Overview of direct oral anticoagulant therapy reversal.
Strategies for the management of bleeding complications and facilitation of an invasive procedure in patients receiving direct oral anticoagulants (DOACs) are reviewed. The DOACs provide clinical advantages versus vitamin K antagonists, including fixed dosing with no routine coagulation monitoring and evidence of a lower risk of bleeding. However, as with all anticoagulants, there is a risk of bleeding complications in patients receiving DOACs, so urgent reversal of their anticoagulant activity may be required for spontaneous or traumatic bleeding events and in patients undergoing emergency invasive procedures. Reversal strategies are dependent on the anticoagulant involved, the location and severity of the bleeding, and/or the urgency of the invasive procedure. The recently approved specific reversal agent for dabigatran, idarucizumab, together with other reversal agents in development will hopefully allow for the emergent reversal of DOACs, without increasing the underlying risk of thrombosis. However, research is required to determine the optimal use of these reversal agents, in terms of choice of agent, dosing, and concomitant management. A systematic approach to their implementation in hospitals is also required to ensure that physicians, nurses, and pharmacists receive appropriate education and have the necessary protocols and guidelines to manage these clinical situations. Reversal strategies in patients receiving a DOAC need to be tailored to the anticoagulant involved as well as the urgency and severity of the clinical situation. Reversal agents should help facilitate the urgent reversal of anticoagulation in patients with emergency bleeding or who require urgent surgery, though research and education are required to ensure the optimal use of these agents.


Sensitivity of system A and ASC transport activities to thiol-group-modifying reagents in rat liver plasma-membrane vesicles. Evidence for a direct binding of N-ethylmaleimide and iodoacetamide on A and ASC carriers.
1. In the present study we have examined the sensitivity of A and ASC amino-acid-carrier activities in rat liver plasma-membrane vesicles to the thiol-group modifying reagents N-ethylmaleimide (NEM) and iodoacetamide (IA). To this end, the different Na(+)-dependent entities involved in alanine transport were assessed. 2. NEM inactivated Na(+)-dependent alanine transport as a result of the inhibition of both system A and ASC transport activities. The functional sensitivity of system A to NEM was greater than that of system ASC. 3. The presence of L-alanine (10 mM) during the exposure of vesicles to NEM afforded partial protection to system A, but not to the ASC, carrier. This effect was specific, since the presence of L-phenylalanine (10 mM) did not cause any protection. 4. Na+ did not protect A or ASC carriers against NEM inactivation; however, the presence of Na+ (100 mM-NaCl) and L-alanine (10 mM) during the exposure of the vesicles to NEM protected against inactivation of system A and ASC transport activities. The extent of protection was greater in the case of the system ASC transport activity than in the case of the A carrier. 5. IA also diminished Na(+)-dependent alanine transport by inhibition of A and ASC transport activities. Sodium and L-alanine afforded protection to both A and ASC transport activities from the inhibitory action of IA. The extent of protection induced by substrates was similar for both carriers. 6. It is concluded that there is one, or several, free thiol groups in A and ASC carriers, the integrity of which is essential for transport activity. Sensitivity to thiol-group-specific reagents and the pattern of protection with substrates against inactivation is different in A and ASC carriers. That suggests the existence of topological dissimilarities regarding the thiol-group containing site(s) in A and ASC amino acid carriers.


Introduction of the DSM-5 levels of Personality Functioning Questionnaire.
With the introduction in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) of a hybrid system of personality disorder assessment, the ability to assess patients' traits, as well as their level of personality functioning, has become increasingly important. To assess this criterion, the DSM-5 Levels of Personality Functioning Questionnaire (DLOPFQ) was developed. The DLOPFQ assesses individuals' self-impairments and other impairments in several domains (self-direction, identity, empathy, and intimacy) and across 2 contexts (work/school and relationships). A sample of 140 psychiatric and medical outpatients was administered the DLOPFQ and several other measures to assess its reliability and construct, incremental, and discriminant validity. The internal consistency and convergence with validation measures yielded generally meaningful and expected results. Several DLOPFQ scales and subscales were significantly correlated with measures of DSM-5 trait domains and levels of personality functioning. DLOPFQ scales also correlated with self-reported ratings of overdependence, detachment, healthy dependency, and overall mental health and well-being. The DLOPFQ also predicted interpersonal and general functioning beyond DSM-5 trait domains. These results support the reliability and validity of the DLOPFQ, which appears to be suitable for clinical use and warrants ongoing study. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Alpha-galactosyl epitope-mediated activation of porcine aortic endothelial cells: type II activation.
Xenoreactive natural antibodies (XNAs) and complement mediate hyperacute rejection of discordant xenografts. Inhibition of complement alone results in some prolongation of graft survival, but delayed xenograft rejection still precludes long-term graft survival. In vitro data provide evidence for the direct proinflammatory activation of endothelial cells (ECs) by XNAs. These antibodies are primarily directed against galactose alpha(1-3)-galactose (alpha-gal), the major xenoantigen in the pig to primate xenotransplant model. Previous studies have shown EC activation by XNAs but failed to address the question of whether alpha-gal-specific ligands can induce EC activation. The aim of this study was to investigate whether agonist binding to the alpha-gal epitope by alpha-gal-specific lectins as compared with XNAs or elicited xenoreactive antibodies can directly elicit type II porcine aortic EC (PAEC) activation (i.e., activation that requires protein synthesis). The tetravalent, alpha-gal-binding Bandeiraea simplicifolia lectin I (BS-I), the wholly alpha-gal-specific BS-I isolectin B4, and elicited primate anti-pig xenoreactive antibodies (decomplemented cynomolgus monkey anti-porcine serum) induced E-selectin protein expression in PAECs. This induction was alpha-gal-specific, as preincubation with synthetic alpha-gal carbohydrate or adsorption of lectin or serum to rabbit, but not human, red blood cells removed the activating component. E-selectin expression, induced by BS-I, was inhibited in the presence of genistein, a tyrosine kinase inhibitor, and by mepacrine, an inhibitor of phospholipase A2. Human and primate XNAs lacked this activity when tested at relevant concentrations; however, stimulation of PAECs with affinity-purified human XNA (IgM and IgG) resulted in slightly increased interleukin-8 and P-selectin mRNA levels but had no apparent effects on E-selectin transcription. BS-I strongly induced E-selectin, P-selectin, intercellular adhesion molecule-1, and interleukin-8 mRNA in an NF-kappaB-dependent manner. Several agonists that specifically bind to alpha-gal can evoke type II EC activation. Hence, anti-Gal antibodies may contribute directly to xenograft rejection in the absence of complement activation.


Pharmacological evaluation of fruits of Terminalia belerica Roxb. for antiulcer activity.
The antiulcer activity of 70% methanolic extract of fruits of Terminalia belerica Roxb. (Combretaceae) was evaluated using ethanol induced, aspirin induced, cold stress restraint and pylorus ligated ulcer in rats. The methanolic extract (100, 250, 500, 1000 mg/kg, p.o.) significantly suppressed the peptic ulcer induced by ethanol. Methanolic extract at a dose of 500 mg/kg showed, promising antiulcer activity hence, this dose was selected for further evaluation of antiulcer studies. The methanolic extract (500 mg/kg) showed significant (p<0.05) reduction in gastric volume, free acidity, total acidity, ulcer index, protein and pepsin content and increase in mucus content in pylorus ligated rats as compared to control. Treatment with Terminalia belerica methanolic extract further provided significant antiulcer protection against aspirin induced ulcers but not in cold stress restraint model. These results suggested that the 70% methanolic extract of Terminalia belerica increased resistance to necrotizing agents, providing a direct protective effect on the gastric mucosa and exhibited antiulcer effect.


Reversible folding of human peripheral myelin protein 22, a tetraspan membrane protein.
Misfolding of the α-helical membrane protein peripheral myelin protein 22 (PMP22) has been implicated in the pathogenesis of the common neurodegenerative disease known as Charcot-Marie-Tooth disease (CMTD) and also several other related peripheral neuropathies. Emerging evidence suggests that the propensity of PMP22 to misfold in the cell may be due to an intrinsic lack of conformational stability. Therefore, quantitative studies of the conformational equilibrium of PMP22 are needed to gain insight into the molecular basis of CMTD. In this work, we have investigated the folding and unfolding of wild type (WT) human PMP22 in mixed micelles. Both kinetic and thermodynamic measurements demonstrate that the denaturation of PMP22 by n-lauroyl sarcosine (LS) in dodecylphosphocholine (DPC) micelles is reversible. Assessment of the conformational equilibrium indicates that a significant fraction of unfolded PMP22 persists even in the absence of the denaturing detergent. However, we find the stability of PMP22 is increased by glycerol, which facilitates quantitation of thermodynamic parameters. To our knowledge, this work represents the first report of reversible unfolding of a eukaryotic multispan membrane protein. The results indicate that WT PMP22 possesses minimal conformational stability in micelles, which parallels its poor folding efficiency in the endoplasmic reticulum. Folding equilibrium measurements for PMP22 in micelles may provide an approach to assess the effects of cellular metabolites or potential therapeutic agents on its stability. Furthermore, these results pave the way for future investigation of the effects of pathogenic mutations on the conformational equilibrium of PMP22.


A Drosophila model of Friedreich ataxia with CRISPR/Cas9 insertion of GAA repeats in the frataxin gene reveals in vivo protection by N-acetyl cysteine.
Friedreich ataxia (FA) is caused by GAA repeat expansions in the first intron of FXN, the gene encoding frataxin, which results in decreased gene expression. Thanks to the high degree of frataxin conservation, the Drosophila melanogaster fruitfly appears as an adequate animal model to study this disease and to evaluate therapeutic interventions. Here, we generated a Drosophila model of FA with CRISPR/Cas9 insertion of approximately 200 GAA in the intron of the fly frataxin gene fh. These flies exhibit a developmental delay and lethality associated with decreased frataxin expression. We were able to bypass preadult lethality using genetic tools to overexpress frataxin only during the developmental period. These frataxin-deficient adults are short-lived and present strong locomotor defects. RNA-Seq analysis identified deregulation of genes involved in amino-acid metabolism and transcriptomic signatures of oxidative stress. In particular, we observed a progressive increase of Tspo expression, fully rescued by adult frataxin expression. Thus, Tspo expression constitutes a molecular marker of the disease progression in our fly model and might be of interest in other animal models or in patients. Finally, in a candidate drug screening, we observed that N-acetyl cysteine improved the survival, locomotor function, resistance to oxidative stress and aconitase activity of frataxin-deficient flies. Therefore, our model provides the opportunity to elucidate in vivo, the protective mechanisms of this molecule of therapeutic potential. This study also highlights the strength of the CRISPR/Cas9 technology to introduce human mutations in endogenous orthologous genes, leading to Drosophila models of human diseases with improved physiological relevance.


