Illegal behaviors in problem gambling: analysis of data from a gambling helpline.
Problem gambling behaviors, particularly the most severe form, which is pathological gambling (PG), represent an emerging public health problem. Compared with the general population, individuals with problem gambling are more likely to have reports of legal issues, including commission of crimes, arrest, and incarceration. The goal of the present study is to examine the characteristics of individuals seeking help for gambling problems with regard to reports of illegal behavior secondary to gambling. Individuals with gambling problems were identified through use of a 24-hour gambling helpline, and information regarding the identified problem gambler was investigated with respect to reported presence or absence of gambling-related illegal behaviors. Identified gamblers with reported gambling-related illegal behaviors compared with those without such behaviors appeared to experience more severe gambling-related problems. Despite being on average younger, gamblers with acknowledged gambling-related illegal behaviors were more likely to have reports of having problems with multiple forms of gambling, debts to acquaintances, been suicidal secondary to gambling, used alcohol or drugs excessively, and received mental health treatment. Secondary analyses of the subgroup of gamblers with gambling-related illegal behaviors revealed that those with reports of arrest or incarceration secondary to gambling compared with those with gambling-related illegal behaviors but without arrest or incarceration secondary to gambling were more likely to have features similar to those described for individuals with antisocial personality disorder (ASPD). That is, the gambler with reported arrest or incarceration secondary to gambling was more likely to be male, unemployed, single, and have reports of problems with excessive drug or alcohol use. In contrast, the gamblers acknowledging gambling-related illegal behaviors but not arrest or incarceration secondary to gambling were predominantly female and more likely to have reports of problems with non-strategic forms of gambling (e.g., slot machine), owing money to legitimate sources of borrowing, having filed for bankruptcy, and having family problems related to gambling. The findings indicate: (1) individuals with reported legal problems secondary to gambling represent a more ill subpopulation of problem gamblers; and (2) there exist separate subgroups of gamblers with gambling-related illegal behaviors (i.e., those with or without reported arrest or incarceration secondary to gambling) with strikingly different characteristics and possibly different treatment needs. The results of the present study highlight the importance of the identification and treatment of individuals with gambling problems with respect to legal issues.


Genetic variation of the GLUT10 glucose transporter (SLC2A10) and relationships to type 2 diabetes and intermediary traits.
The SLC2A10 gene encodes the GLUT10 facilitative glucose transporter, which is expressed in high amounts in liver and pancreas. The gene is mapped to chromosome 20q12-q13.1, a region that has been shown to be linked to type 2 diabetes. The gene was examined in 61 Danish type 2 diabetic patients, and a total of six variants (-27C-->T, Ala206Thr, Ala272Ala, IVS2 + 10G-->A, IVS4 + 18T-->G, and IVS4 + 26G-->A) were identified and investigated in an association study, which included 503 type 2 diabetic patients and 510 glucose-tolerant control subjects. None of the variants were associated with type 2 diabetes. Interestingly, carriers of the codon 206 Thr allele had 18% lower fasting serum insulin levels (P = 0.002) and 20% lower insulinogenic index (P = 0.03) than homozygous carriers of the Ala allele. These results suggest that variation in the coding region of SLC2A10 does not contribute substantially to the pathogenesis of type 2 diabetes in the examined study population. However, the codon 206 polymorphism may be related to the interindividual variation in fasting and oral glucose-induced serum insulin levels.


Therapeutic Benefits from Nanoparticles: The Potential Significance of Nanoscience in Retinal Degenerative Diseases.
Several nanotechnology podiums have gained remarkable attention in the area of medical sciences, including diagnostics and treatment. In the past decade, engineered multifunctional nanoparticles have served as drug and gene carriers. The most important aspect of translating nanoparticles from the bench to bedside is safety. These nanoparticles should not elicit any immune response and should not be toxic to humans or the environment. Lipid-based nanoparticles have been shown to be the least toxic for in vivo applications, and significant progress has been made in gene and drug delivery employing lipid-based nanoassemblies. Several excellent reviews and reports discuss the general use and application of lipid-based nanoparticles; our review focuses on the application of lipid-based nanoparticles for the treatment of ocular diseases, and recent advances in and updates on their use.


Aboriginal youth's perceptions of traditional and commercial tobacco in Canada.
Although tobacco has played an important role in Aboriginal culture for millennia, its more recent recreational use has resulted in serious health concerns among Aboriginal users. In Canada, First Nations youth have higher smoking rates than non-Aboriginal youth. The goals of this study were to examine Aboriginal youth's perceptions of traditional and commercial tobacco and to evaluate perception changes following workshops on traditional uses of tobacco by Elders and other community members, native plants used in ceremony and health impacts of cigarette smoking. The twenty-five Aboriginal youth participants ranged in age from 9 to 14 years old. Results show that workshops did not alter participants' knowledge about tobacco and ceremonies but those studied were more effective at articulating their knowledge through open-ended questionnaires after the workshops. These participants were able to identify more traditional tobacco uses and plants that could be used in ceremonies instead of commercial tobacco. Culturally appropriate education about traditional and commercial tobacco use may be effective in limiting its misuse by Aboriginal youth.


First siRNA library screening in hard-to-transfect HUVEC cells.
Meaningful RNAi-based data for target gene identification are strongly dependent on the use of a biologically relevant cell type and efficient delivery of highly functional siRNA reagents into the selected cell type. Here we report the use of the Amaxa(R) Nucleofector(R) 96-well Shuttle(R) System for siRNA screening in primary cells. Lonza's Clonetics(R) HUVEC-Human Umbilical Vein Endothelial Cells were transfected with Thermo Scientific Dharmacon siGENOME(R) siRNA Libraries targeting protein kinases and cell cycle related genes and screened for genes important for cell viability. Of the 37 primary hits, down-regulation of 33 led to reduced proliferation or increased cell death, while down-regulation of two allowed for better cell viability. The validated four genes out of the 16 strongest primary hits (COPB2, PYCS, CDK4 and MYC) influenced cell proliferation to varying degrees, reflecting differing importance for survival of HUVEC cells. Our results demonstrate that the Nucleofector(R) 96-well Shuttle(R) System allows the delivery of siRNA libraries in cell types previously considered to be difficult to transfect. Thus, identification and validation of gene targets can now be conducted in primary cells, as the selection of cell types is not limited to those accessible by lipid-mediated transfection.


Novel role for prostaglandin E2 in fish hepatocytes: regulation of glucose metabolism.
Prostaglandin E(2) (PGE(2)) potently activated glycogenolysis and gluconeogenesis in isolated rockfish (Sebastes caurinus) hepatocytes. The average degree of activation for glycogenolysis was 6.4+/-0.67-fold (mean+/-S.E.M.; n=37), and could be as much as 19-fold. Analysis of dose-concentration relationships between glycogenolytic actions and PGE(2) concentrations yielded an EC(50) around 120 nM in hepatocyte suspensions and 2 nM for hepatocytes immobilized on perifusion columns. For the activation of gluconeogenesis (1.74+/-0.14-fold; n=10), the EC(50) for suspensions was 60 nM. Intracellular targets for PGE(2) actions are adenylyl cyclase, protein kinase A and glycogen phosphorylase. Concentrations of cAMP increased with increasing concentrations of PGE(2), and peaked within 2 min of hormone application. In the presence of the phosphodiesterase inhibitor, isobutyl-3-methylxanthine, peak height was increased and peak duration extended. The protein kinase A inhibitor, Rp-cAMPS, counteracted the activation of glycogenolysis by PGE(2), implying that the adenylyl cyclase/protein kinase A pathway is the most important, if not exclusive, route of message transduction. PGE(2) activated plasma membrane adenylyl cyclase and hepatocyte glycogen phosphorylase in a dose-dependent manner. The effects were specific for PGE(2); smaller degrees of activation of glycogenolysis were noted for PGE(1), 11-deoxy PGE(1), 19-R-hydroxy-PGE(2), and prostaglandins of the A, B and Falpha-series. The selective EP(2)-receptor agonist, butaprost, was as effective as PGE(2), suggesting that rockfish liver contains prostaglandin receptors pharmacologically related to the EP(2) receptors of non-hepatic tissues of mammals. Rockfish hepatocytes quickly degraded added PGE(2) (t((1/2))=17-26 min). A similar ability to degrade PGE(2) has been noted in catfish (Ameiurus nebulosus) hepatocytes, but no glycogenolytic or gluconeogenic actions of the hormone are noted for this species. We conclude that PGE(2) is an important metabolic hormone in fish liver, with cAMP-mediated actions on glycogen and glucose metabolism, and probably other pathways regulated by cAMP and protein kinase A. The constant presence of EP(2)-like receptors is a unique feature of the fish liver, with interesting implications for function and evolution of prostaglandin receptors in vertebrates.


Protection against MPTP treatment by an analog of Pro-Leu-Gly-NH2 (PLG, MIF-1)
3(R)-[(2(S)-Pyrrolidinyl-carbonyl)amino]-2-oxo-1-pyrrolidineacetamide (PAOPA) is a peptidomimetic analog of Pro-Leu-Gly-NH2 (PLG or MIF-1) that has previously been demonstrated to be more potent and efficacious that MIF-1 in enhancing dopamine receptor activity. Given the ability of MIF-1 to protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) lesioning in C57 BL/6 mice, the present study was undertaken to evaluate the neuroprotective effect of PAOPA in this model. PAOPA was found to be more potent and efficacious that MIF-1 in sparing dopamine and its metabolite levels following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine administration. Whether the enhanced neuroprotective effect of PAOPA is due to dopamine receptor stimulation, or a result of reduced oxidative stress through normalization of dopamine turnover, remains to be determined.


