, p = 0.4) and physical examination measurements (r = 0.19, p = 0.4), while a non-significant negative linear relationship was found between the inferior vena cava collapsibility index and fluid overload by weight (r = -0.24, p = 0.3). In multiple linear regression models, a combination of three fluid parameters, namely lung ultrasound B-lines, clinical examination and systolic blood pressure, best predicted fluid overload (R 2 = 0.46, p = 0.05). Lung ultrasound may be superior to echocardiographic methods and bioimpedance spectroscopy in detecting volume overload in children with ESRD. Given the practicality and sensitivity of this new technique, it can be adopted alongside clinical examination and blood pressure in the routine assessment of fluid status in children with ESRD.


Relationship of mineral status and intake to periodontal disease.
A periodontal index (PI), based on visual and radiographic data, was used to evaluate the severity of periodontal disease in a group of 80 dental patients. The average PI was 3.1 +/- 1.8 SD (0=no detectable lesions; 8=most severe lesions). The PI was directly related to patient age (r=0.73; P is less than 0.01). The serum concentration of calcium copper, iron, magnesium, manganese, and zinc was measured. Regression of PI on the serum constituents demonstrated that serum copper was linearly related (r-0.64; P is less than 0.001) to the PI. The other serum parameters were not correlated to the PI. Dietary protein, calcium, and vitamin A were slightly related to the PI, based on a 24-hr dietary recall. Dietary trace elements were not related to the PI. These results indicate that the serum copper concentration, in some way is related to the severity of periodontal disease. Possibly the observation is linked to the role of copper in the inflammatory response since inflammation is a characteristic of periodontal disease.


Cytokeratin expression during AFB1-induced carcinogenesis.
The early stages of the carcinogenic process induced by aflatoxin B1 (AFB1) in rat liver during 24 weeks of feeding and the resulting tumours have been studied with respect to cytokeratin (CK) expression. A previously uncharacterized monoclonal antibody, MRCTU/J1, has been shown to recognize rat CK18 and together with antibodies against human CK8, 18 and 19, has been used to examine the possible lineage of tumour cells and also to identify the altered foci that might be most relevant to tumorigenesis. Results suggested that AFB1-induced transformation in liver may occur in more than one cell type, since tumours with the normal hepatocyte CK pattern and those with bile duct or oval cell CK phenotype were identified. Additionally, hepatocytes with a bile duct CK phenotype appeared during the early stages of carcinogenesis. The in vivo pattern of CK expression also appeared to be maintained in one normal and one hepatoma-derived cell line. Overexpression of CKs (particularly of CK19) was a much more selective marker for altered foci, compared to gamma-glutamyltranspeptidase, and was more consistently expressed at high levels in tumours, suggesting that it might be a more reliable way of identifying those cells involved in the transformation process.


Dizinc cation [Zn2](2+) trapped in a homoleptic metalloid coordination environment stabilized by dispersion forces: [Zn2(GaCp*)6][BAr4(F)]2.
The synthesis and characterization of the cationic mixed metal Ga/Zn cluster [Zn2(GaCp*)6](2+) (1) is presented. The reaction of [Zn2Cp*2] with [Ga2Cp*][BAr4(F)] leads to the formation of the novel complex being the first example of a [Zn2](2+) core exclusively ligated by metalloid group-13 organyl-ligands. Compound 1 exhibits two different coordination modes: In the solid state, two of the six GaCp* ligands occupy bridging positions, whereas VT (1)H NMR indicates the coexistence of a second isomer in solution featuring six terminal GaCp* ligands. Quantum chemical calculations have been carried out to assign the gallium and zinc positions; the bonding situation in 1 is characterized and the importance of dispersion forces is discussed.


Studies in the biotransformation of cortisol to cortoic acids in man. III. 21-Oxidation of 4-14C,21-3H-desoxycorticosterone.
The metabolism of (4-14C, 21-3H) desoxycorticosterone (DOC) in man has been studied. DOC, like cortisol, undergoes oxidation at C-21 with the formation of acidic metabolites (2.3-8.2% of the dose). The release of 3H into the body water, either by oxidation or exchange, is approximately twice as great as the actual formation of acidic metabolites (as judged from the recovery of 14C in the urinary acidic fraction), but both parameters were considerably smaller than the corresponding values for cortisol. The principal metabolite isolated from the urine was 21-hydroxypregnanolone, which had an isotope ratio (3H/14C) greater than that of the dose; the pregnanetriols, which were formed in lesser amounts, had significantly lower isotope ratios than the dose. The absence of the 17-hydroxy group in DOC appears to modify C-21 oxidation, decreasing its magnitude and altering the character of the products.


A resolution of the ascertainment sampling problem: IV. Continuous phenotypes.
This paper considers ascertainment corrections for continuous phenotypes. Two main points are considered. The first is a discussion of what ascertainment corrections can be devised to ensure asymptotically unbiased parameter estimates when the nature of the ascertainment procedure is not known. The second is an analysis of the properties of various forms of ascertainment correction possible when the nature of the ascertainment procedure is known. Some ascertainment corrections are thus shown to be valid and others invalid.


The ADRON-RM Instrument Onboard the ExoMars Rover.
This overview presents the physical principles, design, measurement capabilities, and summary of planned operations of the autonomous detector of radiation of neutrons onboard rover at Mars (ADRON-RM) on the surface of Mars. ADRON-RM is a Russian project selected for the joint European Space Agency-Roscosmos ExoMars 2020 landing mission. A compact passive neutron spectrometer, ADRON-RM, was designed to study the abundance and distribution of water and neutron absorption elements (such as Cl, Fe, and others) in the martian subsurface along the path of the ExoMars rover. Key Words: Mars exploration-Surface-Neutron Spectroscopy-Water. Astrobiology 17, 585-594.


Antibacterial biocompatible arginine functionalized mono-layer graphene: No more risk of silver toxicity.
Antibacterial ability is vital in biological approaches as well as functional biomaterials. Besides, cytocompatibility aspect of biologic media, tissue and organs is always concern for appropriate synthesis. From the past, metallic/oxide phases of silver (Ag) material in various macro, micro or nano configurations have been widely used for antibacterial targets. While, background of Ag toxicity within particle, film and composites is posing gradual ion release affected by molecular bounding. Recent researches conducted to control, optimize and neutralize Ag limitations finding the benefits of ideal (∼ 100%) mediation against both Gram-negative and Gram-positive bacteria. Whereas, non-degradable releases history is still a challenge and its longer accumulation may cause to disrupt biostructures and disease risk. Thus, facile development of large-area organic materials with switchable bacteria toxicity and normal cell compatibility function is interesting for concerned approaches. Here, smart positively-charged stable arginine amino acid incorporated mono layer graphene (Arg-EMGr) nanobiocomposite introduced as useful antibacterial and safe bactericidal agent competitive with Ag direct. The immunity characteristic versus Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) comparably assessed with graphene oxide (GO) and different concentrations GO-AgNPs morphology. As cell viability matter, 1,3,5,7-days vitro culture assay shown attachment proliferation and cytotoxicity due to short interaction.


Computational drug repositioning using low-rank matrix approximation and randomized algorithms.
Computational drug repositioning is an important and efficient approach towards identifying novel treatments for diseases in drug discovery. The emergence of large-scale, heterogeneous biological and biomedical datasets has provided an unprecedented opportunity for developing computational drug repositioning methods. The drug repositioning problem can be modeled as a recommendation system that recommends novel treatments based on known drug-disease associations. The formulation under this recommendation system is matrix completion, assuming that the hidden factors contributing to drug-disease associations are highly correlated and thus the corresponding data matrix is low-rank. Under this assumption, the matrix completion algorithm fills out the unknown entries in the drug-disease matrix by constructing a low-rank matrix approximation, where new drug-disease associations having not been validated can be screened. In this work, we propose a drug repositioning recommendation system (DRRS) to predict novel drug indications by integrating related data sources and validated information of drugs and diseases. Firstly, we construct a heterogeneous drug-disease interaction network by integrating drug-drug, disease-disease and drug-disease networks. The heterogeneous network is represented by a large drug-disease adjacency matrix, whose entries include drug pairs, disease pairs, known drug-disease interaction pairs and unknown drug-disease pairs. Then, we adopt a fast Singular Value Thresholding (SVT) algorithm to complete the drug-disease adjacency matrix with predicted scores for unknown drug-disease pairs. The comprehensive experimental results show that DRRS improves the prediction accuracy compared with the other state-of-the-art approaches. In addition, case studies for several selected drugs further demonstrate the practical usefulness of the proposed method. http://bioinformatics.csu.edu.cn/resources/softs/DrugRepositioning/DRRS/index.html. firstname.lastname@example.com or email@example.com. Supplementary data are available at Bioinformatics online.