The microbial landscape in bioturbated mangrove sediment: A resource for promoting nature-based solutions for mangroves.
Globally, soils and sediments are affected by the bioturbation activities of benthic species. The consequences of these activities are particularly impactful in intertidal sediment, which is generally anoxic and nutrient-poor. Mangrove intertidal sediments are of particular interest because, as the most productive forests and one of the most important stores of blue carbon, they provide global-scale ecosystem services. The mangrove sediment microbiome is fundamental for ecosystem functioning, influencing the efficiency of nutrient cycling and the abundance and distribution of key biological elements. Redox reactions in bioturbated sediment can be extremely complex, with one reaction creating a cascade effect on the succession of respiration pathways. This facilitates the overlap of different respiratory metabolisms important in the element cycles of the mangrove sediment, including carbon, nitrogen, sulphur and iron cycles, among others. Considering that all ecological functions and services provided by mangrove environments involve microorganisms, this work reviews the microbial roles in nutrient cycling in relation to bioturbation by animals and plants, the main mangrove ecosystem engineers. We highlight the diversity of bioturbating organisms and explore the diversity, dynamics and functions of the sediment microbiome, considering both the impacts of bioturbation. Finally, we review the growing evidence that bioturbation, through altering the sediment microbiome and environment, determining a 'halo effect', can ameliorate conditions for plant growth, highlighting the potential of the mangrove microbiome as a nature-based solution to sustain mangrove development and support the role of this ecosystem to deliver essential ecological services.


PUFA Supplementation and Heart Failure: Effects on Fibrosis and Cardiac Remodeling.
Heart failure (HF) characterized by cardiac remodeling is a condition in which inflammation and fibrosis play a key role. Dietary supplementation with n-3 polyunsaturated fatty acids (PUFAs) seems to produce good results. In fact, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have anti-inflammatory and antioxidant properties and different cardioprotective mechanisms. In particular, following their interaction with the nuclear factor erythropoietin 2 related factor 2 (NRF2), the free fatty acid receptor 4 (Ffar4) receptor, or the G-protein coupled receptor 120 (GPR120) fibroblast receptors, they inhibit cardiac fibrosis and protect the heart from HF onset. Furthermore, n-3 PUFAs increase the left ventricular ejection fraction (LVEF), reduce global longitudinal deformation, E/e ratio (early ventricular filling and early mitral annulus velocity), soluble interleukin-1 receptor-like 1 (sST2) and high-sensitive C Reactive protein (hsCRP) levels, and increase flow-mediated dilation. Moreover, lower levels of brain natriuretic peptide (BNP) and serum norepinephrine (sNE) are reported and have a positive effect on cardiac hemodynamics. In addition, they reduce cardiac remodeling and inflammation by protecting patients from HF onset after myocardial infarction (MI). The positive effects of PUFA supplementation are associated with treatment duration and a daily dosage of 1-2 g. Therefore, both the European Society of Cardiology (ESC) and the American College of Cardiology/American Heart Association (ACC/AHA) define dietary supplementation with n-3 PUFAs as an effective therapy for reducing the risk of hospitalization and death in HF patients. In this review, we seek to highlight the most recent studies related to the effect of PUFA supplementation in HF. For that purpose, a PubMed literature survey was conducted with a focus on various in vitro and in vivo studies and clinical trials from 2015 to 2021.


The Use of Electronic Health Record Data to Identify Variation in Referral, Consent, and Engagement in a Pediatric Intervention for Overweight and Obesity: A Cross-Sectional Study.
Clinical weight management programs face low participation. The authors assessed whether using electronic health record (EHR) data can identify variation in referral, consent, and engagement in a pediatric overweight and obesity (OW/OB) intervention. Using Epic EHR data collected between August 2020 and April 2021, sociodemographic and clinical diagnostic data (ie, International Classification of Disease [ICD] codes from visit and problem list [PL]) were analyzed to determine their association with referral, consent, and engagement in an OW/OB intervention. Bivariate analyses and multivariable logistic regression modeling were performed, with Bayesian inclusion criterion score used for model selection. Compared with the 581 eligible patients, referred patients were more likely to be boys (60% vs. 54%, respectively; P = 0.04) and have a higher %BMIp95 (119% vs. 112%, respectively; P < 0.01); consented patients were more likely to have a higher %BMIp95 (120% vs. 112%, respectively; P < 0.01) and speak Spanish (71% vs. 59%, respectively; P = 0.02); and engaged patients were more likely to have a higher %BMIp95 (117% vs. 112%, respectively; P = 0.03) and speak Spanish (78% vs. 59%, respectively; P < 0.01). The regression model without either ICD codes or PL diagnoses was the best fit across all outcomes, which were associated with baseline %BMIp95 and health clinic location. Neither visit nor PL diagnoses helped to identify variation in referral, consent, and engagement in a pediatric OW/OB intervention, and their role in understanding participation in such interventions remains unclear. However, additional efforts are needed to refer and engage younger girls with less extreme cases of OW/OB, and to support non-Hispanic families to consent.


Increased placental macrophages and a pro-inflammatory profile in placentas and maternal serum in infants with a decreased growth rate in the third trimester of pregnancy.
There is growing evidence for the role of placental inflammation in the pathophysiology of pregnancy complications including fetal growth restriction (FGR). This study aimed to characterize the inflammatory profile in the maternal circulation and the placenta of infants who were growth restricted and those that were small for gestational age (SGA). Placental villous tissue and maternal serum were obtained from pregnancies where infants were SGA at birth or who had a decreasing growth rate (≥25 centiles) across the third trimester. Immunohistochemical and histological analyses of placental samples were conducted for macrophage number, alongside vascular and cell turnover analysis. Inflammatory profile was analyzed in maternal and placental compartments via ELISAs and multiplex assays. There were significantly more CD163+ macrophages in placentas of infants with a decreased growth rate compared to controls, but not in SGA infants (median 8.6/ nuclei vs 3.8 and 2.9, P = .008 and P = .003, respectively). Uric acid (P = .0007) and IL-8 (P = .0008) were increased in placentas, and S100A8 (P < .0002) was increased in maternal serum of infants with decreased growth rate. No changes in the maternal serum or placental lysates of SGA infants were observed. The evidence of an altered inflammatory profile in infants with a decreasing growth rate, but not in those that were born SGA, provides further evidence that inflammation plays a role in true FGR. It remains unclear whether the increased placental macrophages occur as a direct result, or as a consequence of the pro-inflammatory environment observed in fetal growth restriction.


2018 Update on Pediatric Medical Overuse: A Review.
Efforts to combat medical overuse have gained traction in recent years, but success has been intermittent and shortcomings have been recognized. A commitment to a strong evidence base is needed to more broadly engage clinicians and reduce overuse. A structured MEDLINE search and a manual review of tables of contents from selected high-impact journals was performed to identify original research published in 2017 relevant to pediatric overuse. Articles were scored from low to high for 3 categories: quality of methods, magnitude of potential harm, and number of patients potentially harmed. The top-scoring articles presented in this review highlight examples of safe reductions in treatment intensity, including in the setting of cancer, appendicitis, acute respiratory tract infection, and elective anesthesia. This year's articles also provide cautionary examples of rational interventions adopted without a full understanding of potential harms, including pharmacologic migraine therapies, docosahexaenoic acid supplementation for preterm neonates, tight glycemic control for individuals with critically illness, and prophylactic antibiotics for children with vesicoureteral reflux. The articles represent high-quality, original research from 2017 that may help mitigate overuse. These works should be fundamental to the maturation of the pediatric overuse field.


Erythromycin-resistant group A streptococci in schoolchildren in Pittsburgh.
Resistance to erythromycin has been very uncommon among group A streptococci in the United States. As part of a longitudinal study, we obtained surveillance throat cultures twice monthly and with each new respiratory tract illness from children in kindergarten through grade 8 at one school in Pittsburgh. Screening for resistance to erythromycin and clindamycin was initially accomplished with use of the Kirby-Bauer disk-diffusion test. The minimal inhibitory concentration of resistant isolates was determined by the E test. A double disk-diffusion test was used to characterize the resistance phenotype, and the polymerase-chain-reaction assay was used to identify the resistance gene. The molecular relatedness of strains was determined by field-inversion gel electrophoresis. A total of 1794 throat cultures were obtained from 100 children between October 2000 and May 2001, of which 318 cultures (18 percent) from 60 of the children were positive for group A streptococci. Forty-eight percent of these isolates (153 of 318) were resistant to erythromycin. None were resistant to clindamycin. Results of the double disk-diffusion test indicated the presence of the M phenotype of erythromycin resistance. Molecular typing indicated that the outbreak was due to a single strain of group A streptococci. Of 100 randomly selected isolates of group A streptococci obtained from the community between April and June 2001, 38 were resistant to erythromycin. In January 2001, during a longitudinal study of schoolchildren, we detected the emergence of erythromycin resistance in pharyngeal isolates of group A streptococci. This clonal outbreak also affected the wider community.


