Connectivity mapping uncovers small molecules that modulate neurodegeneration in Huntington's disease models.
Huntington's disease (HD) is a genetic disease caused by a CAG trinucleotide repeat expansion encoding a polyglutamine tract in the huntingtin (HTT) protein, ultimately leading to neuronal loss and consequent cognitive decline and death. As no treatments for HD currently exist, several chemical screens have been performed using cell-based models of mutant HTT toxicity. These screens measured single disease-related endpoints, such as cell death, but had low 'hit rates' and limited dimensionality for therapeutic detection. Here, we have employed gene expression microarray analysis of HD samples--a snapshot of the expression of 25,000 genes--to define a gene expression signature for HD from publically available data. We used this information to mine a database for chemicals positively and negatively correlated to the HD gene expression signature using the Connectivity Map, a tool for comparing large sets of gene expression patterns. Chemicals with negatively correlated expression profiles were highly enriched for protective characteristics against mutant HTT fragment toxicity in in vitro and in vivo models. This study demonstrates the potential of using gene expression to mine chemical activity, guide chemical screening, and detect potential novel therapeutic compounds. Single-endpoint chemical screens have low therapeutic discovery hit-rates. In the context of HD, we guided a chemical screen using gene expression data. The resulting chemicals were highly enriched for suppressors of mutant HTT fragment toxicity. This study provides a proof of concept for wider usage in all chemical screening.


Self-organization of distributedly represented multiple behavior schemata in a mirror system: reviews of robot experiments using RNNPB.
The current paper reviews a connectionist model, the recurrent neural network with parametric biases (RNNPB), in which multiple behavior schemata can be learned by the network in a distributed manner. The parametric biases in the network play an essential role in both generating and recognizing behavior patterns. They act as a mirror system by means of self-organizing adequate memory structures. Three different robot experiments are reviewed: robot and user interactions; learning and generating different types of dynamic patterns; and linguistic-behavior binding. The hallmark of this study is explaining how self-organizing internal structures can contribute to generalization in learning, and diversity in behavior generation, in the proposed distributed representation scheme.


Cosmetic side effects of antiepileptic drugs in adults with epilepsy.
Cosmetic side effects (CSEs) such as weight gain and alopecia are common, undesirable effects associated with several AEDs. The objective of the study was to compare the CSE profiles in a large specialty practice-based sample of patients taking both older and newer AEDs. As part of the Columbia and Yale AED Database Project, we reviewed patient records including demographics, medical history, AED use, and side effects for 1903 adult patients (≥16years of age) newly started on an AED. Cosmetic side effects were determined by patient or physician report in the medical record and included acne, gingival hyperplasia, hair loss, hirsutism, and weight gain. We compared the overall rate of CSEs and intolerable CSEs (ICSEs-CSEs that led to dosage reduction or discontinuation) between different AEDs in both monotherapy and polytherapy. Overall, CSEs occurred in 110/1903 (5.8%) patients and led to intolerability in 70/1903 (3.7%) patients. Weight gain was the most commonly reported CSE (68/1903, 3.6%) and led to intolerability in 63 (3.3%) patients. Alopecia was the second most common patient-reported CSE (36/1903, 1.9%) and was intolerable in 33/1903 (1.7%) patients. Risk factors for CSEs included female sex (7.0% vs. 4.3% in males; p<0.05) and any prior CSE (37% vs. 2.9% in patients without prior CSE; p<0.001). Significantly more CSEs were attributed to valproic acid (59/270; 21.9%; p<0.001) and pregabalin (14/143; 9.8%; p<0.001) than to all other AEDs. Significantly less CSEs were attributed to levetiracetam (7/524; 1.3%; p=0.002). Weight gain was most frequently associated with valproic acid (35/270; 13.0%; p<0.001) and pregabalin (12/143; 8.4%; p<0.001). Hair loss was most commonly reported among patients taking valproic acid (24/270; 8.9%; p<0.001). Finally, gingival hyperplasia was most commonly reported in patients taking phenytoin (10/404; 2.5%; p<0.001). Cosmetic side effects leading to dosage change or discontinuation occurred most frequently with pregabalin and valproic acid compared with all other AEDs (13.3 and 5.6% vs. 2.3%; p<0.001). For patients who had been on an AED in monotherapy (n=677), CSEs and ICSEs were still more likely to be attributed to valproic acid (30.2% and 17.1%, respectively) than to any other AED (both p<0.001). Weight gain and alopecia were the most common patient-reported CSEs in this study, and weight gain was the most likely cosmetic side effect to result in dosage adjustment or medication discontinuation. Particular attention should be paid to pregabalin, phenytoin, and valproic acid when considering cosmetic side effects. Female patients and patients who have had prior CSE(s) to AED(s) were more likely to report CSEs. Knowledge of specific CSE rates for each AED found in this study may be useful in clinical practice.