Endothelial injuries of coronary arteries distal to thrombotic sites: role of adhesive interaction between endothelial P-selectin and leukocyte sialyl LewisX.
Intracoronary thrombus formation is associated with epicardial vasoconstriction distal to the thrombotic site. To investigate the mechanisms of abnormal vasomotor function of the artery distal to the thrombotic site, we studied coronary vessels in dogs with cyclic flow variations (CFVs) of the left anterior descending coronary artery (LAD) stenosis with endothelial injury. Coronary rings isolated from the LAD (proximal, stenotic, and distal sites) and control circumflex coronary arteries were tested for responsiveness to endothelium-dependent (acetylcholine and A23187) and endothelium-independent vasodilators (NaNO2). Endothelium-independent relaxation was intact in all 4 sites. Endothelium-dependent relaxation was intact in the control and proximal sites and impaired in the stenotic sites. Relaxations not only to acetylcholine and A23187 but also to serotonin, ADP, and thrombin were impaired in the distal sites after observing CFVs for 80 minutes. Electron microscopy revealed the loss of endothelial integrity with leukocyte adherence to the endothelium in the distal sites. Immunohistochemical expression of P-selectin on the endothelial cells was more upregulated in the distal site than in the proximal site, and P-selectin mRNA expression was significantly greater in the ischemic region distal to the thrombotic site than in the proximal nonischemic region. PB1.3, a neutralizing monoclonal antibody against P-selectin, and sialyl LewisX (SLeX)-containing oligosaccharide SLeX, a carbohydrate analogue of selectin ligand, preserved endothelial function without affecting CFVs. SLeX-containing oligosaccharide preserved endothelial integrity of the distal site and inhibited P-selectin expression of the distal site. Thus, the adhesive interaction between endothelial P-selectin and leukocyte SLeX may play an important role in endothelial injuries of the coronary artery distal to the thrombotic site.


Proteomic profiling and post-translational modifications in human keratinocytes treated with Mucuna pruriens leaf extract.
Mucuna pruriens (Mp) is a plant belonging to the Fabaceae family, with several medicinal properties among which its potential to treat diseases where reactive oxygen species (ROS) play an important role in the pathogeneses. The aim was to investigate the effects of Mp leaf methanolic extract (MPME) on human keratinocytes protein expression and its role in preventing proteins oxidation after oxidative stress (OS) exposure. The effects of MPME on HaCaT cells protein expression were evaluated treating cells with different concentrations of MPME, with glucose oxidase (GO, source of OS) and with MPME subsequently treated with GO. The protein patterns of treated HaCaT cells are analyzed by two-dimensional gel electrophoresis (2-DE) and compared with that of untreated HaCaT. Immunoblotting was then used to evaluate the role of MPME in preventing the 4-hydroxynonenal protein adducts (4-HNE PAs) formation (marker of OS). Eighteen proteins, identified by mass spectrometry (LC-ESI-CID-MS/MS), were modulated distinctly by MPME in HaCaT. Overall, MPME counteract GO effect, reducing the GO-induced overexpression of several proteins involved in stress response (T-complex protein 1, Protein disulfide-isomerase A3, Protein DJ-1, and Stress-induced-phosphoprotein 1), in cell energy methabolism (Inorganic pyrophosphatase, Triosephosphate isomerase isoform 1, 2-phosphopyruvate-hydratase alpha-enolase, and Fructose-bisphosphate aldolase A isoform 1), in cytoskeletal organization (Cytokeratins 18, 9, 2, Cofilin-1, Annexin A2 and F-actin-capping protein subunit beta isoform 1) and in cell cycle progression (Eukaryotic translation initiation factor 5A-1 isoform B). In addition, MPME decreased the 4-HNE PAs levels, in particular on 2-phosphopyruvate-hydratase alpha-enolase and Cytokeratin 9. Our findings show that MPME might be helpful in the treatment of OS-related skin diseases by preventing protein post-translational modifications (4-HNE PAs).


Antibiotics--their administration, choice and prophylactic use.
A wider range of antibiotics is now available with the advent of two new cephalosporins--cefamandole and cefuroxime, and the first cephamycin--cefoxitin. These drugs may be particularly useful in the treatment of septicaemia, and when combined with ticarcillin are effective against nearly all pathogens. Cefuroxime can be given intramuscularly. Intravenous cotrimoxazole must be given in large doses to achieve therapeutic concentrations in the cerebrospinal fluid. The correct use of antibiotics for prophylaxis demands an accurate knowledge of the organisms likely to cause infection and the most efficient way of using the antibiotics to prevent that infection. There is little argument about the need for prophylaxis to prevent recurrent rheumatic heart disease or tetanus from a contaminated wound. However, in the various areas of surgery a wide range of antibiotics is used not always as a result of controlled trials having proved their value. Non-surgical situations for prophylaxis are briefly reviewed.


[The perinatal transmission of the hepatitis B virus in the Paris area].
HBsAg was detected in 152 pregnant women among 6,605 (2.3%) screened in the prenatal clinics of four hospitals representative of the Paris metropolitan area. In 98% of cases, HBsAg positivity indicated chronic HBV carrier status. Among patients born out of continental France (47% of screened women, 79% of positive women) relative risk of chronic infection was 6 in Asians, 5.5 in Africans, and 4 in French women born in non-continental France. No significant difference in medical history was seen between HBsAg-positive and HBsAg-negative patients, in any of the birthplace groups. In women born out of continental France, number of children and crowding in the home were correlated with HBsAg-positivity; these correlations were not found in French women born in continental France. In non-African, non-Asian women, screening on the basis of medical, social and familial criteria (simulated in this study) would not be effective. Routine screening for HBsAg in pregnancy is advocated. The cost of the prevention of each case of perinatally acquired chronic HBV infection by routine screening followed by prophylactic treatment of a risk neonates was estimated at 180,000 French Francs (35,000 dollars). This approach is the only means of preventing the long-term life-threatening complications of chronic HBV infection in the 600 neonates born each year in France to HBsAg-positive mothers.


The association of visual field deficits and visuo-spatial neglect in acute right-hemisphere stroke patients.
Visuo-spatial neglect (VSN) after stroke is associated with a poor prognosis for rehabilitation. The co-existence of a visual field deficit (VFD) with VSN may be associated with impaired visuo-spatial functioning and thereby poor functional outcome. To determine whether the presence of a VFD (i) exacerbates VSN and (ii) influences recovery of VSN. A prospective study of consecutive acute (<7 days), right-hemisphere stroke patients who were able to undergo detailed assessment of visuo-spatial functioning and visual fields. Clinical assessment and a standardized neuropsychological test was administered by one observer, followed by independent assessment of visual fields by a second observer. Patients were followed up for 12 weeks with 4-weekly re-assessments. 44 consecutive patients (23 women) with a first in a lifetime, acute hemisphere stroke were recruited. Twenty had VSN and VFD, seven VSN only, one VFD only and 17 had normal visual fields and no neglect. The finding of a VFD was significantly associated with the presence of VSN (P<0.0001). Patients with both VFD and VSN had a significantly lower score on the behavioural inattention test. One month post-stroke, this difference was no longer significant. Recovery of VSN and VFD was maximal in the first month, however VSN recovery continued for up to 12 weeks. Patients with VSN and a VFD on admission had a greater mortality at 1 and 3 months. The presence of a VFD does appear to exacerbate neglect in the acute stroke patient; this effect is no longer seen after 1 month. Recovery of VSN continues independent of a VFD. Patients with neglect and a VFD have an increased mortality, probably because of greater neurological impairment.


Prediction of viral filtration performance of monoclonal antibodies based on biophysical properties of feed.
Controlling viral contamination is an important issue in the process development of monoclonal antibodies (MAbs) produced from mammalian cell lines. Virus filtration (VF) has been demonstrated to be a robust and effective clearance step which can provide ≥4 logs of reduction via size exclusion. The minimization of VF area by increasing flux and filter loading is critical to achieving cost targets as VFs are single use and often represent up to 10% of total purification costs. The research presented in this publication describes a development strategy focused on biophysical attributes of product streams that are directly applicable to VF process performance. This article summarizes a case study where biophysical tools (high-pressure size exclusion chromatography, dynamic light scattering, and absolute size exclusion chromatography) were applied to a specific MAb program to illustrate how changes in feed composition (pH, sodium chloride concentration, and buffer salt type) can change biophysical properties which correlate with VF performance. The approach was subsequently refined and expanded over the course of development of three MAbs where performance metrics (i.e., loading and flux) were evaluated for two specific virus filters (Viresolve Pro and Planova 20N) during both unspiked control runs and virus clearance experiments. The analyses of feed attributes can be applied to a decision tree to guide the recommendation of a VF filter and operating conditions for use in future MAb program development. The understanding of the biophysical properties of the feed can be correlated to virus filter performance to significantly reduce the mass of product, time, and costs associated with virus filter step development.


Forgiving, fast and slow: validity of the implicit association test for predicting differential response latencies in a transgression-recall paradigm.
This study examined the role of automaticity in forgiving a real-life offense. As an alternative to self-report, an Implicit Association Test (IAT) of forgiveness was developed. Implicit (IAT-measured) and explicit (self-reported) forgiveness predicted shorter response times of state forgiveness ratings. The forgiveness IAT was highly reliable, moderately stable over time, and demonstrated incremental validity. Results suggest that the newly introduced forgiveness IAT could advance personality research beyond what is known from self-report measures, further corroborating the notion of implicit forgiveness. Implications for personality assessment are discussed.


Digitized holography: modern holography for 3D imaging of virtual and real objects.
Recent developments in computer algorithms, image sensors, and microfabrication technologies make it possible to digitize the whole process of classical holography. This technique, referred to as digitized holography, allows us to create fine spatial three-dimensional (3D) images composed of virtual and real objects. In the technique, the wave field of real objects is captured in a wide area and at very high resolution using the technique of synthetic aperture digital holography. The captured field is incorporated in virtual 3D scenes including two-dimensional digital images and 3D polygon mesh objects. The synthetic field is optically reconstructed using the technique of computer-generated holograms. The reconstructed 3D images present all depth cues like classical holograms but are digitally editable, archivable, and transmittable unlike classical holograms. The synthetic hologram printed by a laser lithography system has a wide viewing zone in full-parallax and give viewers a strong sensation of depth, which has never been achieved by conventional 3D systems. A real hologram as well as the details of the technique is presented to verify the proposed technique.