The root epidermis-specific pea gene RH2 is homologous to a pathogenesis-related gene.
Two-dimensional gel electrophoresis of pea root and root hair proteins revealed the existence of at least 10 proteins present at elevated levels in root hairs. One of these, named RH2, was isolated and a partial amino acid sequence was determined from two tryptic peptides. Using this sequence information oligonucleotides were designed to isolate by PCR an RH2 cDNA clone. In situ hybridization studies with this cDNA clone showed that rh2 is not only expressed in root hairs, but also in root epidermal cells lacking these tubular outgrowths. During post-embryonic development the gene is switched on after the transition of protoderm into epidermis and since rh2 is already expressed in a globular pea embryo in the protoderm at the side attached to the suspensor, we conclude that the expression of rh2 is developmentally regulated. At the amino acid level RH2 is 95% homologous to the pea PR protein I49a. These gene encoding I49a is induced in pea pods upon inoculation with the pathogen Fusarium solani [12]. We postulate that rh2 contributes to a constitutive defence barrier in the root epidermis. A similar role has been proposed for chalcone synthase (CHS) and chitinase, pathogenesis-related protein that are also constitutively present in certain epidermal tissues.


Taxonomy and Phylogeny of Peronospora Species (Oomycota) Parasitic to Stellaria and Pseudostellaria in Korea, with the Introduction of Peronospora casparyi sp. nov.
The genus Peronospora, an obligate biotrophic group belonging to Oomycota, causes serious damage to a variety of wild and ornamental plants, as well as cultivated crops, such as beet, rose, spinach, and tobacco. To investigate the diversity of Peronospora species parasitic to Stellaria and Pseudostellaria (Caryophyllaceae) plants in Korea, we performed a morphological analysis on dried herbarium specimens and molecular phylogenetic inferences based on internal transcribed spacer rDNA and cox2 mitochondrial DNA sequences. As a result, it was confirmed that there are four species of Peronospora parasitic to specific species of Stellaria and Pseudostellaria, all of which were hitherto unrecorded in Korea: P. alsinearum (ex Stellaria media), P. stellariae-aquaticae (ex Stellaria aquatica), P. stellariae-uliginosae (ex Stellaria alsine), and P. pseudostellariae (ex Pseudostellaria palibiniana). In addition, Peronospora specimens parasitic to Pseudostellaria davidii differed morphologically from P. pseudostellariae owing to the large and ellipsoidal conidia; this morphological discrepancy was also validated by the high genetic divergence between the two species. Peronospora casparyi sp. nov. is described and illustrated here.


SiMa Cells for a Serotype Specific and Sensitive Cell-Based Neutralization Test for Botulinum Toxin A and E.
Botulinum toxins (BoNTs), of which there are seven serotypes, are among the most potent neurotoxins, with serotypes A, B and E causing human botulism. Antitoxins form the first line of treatment for botulism, and functional, highly sensitive in vitro methods for toxin neutralization are needed to replace the current in vivo methods used for determination of antitoxin potency. In this preliminary proof of concept study, we report the development of a neutralization test using the neuroblastoma SiMa cell line. The assay is serotype specific for either BoNT/A or BoNT/E, which both cleave unique sequences on SNAP-25 within SiMa cells. The end point is simple immunodetection of cleaved SNAP-25 from cell lysates with antibodies detecting only the newly exposed sequence on SNAP-25. Neutralizing antibodies prevent the toxin-induced cleavage of SNAP-25. The toxin neutralization assay, with an EC50 of ~2 mIU/mL determined with a standardized reference antiserum, is more sensitive than the mouse bioassays. Relevance was demonstrated with commercial and experimental antitoxins targeting different functional domains, and of known in vivo neutralizing activities. This is the first report describing a simple, specific, in vitro cell-based assay for the detection of neutralizing antibodies against BoNT/A and BoNT/E with a sensitivity exceeding that of the mouse bioassay.


Telemedicine: a child psychiatry case report.
The topic addressed is the use of telemedicine involving a rural health care setting and child clinical services provided through an innovative model of telehealth applications. Telehealth technology and services have gained the attention of both scientists and practitioners examining trends and models of health care delivery for underserved populations and in situations where consultation with a team of professionals may benefit service providers in rural communities. Examined is an innovative model of telehealth care delivery through a rural school system in an underserved regional setting. The goals of this model are offered, as are a number of applications within the broad spectrum of services utilizing telehealth. Changing patterns in clinically based health care delivery in a managed care environment are discussed.


Cirrhotics with variceal hemorrhage: the importance of the time interval between admission and the start of analysis for survival and rebleeding rates.
The importance of variable time of entry for analysis of survival following variceal bleeding has recently been disputed. In a study of 194 cirrhotic patients with bleeding esophageal varices in whom 2-day mortality was 3%, statistically significant differences in both survival and rebleeding rates were obtained by shifting the starting point for analysis of survival by 2 weeks following admission to hospital or by 5 days for the analysis of rebleeding. In addition, the curve of hazard function for death or for failure to control bleeding following admission clearly showed that any change in entry time in a study of variceal bleeding would introduce bias and alter survival or rebleeding rates. Thus, the starting point for analysis following variceal hemorrhage is an important confounding variable when calculating both survival and rebleeding. It should always be taken into account, particularly in clinical trials, which are often performed in centers where patients are referred from other hospitals at different times following bleeding.


[Laparoscopic living donor nephrectomy for kidney transplantation].
Globally, the number of patients with end-stage renal disease (ESRD) has been steadily increasing every year. These patients need renal replacement therapy which includes peritoneal dialysis, programmed hemodialysis and kidney transplantation, which is the ultimate form of replacement. Living donor kidney transplantation has significant advantages over cadaveric kidney transplantation regarding the long-term functional results of renal transplant and the survival of recipients. Open nephrectomy, previously used to retrieve a healthy kidney from a donor is associated with traumatic access and a long recovery period of the donor. With the development of minimally invasive technologies, laparoscopic living donor nephrectomy has been developed, which currently is the best method for retrieving transplant kidney. The review outlines the characteristics and outcomes of laparoscopic living donor nephrectomy for kidney transplantation.


Engaging families in child welfare services: an evidence-based approach to best practice.
Successfully engaging clients in the helping process is a critical task for child welfare practitioners. Drop-out and noncompliance rates in child welfare services are high and lead to high rates of removal of children from their families and to eventual termination of parental rights. Although no known interventions guarantee treatment compliance, this review of the empirical literature delineates critical components of engagement in child welfare services. Effective engagement strategies, including service components and caseworker qualities and behaviors, are identified as contributing to the positive case outcomes of treatment compliance, family preservation, and placement prevention. The unique needs of neglectful parents are also examined, with recommendations for practice.


Energy considerations in a model of nematode sperm crawling.
In this paper we propose a mathematical model for nematode sperm cell crawling. The model takes into account both force and energy balance in the process of lamellipodium protrusion and cell nucleus drag. It is shown that by specifying the (possibly variable) efficiency of the major sperm protein biomotor one completely determines a self-consistent problem of the lamellipodium-nucleus motion. The model thus obtained properly accounts for the feedback of the load on the lamellipodium protrusion, which in general should not be neglected. We study and analyze the steady crawling state for a particular efficiency function and find that all nonzero modes, up to a large magnitude, are linearly asymptotically stable, thus reproducing the experimental observations of the long periods of steady crawling exhibited by the nematode sperm cells.


Intramolecular Hydrogen Bonding Involving Organic Fluorine: NMR Investigations Corroborated by DFT-Based Theoretical Calculations.
The combined utility of many one and two dimensional NMR methodologies and DFT-based theoretical calculations have been exploited to detect the intramolecular hydrogen bond (HB) in number of different organic fluorine-containing derivatives of molecules, viz. benzanilides, hydrazides, imides, benzamides, and diphenyloxamides. The existence of two and three centered hydrogen bonds has been convincingly established in the investigated molecules. The NMR spectral parameters, viz., coupling mediated through hydrogen bond, one-bond NH scalar couplings, physical parameter dependent variation of chemical shifts of NH protons have paved the way for understanding the presence of hydrogen bond involving organic fluorine in all the investigated molecules. The experimental NMR findings are further corroborated by DFT-based theoretical calculations including NCI, QTAIM, MD simulations and NBO analysis. The monitoring of H/D exchange with NMR spectroscopy established the effect of intramolecular HB and the influence of electronegativity of various substituents on the chemical kinetics in the number of organic building blocks. The utility of DQ-SQ technique in determining the information about HB in various fluorine substituted molecules has been convincingly established.


The influence of forgiveness and apology on cardiovascular reactivity and recovery in response to mental stress.
To investigate the relation between forgiveness and apology as they relate to cardiovascular reactivity and recovery, 29 men and 50 women were exposed to an interpersonal transgression (i.e., verbal harassment) while performing a serial subtraction task. Participants were categorized into high and low forgiveness groups based on scores on the forgiving personality scale. Following the task, approximately half of the participants received an apology from the experimenter for his/her comments during the task. Although no group differences in cardiovascular reactivity were observed during the serial subtraction task, persons high in forgiveness displayed more rapid diastolic and mean arterial blood pressure recovery than persons low in forgiveness. In response to the apology, participants displayed greater high frequency heart rate variability recovery compared to those who did not receive an apology. A significant apology x sex interaction was observed for diastolic blood pressure and mean arterial blood pressure. Women who received an apology exhibited faster recovery from the transgression than women who did not receive an apology. In contrast, men who received an apology exhibited delayed recovery from the transgression compared to men who did not receive an apology. These results indicate that there are potentially healthful benefits to forgiveness and apology, but the relation is influenced by situation and by sex.