Prospective trial of a predictive algorithm to transplant cadaver kidneys into highly sensitized patients.
The difficulty of transplanting sensitized patients increases proportionally to the panel reactive antibody (PRA) titer. Because of the high likelihood of a positive final crossmatch, these patients are often excluded from a prospective transplant unless there is a 0 HLA-A, -B, -DR mismatch. To address this problem, we developed a computerized algorithm, termed the Kentucky Antibody Testing System (KATS), that predicts class I HLA antigens that would be both "unacceptable" and "acceptable" to the recipient. This report describes the results of a prospective trial among voluntarily participating centers that agreed to share kidneys based on the KATS predictions for patients whose PRA exceeded 40%. The results of three antibody screens on each patient were compared with the HLA phenotypes of the cells in the panel in 2x2 tables with calculation of chi-square and correlation coefficient statistics. Private, broad, and public antigens were identified and a list of acceptable and unacceptable antigens were entered into the UNOS computer for each patient listed in the KATS sharing algorithm. Of the total 418 patients meeting the inclusion/exclusion criteria, the largest single group submitted was Black-not-of-Hispanic-origin females. The mean PRA of the patients was 72%. The first transplant via KATS allocation was performed on March 8, 1997. Between that time and the last transplant on July 31, 2000, 145 kidneys were offered to the participating centers and 48 transplants were performed. Of the many reasons listed for not accepting an offer or not transplanting the shared kidney into its intended recipient, only two occurred because of a positive T cell crossmatch and six because of a positive B cell crossmatch. As compared to all other high PRA patients within Southeastern Organ Procurement Foundation who were transplanted during the study period, they were more likely to be non-Caucasian, to be less well matched for private HLA-A, B, and DR antigens, and to have waited for a longer time than the other groups. Although there was a higher incidence of delayed graft function, there was no significant difference in cold ischemia, rejection episodes, or patient or graft survival. We conclude that KATS, or some other system to prospectively identify a list of acceptable and unacceptable HLA antigens, could improve the access of highly sensitized patients to a successful kidney transplant.


Context-dependent regulation of cutaneous immunological responses by TGFbeta1 and its role in skin carcinogenesis.
Transforming growth factor beta1 (TGFbeta1) signaling plays a critical role in skin carcinogenesis. While most studies have focused on TGFbeta1 signaling and response in keratinocytes, it is now becoming clear that the interaction of keratinocyte-derived TGFbeta1 with cells of the immune system has an equally important role in tumor development. Tumors form within the context of innate and adaptive immune responses and studies in skin and skin carcinogenesis models have provided important insight into the impact of context-dependent pro-inflammatory and immunosuppressive actions of TGFbeta1 on tumor development. Indeed, the paradigm of TGFbeta1 duality is clearly evident in its ability to both promote and inhibit inflammatory responses. Recent studies have begun to shed new light on the molecular basis for these actions and to provide insight into how these may contribute to context-dependent effects of TGFbeta1 on carcinogenesis in the skin and other epithelial tissues.