MIR@NT@N: a framework integrating transcription factors, microRNAs and their targets to identify sub-network motifs in a meta-regulation network model.
To understand biological processes and diseases, it is crucial to unravel the concerted interplay of transcription factors (TFs), microRNAs (miRNAs) and their targets within regulatory networks and fundamental sub-networks. An integrative computational resource generating a comprehensive view of these regulatory molecular interactions at a genome-wide scale would be of great interest to biologists, but is not available to date. To identify and analyze molecular interaction networks, we developed MIR@NT@N, an integrative approach based on a meta-regulation network model and a large-scale database. MIR@NT@N uses a graph-based approach to predict novel molecular actors across multiple regulatory processes (i.e. TFs acting on protein-coding or miRNA genes, or miRNAs acting on messenger RNAs). Exploiting these predictions, the user can generate networks and further analyze them to identify sub-networks, including motifs such as feedback and feedforward loops (FBL and FFL). In addition, networks can be built from lists of molecular actors with an a priori role in a given biological process to predict novel and unanticipated interactions. Analyses can be contextualized and filtered by integrating additional information such as microarray expression data. All results, including generated graphs, can be visualized, saved and exported into various formats. MIR@NT@N performances have been evaluated using published data and then applied to the regulatory program underlying epithelium to mesenchyme transition (EMT), an evolutionary-conserved process which is implicated in embryonic development and disease. MIR@NT@N is an effective computational approach to identify novel molecular regulations and to predict gene regulatory networks and sub-networks including conserved motifs within a given biological context. Taking advantage of the M@IA environment, MIR@NT@N is a user-friendly web resource freely available at http://mironton.uni.lu which will be updated on a regular basis.


The Role of Autophagy in Type 2 Diabetic Kidney Disease Management.
Diabetic kidney disease (DKD), or diabetic nephropathy (DN), is one of the most prevalent complications of type 2 diabetes mellitus (T2DM) and causes severe burden on the general welfare of T2DM patients around the world. While several new agents have shown promise in treating this condition and potentially halting the progression of the disease, more work is needed to understand the complex regulatory network involved in the disorder. Recent studies have provided new insights into the connection between autophagy, a physiological metabolic process known to maintain cellular homeostasis, and the pathophysiological pathways of DKD. Typically, autophagic activity plays a role in DKD progression mainly by promoting an inflammatory response to tissue damage, while both overactivated and downregulated autophagy worsen disease outcomes in different stages of DKD. This correlation demonstrates the potential of autophagy as a novel therapeutic target for the disease, and also highlights new possibilities for utilizing already available DN-related medications. In this review, we summarize findings on the relationship between autophagy and DKD, and the impact of these results on clinical management strategies.


[Modeling hyperparathyroid osteodystrophy in animals].
For the investigation of roentgenologic signs of osteodystrophy, especially of their early signs and symptoms, an animal model is proposed. The roentgenologic, micro-roentgenographic and pathomorphologic skeletal image of the disease can be studied in this model over all stages, which is useful for the understanding of pathologic transformations of bone structure in hyperparathyroid osteodystrophy. Especially micro-roentgenography is able to visualize changes of bone architecture in the early stages of the disease.


Differing perceptions of quality of life in patients with prostate cancer and their doctors.
As the number of prostate cancer survivors increases, urologists must recognize their quality of life impairment. In the past physician ratings of patient symptoms did not correlate with patient self-assessments. We determined if urologists have improved their reporting of patient health related quality of life. We also investigated if urologists assessed health related quality of life more accurately in the short or long term. We identified 1,366 men from CaPSURE™, a national, prospective cohort, who had undergone prostatectomy, brachytherapy or external beam radiation therapy. At each visit urologists assessed fatigue, pain, and sexual, urinary and bowel dysfunction. Participants independently completed the SF-36™ and the UCLA-PCI. We contrasted the frequency of impairment reported by physicians and participants in select health related quality of life domains in the short (less than 1 year) and long (greater than 2 years) term. We also compared physician-patient concordance between the periods 1995 to 2000 and 2001 to 2007. In short-term and long-term followup, and for the 1995 to 2000 and 2001 to 2007 cohorts, physician and participant assessments differed in all analyzed domains. Urologists noted impairment in urinary and sexual function more often than fatigue or pain. Disagreement between physician and participant ratings did not vary dramatically from short-term to long-term followup, or from the earlier to the later cohort. In men treated for localized prostate cancer physician ratings of symptoms do not correlate well with patient self-assessments of health related quality of life. Physician reporting did not improve over time. It is increasingly important to recognize and address impairments in quality of life from prostate cancer and its treatment.


Nascent craft specialization in the Pre-Pottery Neolithic A? Bead making at Shubayqa 6 (northeast Jordan).
The emergence of craft specialisation is a key area of interest for archaeologists investigating the socio-economic history and development of past societies. In southwest Asia, as elsewhere, the origins of craft specialisation have been associated with the emergence of surplus food production, households and social stratification. We present evidence for nascent skilled production of green stone beads at the Pre-Pottery Neolithic A (PPNA) site Shubayqa 6, northeast Jordan. Thousands of pieces of debitage, roughouts and finished beads exhibit signs of standardised production that was probably geared towards exchange. This hints towards incipient skilled craft production that was likely part-time and seasonal. We therefore argue that the appearance of specialist artisans in this autonomous and non-hierarchical society has no correlation with surplus food production, households, or social stratification.


Discovery and Computational Rationalization of Diminishing Alternation in [n]Dendralenes.
The [n]dendralenes are a family of acyclic hydrocarbons which, by virtue of their ability to rapidly generate structural complexity, have attracted significant recent synthetic attention. [3]Dendralene through [8]dendralene have been previously prepared but no higher member of the family has been reported to date. Here, we describe the first chemical syntheses of the "higher" dendralenes, [9]dendralene through [12]dendralene. We also report a detailed investigation into the spectroscopic properties and chemical reactivity of the complete family of fundamental hydrocarbons, [3]dendralene to [12]dendralene. These studies reveal the first case of diminishing alternation in behavior in a series of related structures. We also report a comprehensive series of computational studies, which trace this dampening oscillatory effect in both spectroscopic measurements and chemical reactivity to conformational preferences.


Temporal variations and bioaccumulation of heavy metals in different Suaeda salsa marshes of the Yellow River estuary, China.
To understand the temporal variations and bioaccumulation of heavy metals in the coastal marshes, the concentrations of heavy metals (Cr, Ni, Pb, and Cu) in the two Suaeda salsa marshes [middle S. salsa marsh (MM) and low S. salsa marsh (LM)] of the Yellow River estuary were determined from May to November in 2008 by in situ sampling and inductively coupled plasma mass spectrometry (ICP-MS) analysis. Results showed that heavy metal concentrations in S. salsa of MM and LM were generally in the order of Cu > Cr > Pb > Ni, while those in sediments fell in the order of Cr > Ni > Cu > Pb. Heavy metal concentrations of S. salsa in MM and LM were different, and significant differences were observed in stems (F = 4.797, p = 0.046) and litters (F = 6.799, p = 0.026) for Ni. Litter was the main stock of heavy metals, and the allocations of Cr, Ni, and Pb reached 31.25-51.31, 28.49-42.58, and 29.55-66.79 % (in MM) and 36.73-48.60, 41.70-57.87, and 33.30-60.64 % (in LM), respectively. The ratios of roots/leaves (R/L) and roots/stems (R/S) for Cr and Ni in MM were mostly greater than 1, while those ratios in LM were mostly less than 1, indicating that Cr and Ni in S. salsa at LM had greater mobility compared with those at MM. Moreover, the [accumulation factor, AF]plant of Cr, Ni, Cu, and Pb in LM, especially [AF]root and [AF]stem of Cr and [AF]litter of Ni, was also higher than that in MM. These indicated that S. salsa grown in LM was more suitable for potential biomonitor or phytoremediation of Cr, Ni, Cu, and Pb if intertidal sediments were seriously contaminated with an increase of pollutant loading (especially heavy metals) in the Yellow River estuary. The use of biomonitor (S. salsa) living and growing in LM could yield valuable information not only on the presence of anthropogenic stressors, but, more importantly, on the adverse influence the stressors are having on the environment.


Experimental Schistosomiasis japonica in the pig: immunohistology of the hepatic egg granuloma.
Use of the pig as an animal model in schistosomiasis research is increasing, but knowledge of the porcine immune response to schistosome infection is still very limited. We investigated the immunohistology of different maturation stages of the Schistosoma japonicum egg granuloma in pigs. Liver sections from pigs experimentally infected with S.japonicum for 9, 12 or 21 weeks were examined by immunohistochemistry using a three-step streptavidin-biotin-complex/immunoperoxidase method or a two-step alkaline phosphatase-mediated system. All granulomas showed marked expression of major histocompatibility complex (MHC) class II in epithelioid macrophages and were dominated by T lymphocytes, comprising both CD4+ and CD8+ phenotypes, with consistently higher proportions noted for CD8+ cells. B lymphocytes, as identified by expression of CD21, were confined to lymphoid nodular structures primarily associated with mature granulomas. Early and mature granulomas contained numerous immunoglobulin (Ig)G+ plasma cells. Significant differences in immunohistology related to duration of infection were not observed. The results indicate that all stages of the hepatic S.japonicum egg granuloma in the pig manifests MHC class II-dependent CD4+ T cell activity concomitant with infiltration of CD8+ T cells. B cell activity preceding the effector cell stage appears to occur in granuloma-associated lymphoid nodules, whereas antibody, mainly IgG, is produced within the granuloma.


Rosacea is associated with Helicobacter pylori: a systematic review and meta-analysis.
Rosacea is a common skin disease characterized by facial erythema, telangiectasia, papules and pustules. Helicobacter pylori infection has been suggested to play a role in the etiopathogenesis of rosacea. To systematically review and meta-analyse the relationship between rosacea and infection with Helicobacter pylori. A literature search was performed using PubMed, EMBASE and Web of Science. Data extraction and analyses were performed on descriptive data. Study quality was assessed using the Newcastle-Ottawa Scale. Random-effects models with DerSimonian-Laird methods were utilized to estimate pooled odds ratios (ORs), with 95% confidence intervals (95% CIs). Heterogeneity of results was assessed using I² statistics. A total of 454 articles were identified and 42 full-text articles were chosen for further review. Fourteen studies were included in the quantitative meta-analysis, comprising a total of 928 rosacea patients and 1527 controls. The overall association between Helicobacter pylori infection and rosacea was non-significant (OR 1.68, 95% CI 1.00-2.84, P = 0.052), but analysis restricted to C-urea breath test showed a significant association (OR 3.12, 95% CI 1.92-5.07, P < 0.0001). Effect of eradication treatment on rosacea symptoms was assessed in seven studies, but without significant effect (RR 1.28, 95% CI 0.98-1.67, P = 0.069). This meta-analysis found weak associations between rosacea and Helicobacter pylori infection as well as an effect of Helicobacter pylori therapy on rosacea symptoms, albeit that these did not reach statistical significance. Whether a pathogenic link between the two conditions exists, or whether Helicobacter pylori infection represents a proxy for other factors remains unknown.