Amoxapine--an antidepressant with some neuroleptic properties? A review of its chemistry, animal pharmacology and toxicology, human pharmacology, and clinical efficacy.
Amoxapine, a new antidepressant, exhibits both antidepressant and neuroleptic effects in laboratory animals and in humans. Evidence from human studies (extrapyramidal reactions, hyperprolactinemia, and galactorrhea), animal screening tests, and neurochemical experiments support the contention that amoxapine or a metabolite occasionally produces neuroleptic-like effects. Amoxapine's neuroleptic activity may derive from 7-hydroxy-amoxapine, a minor metabolite in humans, which exhibits significant dopamine receptor-blocking activity. Evidence for an early antidepressant effect of amoxapine in the treatment of depressive illness has not been consistently demonstrated. In comparable doses (roughly twice the dose of imipramine or amitriptyline), amoxapine appears to be similar to reference antidepressants in efficacy, unwanted effects, and acute toxicity.


Ocular ultrasound as an easy applicable tool for detection of Terson's syndrome after aneurysmal subarachnoid hemorrhage.
Intraocular hemorrhage in patients suffering from aneurysmal subarachnoid hemorrhage is known as Terson's syndrome and is an underestimated but common pathology. We therefore designed a prospective single-blinded study to evaluate the validity of ocular ultrasound compared to the gold standard indirect funduscopy in the diagnosis of Terson's syndrome. Fifty-two patients (104 eyes in total) suffering from aneurysmal subarachnoid hemorrhage were enrolled in this study. Two investigators independently performed a single-blinded ocular ultrasound using a standard intensive care ultrasound system to detect an intraocular hemorrhage. Indirect funduscopy following iatrogenic mydriasis served as the gold standard for confirmation or exclusion of an intraocular hemorrhage. Statistical analyses were performed to evaluate the sensitivity and specificity, positive and negative predictive values of the method as well as the learning curve of ocular ultrasound. Indirect funduscopy detected Terson's syndrome in 11 of 52 (21.2%) respectively in 21 of 104 (20.2%) eyes in patients suffering from subarachnoid hemorrhage. Sensitivity and specificity increased with the number of ocular ultrasound examinations for both investigators, reaching 81.8% and 100% respectively. Positive and negative predictive values were different for both investigators (63.6% vs. 100% positive and 100% vs. 95.7% negative) but were both correlated to the amount of intraocular hemorrhage. A low Glasgow Coma scale (p = 0.015) and high Hunt & Hess grade (p = 0.003) was associated with a higher rate of Terson's syndrome. Ocular ultrasound using standard ultrasound equipment has been confirmed as a reliable, easy-to-handle bedside screening tool for detecting Terson's syndrome. Nevertheless funduscopy remains the gold standard to detect Terson's syndrome.


Endoscopic submucosal dissection of a rare antral mass: gastric fibrolipoma.
A 60-year-old man was hospitalized with upper abdominal pain for 2-month duration. He had no previous history of symptoms related to his upper gastrointestinal tract. Physical examination was unremarkable. A semicircular round mass with smooth surface was found at the gastric antrum measuring 2.0 × 2.0-cm by gastroscopy. Endoscopic ultrasound (EUS) demonstrated a heterogeneous hyperechoic mass originated from the submucosal layer. Abdominal computed tomography (CT) revealed a fat-density mass with internal mixed density at the gastric antrum. His complete blood test, liver, renal, coagulation function and tumor markers were within reference values. Endoscopic submucosal dissection (ESD) was performed. Postoperative histopathology examination revealed the tumor was composed of closely arranged proliferating mature adipocytes with consistent size and shape. Collagen fiber hyperplasia between adipocytes was found in focal area. The patient was diagnosed as gastric fibrolipoma. He was discharged home uneventful and no symptoms was found in 12 months follow-up.


The challenge of poliomyelitis in tropical Africa.
Recent surveys of lameness attributable to poliomyelitis in several African countries have revealed rates of residual paralysis of two to 11 per 1,000 children, a prevalence implying annual incidence rates for poliomyelitis of 10-55 cases per 100,000 population. Factors responsible for this high incidence are the unsanitary conditions present in many African communities, especially rural and periurban areas. Whereas poliomyelitis is endemic in Africa and paralysis occurs early in childhood, the apprehension and alarm that is evident in the developed world is almost absent in Africa. Hence, the administrative and economic resources for massive programs of poliomyelitis vaccination are lacking. Lameness prevalence rates of 2.6 per 1,000 children in certain areas of Africa illustrate that the programs of immunization against poliomyelitis should be intensified.


Novel Naja atra cardiotoxin 1 (CTX-1) derived antimicrobial peptides with broad spectrum activity.
Naja atra subsp. atra cardiotoxin 1 (CTX-1), produced by Chinese cobra snakes, belonging to Elapidae family, is included in the three-finger toxin family and exerts high cytotoxicity and antimicrobial activity too. Using as template mainly the tip and the subsequent β-strand of the first "finger" of this toxin, different sequences of 20 amino acids linear peptides have been designed in order to avoid toxic effects but to maintain or even strengthen the partial antimicrobial activity already seen for the complete toxin. As a result, the sequence NCP-0 (Naja Cardiotoxin Peptide-0) was designed as ancestor and subsequently 4 other variant sequences of NCP-0 were developed. These synthesized variant sequences have shown microbicidal activity towards a panel of reference and field strains of Gram-positive and Gram-negative bacteria. The sequence named NCP-3, and its variants NCP-3a and NCP-3b, have shown the best antimicrobial activity, together with low cytotoxicity against eukaryotic cells and low hemolytic activity. Bactericidal activity has been demonstrated by minimum bactericidal concentration (MBC) assay at values below 10 μg/ml for most of the tested bacterial strains. This potent antimicrobial activity was confirmed even for unicellular fungi Candida albicans, Candida glabrata and Malassezia pachydermatis (MBC 50-6.3 μg/ml), and against the fast-growing mycobacteria Mycobacterium smegmatis and Mycobacterium fortuitum. Moreover, NCP-3 has shown virucidal activity on Bovine Herpesvirus 1 (BoHV1) belonging to Herpesviridae family. The bactericidal activity is maintained even in a high salt concentration medium (125 and 250 mM NaCl) and phosphate buffer with 20% Mueller Hinton (MH) medium against E. coli, methicillin resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa reference strains. Considering these in vitro obtained data, the search for active sequences within proteins presenting an intrinsic microbicidal activity could provide a new way for discovering a large number of novel and promising antimicrobial peptides families.


Lung derived surface active material (SAM) inhibits natural killer cell tumor cytotoxicity.
Natural killer cells have been identified in lung tissue but have been found to have significantly less tumor cytotoxicity than natural killer cells found elsewhere in the body. The natural killer cells in the lung are still functional, since their killing can be enhanced by Interleukin-2. The surface active material (SAM) of lung lining fluid has been shown to have immunomodulating activity, including the suppression of lymphocyte blast transformation and enhancement of macrophage tumor cytotoxicity. We studied the effect of SAM purified from lung lining fluid on natural killer cell tumor cytotoxicity. SAM markedly inhibited tumor killing (Percent cytotoxicity of cells alone: 41 +/- 7.3% (mean +/- SEM); cells plus SAM: 10 +/- 9.7%, p less than 0.02). Further studies demonstrated that some phospholipids, including phosphatidylcholine, the major phospholipid of SAM, also significantly inhibited natural killer cell tumor killing. This inhibition of natural killer cell tumor cytotoxicity could be reversed by the addition of Interleukin-2 to the natural killer cells. These studies demonstrated that the surfactant found in lung lining fluid significantly inhibited natural killer cell tumor cytotoxicity and this effect could be reversed by Interleukin-2.


Racial/ethnic differences in the self-reported use of screening mammography.
The efficacy of mammography in reducing breast cancer mortality among women 50-69 years of age has been demonstrated in randomized controlled studies, but many women, especially ethnic minorities, have not been receiving regular mammographic screening. The current study investigated racial/ethnic differences in mammography use and their association with demographic characteristics and other factors. The study population consisted of 4,444 women aged 40 years and older who participated in the 1996 Medical Expenditure Panel Survey. Outcome measures studied included the self-reporting of mammography within the past two years and past year. Multivariate logistic regression modeling was used to examine the effect of race while controlling for other factors. In the univariate analysis, there was virtually no difference between white, black, and Hispanic women in mammography rates within either one or two years. However, multivariate logistic regression suggested that both blacks and Hispanics were more likely than whites to have received recent mammography, as black women were 31% and Hispanic women were 43% more likely than white women to have had a mammogram within the previous two years. Our results suggest that white women are no longer more likely to receive periodic screening mammography than black and Hispanic women, and in fact, might even be less likely to undergo the procedure. This reversal might indicate, at least in part, that programs and other activities to promote screening mammography among ethnic minority women have been successful and should now be expanded to include other women.