The accessory magnocellular neurosecretory system of the rostral human hypothalamus.
The morphology and neurophysin expression of the magnocellular accessory neuroendocrine system located in the rostral human hypothalamus is investigated in a series of brains obtained at autopsy. The hypothalami were fixed in formalin and embedded in paraffin, or after cryoprotection, frozen for cryostat sectioning. Paraffin sections were either stained with Luxol Fast blue or immunoreacted for neurophysin I or neurophysin II, the precursor molecule for oxytocin and vasopressin. Further, 50-μm-thick serial cryostat sections were immunoreacted with the same antibodies. Both the paraventricular and supraoptic nuclei as well as the hypothalamo-hypophysial tracts exhibited strong immunoreactivity for the neurophysin antibodies. In addition, large collections of immunoreactive accessory magnocellular nuclei and single scattered neurophysin-positive neurons were located in the preoptic region between the paraventricular and supraoptic nucleus among the hypothalamo-hypophysial nerve fibers. In addition, smaller collections of neurophysin-immunoreactive neurons were located in the basal part of this region. Among the accessory magnocellular nuclei, the classical circular nucleus was identified. Accessory magnocellular neurons were often located along the blood vessels and projections of some of these neurons penetrated the vascular endothelium. The accessory magnocellular cell bodies expressed either neurophysin I or neurophysin II immunoreactivity. Summarizing, the accessory magnocellular system in the human brain is large and differs in morphology compared to the system seen in other vertebrates. The neurons of this system contain both vasopressin and oxytocin. Some neurons of the accessory neuronal systems might secrete vasopressin or oxytocin directly into the blood stream.


[The effect of Cd2+ on the respiration and swelling of isolated rat liver mitochondria in incubation media of different compositions].
We studied a change in Cd2+ influence on state 4 and 3 respiration and on 2,4-dinitrophenol (DNP)-stimulated respiration of isolated rat liver mitochondria, after replacement of KCl for equimolar quantities of NaCl, LiCl or for 200 mM sucrose in the incubation medium. Cd2+ concentrations higher than 2.5 microM reduced state 3 respiration or DNP-stimulated one in KCl medium (control), and activated state 4 respiration, however, at Cd2+ concentrations higher than 12.5 microM the respiratory activation became short-term with a following reduction in respiration. Cd2+ influence on respiration of mitochondria increased in the medium with NaCl, which resulted in decreasing the acting Cd2+ concentrations and in activating state 4 respiration more intensely. Cd2+ did not modify respiration of mitochondria in the experiments with LiCl in the medium, except that Cd2+ in concentrations higher than 10 microM decreased the state 4 respiration mitochondria just as in NaCl experiments. On the contrary, Cd2+ influence on respiration of mitochondria was seen decreased in the medium with sucrose. This was displayed in the increase in Cd2+ acting concentrations and in the decrease in activation of state 4 respiration mitochondria, and is associated with the nonosmotic character of these effects. On 20 microM Cd2+ concentration only, an extensive swelling of nonenergized mitochondria was observed in 12.5 mM nitrate media and its increase in series of Li < Na < K < NH4. The initially low swelling of succinate energized mitochondria in KCl medium was increased at raising Cd2+ concentration in this medium. A non-uniform Cd2+ influence on respiration and mitochondrial swelling in the above incubated media may be firstly associated with different effects of K+, Na+, Li+ and sucrose on membrane ion permeability and on Cd+ binding with the membrane, and, secondly, correlated with the action of medium components on energy dependent Cd2+ uptake by mitochondria and on Cd2+ penetration to mitochondrial respiratory enzymes.


Formation of thymine containing dimers in skin exposed to ultraviolet radiation.
Nuclear DNA appears to be the major molecular target for the inhibitory, mutagenic and lethal effects of ultraviolet radiation on cells in culture. Cyclobutyl dimers between adjacent pyrimidine bases, the major photochemical lesions for these effects in prokaryotes, also play a part in UVR effects on eukaryote cells. Pyrimidine dimers have been isolated from in vivo UV-irradiated guinea pig and mouse skin. The wavelength dependence for dimer induction is similar to that for acute skin reactions but no direct causal relationship has been established. Sunlight UVA may induce dimers in skin DNA. Excision of dimers from mouse skin in vivo is deficient as it is for most rodent cells in culture; human cell excision is efficient and the difficulties in interpretation of UV-carcinogenesis results with mice in terms of human skin cancer are therefore, increased.


High glycemic diet and breast cancer occurrence in the Italian EPIC cohort.
There are theoretical reasons for suspecting that a high glycemic index (GI) or glycemic load (GL) diet may increase breast cancer risk, perhaps via an effect on the insulin-like growth factor (IGF) axis. However observational studies have produced inconsistent findings and it is controversial whether breast cancer risk is influenced by the carbohydrate characteristics of the diet. We prospectively investigated the association between dietary GI and GL and breast cancer in the Italian section of the European Prospective Investigation into Cancer and Nutrition (EPIC). Women were recruited from 1993 to 1998 at five centers: Varese and Turin (north Italy), Florence (central Italy), and Ragusa and Naples (south Italy). Participants completed validated food frequency questionnaires from which GI and GL were estimated. Multivariable Cox proportional hazard regression models quantified the association between breast cancer risk and total carbohydrate intake, GI, and GL. During 11 years of follow-up, 879 breast cancer (797 invasive and 82 in situ) cases were indentified. High dietary GL was associated with increased breast cancer risk (RR 1.45, 95% CI = 1.06-1.99; highest vs. lowest quintile; p-trend 0.029), whereas dietary GI and total carbohydrate had no influence. The association was not modified by menopausal status or body mass index. Our data indicate that, in a Mediterranean population characterized by traditionally high and varied carbohydrate intake, a diet high in GL plays a role in the development of breast cancer.


Immunoglobulin V genes in rheumatoid arthritis.
Recent studies have revealed that a broad spectrum of Ig V genes (VH, V kappa, V lambda) contribute to the rheumatoid factor (RF) and non-RF produced by B cells in patients with rheumatoid arthritis (RA). This result contrasts with the restricted V gene use of paraprotein IgM RF. Certain VH and VL genes, however, appear more often in RA than expected from random expression, including some of the paraprotein-associated V genes. Many V genes expressed in RA are the same as those found in the fetal/CD5+ repertoire, suggesting an important role for CD5+ B cells in RA. Somatic mutations suggest that the B-cell response in RA is at least in part antigen-driven, although many Ig in RA have little or no mutation. Improved detection of V-gene polymorphisms now enable study of Ig V gene RFLP in RA.


[The mathematical modelling of the changes in energy metabolism in animal ontogeny].
A mathematical modeling of the intensity of respiration during growth is performed on the basis of equations that contain an intermediate independent variable tau. The model imitates k of peaks and lulls characteristic of the postembryonic period, which are followed by a continuous decrease in the energy level. The form of the equation for tau allowed us to suggest a hypothesis dealing with interactions of k + 1 cell generations, each bearing an imprinting of corresponding stage of development.


Managing resistance to anti-HIV drugs: an important consideration for effective disease management.
Current recommendations for the treatment of HIV-infected patients advise highly active antiretroviral therapy (HAART) consisting of combinations of 3 or more drugs to provide long-term clinical benefit. This is because only a complete suppression of virus replication will be able to prevent virus drug resistance, the main cause of drug failure. Virus drug resistance may remain a cause of concern in patients who have already received suboptimal mono- or bitherapy, or for patients who do not experience complete shut-down of virus replication under HAART. For these patients, replacement of one combination therapy regimen by another at drug failure, taking into account the existing resistance profile, will be needed. The development of new drugs will remain necessary for those patients who have failed to respond to all currently available drugs, as will be the institution of more effective and less toxic HAART regimens.


TAP-inhibiting proteins US6, ICP47 and UL49.5 differentially affect minor and major histocompatibility antigen-specific recognition by cytotoxic T lymphocytes.
CTLs specific for hematopoietic system-restricted minor histocompatibility antigens (mHags) can serve as reagents for cellular adoptive immunotherapy after allogeneic stem cell transplantation (SCT). In the HLA-mismatched setting, CTLs specific for hematopoietic system-restricted mHags expressed solely by the non-self 'allo' HLA molecules could be used to treat relapse after HLA-mismatched SCT. The generation of mHag-specific allo-HLA-restricted CTLs requires antigen-presenting cells (APCs) expressing low numbers of endogenous peptides to avoid co-induction of undesired allo-HLA reactivities. In this study, we exploited viral evasion strategies to generate APCs expressing a controlled set of endogenous peptides. Herpesviruses persist lifelong following primary infection due to expression of viral gene products that hamper T-cell recognition of infected cells. The herpesvirus-derived proteins US6, ICP47 and UL49.5 down-regulate endogenous antigen presentation in human APCs via inhibition of the transporter associated with antigen processing. EBV-transformed B cell lines transduced with retroviral vectors encoding US6, ICP47 or UL49.5 exhibited a stable decrease in cell-surface HLA class I expression and were protected from lysis by mHag-specific CTLs. Exogenous addition of mHag peptide fully restored target cell recognition. UL49.5 showed the most pronounced inhibitory effect, reducing HLA class I expression and mHag-specific lysis up to 99%. UL49.5 also significantly diminished allo-HLA reactivities mediated by allo-HLA-specific CTLs. In conclusion, UL49.5 could be a powerful new tool to study and modulate endogenous antigen presentation.


SREBP1 regulates mitochondrial metabolism in oncogenic KRAS expressing NSCLC.
Cancer cells require extensive metabolic reprograming in order to provide the bioenergetics and macromolecular precursors needed to sustain a malignant phenotype. Mutant KRAS is a driver oncogene that is well-known for its ability to regulate the ERK and PI3K signaling pathways. However, it is now appreciated that KRAS can promote the tumor growth via upregulation of anabolic metabolism. We recently reported that oncogenic KRAS promotes a gene expression program of de novo lipogenesis in non-small cell lung cancer (NSCLC). To define the mechanism(s) responsible, we focused on the lipogenic transcription factor SREBP1. We observed that KRAS increases SREBP1 expression and genetic knockdown of SREBP1 significantly inhibited the cell proliferation of mutant KRAS-expressing cells. Unexpectedly, lipogenesis was not significantly altered in cells subject to SREBP1 knockdown. Carbon tracing metabolic studies showed a significant decrease in oxidative phosphorylation and RNA-seq data revealed a significant decrease in mitochondrial encoded subunits of the electron transport chain (ETC). Taken together, these data support a novel role, distinct from lipogenesis, of SREBP1 on mitochondrial function in mutant KRAS NSCLC.