[Detection of DNA human cytomegalovirus of a molecular methods: hybrid capture DNA CMV by immunocompromised].
Human cytomegalovirus (HCMV), a member of the beta-virus herpes family, is a ubiquitous human pathogen. After a primary infection, HCMV establishes life latency. HCMV rarely causes symptomatic disease in an immunocompetent host, however, it is a major cause of infectious morbidity and mortality in immunocompromised individuals and developing fetuses. The HCMV genome consists of 240 kbp of double stranded DNA. Early diagnosis molecular of CMV infection is important. The objective of this study was to develop a molecular methods: Quantitative Hybrid capture for the detection of DNA CMV. We present results for 200 immunocompromised collected from 1999 to 2003 (122 men and 78 women, whom mean age was 35 years). Our results showed that 25% of women and 36% of men were positif for hybrid capture DNA CMV. This simple test (cold probe) provide quantitative and fast results. Also the efficacity of anti-CMV therapy can be followed. More over, in contrary with pp65-antigenemia assay and CMV PCR, this test can be managed on biopsy sample.


Involvement of somatostatin in breathing control before and after birth, and in perinatal and infant sudden unexplained death.
The distribution of the somatostatin was studied by immunohistochemistry on serial sections of the 56 brain stems from subjects aged from 30 gestational weeks to 12 postnatal months, dying of both known and unknown causes. The unexplained deaths included 13 sudden intrauterine deaths, 4 sudden neonatal deaths and 24 sudden infant deaths. We observed intense somatostatin positivity in the cell bodies and fibres of many brainstem nuclei prevalently involved in the respiratory activity (parabrachial/Kölliker-Fuse complex, locus coeruleus, hypoglossus nucleus, dorsal vagus motor nucleus, tractus solitarii nucleus, ambiguus nucleus, and reticular formation) in stillbirths. Only in 8 foetuses with unexplained death the hypoglossus nucleus was somatostatin-negative. In the postnatal deaths, the immunopositivity was prevalently limited to the ventrolateral and ventral subnuclei of the tractus solitarii nucleus. In 13 sudden infant death victims and in one case of death due to pneumonia, somatostatin-positivity was also present in the hypoglossus nucleus. We concluded that: 1) the somatostatin is an important foetal breathing-inhibitor, but it becomes important for the physiological control of respiration immediately after delivery; 2) functional alterations of the hypoglossal nucleus can occur in both sudden perinatal and infant deaths and contribute to the induction of both fatal breathing movements in foetuses and abnormal ventilatory control in infants leading to irreversible apneic phenomena.


3site Multisubstrate-Bound State of Cytochrome P450cam.
We identified a multisubstrate-bound state, hereby referred as a 3site state, in cytochrome P450cam via integrating molecular dynamics simulation with nuclear magnetic resonance (NMR) pseudocontact shift measurements. The 3site state is a result of simultaneous binding of three camphor molecules in three locations around P450cam: (a) in a well-established "catalytic" site near heme, (b) in a kink-separated "waiting" site along channel-1, and (c) in a previously reported "allosteric" site at E, F, G, and H helical junctions. These three spatially distinct binding modes in the 3site state mutually communicate with each other via homotropic allostery and act cooperatively to render P450cam functional. The 3site state shows a significantly superior fit with NMR pseudo contact shift (PCS) data with a Q-score of 0.045 than previously known bound states and consists of D251 free of salt-bridges with K178 and R186, rendering the enzyme functionally primed. To date, none of the reported cocomplex of P450cam with its redox partner putidaredoxin (pdx) has been able to match solution NMR data and controversial pdx-induced opening of P450cam's channel-1 remains a matter of recurrent discourse. In this regard, inclusion of pdx to the 3site state is able to perfectly fit the NMR PCS measurement with a Q-score of 0.08 and disfavors the pdx-induced opening of channel-1, reconciling previously unexplained remarkably fast hydroxylation kinetics with a koff of 10.2 s-1. Together, our findings hint that previous experimental observations may have inadvertently captured the 3site state as an in vitro solution state, instead of the catalytic state alone, and provided a distinct departure from the conventional understanding of cytochrome P450.