Oncolytic Herpes Simplex Viral Therapy: A Stride toward Selective Targeting of Cancer Cells.
Oncolytic viral therapy, which makes use of replication-competent lytic viruses, has emerged as a promising modality to treat malignancies. It has shown meaningful outcomes in both solid tumor and hematologic malignancies. Advancements during the last decade, mainly genetic engineering of oncolytic viruses have resulted in improved specificity and efficacy of oncolytic viruses in cancer therapeutics. Oncolytic viral therapy for treating cancer with herpes simplex virus-1 has been of particular interest owing to its range of benefits like: (a) large genome and power to infiltrate in the tumor, (b) easy access to manipulation with the flexibility to insert multiple transgenes, (c) infecting majority of the malignant cell types with quick replication in the infected cells and (d) as Anti-HSV agent to terminate HSV replication. This review provides an exhaustive list of oncolytic herpes simplex virus-1 along with their genetic alterations. It also encompasses the major developments in oncolytic herpes simplex-1 viral therapy and outlines the limitations and drawbacks of oncolytic herpes simplex viral therapy.


[Dengue with unusual clinical features in an infant: Case report].
The state of Yucatan, in Mexico, is an endemic area for dengue. During 2015, there was an unpredicted increase in the number of expected cases of dengue fever. To describe and analyse the clinical presentation, progress, and management of a case of dengue infection with non-specific clinical manifestations in an infant which resulted in a dengue shock syndrome. One month old infant admitted to hospital with a generalised rash and a history of being bitten by an insect. He was diagnosed with anaphylaxis based on clinical manifestations and anamnesis. While in hospital, he developed hypotension, tachycardia, anaemia, and respiratory distress. He was transferred to the intensive care unit, but died on the fifth day. He tested positive to dengue virus in the PCR test and for IgG antibodies using Elisa. The basic cause of death was dengue shock syndrome. Dengue fever in young infant infections may be afebrile, so it is important to suspect them appropriately in the presence of a generalised rash, tachycardia, and hypotension, in order to avoid the deadly consequences of dengue shock.


Some concepts relating to the molecular genetic basis of certain MNS blood group antigens.
The unfolding story of genes encoding variant glycophorin molecules is already known to be more complicated than described here. The principles outlined provide a basis for understanding the fundamental events that occur in genes encoding the glycophorins as well as genes encoding unrelated proteins carrying other blood group antigens. Over 20 different genes involving the GYPA and GYPB family have been described. These genes arise from gene rearrangements within a relatively short region. This hot spot of activity has inverted palindromic sequences, which are known to be sites for DNA recombination. Similar structures exist in the major histocompatibility complex (MHC) where allelic diversity is a functional requisite. However, the significance of allelic diversity in the glycophorin gene family is not understood. The GYPA, GYPB and GYPE gene cluster is known to be prone to mutation by radiation because there is a high incidence of somatic mutation events in atomic bomb survivors, in people exposed to accidental radiation, in patients with Bloom's syndrome and in patients receiving radiation therapy. The mutation events were dose dependent: the greater the exposure, the greater proportion of red blood cells exhibited mutations. While it is known that MHC diversity protects against infection, the reason for glycophorin rearrangements remains to be determined.


Technology combo yields high satisfaction rates.
Every technology has weaknesses. Recognize what they are, and complement it with other technology that will help fill in the gaps. Systems that track patients through your ED must be accompanied by electronic patient records for optimal results. No technology will reach its full potential unless you also have revised essential patient care processes. Increase staffing levels during training so that neither patient care nor educational processes will suffer.


Use of first-dose response or plasma renin activity to predict the long-term effect of captopril: identification of triphasic pattern of blood pressure response.
A triphasic pattern of blood pressure response was observed in 13 of 26 patients treated with captopril. Pressure fell after the first dose in all but 3 patients, reaching a minimum by 90 min. Over the next several days, pressure either remained at this level (n = 13) or returned towards control levels (n = 13) even with increased dosage. Blood pressure then fell gradually, usually reaching a level by 10 days that was equal to or slightly lower than that after the first dose. The secondary rise in blood pressure was more frequent and pronounced in patients who had larger initial responses and higher renin levels. The transient increase in pressure could not be explained by failure of blockade of the renin system, since aldosterone excretion was maximally suppressed by the second day of treatment. The data indicate that both the blood pressure response to the first dose and the renin level can be used to predict the long-term response to captopril.


Distal ERBB2 promoter fragment displays specific transcriptional and nuclear binding activities in ERBB2 overexpressing breast cancer cells.
Overexpression of the ERBB2 gene occurs in 30% of human breast cancers and is correlated with poor prognosis. The deregulation is the consequence of an increased transcription level and gene amplification. Several laboratories, including our own, have identified, in the proximal promoter, enhancers implicated in the gene overexpression. However, our previous studies of a 6-kb ERBB2 promoter fragment revealed the presence of repressing fragments, which were able to overcome the effect of the proximal enhancers. These repressing elements were functional in all cell lines, regardless of their endogenous ERBB2 expression level. Here, we show that a distal ERBB2 promoter region restores high transcription rates specifically in ERBB2 overexpressing breast cancer cells. This distal promoter region thus contains enhancers essential for the overexpression of the gene. By EMSA, performed with nuclear extract of cells overexpressing (BT-474) or not (MDA-MB-231) the ERBB2 gene, we show that at least two sequences of the distal promoter region are bound exclusively by BT-474 extract. Further experiments reveal that AP-2 transcription factors contribute to this differential binding activity, by binding recognition sequences located 4500 bp and 4000 bp upstream of the transcription start site. These sites are occupied by AP2 in vivo, as demonstrated by ChIP assay. Inactivation of AP-2 proteins in ERBB2 overexpressing cells reduces the distal promoter activity up to 70%, indicating the AP-2 factors are implicated in the strong distal enhancing effect. Moreover, we identified a 54-bp fragment that is bound specifically by BT-474 nuclear extract. Further experiments did not lead to the identification of the protein responsible for this binding. Our results thus highlight the importance of ERBB2 distal promoter region and further implicate AP-2 in ERBB2 overexpression in breast cancer cells.


The Hidden One: What We Know About Bitter Taste Receptor 39.
Over thousands of years of evolution, animals have developed many ways to protect themselves. One of the most protective ways to avoid disease is to prevent the absorption of harmful components. This protective function is a basic role of bitter taste receptors (TAS2Rs), a G protein-coupled receptor family, whose presence in extraoral tissues has intrigued many researchers. In humans, there are 25 TAS2Rs, and although we know a great deal about some of them, others are still shrouded in mystery. One in this latter category is bitter taste receptor 39 (TAS2R39). Besides the oral cavity, it has also been found in the gastrointestinal tract and the respiratory, nervous and reproductive systems. TAS2R39 is a relatively non-selective receptor, which means that it can be activated by a range of mostly plant-derived compounds such as theaflavins, catechins and isoflavones. On the other hand, few antagonists for this receptor are available, since only some flavones have antagonistic properties (all of them detailed in the document). The primary role of TAS2R39 is to sense the bitter components of food and protect the organism from harmful compounds. There is also some indication that this bitter taste receptor regulates enterohormones and in turn, regulates food intake. In the respiratory system, it may be involved in the congestion process of allergic rhinitis and may stimulate inflammatory cytokines. However, more thorough research is needed to determine the precise role of TAS2R39 in these and other tissues.


Reduced-fat cheddar cheese manufactured using a novel fat removal process.
Normally, reduced-fat Cheddar cheese is made by removal of fat from milk prior to cheese making. Typical aged flavor may not develop when 50% reduced-fat Cheddar cheese is produced by this approach. Moreover, the texture of the reduced-fat cheeses produced by the current method may often be hard and rubbery. Previous researchers have demonstrated that aged Cheddar cheese flavor intensity resides in the water-soluble fraction. Therefore, we investigated the feasibility of fat removal after the aging of Cheddar cheese. We hypothesized the typical aged cheese flavor would remain with the cheese following fat removal. A physical process for the removal of fat from full-fat aged Cheddar cheese was developed. The efficiency of fat removal at various temperatures, gravitational forces, and for various durations of applied forces was determined. Temperature had the greatest effect on the removal of fat. Gravitational force and the duration of applied force were less important at higher temperatures. A positive linear relationship between temperature and fat removal was observed from 20 to 33 degrees C. Conditions of 30 degrees C and 23,500 x g for 5 min removed 50% of the fat. The removed fat had some aroma but little or no taste. The fatty acid composition, triglyceride molecular weight distribution, and melting profile of the fat retained in the reduced-fat cheeses were all consistent with a slight increase in the proportion of saturated fat relative to the full-fat cheeses. The process of fat removal decreased the grams of saturated fat per serving of cheese from 6.30 to 3.11 g. The flavor intensity of the reduced-fat cheeses were at least as intense as the full-fat cheeses.