[An insufficiency of essential trace elements and its manifestations in pathology].
There exist 11 essential elements (iron, copper, iodine, zinc, chromium, cobalt, molybdenum, manganese, nickel, selenium and fluorine) vitally important for human health. The deficiency of these trace elements results in many pathological processes, including such world-wide human diseases, as iron-deficient anemias and other forms of hyposiderosis, endemic hypothyroid goiter, endemic fluorine deficiency states, particularly manifested by dental caries in children and osteoporosis of the advanced age. Selenium deficiency causes endemic cardiopathy in China, is a risk factor for the development and severe course of congestive cardiopathies, as well as myocardial infarction in a number of European countries. Correction of selenium deficiency with sodium selenite reduced to the minimum the mortality rate from endemic cardiopathy in China (Keshan disease) and had a favourable effect on other forms of cardiovascular pathology. Essential trace element deficiency results in many forms of embryonal and fetal pathology and decreases the antitumour resistance in man. New essential trace elements are still being identified and their number reached 19 at present. Development of pathological anatomy of essential trace element deficiency is an important task of modern medicine.


Induction of a cell membrane antigen by equine infectious anemia virus.
Equine fibroblasts persistently infected with equine infectious anemia virus acquire a new cell membrane antigen demonstrable by indirect radioimmunoassay, using infected horse serum as an antibody source.


Multiplex Three-Dimensional Mapping of Macromolecular Drug Distribution in the Tumor Microenvironment.
Macromolecular cancer drugs such as therapeutic antibodies and nanoparticles are well known to display slow extravasation and incomplete penetration into tumors, potentially protecting cancer cells from therapeutic effects. Conventional assays to track macromolecular drug delivery are poorly matched to the heterogeneous tumor microenvironment, but recent progress on optical tissue clearing and three-dimensional (3D) tumor imaging offers a path to quantitative assays with cellular resolution. Here, we apply transparent tissue tomography (T3) as a tool to track perfusion and delivery in the tumor and to evaluate target binding and vascular permeability. Using T3, we mapped anti-programmed cell death protein-ligand 1 (PD-L1) antibody distribution in whole mouse tumors. By measuring 3D penetration distances of the antibody drug out from the blood vessel boundaries into the tumor parenchyma, we determined spatial pharmacokinetics of anti-PD-L1 antibody drugs in mouse tumors. With multiplex imaging of tumor components, we determined the distinct distribution of anti-PD-L1 antibody drug in the tumor microenvironment with different PD-L1 expression patterns. T3 imaging revealed CD31+ capillaries are more permeable to anti-PD-L1 antibody transport compared with the blood vessels composed of endothelium supported by vascular fibroblasts and smooth muscle cells. T3 analysis also confirmed that isotype IgG antibody penetrates more deeply into tumor parenchyma than anti-Her2 or anti-EGFR antibody, which were restrained by binding to their respective antigens on tumor cells. Thus, T3 offers simple and rapid access to 3D, quantitative maps of macromolecular drug distribution in the tumor microenvironment, offering a new tool for development of macromolecular cancer therapeutics.


Link Connectivity and Coverage of Underwater Cognitive Acoustic Networks under Spectrum Constraint.
Extensive attention has been given to the use of cognitive radio technology in underwater acoustic networks since the acoustic spectrum became scarce due to the proliferation of human aquatic activities. Most of the recent studies on underwater cognitive acoustic networks (UCANs) mainly focus on spectrum management or protocol design. Few efforts have addressed the quality-of-service (QoS) of UCANs. In UCANs, secondary users (SUs) have lower priority to use acoustic spectrum than primary users (PUs) with higher priority to access spectrum. As a result, the QoS of SUs is difficult to ensure in UCANs. This paper proposes an analytical model to investigate the link connectivity and the probability of coverage of SUs in UCANs. In particular, this model takes both topological connectivity and spectrum availability into account, though spectrum availability has been ignored in most recent studies. We conduct extensive simulations to evaluate the effectiveness and the accuracy of our proposed model. Simulation results show that our proposed model is quite accurate. Besides, our results also imply that the link connectivity and the probability of coverage of SUs heavily depend on both the underwater acoustic channel conditions and the activities of PUs.


F-18-fluorodeoxyglucose positron emission tomography imaging-assisted management of patients with severe left ventricular dysfunction and suspected coronary disease: a randomized, controlled trial (PARR-2).
We conducted a randomized trial to assess the effectiveness of F-18-fluorodeoxyglucose (FDG) positron emission tomography (PET)-assisted management in patients with severe ventricular dysfunction and suspected coronary disease. Such patients may benefit from revascularization, but have significant perioperative morbidity and mortality. F-18-fluorodeoxyglucose PET can detect viable myocardium that might recover after revascularization. Included were patients with severe left ventricular (LV) dysfunction and suspected coronary disease being considered for revascularization, heart failure, or transplantation work-ups or in whom PET was considered potentially useful. Patients were stratified according to recent angiography or not, then randomized to management assisted by FDG PET (n = 218) or standard care (n = 212). The primary outcome was the composite of cardiac death, myocardial infarction, or recurrent hospital stay for cardiac cause, within 1 year. At 1 year, the cumulative proportion of patients who had experienced the composite event was 30% (PET arm) versus 36% (standard arm) (relative risk 0.82, 95% confidence interval [CI] 0.59 to 1.14; p = 0.16). The hazard ratio (HR) for the composite outcome, PET versus standard care, was 0.78 (95% CI 0.58 to 1.1; p = 0.15); for patients that adhered to PET recommendations for revascularization, revascularization work-up, or neither, HR = 0.62 (95% CI 0.42 to 0.93; p = 0.019); in those without recent angiography, for cardiac death, HR = 0.4 (95% CI 0.17 to 0.96; p = 0.035). This study did not demonstrate a significant reduction in cardiac events in patients with LV dysfunction and suspected coronary disease for FDG PET-assisted management versus standard care. In those who adhered to PET recommendations and in patients without recent angiography, significant benefits were observed. The utility of FDG PET is best realized in this subpopulation and when adherence to recommendations can be achieved.


The JCAH quality initiative: what can hospitals and physicians expect?. Interview by Emily Friedman.
In his new position at the JCAH, Ol'Leary has been shaking things up. His most talked-about move has been to initiate a broad-scale reconfiguration of the Joint Commission's approaches to evaluating quality. The principle element of this effort calls for structural and functional indicators to be augmented by clinical performance and outcome criteria. This is the first of two articles concerning Dr. O'Leary's views of the future of the Joint Commission. In this article, he discusses the new quality assurance initiative; in an article in the September-October issue of Physician Executive, his vision of the future of the JCAH, and the environment in which it will be operating, will be explored.


Renal cell type specificity of cephalosporin-induced cytotoxicity in suspensions of isolated proximal tubular and distal tubular cells.
We have developed an in vitro model for investigation of nephron heterogeneity and cell type-specific patterns of renal injury. To further validate our model and to study biochemical mechanisms of cephalosporin-induced injury, cytotoxicity of three cephalosporins was studied in freshly isolated proximal tubular (PT) and distal tubular (DT) cells from rat kidney. The three cephalosporins [cephaloridine (CPH), cephalexin (CXN), cephalothin (CTN)] were chosen because they exhibit varying degrees of nephrotoxicity in vivo and contain different functional groups. CPH produced greater amounts of lactate dehydrogenase release from PT cells than either CXN or CTN, indicating greater toxicity of CPH, which agrees with in vivo observations. DT cells were not affected by any of the cephalosporins. Thus, the cephem ring is sufficient to produce PT cell injury but the presence of other functional groups modifies toxicity. SKF-525A and alpha-tocopherol protected PT cells from both CPH and CTN, suggesting involvement of cytochrome P-450 metabolism and oxidative stress. Both PT and DT cells exhibited transport of CPH or CXN and transport of CPH into PT cells was inhibitable by probenecid, consistent with action of a specific carrier. Transport alone, therefore, cannot account for the cell type specificity pattern in vitro. Effects on intracellular glutathione status, malondaldehyde formation, and uncoupler-stimulated respiration were also investigated, and these generally correlated with cell type specificity patterns but not always with degree of cytotoxicity. These results validate further the isolated PT and DT cells as in vitro models to study cell type-specific renal injury and show a role for oxidative stress, cytochrome P-450 bioactivation, and mitochondrial dysfunction in cephalosporin-induced PT cell injury.


The Enlightened Brain: Novel Imaging Methods Focus on Epileptic Networks at Multiple Scales.
Epilepsy research is rapidly adopting novel fluorescence optical imaging methods to tackle unresolved questions on the cellular and circuit mechanisms of seizure generation and evolution. State of the art two-photon microscopy and wide-field fluorescence imaging can record the activity in epileptic networks at multiple scales, from neuronal microcircuits to brain-wide networks. These approaches exploit transgenic and viral technologies to target genetically encoded calcium and voltage sensitive indicators to subclasses of neurons, and achieve genetic specificity, spatial resolution and scalability that can complement electrophysiological recordings from awake animal models of epilepsy. Two-photon microscopy is well suited to study single neuron dynamics during interictal and ictal events, and highlight the differences between the activity of excitatory and inhibitory neuronal classes in the focus and propagation zone. In contrast, wide-field fluorescence imaging provides mesoscopic recordings from the entire cortical surface, necessary to investigate seizure propagation pathways, and how the unfolding of epileptic events depends on the topology of brain-wide functional connectivity. Answering these questions will inform pre-clinical studies attempting to suppress seizures with gene therapy, optogenetic or chemogenetic strategies. Dissecting which network nodes outside the seizure onset zone are important for seizure generation, propagation and termination can be used to optimize current and future evaluation methods to identify an optimal surgical strategy.


Septic shock: desperately seeking treatment.
Septic shock results from the dysregulation of the innate immune response following infection. Despite major advances in fundamental and clinical research, patients diagnosed with septic shock still have a poor prognostic outcome, with a mortality rate of up to 50%. Indeed, the reasons leading to septic shock are still poorly understood. First postulated 30 years ago, the general view of septic shock as an acute and overwhelming inflammatory response still prevails today. Recently, the fact that numerous clinical trials have failed to demonstrate any positive medical outcomes has caused us to question our fundamental understanding of this condition. New and sophisticated technologies now allow us to accurately profile the various stages and contributory components of the inflammatory response defining septic shock, and many studies now report a more complex inflammatory response, particularly during the early phase of sepsis. In addition, novel experimental approaches, using more clinically relevant animal models, to standardize and stratify research outcomes are now being argued for. In the present review, we discuss the most recent findings in relation to our understanding of the underlying mechanisms involved in septic shock, and highlight the attempts made to improve animal experimental models. We also review recent studies reporting promising results with two vastly different therapeutic approaches influencing the renin-angiotensin system and applying mesenchymal stem cells for clinical intervention.