Traumatic stress as a mediator of quality of life and burden in informal caregivers of amputees due to diabetic foot: a longitudinal study.
Type 2 diabetes mellitus (DMT2) is frequently associated with complications that can lead to lower limb amputation, being the second major cause of amputation. The amputee becomes dependent on caregivers, who are often unprepared for this new role and face many challenges. Care-giving influences several dimensions of the caregiver's life. This study evaluates the mediating role of traumatic stress in the relationship between caregiver perceived stress and burden/quality of life (QoL) taking also into consideration the duration of caregiving and whether the caregiver receives help in caregiving tasks, in informal caregivers of amputees due to diabetic foot over a 10 month period. The sample consisted of 110 informal caregivers of amputees due to diabetic foot with type 2 diabetes. This longitudinal study evaluates, at three time points, the mediating role of traumatic stress in the relationship between perceived stress and burden/quality of life considering the duration of caregiving and whether the caregiver received help and social support in caregiving tasks. The results showed that the duration of caregiving, help in caregiving and caregiver stress (T1) predicted traumatic stress seven months (T2) after patients' amputation; and traumatic stress (T2) predicted mental QoL and burden, 10 months later (T3). This study emphasizes the importance of intervention in caregivers at the beginning of caregiving, to improve mental QoL and decrease the burden, ten months after amputation.


Combining neuropsychological and structural neuroimaging indicators of conversion to Alzheimer's disease in amnestic mild cognitive impairment.
Morphometric and neuropsychological retrospective studies of amnestic mild cognitive impairment (MCI) have demonstrated that regional atrophies and cognitive impairments may differentiate stable from progressing MCI. No measure has proved helpful prospectively. In this study, twenty five amnestic MCI patients and 25 healthy controls underwent structural MRI and comprehensive neuropsychological assessment. The groups' grey matter volumes were compared with voxel based morphometry and were also correlated with scores obtained on paired associates learning and category fluency tasks. MCI patients had significantly reduced grey matter volume in left mediotemporal and other neocortical regions compared with controls. Atrophy in perirhinal and anterior inferior temporal cortex was associated with poor scores on both category fluency and paired associates learning tasks. After 36 months, 44% of the MCI sample converted to dementia. Converter and non-converter MCI subgroups differed in paired associates learning and in category fluency scores, and showed limited differences in grey matter loss in the hippocampal complex. Variable atrophy in the hippocampus was not a relevant element in the converter/non converter distinction, but converters had significant volumetric reductions in the perhirinal cortex and in other anterior temporal and frontal neocortical areas. A high proportion of converters (91%) could be identified from baseline data using a combination of measures of regional atrophy in left temporal association cortex and poor scores on paired associates learning and category fluency tasks. This combined approach may offer a better option than using each measure alone to prospectively identify individuals at more immediate risk of conversion to dementia in the MCI population. The clinical advantage of this combination of structural MRI and neuropsychological measures in predicting conversion to dementia will need additional prospective validation.


ASFP (Artificial Intelligence based Scoring Function Platform): a web server for the development of customized scoring functions.
Virtual screening (VS) based on molecular docking has emerged as one of the mainstream technologies of drug discovery due to its low cost and high efficiency. However, the scoring functions (SFs) implemented in most docking programs are not always accurate enough and how to improve their prediction accuracy is still a big challenge. Here, we propose an integrated platform called ASFP, a web server for the development of customized SFs for structure-based VS. There are three main modules in ASFP: (1) the descriptor generation module that can generate up to 3437 descriptors for the modelling of protein-ligand interactions; (2) the AI-based SF construction module that can establish target-specific SFs based on the pre-generated descriptors through three machine learning (ML) techniques; (3) the online prediction module that provides some well-constructed target-specific SFs for VS and an additional generic SF for binding affinity prediction. Our methodology has been validated on several benchmark datasets. The target-specific SFs can achieve an average ROC AUC of 0.973 towards 32 targets and the generic SF can achieve the Pearson correlation coefficient of 0.81 on the PDBbind version 2016 core set. To sum up, the ASFP server is a powerful tool for structure-based VS.


The [4Fe-4S] cluster of sulfurtransferase TtuA desulfurizes TtuB during tRNA modification in Thermus thermophilus.
TtuA and TtuB are the sulfurtransferase and sulfur donor proteins, respectively, for biosynthesis of 2-thioribothymidine (s2T) at position 54 of transfer RNA (tRNA), which is responsible for adaptation to high temperature environments in Thermus thermophilus. The enzymatic activity of TtuA requires an iron-sulfur (Fe-S) cluster, by which a sulfur atom supplied by TtuB is transferred to the tRNA substrate. Here, we demonstrate that the Fe-S cluster directly receives sulfur from TtuB through its inherent coordination ability. TtuB forms a [4Fe-4S]-TtuB intermediate, but that sulfur is not immediately released from TtuB. Further desulfurization assays and mutation studies demonstrated that the release of sulfur from the thiocarboxylated C-terminus of TtuB is dependent on adenylation of the substrate tRNA, and the essential residue for TtuB desulfurization was identified. Based on these findings, the molecular mechanism of sulfur transfer from TtuB to Fe-S cluster is proposed.


Linear topology in amorphous metal oxide electrochromic networks obtained via low-temperature solution processing.
Amorphous transition metal oxides are recognized as leading candidates for electrochromic window coatings that can dynamically modulate solar irradiation and improve building energy efficiency. However, their thin films are normally prepared by energy-intensive sputtering techniques or high-temperature solution methods, which increase manufacturing cost and complexity. Here, we report on a room-temperature solution process to fabricate electrochromic films of niobium oxide glass (NbOx) and 'nanocrystal-in-glass' composites (that is, tin-doped indium oxide (ITO) nanocrystals embedded in NbOx glass) via acid-catalysed condensation of polyniobate clusters. A combination of X-ray scattering and spectroscopic characterization with complementary simulations reveals that this strategy leads to a unique one-dimensional chain-like NbOx structure, which significantly enhances the electrochromic performance, compared to a typical three-dimensional NbOx network obtained from conventional high-temperature thermal processing. In addition, we show how self-assembled ITO-in-NbOx composite films can be successfully integrated into high-performance flexible electrochromic devices.


MRI radiomics model for predicting TERT promoter mutation status in glioblastoma.
The presence of TERT promoter mutations has been associated with worse prognosis and resistance to therapy for patients with glioblastoma (GBM). This study aimed to determine whether the combination model of different feature selections and classification algorithms based on multiparameter MRI can be used to predict TERT subtype in GBM patients. A total of 143 patients were included in our retrospective study, and 2553 features were obtained. The datasets were randomly divided into training and test sets in a ratio of 7:3. The synthetic minority oversampling technique was used to achieve data balance. The Pearson correlation coefficients were used for dimension reduction. Three feature selections and five classification algorithms were used to model the selected features. Finally, 10-fold cross validation was applied to the training dataset. A model with eight features generated by recursive feature elimination (RFE) and linear discriminant analysis (LDA) showed the greatest diagnostic performance (area under the curve values for the training, validation, and testing sets: 0.983, 0.964, and 0.926, respectively), followed by relief and random forest (RF), analysis of variance and RF. Furthermore, the relief was the optimal feature selection for separately evaluating those five classification algorithms, and RF was the most preferable algorithm for separately assessing the three feature selectors. ADC entropy was the parameter that made the greatest contribution to the discrimination of TERT mutations. Radiomics model generated by RFE and LDA mainly based on ADC entropy showed good performance in predicting TERT promoter mutations in GBM.


Acute hyperoxia-induced transcriptional response in the mouse RPE/choroid.
Oxidative stress has been studied in the retinal pigmented epithelium (RPE) in vitro but not in vivo. Our purpose, therefore, was to develop an in vivo model of acute oxidative stress in the C57BL/6J mouse. Mice were exposed to > or = 98% oxygen for 0, 2, or 6 h, and amplified total RNA from the RPE/choroid was applied to microarrays examining about 2200 unique genes. Statistical analysis determined that 642 genes, out of a total of 1349 expressed, were significantly downregulated at only 2 h, only 6 h, or both 2 and 6 h, and a single gene, ubiquitin, was upregulated. These genes are involved in all aspects of cellular functions, and there are no major differences among the three groups. The effect of hyperoxia on the RPE/choroid in vivo appears to be very similar to oxidative stress studies performed with an RPE cell line in vitro. All 11 genes identified as being regulated by all three oxidants in our previous study, and were expressed by mouse, were also differentially regulated by hyperoxia. At least for the initial response to an oxidative challenge, the in vitro ARPE-19 cell line is a reasonable model for in vivo studies.


Nurses' Experiences Empowering Hospitalized Patients.
Four focus groups were conducted to explore acute care nurses' experiences empowering patients and the facilitators and barriers they encountered during the process. Thirty-four nurses employed at four hospitals in the Midwestern United States participated in the study between February and April 2015. Facilitators of empowerment included establishing a therapeutic relationship, fostering communication, providing education, respecting patient autonomy, engaging support systems, and lifting spirit/giving hope. Barriers included conflicting information about plans of care, lack of time, fear and anxiety over unfamiliar environments and routines, ineffective or inadequate support systems, lack of/low accountability, and killing the soul. Nurses also described innovative strategies they used to overcome the barriers. The development of future inpatient empowerment interventions needs to focus on the innovative strategies nurses used to overcome barriers in addition to considering the facilitators and barriers to empowerment that nurses identified.


Psip1/Ledgf p75 restrains Hox gene expression by recruiting both trithorax and polycomb group proteins.
Trithorax and polycomb group proteins are generally thought to antagonize one another. The trithorax family member MLL (myeloid/lymphoid or mixed-lineage leukemia) is presumed to activate Hox expression, counteracting polycomb-mediated repression. PC4 and SF2 interacting protein 1 (PSIP1)/p75, also known as LEDGF, whose PWWP domain binds to H3K36me3, interacts with MLL and tethers MLL fusion proteins to HOXA9 in leukaemias. Here we show, unexpectedly, that Psip1/p75 regulates homeotic genes by recruiting not only MLL complexes, but also the polycomb group protein Bmi1. In Psip1(-/-) cells binding of Mll1/2, Bmi1 and the co-repressor Ctbp1 at Hox loci are all abrogated and Hoxa and Hoxd mRNA expression increased. Our data not only reveal a potential mechanism of action for Psip1 in the regulation of Hox genes but also suggest an unexpected interplay between proteins usually considered as transcriptional activators and repressors.