Urinary Biomarkers to Predict Neonatal Acute Kidney Injury: A Review of the Science.
Acute kidney injury (AKI) occurs in approximately 30% of all infants hospitalized in the neonatal intensive care unit. About 40% of very low-birth-weight infants develop AKI, with an estimated mortality rate of 50% to 80%. Very low-birth-weight survivors have twice the risk of developing chronic renal disease later in life compared with their term counterparts. Current diagnostic modalities for AKI include serum creatinine and urine output; however, recent studies suggest that these measures are imprecise, as they may not change until 25% to 50% of renal function is lost. Urinary biomarkers may more accurately identify infants at risk for early AKI development. The purpose of this review is to discuss current research findings related to neonatal AKI risk factors, provide an overview of short- and long-term outcomes, describe innovative diagnostic approaches, and identify future research direction needed to improve prediction and intervention strategies associated with renal impairment.


Exogenous PTH-related protein and PTH improve mineral and skeletal status in 25-hydroxyvitamin D-1alpha-hydroxylase and PTH double knockout mice.
We examined the effect of NH2-terminal fragments of PTHrP and PTH in young mutant mice deficient in both PTH and 1,25-dihydroxyvitamin D. Both proteins prolonged murine survival by increasing serum calcium, apparently by enhancing renal calcium transporter expression. The dominant effect on the skeleton was an increase in both endochondral bone and appositional formation without increased bone resorption. PTH-related protein (PTHrP) was discovered as a hypercalcemic agent responsible for the syndrome of humeral hypercalcemia of malignancy, and PTH is the major protein hormone regulating calcium homeostasis. Both proteins have skeletal anabolic actions when administered intermittently. We examined effects of exogenous PTHrP(1-86) and PTH(1-34) in double null mutant mice deficient in both PTH and 25-hydroxyvitamin D-1alpha-hydroxylase [1alpha(OH)ase] to determine the action of these proteins in the absence of the two major regulators of calcium and skeletal homeostasis. Mice heterozygous for the PTH null allele and for the 1alpha(OH)ase null allele were mated to generate pups homozygous for both null alleles. PTHrP(1-86) and PTH(1-34) were administered subcutaneously starting 4 days after birth. Serum biochemistry and skeletal radiology, histology, and histomorphometry were performed, and indices of bone formation, resorption, and renal calcium transport were determined by real time RT-PCR, Western blot, and immunohistochemical approaches. In the double mutant mice, which die within 3 weeks after birth with severe hypocalcemia, tetany, and skeletal defects, exogenous PTHrP and PTH enhanced survival of the animals by improving serum calcium. Both proteins increased renal calcium transporter expression and long bone length and augmented growth plate chondrocyte proliferation, differentiation, and cartilage matrix mineralization. Cortical and trabecular bone mass was increased with augmented osteoblast number and activity; however, bone resorption was not increased. PTHrP and PTH reduced hypocalcemia by enhancing renal calcium reabsorption but not by increasing bone resorption. The major skeletal effects of exogenous PTHrP and PTH were to increase bone anabolism.


Thyroid FNAC cytology: can we do it better?
This article reviews recent developments in thyroid fine needle aspiration cytology (FNAC). While thyroid nodules are common, carcinoma is comparatively rare. Although histological assessment is used in most studies as the benchmark, the differential diagnosis on cytology or histology is not always reproducible. The literature shows wide variations in criteria for inadequate thyroid FNAC and study inclusion or exclusion criteria. In-clinic assessment of specimen adequacy and in-clinic reporting of thyroid FNAC has become popular although the costs and resource implications of in-clinic thyroid FNAC assessment and reporting are substantial. Many centres continue to use conventional techniques although liquid-based cytology and ultrasound-guided FNAC are gaining in popularity. Standardized categorical systems for FNAC reporting can make results easier to understand for clinicians and give clear indications for therapeutic action. Multidisciplinary case review is also essential, especially when there is diagnostic uncertainty. While currently of limited use, molecular pathology testing holds out some promise for the future.


Down-regulation of hematopoiesis master regulator PU.1 via aberrant methylation in chronic myeloid leukemia.
The PU.1 transcription factor is a crucial regulator of hematopoiesis, and its expression is altered in various leukemic processes. It has been shown that expression of PU.1 is severely impaired in patients with chronic myeloid leukemia (CML), but the mechanism underlying this effect remains unknown. Through bisulfite sequencing, semi-quantitative PCR, and indirect immunofluorescence and Western blot techniques, we found aberrant methylation in the promoter region of transcription factor PU.1 in CML patients both in the chronic and blast crisis phases, as well as in the CML blast K562 cell line. Of these, several CpG sites were more highly methylated in blast crisis than chronic phase, while no methylation of these sites was observed in healthy individuals. Interestingly, CML patients achieved complete cytogenetic remission under imatinib mesylate treatment, but the aberrant methylation status of PU.1 was not reversed. Down-regulation of PU.1 expression at the mRNA and protein levels was also observed in association with aberrant methylation. Thus, for the first time, we have revealed a potential epigenetic modification of PU.1 in CML, which may be responsible for the down-regulation of PU.1. These data suggest that aberrant methylation of PU.1 may play a role in CML pathogenesis, and may therefore serve as a useful biomarker and potential target for demethylating drugs.


Pharmacotherapeutic options for treating brain metastases in non-small cell lung cancer.
Central nervous system (CNS) metastases represent an important cause of morbidity and mortality in non-small cell lung cancer (NSCLC) patients. Local approaches of neurosurgery (usually for single brain lesions), whole brain radiotherapy, and stereotactic radiosurgery are often withheld for the treatment of NSCLC-derived brain metastases (BMs). However, systemic treatment is consistently emerging as an option for patients with asymptomatic BMs, which could allow for delaying cranial radiotherapy at symptomatic/radiological progression. Chemotherapy, monoclonal antibodies, tyrosine-kinase inhibitors (TKIs) for molecularly selected NSCLCs, such as epidermal growth factor receptor (EGFR)-mutant and anaplastic lymphoma kinase (ALK)-rearranged diseases, and immune checkpoint inhibitors are all systemic treatments that have shown activity against NSCLC-derived CNS metastases. Among these, EGFR- and ALK-TKIs will be discussed more in detail owing to their superior efficacy in this context. Up-front systemic treatment should be considered for patients with asymptomatic, multiple BMs, as recently acknowledged by the European Society of Medical Oncology guidelines. Nevertheless, it must be emphasized that the best treatment strategy for NSCLC-derived BMs has to be defined within a multidisciplinary team.


Pigmented epithelium to retinal transdifferentiation and Pax6 expression in larval Xenopus laevis.
This study examines the retinal transdifferentiation (TD) of retinal pigmented epithelium (RPE) fragments dissected from Xenopus laevis larvae and implanted into the vitreous chamber of non-lentectomized host eyes. In these experimental conditions, most RPE implants transformed into polarized vesicles in which the side adjacent to the lens maintained the RPE phenotype, while the side adjacent to the host retina transformed into a laminar retina with the photoreceptor layer facing the cavity of the vesicle and with the ganglionar cell layer facing the host retina. The formation of a new retina with a laminar organization is the result of depigmentation, proliferation and differentiation of progenitor cells under the influence of inductive factors from the host retina. The phases of the TD process were followed using BrdU labelling as a marker of the proliferation phase and using a monoclonal antibody (mAbHP1) as a definitive indicator of retina formation. Pigmented RPE cells do not express Pax6. In the early phase of RPE to retinal TD, all depigmented and proliferating progenitor cells expressed Pax6. Changes in the Pax6 expression pattern became apparent in the early phase of differentiation, when Pax6 expression decreased in the presumptive outer nuclear layer (ONL) of the new-forming retina. Finally, during the late differentiation phase, the ONL, which contains photoreceptors, no longer expressed Pax6, Pax6 expression being confined to the ganglion cell layer and the inner nuclear layer. These results indicate that Pax6 may have different roles during the different phases of RPE to retinal TD, acting as an early retinal determinant and later directing progenitor cell fate.


Chest drainage teaching and training for medical students. Use of a surgical ex vivo pig model.
Implement a constructivist approach in thoracic drainage training in surgical ex vivo pig models, to compare the acquisition of homogeneous surgical skills between medical students. Experimental study, prospective, transversal, analytical, controlled, three steps. Selection, training, evaluation. a) students without training in thoracic drainage; b) without exposure to constructivist methodology. 2) EXCLUSION CRITERIA: a) students developed surgical skills; b) a history of allergy. (N = 312). Two groups participated in the study: A and B. Lecture equal for both groups. Differentiated teaching: group A, descriptive and informative method; group B, learning method based on problems. A surgical ex vivo pig model for training the chest drain was created. Were applied pre and post-test, test goal-discursive and OSATS scale. Theoretical averages: Group A = 9.5 ± 0.5; Group B = 8.8 ± 1.1 (p = 0.006). Medium Practices: Group A = 22.8 ± 1.8; Group B = 23.0 ± 2.8 (p <0.001). Through the constructivist methodology implemented in the thoracic drainage training in surgical ex vivo pig models, has proven the acquisition of surgical skills homogeneous compared among medical students.