QMAC-dRAST for the direct testing of antibiotic susceptibility for Enterobacterales in positive blood-culture broth: a comparison of the performances with the MicroScan system and direct disc diffusion testing methods.
To evaluate the performances of the QMAC-dRAST GN (Gram-negative) kit for rapid antimicrobial sensitivity testing (AST) and two other methods, directly on positive blood-culture broth (PBCB), by comparison with a reference method: the MicroScan method based on broth microdilution on colonies isolated on PBCB subculture. In total, 156 samples were collected prospectively from blood cultures positive for a Gram-negative rod. Each sample was tested with four AST techniques: (i) the QMAC dRAST GN kit, (ii) the disc diffusion (DD) method, (iii) the MicroScan method applied directly to PBCB; and (iv) MicroScan with isolates from PBCB subculture, as a reference. For 124 PBCB containing Enterobacterales, overall essential agreement (EA) and categorical agreement (CA) between the QMAC-dRAST on PBCB and the reference reached 95.7% and 93.5%, respectively. There were 3.0% very major errors (VME), 4.0% major errors (ME) and 2.8% minor errors (mE). A comparison of MicroScan on PBCB and the reference yielded 98.8% EA, 98.5% CA, and rates of 0.6% VME, 0.9% ME and 0.7% mE. The DD method on PBCB gave a CA of 95.8% and rates of 1.7% for VME, 2.0% for ME and 1.9% for mE. Results were obtained more rapidly for QMAC-dRAST (median of 6 h 37 min versus 18 h for the MicroScan and DD methods on PBCB). The QMAC-dRAST system provided rapid results well correlated with the reference method on PBCB containing Enterobacterales. Given the shorter time-to-results, the QMAC-dRAST system constitutes a fast and reliable alternative to conventional AST methods.


Unresectable hepatocellular carcinoma at dawn of immunotherapy era: real-world data from the French prospective CHIEF cohort.
Hepatocellular carcinoma epidemiological data are limited in France. The Epidemio Liver Immunotherapy Tecentriq outcome research (ELITor) retrospective study, based on real-world data from the Carcinome HépatocellulaIrE en France (CHIEF) French cohort of hepatocellular carcinoma patients, aimed to get insight into the treatment patterns, the sociodemographic, clinical, biological, and etiological characteristics, and the quality of life of patients with unresectable hepatocellular carcinoma. Between 1 September 2019 and 4 December 2020, 367 patients from the CHIEF cohort received at least one locoregional (52.8%) chemoembolization or radioembolization or systemic treatment (88.3%) and were selected for ELITor. Most patients had a Barcelona Clinic Liver Cancer (BCLC) C (93.2%) hepatocellular carcinoma stage and were affected by cirrhosis (67.7%). Alcohol was confirmed as the main etiology both as a single etiology (29.1%) and in association with other risk factors (26.9%), mainly metabolic disorders (16.2%).Tyrosine-kinase inhibitors, mainly sorafenib, were the most administered systemic treatments in first line. Patients who received at least one combination of atezolizumab and bevacizumab during the study period ( N = 53) had a better performance status and less portal hypertension frequency than the overall population and more hepatitis B virus infection and fewer metabolic disorders as single etiology. Overall, the global health score before treatment (62.3 ± 21.9) was in line with that of reference cancer patients and worsened in 51.9% of the cases after first-line palliative-intent treatment. This study provided real-life data on advanced hepatocellular carcinoma characteristics and treatment patterns and described the first patients to receive the atezolizumab-bevacizumab combination before it became the new standard of care for advanced hepatocellular carcinoma.


Which Small Molecule? Selecting Chemical Probes for Use in Cancer Research and Target Validation.
Small-molecule chemical "probes" complement the use of molecular biology techniques to explore, validate, and generate hypotheses on the function of proteins in diseases such as cancer. Unfortunately, the poor selection and use of small-molecule reagents can lead to incorrect conclusions. Here, we illustrate examples of poor chemical tools and suggest best practices for the selection, validation, and use of high-quality chemical probes in cancer research. We also note the complexity associated with tools for novel drug modalities, exemplified by protein degraders, and provide advice and resources to facilitate the independent identification of appropriate small-molecule probes by researchers. Validation of biological targets and pathways will be aided by a shared understanding of the criteria of potency, selectivity, and target engagement associated with small-molecule reagents ("chemical probes") that enable that work. Interdisciplinary collaboration between cancer biologists, medicinal chemists, and chemical biologists and the awareness of available resources will reduce misleading data generation and interpretation, strengthen data robustness, and improve productivity in academic and industrial research.


Interleukin 6 response to urinary tract infection in childhood.
This study analyzed the interleukin 6 (IL-6) response in 114 children with suspected urinary tract infection (UTI). Urine and serum samples were obtained at the time of enrollment. There were 90 children with UTI, 41 with and 49 without a temperature > or = 38.5 degrees C. The remaining 24 children did not have bacteriuria; 11 were febrile and 13 were not. The urinary IL-6 concentrations were higher in the children with UTI (mean, 129 units/ml) than in the children without bacteriuria (mean, 7 units/ml, P < 0.01). In contrast the serum IL-6 did not differ between children with or without UTI or between children with or without a temperature > or = 38.5 degrees C. The urinary IL-6 response was higher in children who were infected with P fimbriated Escherichia coli than in other children with UTI (P < 0.05). There was a correlation of urinary IL-6 with the degree of proteinuria, hematuria and urinary leukocyte counts (P < 0.001, P < 0.05, P < 0.05, respectively) but not with serum IL-6, CRP or temperature, and of serum IL-6 to C-reactive protein (P = 0.053) and renal concentrating capacity (P < 0.05). The results demonstrate that infections of the urinary tract activate an IL-6 response in children and that the magnitude of the IL-6 response is influenced by the properties of the infecting strain.


Performance of Tools and Measures to Predict Fluid Responsiveness in Pediatric Shock and Critical Illness: A Systematic Review and Meta-Analysis.
In this systematic review and meta-analysis we asked: Do predictors of fluid responsiveness in children perform comparably: 1) in the PICU as in non-PICU settings? 2) in shock states compared with nonshock states? Additionally, 3) is there an association between preload responsiveness and clinical response? Ovid Medline, PubMed, and Embase databases were searched from inception through May 2022. Included studies reported physiological response to IV fluid administration in humans less than 18 years. Only studies reporting an area under the receiver operating characteristic curve (AUROC) were included for descriptive analysis. Only studies for which a se could be estimated were included for meta-analysis. Title, abstract, full text screening, and extraction were completed by two authors (S.B.W., J.M.W.). Variables extracted included predictors ("tools") and outcome measures ("reference tests") of fluid responsiveness, demographic, and clinical variables. We identified 62 articles containing 204 AUROCs for 55 tools, primarily describing mechanically ventilated children in an operating room or PICU. Meta-analysis across all tools showed poor predictive performance (AUROC, 0.66; 95% CI, 0.63-0.69), although individual performance varied greatly (range, 0.49-0.87). After controlling for PICU setting and shock state, PICU setting was associated with decreased predictive performance (coefficient, -0.56; p = 0.0007), while shock state was associated with increased performance (0.54; p = 0.0006). Effect of PICU setting and shock state on each tool was not statistically significant but analysis was limited by sample size. The association between preload responsiveness and clinical response was rarely studied but results did not suggest an association. Ultrasound measurements were prone to inherent test review and incorporation biases. We suggest three opportunities for further research in fluid responsiveness in children: 1) assessing predictive performance of tools during resuscitation in shock states; 2) separating predictive tool from reference test when using ultrasound techniques; and 3) targeting decreasing time in a shock state, rather than just increase in preload.


A single session of anodal transcranial direct current stimulation applied over the affected primary motor cortex does not alter gait parameters in chronic stroke survivors.
The excitability of some neural circuits involved in walking and affected in individuals with chronic stroke can be modulated during and/or immediately after anodal transcranial direct current stimulation (a-tDCS). This study was designed to investigate the effects of a-tDCS during and immediately after application on leg muscle activity during gait, and on spatiotemporal and kinematic gait parameters in patients with chronic stroke. This study was randomized, sham-controlled and double-blinded with a cross-over design and included 24 individuals with chronic stroke. Each participant underwent one 30-minute session each of effective a-tDCS at 2mA and sham tDCS. In both sessions, the anode was placed over the leg motor cortex of the affected hemisphere and the cathode over the contralateral orbit. Six gait trials were performed before, during and immediately after each effective/sham tDCS session. Electromyographic activity of leg muscles, as well as spatiotemporal (e.g. gait speed) and kinematic (e.g. peak knee flexion and ankle dorsiflexion in the swing phase of gait) gait parameters were recorded. Genotyping for the brain-derived neurotrophic factor (BDNF) Val66Met polymorphism was undertaken since this gene may influence motor skill learning and the effects of tDCS. No significant effects of a-tDCS on gait parameters were found either for the total group or for the Val66Met (N=10) and Val66Val (N=14) subgroups. A single session of a-tDCS delivered to the leg motor cortex did not immediately improve gait parameters in individuals with chronic stroke, regardless of their BDNF genotype.


Cortical evoked response audiometry thresholds and neuroleptic, sedative, hypnotic drugs.
Cortical evoked response audiometry is adequate for approximating hearing threshold levels with frequency specificity when the psychoacoustic responses lack reliability and reproducibility (compensation claim). It is well-known that control of wakefulness is essential for the reliability of slow vertex responses (SVR). Therefore, sedative, hypnotic, and neuroleptic drugs are supposed to have possible adverse effects on the detection of SVR. In contrast, brainstem evoked responses (BER) have proved not to be significantly affected by therapeutic doses of these compounds. The purpose of our study was to assess the reliability of SVR-threshold definition in subjects taking neuroleptic, sedative, and hypnotic drugs. Fifteen subjects examined for occupational hearing loss at the Fund for Occupational Diseases in Brussels and regularly taking one or several of these drugs were compared with 27 comparable controls. In each subject the auditory thresholds were defined with both techniques: SVR (1, 2, and 3 kHz) and BER (clicks). A highly significant difference is observed between the two groups: In the group receiving drugs, the SVR threshold for 3 kHz is 12.1 dB (average) higher than the BER threshold, whereas in the group without drugs, the SVR threshold for 3 kHz is 7.77 dB (average) lower than the BER threshold. In the drug group, large interindividual differences are observed. It may be concluded that the use of neuroleptics, sedatives, and hypnotics renders the auditory threshold definition with SVR completely unreliable. In using SVR for medicolegal threshold definition, controlling the 3-kHz threshold with BER always is necessary.