Neutralizing effect by resin-based materials containing silane-coated glass fillers.
The aim of this study was to evaluate the fluoride release, neutralizing ability and inhibitory effect on secondary caries of resin-based materials containing a silane-coated glass filler. Resin-based materials containing fluoro-boro-alumino-silicate glass coated by 3-methacryloxypropyltrimethoxysilane or 3-aminopropyltrimethoxysilane were used and resin material containing glass filler without coating was used as a control. The fluoride release and pH value after immersion were measured for 10 weeks. The inhibitory effect was also evaluated. During the initial period, the material with 3-aminopropyltrimethoxysilane gave the greater amount of fluoride release and produced a higher pH value compared with the other materials. However, the neutralizing ability of the material with 3-aminopropyltrimethoxysilane filler became weaker with ageing of the specimens. The mean depth of outer lesions was similar among the three materials. Within the limitations of this study, secondary caries around restorations could not be inhibited even for products showing high fluoride releasing and neutralizing ability.


Meiotic and developmental competence of mouse antral oocytes.
Mouse antral oocytes show two different patterns of chromatin organization, defining oocytes with or without chromatin surrounding the nucleolus (SN: surrounded nucleus; NSN: not surrounded nucleus). We have previously shown that upon injection of eCG, NSN antral oocytes shift towards the SN kind of chromatin organization. We hypothesized that these newly formed SN oocytes were those that would have been ovulated after an ovulatory stimulus. The main objective of this study was to investigate the meiotic and developmental competence of these two types of oocytes. SN and NSN antral oocytes were isolated after i.p. administration of eCG + hCG or eCG-only, in vitro-cultured until completion of metaphase II, and inseminated with capacitated spermatozoa; and their development to the 4-cell stage was examined. This study demonstrates 1) that SN and NSN oocytes isolated after injection of eCG + hCG are capable of embryonic development, but not beyond the 2-cell stage; and 2) that SN and NSN oocytes isolated after injection of eCG-only are capable of developing to the 2-cell stage, but a significantly higher number (11.9%) of SN oocytes than NSN oocytes (1.5%) reach the 4-cell stage. SN- and NSN-like oocytes have also been described in the antral compartment of human, rat, monkey, pig, and bovine ovaries. The findings reported in this paper may contribute to improved procedures for in vitro fertilization of humans and farm animals.


1H and 31P nuclear magnetic resonance studies of spermine binding to the Z-DNA form of d(m5CGm5CGm5CG)2. Evidence for decreased spermine mobility.
The binding of spermine to the d(m5CGm5CGm5CG) duplex has been studied by proton and phosphorus nuclear magnetic resonance techniques in order to investigate the mobility and nature of spermine bound to the resulting Z-DNA complex. A characterization of the B to Z transition as a function of increasing spermine concentration demonstrated doubling of the non-exchangeable proton and the phosphorus peaks at a ratio of about 1:1 (spermine/duplex) and a re-simplification of the spectrum at 2:1 (spermine/duplex) where about 90% or the DNA was fully converted into the Z-form. However, some of the Z-DNA proton chemical shifts differed between the 1:1 and 2:1 titration points. Since these differences involved primarily the exchangeable terminal imino and amino protons, they could result from end effects. Discrepancies were generally not observed with the non-terminal proton shifts nor with the phosphorus shifts. These proton and phosphorus chemical shift changes are fully consistent with a B to Z transition. Complexed spermine peaks appear about 0.1 parts per million upfield from the uncomplexed form. The spermine and both the B and Z-DNA hexamer signals are noticeably broadened at the 1:1 ratio but the remaining signals re-sharpen at the 2:1 ratio. Both one-dimensional and two-dimensional studies revealed negative nuclear Overhauser effect (NOE) contacts between each spermine proton. Therefore, spermine has a longer correlation time than that observed for unbounded spermine. These results are contrasted with the positive NOE contacts observed for the B-DNA-spermine complexes reported by Wemmer et al. using the dodecamer d(CGCGAATTCGCG)2 and reported here using the hexamer d(ATGCAT)2. While the mobility of spermine in the Z-DNA complex is significantly less than that of the B-DNA complex, no clear evidence of intermolecular spermine-DNA proton NOE contacts is observed.


Multiexon deletion alleles of ATF6 linked to achromatopsia.
Achromatopsia (ACHM) is an autosomal recessive disease that results in severe visual loss. Symptoms of ACHM include impaired visual acuity, nystagmus, and photoaversion starting from infancy; furthermore, ACHM is associated with bilateral foveal hypoplasia and absent or severely reduced cone photoreceptor function on electroretinography. Here, we performed genetic sequencing in 3 patients from 2 families with ACHM, identifying and functionally characterizing 2 mutations in the activating transcription factor 6 (ATF6) gene. We identified a homozygous deletion covering exons 8-14 of the ATF6 gene from 2 siblings from the same family. In another patient from a different family, we identified a heterozygous deletion covering exons 2 and 3 of the ATF6 gene found in trans with a previously identified ATF6 c.970C>T (p.Arg324Cys) ACHM disease allele. Recombinant ATF6 proteins bearing these exon deletions showed markedly impaired transcriptional activity by qPCR and RNA-Seq analysis compared with WT-ATF6. Finally, RNAscope revealed that ATF6 and the related ATF6B transcripts were expressed in cones as well as in all retinal layers in normal human retina. Overall, our data identify loss-of-function ATF6 disease alleles that cause human foveal disease.


Predicting Survival in Thai Patients After Low Impact Hip Fracture Using Flexible Parametric Modelling: A Retrospective Cohort Study.
Predictive post-hip fracture mortality models have been presented for specific time points (in-hospital, 30-days or 1-year) and most provide marginal predictions based on the patient's risk group. However, the predictive model for individual survival probability following hip fracture is not available. This study aimed to develop a flexible parametric model for predicting individual survival probability for hip fracture patients. In this retrospective study, the medical charts of 765 Thai patients admitted to hospital with a hip fracture resulting from low-impact injury from January 2014 to December 2018 were reviewed. Predictors for all-cause mortality were identified using flexible parametric survival analysis and were used to develop the predictive model. The model was calibrated using a calibration graph and discrimination performance was evaluated using the C-statistic. Internal validity was assessed using bootstrapping. The overall mortality rate of the hip fracture patients was 14%. Predictors significantly associated with survival after hip fracture were age, active malignancy, dementia or Alzheimer's disease, chronic obstructive pulmonary disorder, diabetes mellitus, hemoglobin concentration, eGFR<30 mL/min/1.73m2 and operative treatments. The model-predicted survival was similar to that actually observed in the very low survival group in the first year after hip fracture. In bootstrapping, the apparent C-statistic and the test C-statistic of the reduced model were 0.79 (95% CI 0.77-0.81) and 0.79 (95% CI 0.78-0.80), respectively. The flexible survival model provides good predictive power for individual survival probability at any given time point within the first year after hip fracture and would be an easy to use tool in clinical practice.


Hematoma formation during breast core needle biopsy in women taking antithrombotic therapy.
The purpose of this study was to compare hematoma formation after breast core needle biopsy performed on patients undergoing and those not undergoing concurrent antithrombotic therapy. A prospective assessment of core needle biopsies (stereotactic, ultrasound guided, or MRI guided) performed on patients enrolled between September 2011 and July 2012 formed the basis of this study. Postprocedure mediolateral and craniocaudal mammograms were evaluated for the presence and size of hematomas. Patients were clinically evaluated for complications 24-48 hours after the procedure through telephone call or face-to-face consultation. Needle size, type of biopsy, and presence of hematoma and documented complications were correlated with use of antithrombotic agents (including aspirin, warfarin, clopidogrel, and daily nonsteroidal antiinflammatory medications). No clinically significant hematomas or bleeding complications were found. Eighty-nine of 617 (14.4%) non-clinically significant hematomas were detected on postprocedure mammograms. The probability of development of a non-clinically significant hematoma was 21.6% for patients taking antithrombotics and 13.0% for those not taking antithrombotics. Concurrent antithrombotic therapy and larger needle gauge were significant factors contributing to the probability of hematoma formation. The volume of the hematoma was not related to needle gauge or presence of antithrombotic therapy. No clinically significant hematomas were found. Because there are potential life-threatening risks to stopping antithrombotic therapy before breast biopsy, withholding antithrombotic therapy for core needle breast biopsy is not recommended because the incidence of non-clinically significant hematoma is low.


A two-component system plays an important role in the root-colonizing ability of Pseudomonas fluorescens strain WCS365.
We describe the characterization of a novel Tn5lacZ colonization mutant of the efficiently colonizing Pseudomonas fluorescens strain WCS365, mutant strain PCL1210, which is at least 300- to 1,000-fold impaired in colonization of the potato root tip after co-inoculation of potato stem cuttings with a 1:1 mixture of mutant and parental cells. Similarly, the mutant is also impaired in colonization of tomato, wheat, and radish, indicating that the gene involved plays a role in the ability of P. fluorescens WCS365 to colonize a wide range of plant species. A 3.1-kb DNA fragment was found to be able to complement the observed mutation. The nucleotide sequence of the region around the Tn5lacZ insertion showed three open reading frames (ORFs). The transcriptional start site was determined. The operon is preceded by an integration host factor (IHF) binding site consensus sequence whereas no clear -10 and -35 sequences are present. The deduced amino acid sequences of the first two genes of the operon, designated as colR and colS, show strong similarity with known members of two-component regulatory systems. ColR has homology with the response regulators of the OmpR-PhoB subclass whereas ColS, the product of the gene in which the mutation resides, shows similarity to the sensor kinase members of these two-component systems. Hydrophobicity plots show that this hypothetical sensor kinase has two transmembrane domains, as is also known for other sensor kinases. The product of the third ORF, Orf222, shows no homology with known proteins. Only part of the orf222 gene is present in the colonization-complementing, 3.1-kb region, and it therefore does not play a role in complementation. No experimental evidence for a role of the ColR/ColS two-component system in the suspected colonization traits chemotaxis and transport of exudate compounds could be obtained. The function of this novel two-component system therefore remains to be elucidated. We conclude that colonization is an active process in which an environmental stimulus, through this two-component system, activates a so far unknown trait that is crucial for colonization.