The motivation for beer in rats: effects of ritanserin, naloxone and SR 141716.
Rats were given two weeks of home cage access to either "near-beer" (a beverage that tastes like beer but contains <0.5% ethanol v/v) or near-beer with added ethanol (4.5% v/v), which is simply referred to as "beer". The two groups of rats (near-beer and beer) were then trained on a "lick-based progressive ratio paradigm" in operant chambers in which an ever increasing number of licks had to be emitted for each successive fixed unit of near-beer or beer delivered. Break points (the ratio at which responding ceased) for near-beer and beer were approximately equal under baseline conditions. Rats were then tested for the effects of the 5HT2A/2C receptor antagonist ritanserin (0.625, 2.5 or 10 mg/kg), the opioid receptor antagonist naloxone (0.625, 2.5 or 10 mg/kg) or the cannabinoid CB1 receptor antagonist SR 141716 (0.3, 1 or 3 mg/kg). All three drugs caused a dose-dependent reduction of break-points and locomotor activity in both the beer and near-beer groups. However, the effects of SR 141716 and naloxone, but not ritanserin, on breakpoints were significantly more pronounced on rats drinking beer compared to those drinking near-beer. There were no such differential effects of any of the drugs on locomotor activity across the two groups. These results suggest that both SR 141716 and naloxone differentially affect the motivation to consume alcoholic beverages and may thus have potential as drugs for the treatment of alcohol craving.


Scope of wounds.
The scope of extremity wounds in the current conflict presents surgeons with new lessons to master. Unique to this conflict is a new type of patient, one with multiple and severely injured extremities who is otherwise free of serious injury. These injuries provide challenges to the medical system in terms of the volume and complexity of care, and to the patient and surgeon trying to achieve limb salvage and rehabilitation. These patients present with a combination of high-energy injury, massive evolving tissue destruction, and widespread contamination, resulting in an evolving zone of injury that respects no tissue planes, anatomic boundaries, or normal physiologic rules. We must ensure that our skills and techniques as surgeons evolve faster than do the injuries themselves.


[Cloning and expression of the HER2 gene in MCF-7 cells].
To clone human epidermal growth factor receptor (HER2) gene and establish a MCF-7 cell line with stable co-expression of HER2 and estrogen receptor (ER). The full-length HER2 gene fragment was amplified by RT-PCR from the total RNA extracted from human breast cancer cell line SKBR-3 which over-expressed HER2 protein. The gene fragment was then inserted into the vector pcDNA3.1 to construct the recombinant expression plasmid pcDNA3.1-HER2, which was identified by sequence analysis and transfected into ER-expressing MCF-7 cells via lipofectamine. The positive cell clones were obtained after G418 selection. The MCF-7 cells transfected with HER2-cDNA was assayed by immunocytochemistry, flow cytometry and Western blotting. DNA sequencing results showed that HER2 gene was exactly consistent with the sequence reported in GenBank. The MCF-7 cells transfected with HER2 could highly express HER2 protein as shown by immunocytochemistry, flow cytometry and Western blotting. HER2 gene has been cloned successfully and the MCF-7 cell line co-expressing HER2 and ER is obtained.


Spectral analysis of heart rate variability in athletes.
The objectives of the study were to characterize power spectrum pattern of the heart rate variability and assessment of the relative contributions of sympathetic and parasympathetic cardiac nervous system control in athletes. Thirty-three male athletes, swimming (1), canoeing (10), cycling (6), athletics (4), football (3), roller-skating (2) and volleyball (7) aged 23.4 +/- 5.5 years, with a mean athletic level of 18 hours/week (8-45) and 33 sedentary healthy control subjects were included. Ecg signals were recorded after a period of 15 minutes in supine rest with controlled breathing at 15 cycles/min. Signal acquisition was done at 300 samples/sec. From 512 consecutive heart beats, we calculated mean average, standard deviation, maximum and minimum R-R intervals and, after computing the fast Fourier transform, total spectrum power, low frequency (LF), high frequency (HF) components and its ratio (LF/HF). The average R-R interval was 987.7 +/- 168.8 ms and 762.7 +/- 125.3 ms, the variance was 5.44 and 2.51 ms2 and ratio of R-R interval maximum/minimum (E/I ratio) 1.53 +/- 0.16 and 1.41 +/- 0.16, respectively for athletes and control group. Differences between groups were significant (p < 0.01) for all parameters, with higher variability in the athletes. Both spectral bands (LF and HF) had higher power in athletes (LF = 925 +/- 920 and HF = 2258 +/- 2349 ms2) than in the control group (LF = 442 +/- 446 and HF = 1179 +/- 1542 ms2) (p < 0.01). There were no significant differences for LF/HF ratio, or normalized LF (LF%) or normalized HF (HF%) between groups.(ABSTRACT TRUNCATED AT 250 WORDS)


Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing.
Apple latent spherical virus (ALSV) is an efficient virus-induced gene silencing vector in functional genomics analyses of a broad range of plant species. Here, an Agrobacterium-mediated inoculation (agroinoculation) system was developed for the ALSV vector, and virus-induced transcriptional gene silencing (VITGS) is described in plants infected with the ALSV vector. The cDNAs of ALSV RNA1 and RNA2 were inserted between the cauliflower mosaic virus 35S promoter and the NOS-T sequences in a binary vector pCAMBIA1300 to produce pCALSR1 and pCALSR2-XSB or pCALSR2-XSB/MN. When these vector constructs were agroinoculated into Nicotiana benthamiana plants with a construct expressing a viral silencing suppressor, the infection efficiency of the vectors was 100%. A recombinant ALSV vector carrying part of the 35S promoter sequence induced transcriptional gene silencing of the green fluorescent protein gene in a line of N. benthamiana plants, resulting in the disappearance of green fluorescence of infected plants. Bisulfite sequencing showed that cytosine residues at CG and CHG sites of the 35S promoter sequence were highly methylated in the silenced generation zero plants infected with the ALSV carrying the promoter sequence as well as in progeny. The ALSV-mediated VITGS state was inherited by progeny for multiple generations. In addition, induction of VITGS of an endogenous gene (chalcone synthase-A) was demonstrated in petunia plants infected with an ALSV vector carrying the native promoter sequence. These results suggest that ALSV-based vectors can be applied to study DNA methylation in plant genomes, and provide a useful tool for plant breeding via epigenetic modification.


[Neither propranolol nor the vasodilating beta-blocker carvedilol have a direct effect on coronary resistance vessels].
Seventeen patients with coronary heart disease were included in a double-blind randomized study. They received either 5 mg of carvedilol or 6 mg of propranolol intravenously. Heart rate, aortic pressure, mean coronary sinus pressure and coronary flow (thermodilution) were measured, and coronary resistance and rate-pressure product were calculated before and 15 min after the infusion, which lasted 10 min. Carvedilol lowered significantly (p less than 0.05) heart rate (mean 76 to 69/min), aortic pressure (mean 153/80 to 135/72 mm Hg), rate-pressure product (mean 117 to 93 mm Hg/min) and coronary flow (mean 114 to 94 ml/min). Coronary resistance and coronary flow related to rate-pressure product showed no significant change after carvedilol. Propranolol lowered heart rate (mean 76 to 64/min; p less than 0.05) and rate-pressure product (mean 109 to 96 mm Hg/min; ns). Aortic pressure, coronary flow, coronary resistance, and coronary flow related to rate-pressure product showed no significant change after propranolol. Thus, carvedilol lowered rate-pressure product more markedly than propranolol on account of its acute blood-pressure lowering effect. Neither drug seems to have a direct influence on coronary resistance vessels.


Continuous nonradioactive method for screening trypanosomal trans-sialidase activity and its inhibitors.
Trypanosoma cruzi, the agent of American trypanosomiasis is unable to synthesize sialic acid (SA). Instead of using the corresponding nucleotide sugar as donor of the monosaccharide, the transfer occurs from alpha-2,3-linked SA in the host sialoglycoconjugates to terminal beta-galactopyranosyl units of the parasite mucins. For that purpose, T. cruzi expresses a glycosylphosphatidylinositol-anchored trans-sialidase (TcTS) that is shed into the milieu, being detected in the blood during the acute phase of the infection. The essential role of TcTS in infection and the absence of a similar activity in mammals make this enzyme an attractive target for the development of alternative chemotherapies. However, there is no effective inhibitor toward this enzyme. In vitro, 3'-sialyllactose (SL) as donor and radioactive lactose as acceptor substrate are widely used to measure TcTS activity. The radioactive sialylated product is then isolated by anion exchange chromatography and measured. Here we describe a new nonradioactive assay using SL or fetuin as donor and benzyl beta-d-Fuc-(1-->6)-alpha-d-GlcNAc (1) as acceptor. Disaccharide 1 was easily synthesized by regioselective glycosylation of benzyl alpha-d-GlcNAc with tetra-O-benzoyl-d-fucose followed by debenzoylation. Compound 1 lacks the hydroxyl group at C-6 of the acceptor galactose and therefore is not a substrate for galactose oxidase. Our method relies on the specific quantification of terminal galactose produced by trans-sialylation from the donor to the 6-deoxy-galactose (D-Fuc) unit of 1 by a spectrophotometric galactose oxidase assay. This method may also discriminate sialidase and trans-sialylation activities by running the assay in the absence of acceptor 1.