Combined posterior pelvis and lumbar spine resection for sarcoma.
The oncologic outcome in the treatment of pelvic sarcomas is not comparable with that in the extremity. Particular problems arise when the tumor involves the posterior pelvis and the sacrum or even the lower lumbar spine. Because of the difficulty in achieving local control and the major functional deficits after extensive surgery, some authors suggest conservative, nonsurgical treatment for these patients. The purpose of the current study was to analyze the oncologic outcome of patients who were treated for a pelvic sarcoma necessitating resection of the ilium, part of the sacrum and part of the lower spine, which is defined as extended hemipelvectomy. Between 1979 and 1999, 11 males and seven females with a mean age of 34.5 years (range, 14-67 years) had an extended hemipelvectomy for a sarcoma of the posterior pelvis. The tumors included seven osteogenic sarcomas, six chondrosarcomas, and five fibrosarcomas: 13 were classified as high-grade lesions. The mean size of the tumor was 11 x 9 x 6 cm. In 11 patients, a hemivertebrectomy of L5 and in seven patients of L4 combined with L5 was done to achieve an adequate margin. In four patients, the resection was followed by reconstruction with a vascularized fibula or autograft. At a mean followup of 56 months (range, 3-220 months), 12 of 18 patients were alive and without disease. Six patients died; four of these patients had metastasis to the lung at presentation. The two remaining patients died of complications of adjuvant treatment. The authors conclude from this small series of patients that an aggressive surgical resection for localized, nonmetastatic, high-grade sarcomas of the posterior pelvis may be justified to provide local control and improve survival.


Reduced specificity of functional connectivity in the aging brain during task performance.
The importance of studying connectivity in the aging brain is increasingly recognized. Recent studies have shown that connectivity within the default mode network is reduced with age and have demonstrated a clear relation of these changes with cognitive functioning. However, research on age-related changes in other functional networks is sparse and mainly focused on prespecified functional networks. Using functional magnetic resonance imaging, we investigated age-related changes in functional connectivity during a visual oddball task in a range of functional networks. It was found that compared with young participants, elderly showed a decrease in connectivity between areas belonging to the same functional network. This was found in the default mode network and the somatomotor network. Moreover, in all identified networks, elderly showed increased connectivity between areas within these networks and areas belonging to different functional networks. Decreased connectivity within functional networks was related to poorer cognitive functioning in elderly. The results were interpreted as a decrease in the specificity of functional networks in older participants.


Manipulating cofactor binding thermodynamics in an artificial oxygen transport protein.
We report the mutational analysis of an artificial oxygen transport protein, HP7, which operates via a mechanism akin to that of human neuroglobin and cytoglobin. This protein destabilizes one of two heme-ligating histidine residues by coupling histidine side chain ligation with the burial of three charged glutamate residues on the same helix. Replacement of these glutamate residues with alanine, which is uncharged, increases the affinity of the distal histidine ligand by a factor of 13. Paradoxically, it also decreases heme binding affinity by a factor of 5 in the reduced state and 60 in the oxidized state. Application of a three-state binding model, in which an initial pentacoordinate binding event is followed by a protein conformational change to hexacoordinate, provides insight into the mechanism of this seemingly counterintuitive result: the initial pentacoordinate encounter complex is significantly destabilized by the loss of the glutamate side chains, and the increased affinity for the distal histidine only partially compensates for that. These results point to the importance of considering each oxidation and conformational state in the design of functional artificial proteins.


Bilateral temporal lobe dysplasia and seizure onset associated with biallelic CNTNAP2 variants.
Biallelic CNTNAP2 variants have been associated with Pitt-Hopkins like syndrome. We describe six novel and one previously reported patients from six independent families and review the literature including 64 patients carrying biallelic CNTNAP2 variants. Initial reports highlighted intractable focal seizures and the failure of epilepsy surgery in children, but subsequent reports did not expand on this aspect. In all our patients (n=7), brain MRI showed bilateral temporal grey/white matter blurring with white matter high signal intensity, more obvious in the T2-FLAIR sequences, consistent with bilateral temporal lobe dysplasia. All patients had focal seizures with temporal lobe onset and semiology, which were recorded in five, showing bilateral independent temporal onset in four. Epilepsy was responsive to anti-seizure medications in two patients (2/7, 28.5%), and pharmaco-resistant in five (5/7, 71.5%). Splice-site variants identified in five patients (5/7, 71.5%) were the most common mutational finding. Our observation expands the phenotypic and genetic spectrum of biallelic CNTNAP2 alterations focusing on the neuroimaging features and provides evidence for an elective bilateral anatomoelectroclinical involvement of the temporal lobes in the associated epilepsy with relevant implications on clinical management.


Electronic structure of Fe- vs. Ru-based dye molecules.
In order to explore whether Ru can be replaced by inexpensive Fe in dye molecules for solar cells, the differences in the electronic structure of Fe- and Ru-based dyes are investigated by X-ray absorption spectroscopy and first-principles calculations. Molecules with the metal in a sixfold, octahedral N cage, such as tris(bipyridines) and tris(phenanthrolines), exhibit a systematic downward shift of the N 1s-to-π* transition when Ru is replaced by Fe. This shift is explained by an extra transfer of negative charge from the metal to the N ligands in the case of Fe, which reduces the binding energy of the N 1s core level. The C 1s-to-π* transitions show the opposite trend, with an increase in the transition energy when replacing Ru by Fe. Molecules with the metal in a fourfold, planar N cage (porphyrins) exhibit a more complex behavior due to a subtle competition between the crystal field, axial ligands, and the 2+ vs. 3+ oxidation states.


Interactions between endocarditis-derived Streptococcus gallolyticus subsp. gallolyticus isolates and human endothelial cells.
Streptococcus gallolyticus subsp. gallolyticus is an important causative agent of infective endocarditis (IE) but the knowledge on virulence factors is limited and the pathogenesis of the infection is poorly understood. In the present study, we established an experimental in vitro IE cell culture model using EA.hy926 and HUVEC cells to investigate the adhesion and invasion characteristics of 23 Streptococcus gallolyticus subsp. gallolyticus strains from different origins (human IE-derived isolates, other human clinical isolates, animal isolates). Adhesion to eight components of the extracellular matrix (ECM) and the ability to form biofilms in vitro was examined in order to reveal features of S. gallolyticus subsp. gallolyticus endothelial infection. In addition, the strains were analyzed for the presence of the three virulence factors gtf, pilB, and fimB by PCR. The adherence to and invasion characteristics of the examined S. gallolyticus subsp. gallolyticus strains to the endothelial cell line EA.hy926 differ significantly among themselves. In contrast, the usage of three different in vitro models (EA.hy926 cells, primary endothelial cells (HUVECs), mechanical stretched cells) revealed no differences regarding the adherence to and invasion characteristics of different strains. Adherence to the ECM proteins collagen I, II and IV revealed the highest values, followed by fibrinogen, tenascin and laminin. Moreover, a strong correlation was observed in binding to these proteins by the analyzed strains. All strains show the capability to adhere to polystyrole surfaces and form biofilms. We further confirmed the presence of the genes of two known virulence factors (fimB: all strains, gtf: 19 of 23 strains) and demonstrated the presence of the gene of one new putative virulence factor (pilB: 9 of 23 strains) by PCR. Our study provides the first description of S. gallolyticus subsp. gallolyticus adhesion and invasion of human endothelial cells, revealing important initial information of strain variability, behaviour and characteristics of this as yet barely analyzed pathogen.


Culture, personal experience and agency.
In this article, we explore what we perceive to be a gap between agency as articulated in practice theories and agency as personally experienced. The gap is not created by a turn to practice in theorizing, but by the tendency to produce theoretical representations that silence the particularity of experience and the diversity of voices in experience. In exploring the gap, we identify aspects of practice theories that explicitly commit to theoretical representation over personal experience and describe Bakhtin's commitment to action and personal experience as an alternative. In order to exemplify Bakhtin's approach in practice, we then present an analysis of one artist-teacher's experience of her own agency in making art and in teaching. Finally, we comment on what a commitment to representational theorizing does to accounts of an artist's activities and personal experience.


Investigation of upper gastrointestinal symptoms in patients with AIDS.
Double contrast barium radiology and upper gastrointestinal endoscopy were compared prospectively on 45 occasions in patients infected with HIV who presented with upper gastrointestinal symptoms. In 40 cases, a definite pathological diagnosis was reached and in four cases no organic basis for symptoms could be found. A correct and complete diagnosis was made on visual endoscopic criteria in 43 cases (95.5%) but in only 14 cases (31.1%) from barium studies alone. Radiology showed no abnormality in 22 cases where pathological changes were documented (oesophageal candidiasis in 21 cases). Where pathological confirmation of diagnosis existed (40 cases), endoscopy (without pathological support) had a sensitivity of 97.5% and a specificity of 100% compared with the sensitivity and specificity of 25 and 100% for barium studies. The difference between the sensitivities of the two methods was highly significant (P less than 0.005). The combination of oral candidiasis and upper gastrointestinal symptoms without dysphagia or weight loss was so strongly associated with uncomplicated oesophageal candidiasis (negative predictive value 93%; P less than 0.025), that this is supported as a basis for therapy without the need for further investigation, although if upper gastrointestinal investigation is required, endoscopy should be the method of choice.