'More people talk to you when you have a dog' - dogs as catalysts for social inclusion of people with intellectual disabilities.
Research has shown Australian group homes, and supported living options, fail to support people with intellectual disabilities (IDs) to develop social connections. This pilot study evaluates the effectiveness of a visiting dog walking program to facilitate encounters with other community members. Sixteen adults with IDs were assigned to one of two groups, matched on key characteristics. Group 1 had 14, 1-hour outings in the community with a dog and their handler; Group 2 had 14 outings with a handler alone, followed by an additional five outings with a handler and a dog. Within and between group differences were analysed according to number of encounters when a dog was present and absent. Qualitative data provided insights into the nature of these encounters. The number of encounters was significantly higher when a dog was present than when participants went out into the community with a handler alone. This pattern was reflected in the qualitative data, which also suggested the presence of a dog helped to break social norms about speaking to strangers and discourage disrespect towards people with IDs. A dog walking program has the potential to encourage convivial encounters, which in the long term could be catalysts to help people with IDs build social connections in their communities; this should be further explored.


Unveiling the Origin of Multidomain Structures in Compositionally Modulated Cylindrical Magnetic Nanowires.
CoNi/Ni multisegmented cylindrical nanowires were synthesized via an electrochemical route. The wires are 140 nm in diameter, with 1000 nm long Ni segments and CoNi segments between 600 and 1400 nm in length. The magnetic configuration was imaged by XMCD-PEEM in the demagnetized state and at remanence after magnetizing axially and perpendicularly. Ni segments, with cubic crystal symmetry, show an axial magnetic configuration with a small curling component at the surface. In turn, CoNi segments, with hexagonal crystal symmetry and a strong magnetocrystalline anisotropy perpendicular to the nanowires, show a single vortex state in the shorter segments and multivortex or multitransverse magnetic configurations in medium and long segments, respectively. A detailed study by micromagnetic simulations reveals that the magnetic configuration is determined mainly by the coupling between soft Ni and harder CoNi segments. For short CoNi segments, Ni segments are magnetostatically coupled and the chirality of the single vortex formed in CoNi remains the same as that of the curling in neighboring Ni segments. For longer CoNi segments, the remanent state is either the multivortex or multitransverse state depending on whether the previously applied field was parallel or perpendicular to the magnetocrystalline axis. The results point out the relevance of the cylindrical geometry to promote the occurrence of complex magneto-chiral effects and provide key information for the design of cylindrical magnetic nanowires for multiple applications.


Prognostic value of hemodynamic indices in patients with sepsis after fluid resuscitation.
Sepsis usually causes hemodynamic abnormalities. Hemodynamic index is one of the factors to identify the severity of sepsis and an important parameter to guide the procedure of fluid resuscitation. The present study investigated whether the assessment of hemodynamic indices can predict the outcomes of septic patients undergoing resuscitation therapy. To evaluate the prognostic value of hemodynamic indices in patients with sepsis after fluid resuscitation. A retrospective study was conducted in 120 patients with sepsis at Hainan General Hospital/Hainan Affiliated Hospital of Hainan Medical University between October 2016 and October 2019. All patients were treated with sodium chloride combined with dextran glucose injection for fluid resuscitation. Patients' hemodynamic parameters were monitored, including heart rate (HR), cardiac index (CI), systemic vascular resistance index (SVRI), mean arterial pressure (MAP), central venous pressure (CVP), and central venous oxygen saturation. The prognostic value of hemodynamic indices was determined based on the prognosis status. During fluid resuscitation, 86 patients developed septic shock and 34 did not. Ninety-nine patients survived and 21 patients died at 28 d after the treatment. Heart rate, CI, mean arterial pressure, SVRI, and CVP were higher in patients with septic shock and patients who died from septic shock than in non-shock patients and patients who survived, and central venous oxygen saturation was lower in patients with shock and patients who died than in non-shock patients and the survivors (P < 0.05). When prognosis was considered as a dependent variable and hemodynamic parameters was considered as independent variables, the results of a logistic regression analysis showed that CI, SVRI, and CVP were independent risk factors for septic shock, and CI was an independent risk factor for 28-d mortality (P < 0.05). Hemodynamic indices can be used to evaluate the prognosis of septic patients after fluid resuscitation.


[Hyperammonaemic encephalopathy due to valproic acid].
One of the less frequent idiosyncratic side effects of valproic acid (VPA) is encephalopathy. Here we report one case. An 83-year-old female with no relevant past history, who received treatment with VPA following a post-traumatic subarachnoid haemorrhage and two convulsive seizures. A few days later, she was admitted to the Emergency Department because of a progressive clinical picture of mental slowness, nauseas and apathy. The systemic examination was normal. Neurologically, the most striking features were inattention and disorientation, despite her having a good level of consciousness, and mental confusion. Levels of VPA were within the below-therapeutic range and the basic lab findings (including hepatic profile) were normal, except for hyperammonaemia. Neuroimaging studies and cerebrospinal fluid analysis were also normal. An electroencephalogram (EEG) showed signs of severe diffuse encephalopathy with slow, triphasic waves and a non-convulsive epileptic status was therefore ruled out. After withdrawing the VPA, the patient's condition improved until her basal situation was reached in 48 hours and the EEG became normal, as did her ammonium levels. When faced with a patient who has recently been taking VPA and who presents a clinical picture of mental confusion, the possibility of encephalopathy due to said drug must be taken into consideration.


Fecal indicator bacteria and zoonotic pathogens in marine snow and California mussels (Mytilus californianus).
Pollution of nearshore waters with disease-causing microorganisms impacts ecosystems health through illness and deaths in people and wildlife, as well as negative socioeconomic consequences of impaired marine resources. Insight on pathogen ecology in coastal habitats is crucial for accurately mitigating inputs and impacts of microbial pollution. Three objectives were addressed to (i) compare fecal pollution in proximity to (a) freshwater runoff, and (b) endemic marine wildlife; (ii) evaluate presence and magnitude of fecal microorganisms in marine snow and mussels and (iii) determine if pathogens in mussels and FIB levels in seawater or mussels are correlated. Sampling during the wet season, proximity to freshwater, and FIB levels in mussel homogenates (but not seawater) were associated with pathogen presence in mussels. Pathogens and FIB were enriched in aggregate-rich fractions, further supporting an important role of marine snow in pathogen transmission. The lack of association between FIB in surrounding waters and presence of pathogens in mussels calls into question current regulations for insuring safe seafood to consumers in the United States, and alternative monitoring approaches such as direct testing for select pathogens should be further evaluated.


The effect of some epsilon-aminocaproic acid derivatives on platelet responses.
epsilon-Aminocaproic acid (EACA) is a synthetic low molecular drug with antifibrinolytic activity. However, treatment with this drug can be incidentally associated with an increased thrombotic tendency. The aim of the present work was to test synthetic EACA derivatives for their antiplatelet activities. We investigated the effect of three EACA derivatives with antifibrinolytic activity: I. epsilon-aminocaproyl-L-leucine hydrochloride (HCl*H-EACA-L-Leu-OH), II. epsilon-aminocaproyl-L-(S-benzyl)-cysteine hydrochloride (HCl*H-EACA-L-Cys(S-Bzl)-OH) and III. epsilon-aminocaproyl-L-norleucine (H-EACA-L-Nle-OH) on platelet responses (aggregation and adhesion) and on their integrity. It was found that: 1. as judged by LDH release test, none of the tested compounds, up to 20 mM, was toxic to platelets, 2. in comparison with EACA, all the synthetic derivatives inhibited much stronger the ADP- and collagen-induced aggregation of platelets suspended in plasma (platelet rich plasma) and aggregation of these cells in whole blood, 3. EACA and its derivatives exerted a similar inhibitory effect on the thrombin-induced adhesion of platelets to fibrinogen-coated surfaces. Since platelet activation and blood coagulation are tightly associated processes, the antiplatelet properties of EACA derivatives are expected to indicate reduced thrombotic properties of these derivatives compared to EACA.


Sterically hindered Re- and Mn-CO2 reduction catalysts for solar energy conversion.
Novel molecular Re and Mn tricarbonyl complexes bearing a bipyridyl ligand functionalised with sterically hindering substituents in the 6,6'-position, [M(HPEAB)(CO)3(X)] (M/X = Re/Cl, Mn/Br; HPEAB = 6,6'-{N-(4-hexylphenyl)-N(ethyl)-amido}-2,2'-bipyridine) have been synthesised, fully characterised including by single crystal X-ray crystallography, and their propensity to act as catalysts for the electrochemical and photochemical reduction of CO2 has been established. Controlled potential electrolysis showed that the catalysts are effective for electrochemical CO2-reduction, yielding CO as the product (in MeCN for the Re-complex, in 95 : 5 (v/v) MeCN : H2O mixture for the Mn-complex). The recyclability of the catalysts was demonstrated through replenishment of CO2 within solution. The novel catalysts had similar reduction potentials to previously reported complexes of similar structure, and results of the foot-of-the-wave analysis showed comparable maximum turnover rates, too. The tentative mechanisms for activation of the pre-catalysts were proposed on the basis of IR-spectroelectrochemical data aided by DFT calculations. It is shown that the typical dimerisation of the Mn-catalyst was prevented by incorporation of sterically hindering groups, whilst the Re-catalyst undergoes the usual mechanism following chloride ion loss. No photochemical CO2 reduction was observed for the rhenium complex in the presence of a sacrificial donor (triethylamine), which was attributed to the short triplet excited state lifetime (3.6 ns), insufficient for diffusion-controlled electron transfer. Importantly, [Mn(HPEAB)(CO)3Br] can act as a CO2 reduction catalyst when photosensitised by a zinc porphyrin under red light irradiation (λ > 600 nm) in MeCN : H2O (95 : 5); there has been only one reported example of photoactivating Mn-catalysts with porphyrins in this manner. Thus, this work demonstrates the wide utility of sterically protected Re- and Mn-diimine carbonyl catalysts, where the rate and yield of CO-production can be adjusted based on the metal centre and catalytic conditions, with the advantage of suppressing unwanted side-reactions through steric protection of the vacant coordination site.