Structure-Based Model of RNA Pseudoknot Captures Magnesium-Dependent Folding Thermodynamics.
We develop a simple, coarse-grained approach for simulating the folding of the Beet Western Yellow Virus (BWYV) pseudoknot toward the goal of creating a transferable model that can be used to study other small RNA molecules. This approach combines a structure-based model (SBM) of RNA with an electrostatic scheme that has previously been shown to correctly reproduce ionic condensation in the native basin. Mg2+ ions are represented explicitly, directly incorporating ion-ion correlations into the system, and K+ is represented implicitly, through the mean-field generalized Manning counterion condensation theory. Combining the electrostatic scheme with a SBM enables the electrostatic scheme to be tested beyond the native basin. We calibrate the SBM to reproduce experimental BWYV unfolding data by eliminating overstabilizing backbone interactions from the molecular contact map and by strengthening base pairing and stacking contacts relative to other native contacts, consistent with the experimental observation that relative helical stabilities are central determinants of the RNA unfolding sequence. We find that this approach quantitatively captures the Mg2+ dependence of the folding temperature and generates intermediate states that better approximate those revealed by experiment. Finally, we examine how our model captures Mg2+ condensation about the BWYV pseudoknot and a U-tail variant, for which the nine 3' end nucleotides are replaced with uracils, and find our results to be consistent with experimental condensation measurements. This approach can be easily transferred to other RNA molecules by eliminating and strengthening the same classes of contacts in the SBM and including generalized Manning counterion condensation.


Radiofrequency ablation versus surgical resection for the treatment of hepatocellular carcinoma conforming to the Milan criteria: systemic review and meta-analysis.
Radiofrequency ablation (RFA) is a promising ablation technique and has become one of the best alternatives for hepatocellular carcinoma (HCC) patients. But whether RFA or surgical resection (SR) is the better treatment for HCC conforming to the Milan criteria has long been debated. A meta-analysis of trials that compared RFA versus SR was conducted regarding the survival rate and recurrence rate. Pooled odds ratios (OR) with 95% confidence intervals (95% CI) were calculated using fixed or random effects models. Nineteen studies, comprising 2 randomized controlled trials and 17 non-randomized controlled trials, were included with a total of 2895 patients. The 5 years overall survival rate for SR group was significantly higher than that for RFA group. In the SR group, the local recurrence rate was significantly lower when compared with the RFA group. This meta-analysis yielded no significant differences between laparoscopic RFA and SR in 5-year overall survival rate. In conclusion, surgical resection remains the better choice of treatment for HCC conforming to the Milan criteria, whereas RFA should be considered as an effective alternative treatment when surgery is not feasible. As for RFA technique, laparoscopic approach may be more effective than percutaneous approach for HCC conforming to Milan criteria.


The LloydRelaxer: An Approach to Minimize Scaling Effects for Multivariate Projections.
Star Coordinates are a popular projection technique in order to analyze and to disclose characteristic patterns of multidimensional data. Unfortunately, the shape, appearance, and distribution of such patterns are strongly affected by the given scaling of the data and can mislead the projection-based data analysis. In an extreme case, patterns might be more related to the choice of scaling than to the data themselves. Thus, we present the LloydRelaxer : a tool to minimize scaling-based effects in Star Coordinates. Our algorithm enforces a scaling configuration for which the data explains the observed patterns better than any scaling of them could do. It does so by an iterative minimizing and optimization process based on Voronoi diagrams and on the Lloyd relaxation within the projection space. We evaluate and test our approach by real benchmark multidimensional data of the UCI data repository.


Influence of abutment shape on peri-implant tissue conditions: A randomized clinical trial.
To analyze the influence of 3-mm high abutments with different shapes (cylindrical abutment vs. wide abutment) on marginal bone-level changes (bone loss and bone remodeling). The influence of abutment shape on implant success, probing pocket depth (PPD), and bleeding on probing (BoP) was studied as secondary objectives. Patients with a partially edentulous area requiring fixed dental prostheses by two implants in the posterior mandible or maxilla were included. The implants were 1 mm subcrestally placed, and osseointegration healing was submerged. Three-mm high abutments with two different shapes were randomly placed in second-stage surgery: cylindrical abutments (cylindrical group) and wide abutments (wide group). Marginal bone-level changes were measured using parallelized periapical radiographs at abutment placement, at definitive prosthesis placement, and at 1, 3, 6, and 12 months after loading. PPD and BoP were likewise measured at the control visits. Sixty-four dental implants in 25 patients were included. Statistically significant differences were found in bone-level changes. The cylindrical group exhibited less mean marginal bone remodeling (MBR) and marginal bone loss (MBL) than the wide group (p < .05). Moreover, the cylindrical group showed significantly less BoP (p < .05). Abutment shape had a significant influence upon marginal bone-level changes during the first 12 months. Cylindrical abutments caused less MBR and MBL than wide abutments. More clinical studies involving longer follow-ups and analyzing other abutment modifications are needed to improve our understanding of how abutments can affect peri-implant tissue stability.