Intraspinal temperatures during regional deep hyperthermia induced by electromagnetic radiation in the Macaca mulatta.
A capacitive ring system operating at 27 MHz was tested for temperature distribution in the abdominal cavity of the living Rhesus monkey. Catheters were introduced through the abdomen and their location checked after the experiment. During two of these experiments a catheter was also introduced into the intraspinal canal. Temperature distribution was rather homogeneous within the abdominal cavity. Core temperature remained 3-5 degrees C below intra-abdominal temperatures. The intraspinal temperature increase was of the same magnitude compared with intra-abdominal temperature increase. Similar findings in other species have been reported by others. It is concluded that during non-invasive induction of regional hyperthermia by electromagnetic radiation, where a high level of energy is directed close to the spine, the intraspinal temperatures may be increased as high as the temperatures within the surrounding tissues. The central nervous system may be relatively sensitive to hyperthermia and hyperthermia may sensitize the spinal cord to radiation. Therefore, when hyperthermia is targeted at areas near the spine, either alone or in combination with radiotherapy applied to the tolerance limit of the spinal cord, these findings will have to be taken into consideration.


Early mobilization after flexor tendon grafting for isolated profundus tendon lesions.
After reconstruction of a flexor profundus tendon with a free graft passed through or around an intact superficialis decussation, impaired function may result if adherences develop. In an attempt to avoid adherences, early mobilization using Kleinert's method was applied in 20 grafts. At follow-up, on average 36 months after operation, all the patients had improved ROM except one patient with tendinous infection one year postoperatively. On average the gain in DIP-joint flexion was 34 degrees, while six patients had a loss of 17 degrees and 12 patients a gain of 22 degrees in PIP-joint motion. Only nine patients required a few weeks of occupational therapy. It is concluded that early mobilization a.m. Kleinert after tendon grafting is a safe method for prevention of loss of PIP-joint motion due to the formation of adherences.


Improving the performance of an active staggered window with multiple resonant absorbers.
The active noise control (ANC) technique has been applied in staggered windows to improve the noise reduction at low frequencies. The control performance of such a system deteriorates significantly at some frequencies where the secondary source cannot radiate effectively due to the reflection at the boundaries of the staggered window. A resonant absorber consisting of a perforated panel and coiled up tubes is proposed to solve the problem. By designing a combination of different absorbers, a proper sound absorption coefficient is achieved around the ineffective frequency. Numerical simulations show that the active sound power reduction increases by 13.5 dB at the frequency with the absorbers attached on one end of the staggered window, and the overall sound power reduction between 100 and 500 Hz increases from 25.9 to 31.2 dB. Attaching the sound absorbers elsewhere in the upstream of the secondary source, for example, on the side walls of the duct also works. The active sound power reduction at 435 Hz increases by 6.3 dB after attaching the absorbers in the experiments, and the noise reduction increment at the evaluation point is 13.6 dB, which agrees with simulation results and demonstrates the feasibility of the proposed sound absorbers.


Targeting the oral plaque microbiome with immobilized anti-biofilm peptides at tooth-restoration interfaces.
Recurrent caries, the development of carious lesions at the interface between the restorative material and the tooth structure, is highly prevalent and represents the primary cause for failure of dental restorations. Correspondingly, we exploited the self-assembly and strong antibiofilm activity of amphipathic antimicrobial peptides (AAMPs) to form novel coatings on dentin that aimed to prevent recurrent caries at susceptible cavosurface margins. AAMPs are alternative to traditional antimicrobial agents and antibiotics with the ability to target the complex and heterogeneous organization of microbial communities. Unlike approaches that have focused on using these AAMPs in aqueous solutions for a transient activity, here we assess the effects on microcosm biofilms of a long-acting AAMPs-based antibiofilm coating to protect the tooth-composite interface. Genomewise, we studied the impact of AAMPs coatings on the dental plaque microbial community. We found that non-native all D-amino acids AAMPs coatings induced a marked shift in the plaque community and selectively targeted three primary acidogenic colonizers, including the most common taxa around Class II composite restorations. Accordingly, we investigated the translational potential of our antibiofilm dentin using multiphoton pulsed near infra-red laser for deep bioimaging to assess the impact of AAMPs-coated dentin on plaque biofilms along dentin-composite interfaces. Multiphoton enabled us to record the antibiofilm potency of AAMPs-coated dentin on plaque biofilms throughout exaggeratedly failed interfaces. In conclusion, AAMPs-coatings on dentin showed selective and long-acting antibiofilm activity against three dominant acidogenic colonizers and potential to resist recurrent caries to promote and sustain the interfacial integrity of adhesive-based interfaces.


Subventricular zone/white matter microglia reconstitute the empty adult microglial niche in a dynamic wave.
Microglia, the brain's resident myeloid cells, play central roles in brain defense, homeostasis, and disease. Using a prolonged colony-stimulating factor 1 receptor inhibitor (CSF1Ri) approach, we report an unprecedented level of microglial depletion and establish a model system that achieves an empty microglial niche in the adult brain. We identify a myeloid cell that migrates from the subventricular zone and associated white matter areas. Following CSF1Ri, these amoeboid cells migrate radially and tangentially in a dynamic wave filling the brain in a distinct pattern, to replace the microglial-depleted brain. These repopulating cells are enriched in disease-associated microglia genes and exhibit similar phenotypic and transcriptional profiles to white-matter-associated microglia. Our findings shed light on the overlapping and distinct functional complexity and diversity of myeloid cells of the CNS and provide new insight into repopulating microglia function and dynamics in the mouse brain.


Seasonal variations in rhino-cerebral Mucor infection.
Rhino-orbito-cerebral mucormycosis (ROCM) is an uncommon, potentially lethal infection that occurs mostly in immunocompromised hosts. The seasonal occurrence of ROCM was studied in two of Israel's largest medical centers. A total of 36 patients were seen during a 25-year period in this retrospective bi-institutional cohort study. Meteorologic data were obtained and analyzed for a possible linkage of infection with precipitation and temperature. Twenty-one of the 36 patients (58%) presented between the months of August and October, and 27 patients (75%) presented between the months of August and December. A peak incidence of ROCM was noted in the month of September (13/36 patients; 36%). No association was noted between meteorologic conditions and the incidence of ROCM infection. A consistent seasonal peak was observed in ROCM. Heightened awareness is important for early recognition and treatment of this disease.


T cell exosome-derived miR-142-3p impairs glandular cell function in Sjögren's syndrome.
Sjögren's syndrome (SS) is a systemic autoimmune disease that mainly affects exocrine salivary and lacrimal glands. Local inflammation in the glands is thought to trigger glandular dysfunction and symptoms of dryness. However, the mechanisms underlying these processes are incompletely understood. Our work suggests T cell exosome-derived miR-142-3p as a pathogenic driver of immunopathology in SS. We first document miR-142-3p expression in the salivary glands of patients with SS, both in epithelial gland cells and within T cells of the inflammatory infiltrate, but not in healthy volunteers. Next, we show that activated T cells secreted exosomes containing miR-142-3p, which transferred into glandular cells. Finally, we uncover a functional role of miR-142-3p-containing exosomes in glandular cell dysfunction. We find that miR-142-3p targets key elements of intracellular Ca2+ signaling and cAMP production - sarco(endo)plasmic reticulum Ca2+ ATPase 2b (SERCA2B), ryanodine receptor 2 (RyR2), and adenylate cyclase 9 (AC9) - leading to restricted cAMP production, altered calcium signaling, and decreased protein production from salivary gland cells. Our work provides evidence for a functional role of the miR-142-3p in SS pathogenesis and promotes the concept that T cell activation may directly impair epithelial cell function through secretion of miRNA-containing exosomes.


TriATNE: Tripartite Adversarial Training for Network Embeddings.
Existing network embedding algorithms based on generative adversarial networks (GANs) improve the robustness of node embeddings by selecting high-quality negative samples with the generator to play against the discriminator. Since most of the negative samples can be easily discriminated from positive samples in graphs, their poor competitiveness weakens the function of the generator. Inspired by the sales skills in the market, in this article, we present tripartite adversarial training for network embeddings (TriATNE), a novel adversarial learning framework for learning stable and robust node embeddings. TriATNE consists of three players: 1) producer; 2) seller; and 3) customer. The producer strives to learn the representation of each sample (node pair), making it easy for the customer to differentiate between the positive and the negative, while the seller tries to confuse the customer by selecting realistic-looking samples. The customer, a biased evaluation metric, provides feedback for training the producer and the seller. To further enhance the robustness of node embedding, we model the customer as a two-layer neural network, where each unit in the hidden layer can be regarded as a customer with different preferences. TriATNE also plays against the producer by adjusting the weight of each customer. We test the performance of TriATNE on two common tasks: classification as well as link prediction. The experimental results on various publicly available datasets show that TriATNE can exploit the network structure well.