Airway management in bariatric surgery patients, our experience in Qatar: A prospective observational cohort study.
Background: Obesity has always been considered a criterion of difficult airway management, and many authors have tackled this subject. We are presenting our experience in airway management in obese patients undergoing bariatric surgery in Qatar and comparing the results with previous studies. Objective: The primary objective of this study was to explore the relationship between difficult mask ventilation and difficult intubation. The secondary objective was to identify other factors that may play a role in either difficulty such as gender, associated comorbidities, and the skill and experience of anesthetists. Design: This study was a prospective observational cohort study. Sample: A total of 401 patients were selected for various elective bariatric surgery in Hamad General Hospital, including 130 males and 271 females with an average body mass index(BMI) of 46.03 kg m- 2. Results: We used Pearson Chi-Square and Yates corrected Chi-square statistical tests in our statistical analysis. Neck circumference had a p value of 0.001 in both genders. The male gender had a p value of 0.052 and 0.012 in mask ventilation and difficult intubation, respectively. The Mallampati score had a p value of 0.56 and 0.006 in mask ventilation and intubation, respectively. In general, neck circumference, Mallampati score, gender, obstructive sleep apnea, and diabetes mellitus had greater negative effects on airway management than BMI alone. Conclusion: It was hard to intubate 25% of patients who had difficult mask ventilation (DMV). All DMV and 20 out of 23 of difficult intubation patients were in the high BMI group ( ≥ 40). Neck circumference, Mallampati score, and male gender were major independent factors; however, other factors, such as obstructive sleep apnea, and diabetes mellitus, should be kept in mind as additional risks.


Predicting different dimensions of schizophrenia-related stigma in Israel: Exploring the applicability of the common sense model of illness.
The common sense model of illness is a promising conceptual framework that can promote our understanding of the predictors of schizophrenia-related public stigma. Because stigma is a multidimensional phenomenon, studies on schizophrenia-related stigma need to account for the origins of the various aspects of this phenomenon. This study explored which common sense model of illness components (cognitive and emotional) predicted three distinct indicators of stigma (stereotypes and discrimination on the individual and structural levels). A nonprobability sample of 149 students from one of the largest universities in Israel was drawn. Data were collected via a self-reported questionnaire. We found that five of the nine common sense model of illness variables predicted schizophrenia stereotypes, whereas very few predicted discrimination. Additionally, we found that greater belief in the effectiveness of schizophrenia treatment was associated with lower stereotypes and social-level discrimination. The less the perceived consequences of schizophrenia (the extent to which the illness is perceived to affect one's life), the lower the stereotypes and discrimination (individual and social). The perceived cause of schizophrenia and attribution of personal control over the illness did not predict any stigma dimension. The findings suggest that the common sense model of illness is more suitable for predicting stereotypes than discrimination. More research is needed to understand the unique drivers of different dimensions of stigma. The findings also imply the need to design separate programs to address different aspects of schizophrenia-related stigma.


Examinations of tRNA Range of Motion Using Simulations of Cryo-EM Microscopy and X-Ray Data.
We examined tRNA flexibility using a combination of steered and unbiased molecular dynamics simulations. Using Maxwell's demon algorithm, molecular dynamics was used to steer X-ray structure data toward that from an alternative state obtained from cryogenic-electron microscopy density maps. Thus, we were able to fit X-ray structures of tRNA onto cryogenic-electron microscopy density maps for hybrid states of tRNA. Additionally, we employed both Maxwell's demon molecular dynamics simulations and unbiased simulation methods to identify possible ribosome-tRNA contact areas where the ribosome may discriminate tRNAs during translation. Herein, we collected >500 ns of simulation data to assess the global range of motion for tRNAs. Biased simulations can be used to steer between known conformational stop points, while unbiased simulations allow for a general testing of conformational space previously unexplored. The unbiased molecular dynamics data describes the global conformational changes of tRNA on a sub-microsecond time scale for comparison with steered data. Additionally, the unbiased molecular dynamics data was used to identify putative contacts between tRNA and the ribosome during the accommodation step of translation. We found that the primary contact regions were H71 and H92 of the 50S subunit and ribosomal proteins L14 and L16.


Pre-treatment growth and IGF-I deficiency as main predictors of response to growth hormone therapy in neural models.
Mathematical models have been applied in prediction of growth hormone treatment effectiveness in children since the end of 1990s. Usually they were multiple linear regression models; however, there are also examples derived by empirical non-linear methods. Proposed solution consists in application of machine learning technique - artificial neural networks - to analyse this problem. This new methodology, contrary to previous ones, allows detection of both linear and non-linear dependencies without assuming their character a priori The aims of this work included: development of models predicting separately growth during 1st year of treatment and final height as well as identification of important predictors and in-depth analysis of their influence on treatment's effectiveness. The models were derived on the basis of clinical data of 272 patients treated for at least 1 year, 133 of whom have already attained final height. Starting from models containing 17 and 20 potential predictors, respectively for 1st year and final height model, we were able to reduce their number to 9 and 10. Basing on the final models, IGF-I concentration and earlier growth were indicated as belonging to most important predictors of response to GH therapy, while results of GH secretion tests were automatically excluded as insignificant. Moreover, majority of the dependencies were observed to be non-linear, thus using neural networks seems to be reasonable approach despite it being more complex than previously applied methods.


Protection against Plasmodium berghei yoelii in chloroquine- and pyrimethamine-treated mice.
The antimalarial drugs chloroquine and pyrimethamine were observed to afford protection to mice treated with these agents. This protection was observed in mice when given a subsequent challenge infection after they had been radically cured of P. b. yoelii infection.


Exceeding the Ordinary: A Framework for Examining Teams Across the Extremeness Continuum and Its Impact on Future Research.
Work teams increasingly face unprecedented challenges in volatile, uncertain, complex, and often ambiguous environments. In response, team researchers have begun to focus more on teams whose work revolves around mitigating risks in these dynamic environments. Some highly insightful contributions to team research and organizational studies have originated from investigating teams that face unconventional or extreme events. Despite this increased attention to extreme teams, however, a comprehensive theoretical framework is missing. We introduce such a framework that envisions team extremeness as a continuous, multidimensional variable consisting of environmental extremeness (i.e., external team context) and task extremeness (i.e., internal team context). The proposed framework allows every team to be placed on the team extremeness continuum, bridging the gap between literature on extreme and more traditional teams. Furthermore, we present six propositions addressing how team extremeness may interact with team processes, emergent states, and outcomes using core variables for team effectiveness and the well-established input-mediator-output-input model to structure our theorizing. Finally, we outline some potential directions for future research by elaborating on temporal considerations (i.e., patterns and trajectories), measurement approaches, and consideration of multilevel relationships involving team extremeness. We hope that our theoretical framework and theorizing can create a path forward, stimulating future research within the organizational team literature to further examine the impact of team extremeness on team dynamics and effectiveness.


Defining the Content for a Quality and Safety in Surgery Curriculum Using a Nominal Group Technique.
There is growing awareness of the need to provide surgical residents with training in quality and safety. Previous studies have revealed a need for a formal curriculum, but the content and structure of such a curriculum has not been defined. Our objective was to develop a surgery resident curriculum using a consensus, team-building approach. This study consisted of moderated, structured focus groups using a nominal group technique to guide discussion. Participants generated rank lists of topics to be included and answered questions regarding structure and design of teaching and assessment modalities. Two separate focus groups among 9 surgical residents and 10 faculty experts in quality and safety were held in conjunction with the American College of Surgeons Quality and Safety Conference in July 2017. A total of 16 institutions were represented. A total of 35 topics were initially proposed by the resident group and a total of 41 topics were proposed by the expert group. After discussion, each group reached consensus on a final list of 9 topics. Most topics in the final lists fell into the broad areas of improvement science and nontechnical skills. Residents indicated that most topics were, on average, poorly covered by their current training program, however, a wide range was noted within each topic. Faculty indicated a preference for didactic instructional methods and assessment using multiple-choice questions. Quality and safety are integral components of surgical training. Learners and experts agreed that topics within the domains of improvement science and nontechnical skills should be included in a formal curriculum. Learners reported wide variation on how well these topics are currently included in graduate medical education training programs.


Quantitative atlas of blood-brain barrier transporters, receptors, and tight junction proteins in rats and common marmoset.
The purpose of this study was to determine the protein amounts of blood-brain barrier (BBB) permeability-related transporters, receptors, and tight junction proteins in Sprague Dawley and Wistar rats and common marmoset, and also to investigate inter-species and inter-strain differences across rodents and primates. Quantification of target proteins in isolated brain capillaries was conducted by liquid chromatography-tandem mass spectrometry-based quantitative targeted absolute proteomics, with in silico peptide selection. Most target proteins showed inter-rodent, inter-primate species, and inter-rat strain differences of less than 2-fold. Comparison of rat and human BBB showed that P-glycoprotein, multidrug resistance-associated protein 4, monocarboxylate transporter 1, l-type amino acid transporter, and organic anion transporter 3 exhibited differences of more than two-fold in protein abundance, whereas the amounts of breast cancer resistance protein, glucose transporter 1, and insulin receptor were similar in rat and human. In contrast, the differences between marmoset and human BBB were less than 2-fold for almost all measured proteins. Thus, the molecular basis of BBB functions may be similar in marmoset and human, whereas that of rats shows significant differences. The marmoset may be a good model to access in vivo human BBB permeability characteristics, as an alternative to rat and macaque monkey.


The Psychological Consequences of Disability over the Life Course: Assessing the Mediating Role of Perceived Interpersonal Discrimination.
We examine whether perceived interpersonal discrimination mediates the association between disability and psychological well-being (depression, negative and positive affect) and how these processes differ across the life course. Data are from two waves (2004-2006; 2013-2014) of the Midlife in the United States (MIDUS; N = 2,503). Perceived discrimination accounts for 5% to 8% of the association between disability and the three mental health outcomes. Moderated mediation analyses reveal significant age differences; perceived discrimination is a stronger explanatory mechanism among midlife (ages 40-64) relative to older (age 65+) adults. Disability stigma takes a heightened psychological toll at midlife, a life stage when adults are expected to be able-bodied and interact with a diverse social network, which may be a source of interpersonal mistreatment. Among older adults, for whom impairment is expected and common, the psychological impact of disability may operate through other pathways. We discuss implications for research and practice.