Antifungal and antibacterial activity of densely dispersed silver nanospheres with homogeneity size which synthesized using chicory: An in vitro study.
With increase in isolation of multi and extensive drug resistance hospital pathogens (MDR, XDR) in burn centers of many hospitals in the world, attempt to use nanomaterials for treatment of burn-infected patients is the focus of researches all around the world. In the present investigation silver nanospheres (Ag NSs) has been synthesized by chicory seed exudates (CSE). The various parameters influencing the mechanism of Ag NSs synthesis including temperature, concentration, pH and time were studied. Greener Ag NSs were formed when the reaction conditions were altered with respect to pH, concentration of AgNO3 and incubation temperature. Finally, we evaluated antimicrobial activity of silver nanospheres biosynthesized by chicory (Cichodrium intybus) against most prevalent burn bacteria pathogens Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, and fungus Fusarium solani. The UV visible spectroscopy, X-Ray diffraction (XRD), dynamic light scattering (DLS) used for primary screening of physicochemical properties. The transmission electron microscopy (TEM) images showed the Ag NSs (with globular shape) with a size less than 25nm that they have the same size about 8nm (more than 97% are 8nm). Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Ag NSs against the standard strains of A. baumannii, P. aeruginosa and K. pneumonia showed a relatively high inhibitory and bactericidal activity (MIC 1.56μg/mL and MBC 3.12μg/mL) of the nanoparticles and F. solani cultures. In antifungal tests, the lowest level of zone of inhibition was observed at a concentration of 5μg/mL synthesized silver nanospheres with the 7% inhibition of growth. Ag NSs have high antimicrobial activity against three common burn bacteria pathogens and fungus F. solani. Therefore, Ag NSs can be used to prevent burn infection and for wound healing.


Follow-up of serotype distribution and antimicrobial susceptibility of Streptococcus pneumoniae in child carriage after a PCV13-to-PCV10 vaccine switch in Belgium.
A three year pneumococcal carriage study was set up in Belgium when the vaccination programme switched from a 13-valent (PCV13) to a 10-valent (PCV10) vaccine. We compared the first follow-up period (October 2016 - June 2017, year 2, Y2) for nasopharyngeal carriage, serotype distribution and antimicrobial susceptibility of S. pneumoniae with the baseline (January-July 2016, year 1, Y1). A single nasopharyngeal swab was taken in children (6-30 months), either attending one of the 112 day-care centres (DCCs), or visiting one of the 21 physicians for an acute otitis media (AOM). S. pneumoniae were cultured, screened for antimicrobial susceptibility, and serotyped. In Y2, 1218 samples were collected. The majority of the Y2-children (>85%) was vaccinated appropriately for their age. Children in Y2 received either PCV13 only (DCC: 23.5%; AOM: 24.6%), PCV10 only (DCC: 29.8%; AOM: 37.7%), or a mix of both vaccines (DCC: 31.9%; AOM: 25.4%). Pneumococcal carriage rates were high (Y2, DCC: 68.2%; AOM: 64.8%). Among carriers, prevalence of PCV13 serotypes was low (Y2 vs Y1, DCC: 3.5% vs 5.4%; AOM: 7.6% vs 7.7%). Although prevalence of PCV13-non-PCV10 serotypes did not increase significantly compared to Y1 (Y2 vs Y1, DCC: 1.6% vs 0.9%; Y2 vs Y1, AOM: 5.1% vs 0.0%), the proportion of serotypes 3, 6A, 19A among PCV13 serotype carriers in DCC was significantly higher in Y2 (46.2% vs Y1: 16.0%, p-value = 0.034). Serotypes 23B and 15B were the predominant non-vaccine serotypes (Y2). Among detected strains, non-susceptibility to at least one of five antibiotics tested (penicillin, tetracycline, erythromycin, levofloxacin, cotrimoxazole) was comparable to Y1 (Y2 vs Y1, DCC: 41.3% vs 42.4%; AOM: 49.4% vs 48.1%). After completion of the PCV13-to-PCV10 vaccine switch in Belgium, the proportion of PCV13-non-PCV10 serotypes (mainly 19A) significantly increased among PCV13 serotype carriers in DCC, stressing the need for strengthened surveillance as the PCV10-vaccinated population grows.