Comprehensive exon array data processing method for quantitative analysis of alternative spliced variants.
Alternative splicing of pre-mRNA generates protein diversity. Dysfunction of splicing machinery and expression of specific transcripts has been linked to cancer progression and drug response. Exon microarray technology enables genome-wide quantification of expression levels of the majority of exons and facilitates the discovery of alternative splicing events. Analysis of exon array data is more challenging than the analysis of gene expression data and there is a need for reliable quantification of exons and alternatively spliced variants. We introduce a novel, computationally efficient methodology, Multiple Exon Array Preprocessing (MEAP), for exon array data pre-processing, analysis and visualization. We compared MEAP with existing pre-processing methods, and validation of six exons and two alternatively spliced variants with qPCR corroborated MEAP expression estimates. Analysis of exon array data from head and neck squamous cell carcinoma (HNSCC) cell lines revealed several transcripts associated with 11q13 amplification, which is related with decreased survival and metastasis in HNSCC patients. Our results demonstrate that MEAP produces reliable expression values at exon, alternatively spliced variant and gene levels, which allows generating novel experimentally testable predictions.


Lipid A fractions analyzed by a technique involving thin-layer chromatography and enzyme-linked immunosorbent assay.
A modified enzyme-linked immunosorbent assay (ELISA), with alkaline phosphatase as enzyme, was used for the study of antigenicity of lipid A fractions directly on thin-layer chromatographic (TLC) plates. For visualization a gel slab containing the enzyme substrate was placed on the plate containing enzyme-conjugated antibodies. The plate was read by a thin-layer chromatogram spectrophotometer. The immunoassay was both highly specific and quite sensitive. Sensitivity was superior to levels obtained by staining the plate with molybdenum blue (for phosphate) or orcinol (for carbohydrate). Fractions of lipid A from Escherichia coli 0111, Shigella flexneri or Salmonella minnesota R595, after being separated by thin-layer chromatography, were analyzed using rabbit anti-(lipid A) serum. Patterns obtained by scanning the same plates for phosphate staining and for the TLC-ELISA corresponded well. For comparison with TLC-ELISA, an inhibition assay was run using a tube ELISA. The tube ELISA, run in aqueous medium, showed that fractions 6-8 (those having the highest RF values) had the least activities. In contrast, TLC-ELISA did not detect large differences between fractions 2-7. This discrepancy probably reflected limited aqueous solubility of fractions 6 and 7. We conclude that TLC-ELISA might reveal antigenic activities of lipids that could be missed by other methods. The data suggested that all fractions, except for fraction 8, were similar in their antigenicity by TLC-ELISA. Differences in antigenicity between the fractions occurred when the fractions were tested in free form in an aqueous environment and these differences possibly could have been due to different solubilities of individual fractions.


Is improvement in the Healthy Food Intake Index (HFII) related to a lower risk for gestational diabetes?
The aim was to analyse whether changes in the Healthy Food Intake Index (HFII) during pregnancy are related to gestational diabetes (GDM) risk. The 251 pregnant women participating had a pre-pregnancy BMI≥30 kg/m2 and/or a history of GDM. A 75 g oral glucose tolerance test (OGTT) was performed during the first and second trimesters of pregnancy for assessment of GDM. A normal OGTT result at first trimester was an inclusion criterion for the study. FFQ collected at first and second trimesters served for calculating the HFII. A higher HFII score reflects higher adherence to the Nordic Nutrition Recommendations (NNR) (score range 0-17). Statistical methods included Student's t test, Mann-Whitney U test, Fisher's exact test and linear and logistic regression analyses. The mean HFII at first trimester was 10·1 (95 % CI 9·7, 10·4) points, and the mean change from the first to the second trimester was 0·35 (95 % CI 0·09, 0·62) points. The range of the HFII changes varied from -7 to 7. The odds for GDM decreased with higher HFII change (adjusted OR 0·83 per one unit increase in HFII; 95 % CI 0·69, 0·99; P=0·043). In the analysis of the association between HFII-sub-indices and GDM, odds for GDM decreased with higher HFII-Fat change (fat percentage of milk and cheese, type of spread and cooking fats) but it was not significant in a fully adjusted model (P=0·058). Dietary changes towards the NNR during pregnancy seem to be related to a lower risk for GDM.


Bacterial vaginosis in a cohort of Danish pregnant women: prevalence and relationship with preterm delivery, low birthweight and perinatal infections.
To determine the prevalence of bacterial vaginosis (BV) in the second trimester of pregnancy in a Danish population using the Schmidt criteria and to examine whether BV was associated with subsequent preterm delivery, low birthweight or perinatal infections. Prospective cohort study. Department of Obstetrics and Gynaecology at a University Hospital, Denmark. Three thousand five hundred and forty pregnant women aged 18 years or more. A smear from the vagina was obtained from all women, air-dried and stored for subsequent diagnosis of BV. After rehydration with isotonic saline, the smear was examined in a phase-contrast microscope at 400x, and the numbers of lactobacilli morphotypes and small bacterial morphotypes were counted. A score for BV was calculated according to the method described by Schmidt. The outcome of pregnancy from 20 weeks of gestation was examined in the 3262 singleton pregnant women who were included in this study before 20 weeks of gestation. The relationship between BV and adverse outcome of pregnancy was examined by univariate and multivariate analyses. Prevalence of BV, preterm delivery (<37 weeks), low birthweight (<2500 g), preterm delivery of a low-birthweight infant and clinical chorioamnionitis. The prevalence of BV was 16%, and the rate of preterm delivery was 5.2% in the study population of 3262 singleton pregnant women who were included before 20 weeks of gestation. Mean birthweight was significantly lower in infants of women with BV than in infants of women without BV (3408 versus 3511 g, P < 0.01). Univariate analyses showed that BV was marginally associated with preterm delivery but significantly associated with low birthweight, preterm delivery of a low birthweight infant, indicated preterm delivery and clinical chorioamnionitis. Multivariate analyses, which adjusted for previous miscarriage, previous preterm delivery, previous conisation, smoking, gestational diabetes, fetal death and preterm premature rupture of membranes, showed that BV was significantly associated with low birthweight (OR 1.95, 95% CI 1.3-2.9), preterm delivery of a low-birthweight infant (OR 2.5, 95% CI 1.6-3.9), indicated preterm delivery (OR 2.4, 95% CI 1.4-4.1) and clinical chorioamnionitis (OR 2.7, 95% CI 1.4-5.1). The prevalence of BV determined using the Schmidt criteria in the early second trimester of pregnancy was similar to that found in similar studies. The presence of BV before 20 weeks of gestation was an independent risk factor for delivery of an infant with low birthweight, preterm delivery of a low-birthweight infant, indicated preterm delivery and clinical chorioamnionitis.


[Cerebral fat emboli after elective total hip replacement].
Fat embolism is a well known complication to fractures of long bones, but is less often seen after surgery. The symptoms vary from non- to total pulmonary and circulatory collapse and also include cerebral affection. Cerebral fat embolism is less common, and with varying presentation. This case report describes a case of fatal cerebral fat embolism after total hip replacement in a 75-year-old male. Diagnostics can be difficult but diffusion-weighted MRI can detect infarctions at an early stage. Treatment is symptomatic, and there is often total spontaneous remission of symptoms.


Reversible Dual-Covalent Molecular Locking of the 14-3-3/ERRγ Protein-Protein Interaction as a Molecular Glue Drug Discovery Approach.
Molecules that stabilize protein-protein interactions (PPIs) are invaluable as tool compounds for biophysics and (structural) biology, and as starting points for molecular glue drug discovery. However, identifying initial starting points for PPI stabilizing matter is highly challenging, and chemical optimization is labor-intensive. Inspired by chemical crosslinking and reversible covalent fragment-based drug discovery, we developed an approach that we term "molecular locks" to rapidly access molecular glue-like tool compounds. These dual-covalent small molecules reversibly react with a nucleophilic amino acid on each of the partner proteins to dynamically crosslink the protein complex. The PPI between the hub protein 14-3-3 and estrogen-related receptor γ (ERRγ) was used as a pharmacologically relevant case study. Based on a focused library of dual-reactive small molecules, a molecular glue tool compound was rapidly developed. Biochemical assays and X-ray crystallographic studies validated the ternary covalent complex formation and overall PPI stabilization via dynamic covalent crosslinking. The molecular lock approach is highly selective for the specific 14-3-3/ERRγ complex, over other 14-3-3 complexes. This selectivity is driven by the interplay of molecular reactivity and molecular recognition of the composite PPI binding interface. The long lifetime of the dual-covalent locks enabled the selective stabilization of the 14-3-3/ERRγ complex even in the presence of several other competing 14-3-3 clients with higher intrinsic binding affinities. The molecular lock approach enables systematic, selective, and potent stabilization of protein complexes to support molecular glue drug discovery.