Mid-range inhomogeneity of eukaryotic genomes.
Multicellular eukaryotic genomes are replete with nonprotein coding sequences, both within genes (introns) and between them (intergenic regions). Excluding the well-recognized functional elements within these sequences (ncRNAs, transcription factor binding sites, intronic enhancers/silencers, etc.), the remaining portion is made up of so-called "dark" DNA, which still occupies the majority of the genome. This dark DNA has a profound nonrandomness in its sequence composition seen at different scales, from a few nucleotides to regions that span over hundreds of thousands of nucleotides. At the mid-range scale (from 30 up to 10,000 nt), this nonrandomness is manifested in base compositional extremes detected for each of four nucleotides (A, G, T, or C) or any of their combinations. Examples of such compositional nonrandomness are A-rich, purine-rich, or G+T-rich regions. Almost every combination of nucleotides has such enriched regions. We refer to these regions as being "inhomogeneous". These regions are associated with unusual DNA conformations and/or particular DNA properties. In particular, mid-range inhomogeneous regions have complex arrangements relative to each other and to specific genomic sites, such as centromeres, telomeres, and promoters, pointing to their important role in genomic functioning and organization.


Prognostic Factors for Prostate Cancer Endpoints Following Biochemical Failure: A Review of the Literature.
In the setting of biochemical failure (BCF) following primary treatment for prostate cancer, additional discrimination between clinically significant and non-clinically significant biochemical recurrence is critical in defining robust surrogate endpoints for prostate cancer and guiding salvage management decisions. We reviewed the literature to determine which prognostic factors are most significant for predicting prostate cancer-specific survival (PCSS), metastases-free survival (MFS), and/or overall survival (OS) after BCF. A search of PubMed from 1980 to 2013 yielded 999 studies that examined prognostic factors predictive for PCSS, MFS, and/or OS in prostate cancer patients with BCF following primary treatment. Eligibility criteria for inclusion were: 1) examined a prostate cancer population in the setting of BCF without overt clinical relapse following primary treatment with radical prostatectomy or radiotherapy; 2) based analyses on patient parameters obtained prior to the initiation of salvage therapies; and 3) determined clinical prognostic factors that were significant prognostic measures for at least one of three clinically relevant endpoints: OS, PCS, or MFS. Nineteen eligible studies reported on 8,040 patients that experienced BCF from 1981-2013. The initial primary therapy was variable: radical prostatectomy alone (n=8), radiotherapy alone (n=4), radiotherapy/radical prostatectomy ± adjuvant therapy (n=5), and multiple treatment arms (n=2). There was also heterogeneity in which outcomes were assessed: PCSS (n=14), MFS (n=7), and OS (n=5). The prognostic factors most commonly found to be significant on multivariate analyses were PSA doubling time (PSADT), time to biochemical failure (TTBF), pathological Gleason score (pGS), and age. Risk stratification in prostate cancer post-BCF is challenging because of limited predictive modeling that can determine which patients will optimally benefit from salvage therapy. Our systematic literature review has identified PSADT, TTBF, pGS, and age as the leading prognostic factors for the prediction of PCSS, MFS, and OS after BCF. We plan to leverage the Canadian ProCaRS database to perform predictive modeling using the putative findings in the present study in order to propose potential evidence-based surrogate endpoints for prostate cancer in the setting of BCF.


Assessing smoking status in children, adolescents and adults: cotinine cut-points revisited.
To reassess saliva cotinine cut-points to discriminate smoking status. Cotinine cut-points that are in use were derived from relatively small samples of smokers and non-smokers 20 or more years ago. It is possible that optimal cut-points may have changed as prevalence and exposure to passive smoking have declined. Cross-sectional survey of the general population, with assessment of self-reported smoking and saliva cotinine. A total of 58 791 respondents aged 4 years and older in the Health Survey for England for the years 1996-2004 who provided valid saliva cotinine specimens. Saliva cotinine concentrations, demographic variables, self-reported smoking, presence or absence of smoking in the home, a composite index of social disadvantage derived from occupation, housing tenure and access to a car. A cut-point of 12 ng/ml performed best overall, with specificity of 96.9% and sensitivity of 96.7% in discriminating confirmed cigarette smokers from never regular smokers. This cut-point also identified correctly 95.8% of children aged 8-15 years smoking six or more cigarettes a week. There was evidence of substantial misreport in claimed ex-smokers, especially adolescents (specificity 72.3%) and young adults aged 16-24 years (77.5%). Optimal cut-points varied by presence (18 ng/ml) or absence (5 ng/ml) of smoking in the home, and there was a gradient from 8 ng/ml to 18 ng/ml with increasing social disadvantage. The extent of non-smokers' exposure to other people's tobacco smoke is the principal factor driving optimal cotinine cut-points. A cut-point of 12 ng/ml can be recommended for general use across the whole age range, although different cut-points may be appropriate for population subgroups and in societies with differing levels of exposure to secondhand smoke.


The gastrointestinal aspects of halitosis.
Halitosis is a common human condition for which the exact pathophysiological mechanism is unclear. It has been attributed mainly to oral pathologies. Halitosis resulting from gastrointestinal disorders is considered to be extremely rare. However, halitosis has often been reported among the symptoms related to Helicobacter pylori infection and gastroesophageal reflux disease. To retrospectively review the experience with children and young adults presenting with halitosis to a pediatric gastroenterology clinic. A retrospective chart review of patients diagnosed with halitosis as a primary or secondary symptom was conducted. All endoscopies were performed by the same endoscopist. A total of 94 patients had halitosis, and of the 56 patients (59.6%) who were recently examined by a dental surgeon, pathology (eg, cavities) was found in only one (1.8%). Pathology was found in only six of 27 patients (28.7%) who were assessed by an otolaryngology surgeon. Gastrointestinal pathology was found to be very common, with halitosis present in 54 of the 94 (57.4%) patients. The pathology was noted regardless of dental or otolaryngological findings. Most pathologies, both macroscopically and microscopically, were noted in the stomach (60% non-H pylori related), followed by the duodenum and the esophagus. Fifty-two of 90 patients (57.8%) were offered a treatment based on their endoscopic findings. Of the 74 patients for whom halitosis improvement data were available, some improvement was noted in 24 patients (32.4%) and complete improvement was noted in 41 patients (55.4%). Gastrointestinal pathology was very common in patients with halitosis regardless of dental or otolaryngological findings, and most patients improved with treatment.


A Preliminary Study on Cranio-Facial Characteristics Associated with Minor Neurological Dysfunctions (MNDs) in Children with Autism Spectrum Disorders (ASD).
Background. Craniofacial anomalies and minor neurological dysfunction (MNDs) have been identified, in literature, as risk factors for neurodevelopmental disorders. They represent physical indicators of embryonic development suggesting a possible contributory role of complications during early, even pre-conceptional, phases of ontogeny in autism spectrum disorders (ASD). Limited research has been conducted about the co-occurrence of the two biomarkers in children with ASD. This study investigates the associative patterns of cranio-facial anomalies and MNDs in ASD children, and whether these neurodevelopmental markers correlate with intensity of ASD symptoms and overall functioning. Methods. Caucasian children with ASD (n = 33) were examined. Measures were based on five anthropometric cranio-facial indexes and a standardized and detailed neurological examination according to Touwen. Relationships between anthropometric z-scores, MNDs and participant characteristics (i.e., age, cognitive abilities, severity of autistic symptoms measured using the Childhood Autism Rating Scale (CARS) checklist) were assessed. Results. With respect to specific MNDs, significant positive correlations were found between Cephalic Index and Sensory deficits (p-value < 0.001), which did not correlate with CARS score. Importantly, CARS score was positively linked with Intercanthal Index (p-value < 0.001), and negatively associated with posture and muscle tone (p-value = 0.027) and Facial Index (p-value = 0.004). Conclusion. Our data show a link between a specific facial phenotype and anomalies in motor responses, suggesting early brain dysmaturation involving subcortical structures in cerebro-craniofacial development of autistic children. This research supports the concept of a "social brain functional morphology" in autism spectrum disorders.


Importance of Dietary Changes During the Coronavirus Pandemic: How to Upgrade Your Immune Response.
The new coronavirus pandemic continues to spread causing further public health, social, and economic issues. The disparities in the rates of death between countries poses questions about the importance of lifestyle habits and the immune status of populations. An exploration of dietary habits and COVID-19-related death might unravel associations between these two variables. Indeed, while both nutritional excess and deficiency are associated with immunodeficiency, adequate nutrition leading to an optimally functioning immune system may be associated with better outcomes with regards to preventing infection and complications of COVID-19, as well as developing a better immune response to other pathogenic viruses and microorganisms. This article outlines the key functions of the immune system and how macronutrients, micronutrients, and metabolites from the gut microbiome can be essential in the development of an efficient immune system. In addition, the effects of intermittent fasting on the inflammatory state as well as metabolic parameters will be discussed.


Population-level impact of infant 10-valent pneumococcal conjugate vaccination on adult pneumonia hospitalisations in Finland.
Limited data are available on population-level herd effects of infant 10-valent pneumococcal conjugate vaccine (PCV10) programmes on pneumonia. We assessed national trends in pneumococcal and all-cause pneumonia hospitalisations in adults aged ≥18 years, before and after infant PCV10 introduction in 2010. Monthly hospitalisation rates of International Statistical Classification of Diseases, 10th revision (ICD-10)-coded primary discharge diagnoses compatible with pneumonia from 2004-2005 to 2014-2015 were calculated with population denominators from the population register. Trends in pneumonia before and after PCV10 introduction were assessed with interrupted time-series analysis. Rates during the PCV10 period were estimated from adjusted negative binomial regression model and compared with those projected as continuation of the pre-PCV10 trend. All-cause hospitalisations were assessed for control purposes. Before PCV10, the all-cause pneumonia rate in adults aged ≥18 years increased annually by 2.4%, followed by a 4.7% annual decline during the PCV10 period. In 2014-2015, the overall all-cause pneumonia hospitalisation rate was 109.3/100 000 (95% CI 96.5 to 121.9) or 15.4% lower than the expected rate. A significant 6.7% decline was seen in persons aged ≥65 years (131.5/100 000), which translates to 1456 fewer pneumonia hospitalisations annually. In comparison, hospitalisations other than pneumonia decreased by 3.5% annually throughout the entire study period. These national data suggest that herd protection from infant PCV10 programme has reversed the increasing trend and substantially decreased all-cause pneumonia hospitalisations in adults, particularly the elderly.