Clinical classification and prognosis of isolated right-sided infective endocarditis.
From an epidemiologic point of view, right-sided infective endocarditis (RSIE) affects different types of patients: intravenous drug users (IDUs), cardiac device carriers (pacemakers and implantable automatic defibrillators), and the "3 noes" endocarditis group: no left-sided, no IDUs, no cardiac devices. Our objective is to describe and compare the clinical profile and outcome of these groups of patients. Every episode of infective endocarditis (IE) consecutively diagnosed in 3 tertiary centers from 1996 to 2012 was included in an ongoing multipurpose database. We assessed 85 epidemiologic, clinical, echocardiographic, and outcome variables in patients with isolated RSIE. A bivariated comparative analysis between the 3 groups was conducted.Among 866 IE episodes, 121 were classified as isolated RSIE (14%): 36 IDUs (30%), 65 cardiac device carriers (54%), and 20 "3 noes" group (16%). IDUs were mainly young men (36 ± 7 years) without previous heart disease, few comorbidities, and frequent previous endocarditis episodes (28%). Human immunodeficiency virus infection was frequent (69%). Cardiac device carriers were older (66 ± 15 years) and had less comorbidities (34%). Removal of the infected device was performed in 91% of the patients without any death. The "3 noes" endocarditis group was composed mainly by middle-age men (56 ± 18 years), health care related infections (50%), and had many comorbidities (75%). Whereas Staphylococcus aureus were the most frequent cause in IDUs (72% vs 34% in device carriers and 34% in the "3 noes" group, P = 0.001), coagulase negative Staphylococci predominated in the device carriers (58% vs 11% in drug users and 35% in the "3 noes", P < 0.001). Significant differences in mortality were found (17% in drug users, 3% in device carriers, and 30% in the "3 noes" group; P < 0.001). These results suggest that RSIE should be separated into 3 groups (IDUs, cardiac device carriers, and the "3 noes") and considered as independent entities as there are relevant epidemiologic, clinical, microbiological, echocardiographic, and prognostic differences among them.


Core structure of S2 from the human coronavirus NL63 spike glycoprotein.
Human coronavirus NL63 (HCoV-NL63) has recently been identified as a causative agent of acute respiratory tract illnesses in infants and young children. The HCoV-NL63 spike (S) protein mediates virion attachment to cells and subsequent fusion of the viral and cellular membranes. This viral entry process is a primary target for vaccine and drug development. HCoV-NL63 S is expressed as a single-chain glycoprotein and consists of an N-terminal receptor-binding domain (S1) and a C-terminal transmembrane fusion domain (S2). The latter contains two highly conserved heptad-repeat (HR) sequences that are each extended by 14 amino acids relative to those of the SARS coronavirus or the prototypic murine coronavirus, mouse hepatitis virus. Limited proteolysis studies of the HCoV-NL63 S2 fusion core identify an alpha-helical domain composed of a trimer of the HR segments N57 and C42. The crystal structure of this complex reveals three C42 helices entwined in an oblique and antiparallel manner around a central triple-stranded coiled coil formed by three N57 helices. The overall geometry comprises distinctive high-affinity conformations of interacting cross-sectional layers of the six helices. As a result, this structure is unusually stable, with an apparent melting temperature of 78 degrees C in the presence of the denaturant guanidine hydrochloride at 5 M concentration. The extended HR regions may therefore be required to prime the group 1 S glycoproteins for their fusion-activating conformational changes during viral entry. Our results provide an initial basis for understanding an intriguing interplay between the presence or absence of proteolytic maturation among the coronavirus groups and the membrane fusion activity of their S glycoproteins. This study also suggests a potential strategy for the development of improved HCoV-NL63 fusion inhibitors.