[The un-healing cause of its management after operation of thoracolumbar tuberculosis].
To analyze the un-healing cause and management after operation of thoracolumbar tuberculosis. From January 2008 to December 2011, the data of 12 patients with postoperative un-healing thoracolumbar tuberculosis were retrospectively analyzed. There were 5 males and 7 females, with an average age of 51.3 years old (ranged, 42 to 65). All the patients occurred different degree of vertebral destruction, abscess-formation, kyphosis and nerve functional injury at the first operation. Among them, 11 patients underwent debridement and fusion with autogenous iliac bone grafting and internal fixation, 1 patient underwent debridement and fusion with autogenous iliac bone grafting. At 2-6 months after operation, internal fixation loosening, fusion failure, abscess-ormation, or erythrocyte sedimentation rate increasing occurred in the patients. For the patients to adjust anti-tuberculosis scheme, 2 patients were treated with puncture to multiple abscess combining with rifampicin local injection; 10 patients were reoperated with debridement and internal fixation adjusting. At 1-2.5 years (mean 1.8 years) after follow-up, 9 cases were cured finally. Re-admission had 3 cases because of re-occurred erythrocyte sedimentation rate increasing and abscess-formation at the 2-4 months after operation, whose bacterial culture showed more than 2 kinds of drug fast, and the treatment effect was still not ideal after adjusting anti-tuberculosis scheme. Insufficient anti-tuberculosis and bad nutritional status before operation,incomplete debridement and unreasonable fixation style during operation, inadequate drainage, irregular anti-tuberculosis and drug fast of Bacillus tuberculosis after operation are main reasons leading to un-healing of tuberculosis. Therefore, actively improving malnutrition, making individual operation plan before operation, ensuring complete debridement, rebuilding spinal stability, removing the compression of spinal cord, combining with postoperative effective, enough, regular combination chemotherapy are important guarantee to cure of spinal tuberculosis.


Biosynthesis of thiamin in Bacillus subtilis. Isolation of mutants accumulating 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate.
Thiamin-deficient mutants of Bacillus subtilis were characterized by their growth responses to the pyrimidine and thiazole moieties of the vitamin molecule and by cross-feeding tests. All mutants growing on the thiazole moiety and all mutants with an absolute requirement for thiamin fed all those growing on the pyrimidine moiety. No other cross-feeding effects were observed. From the culture fluid of a mutant growing on the thiazole moiety, two compounds were isolated which supported growth of mutants requiring the pyrimidine moiety. These compounds were identified by chromatographic, bioautographic and spectrophotometric procedures as 4-amino-5-hydroxymethyl-2-methylpyrimidine and its monophosphate derivative.


An automatic tool to analyze and cluster macromolecular conformations based on self-organizing maps.
Sampling the conformational space of biological macromolecules generates large sets of data with considerable complexity. Data-mining techniques, such as clustering, can extract meaningful information. Among them, the self-organizing maps (SOMs) algorithm has shown great promise; in particular since its computation time rises only linearly with the size of the data set. Whereas SOMs are generally used with few neurons, we investigate here their behavior with large numbers of neurons. We present here a python library implementing the full SOM analysis workflow. Large SOMs can readily be applied on heavy data sets. Coupled with visualization tools they have very interesting properties. Descriptors for each conformation of a trajectory are calculated and mapped onto a 3D landscape, the U-matrix, reporting the distance between neighboring neurons. To delineate clusters, we developed the flooding algorithm, which hierarchically identifies local basins of the U-matrix from the global minimum to the maximum. The python implementation of the SOM library is freely available on github: https://github.com/bougui505/SOM. email@example.com or firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


The CLV3 Homolog in Setaria viridis Selectively Controls Inflorescence Meristem Size.
The CLAVATA pathway controls meristem size during inflorescence development in both eudicots and grasses, and is initiated by peptide ligands encoded by CLV3/ESR-related (CLE) genes. While CLV3 controls all shoot meristems in Arabidopsis, evidence from cereal grasses indicates that different meristem types are regulated by different CLE peptides. The rice peptide FON2 primarily controls the size of the floral meristem, whereas the orthologous peptides CLE7 and CLE14 in maize have their most dramatic effects on inflorescence and branch meristems, hinting at diversification among CLE responses in the grasses. Setaria viridis is more closely related to maize than to rice, so can be used to test whether the maize CLE network can be generalized to all members of subfamily Panicoideae. We used CRISPR-Cas9 in S. viridis to knock out the SvFON2 gene, the closest homolog to CLV3 and FON2. Svfon2 mutants developed larger inflorescence meristems, as in maize, but had normal floral meristems, unlike Osfon2, suggesting a panicoid-specific CLE network. Vegetative traits such as plant height, tiller number and leaf number were not significantly different between mutant and wild type plants, but time to heading was shorter in the mutants. In situ hybridization showed strong expression of Svfon2 in the inflorescence and branch meristems, consistent with the mutant phenotype. Using bioinformatic analysis, we predicted the co-expression network of SvFON2 and its signaling components, which included genes known to control inflorescence architecture in maize as well as genes of unknown function. The similarity between SvFON2 function in Setaria and maize suggests that its developmental specialization in inflorescence meristem control may be shared among panicoid grasses.


Online recruitment of youth for mHealth studies.
Social networking sites and apps have emerged as an opportunity to engage in research young men who have sex with men (YMSM) at risk of HIV infection who may not be otherwise reached by in-person recruitment efforts. This paper highlights lessons learned, best practices and on-going recruitment challenges in the iTech network of the NIH Adolescent Trials Network for HIV Interventions. Recruitment was conducted for four randomized controlled trials (RCTs) of mHealth HIV prevention interventions for YMSM living in 10 US cities. Advertising was purchased on Facebook, Instagram, Snapchat, Twitter and Grindr. Users who clicked on banner ads were taken directly to a study-specific eligibility screener and if eligible, were asked to provide contact information for follow-up by respective study site staff. Ad and screening metrics (impressions, clicks, cost per click (CPC), click-through rate (CTR), number screened, number eligible, number who provided contact information and cost per eligible contact) were compared across platforms, studies and geographic areas (where available). Screening metrics were also calculated for in-person recruitment efforts. Grindr and Snapchat ads produced the highest CTRs as compared to Facebook ads. However, these ads had the lowest proportions of users who initiated eligibility screeners and ultimately Facebook ads yielded the lowest cost per eligible contact across studies. Instagram ads yielded the highest proportions of eligible contacts who were racial/ethnic minorities and under the age of 18. Geographic variability in cost per eligible contact was observed for studies with identical eligibility criteria running concurrently in different regions, driven by both advertising costs and the screening and eligibility rates. Despite lower eligibility rates, the total numbers of eligible contacts were higher for online advertising campaigns, as compared to other recruitment efforts, for all studies except P3. Ads recruiting for P3 had the highest cost per eligible contact, likely due to this study having the most stringent eligibility criteria of the studies described. We implemented a successful online advertising strategy to recruit YMSM at high risk for HIV infection into four RCTs of mHealth interventions. This report provides a framework for evaluation of data from future online recruitment efforts across platforms and geographic areas, regardless of inevitable changes in the digital marketing space.


Disturbance of the bacterial cell wall specifically interferes with biofilm formation.
In nature, bacteria communicate via chemical cues and establish complex communities referred to as biofilms, wherein cells are held together by an extracellular matrix. Much research is focusing on small molecules that manipulate and prevent biofilm assembly by modifying cellular signalling pathways. However, the bacterial cell envelope, presenting the interface between bacterial cells and their surroundings, is largely overlooked. In our study, we identified specific targets within the biosynthesis pathways of the different cell wall components (peptidoglycan, wall teichoic acids and teichuronic acids) hampering biofilm formation and the anchoring of the extracellular matrix with a minimal effect on planktonic growth. In addition, we provide convincing evidence that biofilm hampering by transglycosylation inhibitors and D-Leucine triggers a highly specific response without changing the overall protein levels within the biofilm cells or the overall levels of the extracellular matrix components. The presented results emphasize the central role of the Gram-positive cell wall in biofilm development, resistance and sustainment.


[LONI & Co: about the epistemic specificity of digital spaces of knowledge in cognitive neuroscience].
In the neurosciences digital databases more and more are becoming important tools of data rendering and distributing. This development is due to the growing impact of imaging based trial design in cognitive neuroscience, including morphological as much as functional imaging technologies. As the case of the 'Laboratory of Neuro Imaging' (LONI) is showing, databases are attributed a specific epistemological power: Since the 1990s databasing is seen to foster the integration of neuroscientific data, although local regimes of data production, -manipulation and--interpretation are also challenging this development. Databasing in the neurosciences goes along with the introduction of new structures of integrating local data, hence establishing digital spaces of knowledge (epistemic spaces): At this stage, inherent norms of digital databases are affecting regimes of imaging-based trial design, for example clinical research into Alzheimer's disease.


[Hepatocyte growth factor therapy for amyotrophic lateral sclerosis].
Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder characterized by the death of upper and lower motor neurons. Approximately 20% of familial ALS cases are caused by mutations in the superoxide dismutase 1 (SOD1) gene. We generated rats that express a human SOD1 transgene with two different ALS-associated mutations and found that these rats develop remarkable motor neuron degeneration and paralysis. This rat model, because of the larger size of the animals as compared to ALS-affected mice, will facilitate studies involving manipulation of the cerebrospinal fluid (CSF) (e.g., implantation of intrathecal catheters for chronic therapeutic studies; CSF sampling) or spinal cord (e.g., direct administration of viral- and cell-mediated therapies). The hepatocyte growth factor (HGF) is one of the most potent survival-promoting factors for motor neurons. To examine its protective effect on motor neurons and its therapeutic potential, we administered human recombinant HGF (hrHGF) to the transgenic rats, by continuous intrathecal delivery, for 4 weeks from the onset of paralysis. Intrathecal administration of hrHGF attenuated motor neuron degeneration and prolonged the duration of the disease 62.7% compared with the contrast group. Our results indicated the therapeutic efficacy of continuous intrathecal administration of hrHGF in ALS rats. To explore the potential use of this treatment strategy in humans, we induced a contusive cervical spinal cord injury in the common marmoset, a primate, and then administered hrHGF intrathecally. The intrathecal administration of hrHGF promoted functional recovery. These projects have been supported by the "Super Special Consortium for Supporting the Development of Cutting-edge Medical Care" (tokku), a special program organized by the Cabinet Office of the Japanese government (research representative: Hideyuki Okano, M.D., Ph.D., Professor at Keio University).