Hair Segmentation Using Heuristically-Trained Neural Networks.
We present a method for binary classification using neural networks (NNs) that performs training and classification on the same data using the help of a pretraining heuristic classifier. The heuristic classifier is initially used to segment data into three clusters of high-confidence positives, high-confidence negatives, and low-confidence sets. The high-confidence sets are used to train an NN, which is then used to classify the low-confidence set. Applying this method to the binary classification of hair versus nonhair patches, we obtain a 2.2% performance increase using the heuristically trained NN over the current state-of-the-art hair segmentation method.


Biomolecular condensate phase diagrams with a combinatorial microdroplet platform.
The assembly of biomolecules into condensates is a fundamental process underlying the organisation of the intracellular space and the regulation of many cellular functions. Mapping and characterising phase behaviour of biomolecules is essential to understand the mechanisms of condensate assembly, and to develop therapeutic strategies targeting biomolecular condensate systems. A central concept for characterising phase-separating systems is the phase diagram. Phase diagrams are typically built from numerous individual measurements sampling different parts of the parameter space. However, even when performed in microwell plate format, this process is slow, low throughput and requires significant sample consumption. To address this challenge, we present here a combinatorial droplet microfluidic platform, termed PhaseScan, for rapid and high-resolution acquisition of multidimensional biomolecular phase diagrams. Using this platform, we characterise the phase behaviour of a wide range of systems under a variety of conditions and demonstrate that this approach allows the quantitative characterisation of the effect of small molecules on biomolecular phase transitions.


Impact of Rapid Molecular Detection of Respiratory Viruses on Clinical Outcomes and Patient Management.
To determine if rapid molecular testing for respiratory viruses in patients with respiratory illnesses can provide advantages to patients and hospitals, rigorous investigations on the impacts of using these assays are required. Well-conducted studies are needed to inform decisions about implementation of new rapid assays to replace standard molecular testing or to initiate testing in laboratories that are currently not doing molecular tests for respiratory viruses due to the complex nature of standard panels. In this issue of the Journal of Clinical Microbiology, N. Wabe et al. (J Clin Microbiol 57:e01727-18, 2019, https://doi.org/10.1128/JCM.01727-18) report the results of their evaluation of the impact of using a rapid molecular test for influenza A/influenza B and RSV on outcomes for adults hospitalized with respiratory illness. The median time from admission to test result of the rapid test was 7.5 h compared to 40.3 h for the standard PCR assay. Compared to the use of the standard molecular assay, use of a rapid test significantly shortened time in the hospital and reduced the number of other microbiology tests performed. The authors concluded that rapid PCR testing of adults hospitalized with respiratory illnesses could provide benefits to both the patients and the hospital. Patients were able to leave the hospital earlier and a greater proportion of them had received their test results before discharge, which would allow appropriate treatment to be provided more quickly.


Children who are deaf or hard of hearing in inclusive educational settings: a literature review on interactions with peers.
This review is conducted to describe how children who are deaf or hard of hearing (D/HH) interact with hearing peers in inclusive settings, illustrate the difficulties and challenges faced by them in interacting with peers, and identify effective interventions that promote their social interaction in inclusive education. A systematic search of databases and journals identified 21 papers that met the inclusion criteria. Two broad themes emerged from an analysis of the literatures, which included processes and outcomes of interactions with peers and intervention programs. The research indicates that children who are D/HH face great difficulties in communicating, initiating/entering, and maintaining interactions with hearing peers in inclusive settings. The co-enrollment and social skills training programs are considered to be effective interventions for their social interaction. Communication abilities and social skills of children who are D/HH, responses of children with normal hearing, and the effect of environment are highlighted as crucial aspects of social interactions. In addition, future research is needed to study the interaction between children who are D/HH and hearing peers in natural settings, at different stages of school life, as well as improving social interaction and establishing an inclusive classroom climate for children who are D/HH.


The Supercharged Pedicled Jejunal Flap for Total Esophageal Reconstruction: A Retrospective Review of 100 Cases.
When gastric pull-up is unsuccessful or unsuitable for total esophageal reconstruction, a supercharged pedicled jejunum can be used to reestablish gastrointestinal continuity. The authors reviewed their technique and outcomes of the supercharged pedicled jejunum for total esophageal reconstruction. A retrospective review of a prospectively maintained database was performed of 100 patients who underwent supercharged pedicled jejunum for total esophageal reconstruction between 2000 and 2017 at the Texas Medical Center. Patient characteristics, technical details, and outcomes were analyzed. Mean patient age was 59.5 ± 11.4 years. Forty-two patients (42 percent) had surgical complications (18 percent at the recipient site, 13 percent at the donor site, and 11 percent at both). Medical complications occurred in 28 patients (28 percent). A major surgical complication occurred in 20 patients (20 percent). The average length of stay was 15 days (range, 6 to 152 days). At last follow-up, 20 patients (20 percent) had metastatic disease and six (6 percent) had local recurrence. Fifty-four patients (54 percent) died during the follow-up period. Of 79 patients with follow-up longer than 6 months, 68 (86 percent) tolerated a solid or soft oral diet, with a 16 percent tube feed-dependence rate. Overall survival at 1, 3, and 5 years was 78.8, 53.7, and 33.1 percent, respectively. The median survival time was 38.7 months. The authors present their experience with 100 supercharged pedicled jejunums for total esophageal reconstruction. Functional outcomes are comparable to, or better than, other salvage modalities. With careful multidisciplinary planning and meticulous, well-orchestrated surgical technique, swallowing function can be restored to provide quality of life in patients with few remaining surgical options.


Primary culture of adult rat heart myocytes.
Cultured primary adult rodent heart cells are an important model system for cardiovascular research. Nevertheless, establishment of robust, viable cultured adult myocytes can be a technically challenging, rate-limiting step for many researchers. Here we described a protocol to obtain a high yield of adult rat heart myocytes that remain viable in culture for several days. The heart is isolated and perfused with collagenase and protease under low Ca2+ conditions to recover single myocytes. Ca2+-tolerant cells are obtained by stepwise increases in extracellular Ca2+ concentration in three subsequent wash steps. Cells are filtered, resuspended in culture medium, and plated on laminin coated slips. Cultured myocytes obtained using this protocol are viable for up to four days and are suitable for most experiments including electrophysiology, biochemistry, imaging and molecular biology.


The intestinal intermediate filament network responds to and protects against microbial insults and toxins.
The enrichment of intermediate filaments in the apical cytoplasm of intestinal cells is evolutionarily conserved, forming a sheath that is anchored to apical junctions and positioned below the microvillar brush border, which suggests a protective intracellular barrier function. To test this, we used Caenorhabditiselegans, the intestinal cells of which are endowed with a particularly dense intermediate filament-rich layer that is referred to as the endotube. We found alterations in endotube structure and intermediate filament expression upon infection with nematicidal B.thuringiensis or treatment with its major pore-forming toxin crystal protein Cry5B. Endotube impairment due to defined genetic mutations of intermediate filaments and their regulators results in increased Cry5B sensitivity as evidenced by elevated larval arrest, prolonged time of larval development and reduced survival. Phenotype severity reflects the extent of endotube alterations and correlates with reduced rescue upon toxin removal. The results provide in vivo evidence for a major protective role of a properly configured intermediate filament network as an intracellular barrier in intestinal cells. This notion is further supported by increased sensitivity of endotube mutants to oxidative and osmotic stress.


Altered extracellular striatal in vivo biotransformation of the opioid neuropeptide dynorphin A(1-17) in the unilateral 6-OHDA rat model of Parkinson's disease.
The in vivo biotransformation of dynorphin A(1-17) (Dyn A) was studied in the striatum of hemiparkinsonian rats by using microdialysis in combination with nanoflow reversed-phase liquid chromatography/electrospray time-of-flight mass spectrometry. The microdialysis probes were implanted into both hemispheres of unilaterally 6-hydroxydopamine (6-OHDA) lesioned rats. Dyn A (10 pmol microl(-1)) was infused through the probes at 0.4 microl min(-1) for 2 h. Samples were collected every 30 min and analyzed by mass spectrometry. The results showed for the first time that there was a difference in the Dyn A biotransformation when comparing the two corresponding sides of the brain. Dyn A metabolites 1-8, 1-16, 5-17, 10-17, 7-10 and 8-10 were detected in the dopamine-depleted striatum but not in the untreated striatum. Dyn A biotransformed fragments found in both hemispheres were N-terminal fragments 1-4, 1-5, 1-6, 1-11, 1-12 and 1-13, C-terminal fragments 2-17, 3-17, 4-17, 7-17 and 8-17 and internal fragments 2-5, 2-10, 2-11, 2-12, and 8-15. The relative levels of these fragments were lower in the dopamine-depleted striatum. The results imply that the extracellular in vivo processing of the dynorphin system is being disturbed in the 6-OHDA-lesion animal model of Parkinson's disease.


Glucose sensitivity of mouse olfactory bulb neurons is conveyed by a voltage-gated potassium channel.
The olfactory bulb has recently been proposed to serve as a metabolic sensor of internal chemistry, particularly that modified by metabolism. Because the voltage-dependent potassium channel Kv1.3 regulates a large proportion of the outward current in olfactory bulb neurons and gene-targeted deletion of the protein produces a phenotype of resistance to diet-induced obesity in mice, we hypothesized that this channel may play a role in translating energy availability into a metabolic signal. Here we explored the ability of extracellular glucose concentration to modify evoked excitability of the mitral neurons that principally regulate olfactory coding and processing of olfactory information. Using voltage-clamp electrophysiology of heterologously expressed Kv1.3 channels in HEK 293 cells, we found that Kv1.3 macroscopic currents responded to metabolically active (d-) rather than inactive (l-) glucose with a response profile that followed a bell-shaped curve. Olfactory bulb slices stimulated with varying glucose concentrations showed glucose-dependent mitral cell excitability as evaluated by current-clamp electrophysiology. While glucose could be either excitatory or inhibitory, the majority of the sampled neurons displayed a decreased firing frequency in response to elevated glucose concentration that was linked to increased latency to first spike and decreased action potential cluster length. Unlike modulation attributed to phosphorylation, glucose modulation of mitral cells was rapid, less than one minute, and was reversible within the time course of a patch recording. Moreover, we report that modulation targets properties of spike firing rather than action potential shape, involves synaptic activity of glutamate or GABA signalling circuits, and is dependent upon Kv1.3 expression. Given the rising incidence of metabolic disorders attributed to weight gain, changes in neuronal excitability in brain regions regulating sensory perception of food are of consequence.