Dental Pulp Stem Cells for Bone Tissue Engineering: A Literature Review.
Bone tissue engineering (BTE) is a promising approach for repairing and regenerating damaged bone tissue, using stem cells and scaffold structures. Among various stem cell sources, dental pulp stem cells (DPSCs) have emerged as a potential candidate due to their multipotential capabilities, ability to undergo osteogenic differentiation, low immunogenicity, and ease of isolation. This article reviews the biological characteristics of DPSCs, their potential for BTE, and the underlying transcription factors and signaling pathways involved in osteogenic differentiation; it also highlights the application of DPSCs in inducing scaffold tissues for bone regeneration and summarizes animal and clinical studies conducted in this field. This review demonstrates the potential of DPSC-based BTE for effective bone repair and regeneration, with implications for clinical translation.


Immunohistochemical analysis of nm23/NDP kinase expression in human lung adenocarcinoma: association with tumor progression in Clara cell type.
Levels of nm23-H1/nucleoside diphosphate kinase (NDP kinase) expression have been reported to correlate inversely with metastatic potential in some tumors but not in others. Whether or not nm23 gene product is associated with metastatic potential in lung cancer is not clear as yet. We therefore immunohistochemically examined the expression of nm23 gene products in primary lung adenocarcinomas according to cytologic subtypes in order to clarify the association of its expression with the clinical features of the disease. Seventy-two (64.9%) of the 111 lung adenocarcinomas were positive for nm23 protein. In lung adenocarcinoma of Clara cell type, high levels of nm23 expression were associated with advanced pathologic stage, positive lymph node status, and poorer prognosis (P < 0.05). However, no correlation with clinical outcome was observed in other cell types. Our data suggest that higher levels of nm23 expression are associated with tumor progression in lung adenocarcinoma of Clara cell type.


Use of Intrawound Antibiotics in Orthopaedic Surgery.
Intrawound antibiotics have been used in orthopaedic surgery procedures to prevent the development of surgical site infections. Local delivery of antibiotic powder has demonstrated a notable reduction in surgical site infection in rabbit and rodent in vivo models, as well as in other surgical fields, including vascular, colorectal, cardiothoracic, and dermatologic surgery. Intrawound antibiotic powder has been used in many orthopaedic applications, including spine surgery, total joint arthroplasty, trauma, foot and ankle reconstruction, and elbow surgery. Although the theory behind the use of intrawound antibiotic powder is promising, it has potential adverse effects, including antibiotic resistance, circulatory collapse, and decreased bone healing. In addition, most studies in the orthopaedic literature on the use of intrawound antibiotic powder are retrospective in nature.


Rural-urban differences in social and emotional protective factors and their association with child health and flourishing.
Children in rural communities experience higher mortality rates and less access to health care services than those in urban communities. Protective factors like social support also vary by geography, but their contribution to differences in child health is understudied. Understanding geographic variation in protective health factors could provide insight into their impact on health and guide future intervention strategies. To examine protective factors' association with child flourishing and child health status in rural and urban communities. Publicly available data from the National Survey of Children's Health, 2018-2021, with nonmissing geographic data (N = 150,493) were used to assess the relationship between protective factors and child flourishing and health by rurality. Multivariate survey-weighted probit models examined these relationships, adjusting for child and caregiver characteristics. More than a third of children were not flourishing, according to parental report. An estimated 62% of rural children were flourishing compared to 66% of urban children (P<0.001). Urban caregivers also were more likely to report better adult mental and physical health status. Nevertheless, rural children were reported to receive more social support than urban children, while their caregivers reported more emotional support and living in supportive and safe neighborhoods (P<0.001). Rural caregivers reported more support from places of worship and less from counselors/other mental health care providers than urban caregivers. Despite higher reported caregiver emotional support and child social support, fewer rural children are flourishing. Health systems and community organizations able to leverage these existing social and emotional protective factors in rural communities could help close this gap.


Sarcocystis suihominis detected for the first time from pigs in Japan.
Sarcoystis suihominis was detected for the first time in Japan from the heart and diaphragm of 5 out 600 older culled breeding pigs slaughtered in Saitama Prefecture, Japan. Fresh cysts were 1,080-2,040 x 106-170 microns in size. Bradysoites measured 15 x 4 microns on average. The cyst wall was usually observed thick, 4-6 microns, and striated, but occasionally thin and smooth according to the difference in sectioning angle and in portion of cysts. Scanning electron microscopy showed that many palisade-like villar protrusions, 6-6 x 0.3-0.5 microns in size, were closely folded onto the surface of cyst. A small number of microtubules were seen in the core of protrusion. No dogs nor domestic cats fed with 20 fresh cysts each excreted oocysts or sporocysts in the feces throughout the experimental period of 30 days.


[Surgical treatment of otosclerosis].
A significant step forward in otosclerosis surgery was made in 1956 with the advent of stapedectomy. This led to a significant reduction in surgical complications and to a high level of patient satisfaction. Hearing aids are the alternative to surgery, and have themselves undergone considerable technical improvements. In advanced otosclerosis, cochlear implants can improve hearing when stapes surgery and a conventional hearing aid are inadequate. These advances are modifying the surgical indications.


Indocyanine green near-infrared lymphangiography for evaluation of effectiveness of edema fluid flow under therapeutic compression.
The commonly used modalities for therapy of limb lymphedema are manual lymphatic drainage, manual devices moving edema fluid and intermittent pneumatic compression (IPC). What seems to be necessary for validation of the effect of the compression procedure is imaging of the mobilized moving edema fluid. Picture of edema fluid flow would allow the therapist to use force adjusted to the tissue volume and stiffness differing in various limb regions as well as identify sites of abundant accumulation of fluid requiring more compression. The purpose of the present study was to visualize tissue edema fluid flow during manual drainage, Linforoll massage, IPC and bandaging. To obtain data how high compression pressures should be used to mobilize indocyanine green (ICG)-stained fluid, concomitantly tissue fluid pressure measurements were performed. The following observations were obtained: (1) the possibility of real-time observation of edema fluid movement using various compression modalities, (2) the threshold pressures necessary to move edema fluid to be over 80 mm Hg in the compression device and over 40 mm Hg in the tissue fluid and (3) inefficacy of compression in some cases despite applying high compression force. These observations point to the need of ICG lymphangiography before compression therapy in each patient. The images observed during the compression procedure give an insight into the distribution of edema fluid, sites of its accumulation and efficacy of applied external force on fluid mobilization.


[Combination treatment of atrial arrhythmias with quinidine-verapamil].
Between 1977 and 1979, a combined antiarrhythmic therapy with Quinidine and Verapamil was used to treat 66 patients (aged 16 to 69 years) with chronic atrial arrhythmias of different etiology. In 60% of the 43 cases with atrial fibrillation the combined therapy was effective in reversing atrial fibrillation to regular sinus rhythm. The frequency of successful responses to Quinidine - Verapamil amounted to more than twice compared with earlier therapeutic results with Quinidine only. In 65% of the 23 cases with atrial extrasystoles or atrial tachycardias a significant reduction of the ectopics could be achieved or a regular sinus rhythm could be established. Thus, the combined therapy with Quinidine - Verapamil is very effective in abolishing chronic atrial arrhythmias, especially atrial fibrillation.


Isolation and characterization of I(Kr) in cardiac myocytes by Cs+ permeation.
Isolation of the rapidly activating delayed rectifier potassium current (I(Kr)) from other cardiac currents has been a difficult task for quantitative study of this current. The present study was designed to separate I(Kr) using Cs+ in cardiac myocytes. Cs+ have been known to block a variety of K+ channels, including many of those involved in the cardiac action potential such as inward rectifier potassium current I(K1) and the transient outward potassium current I(to). However, under isotonic Cs+ conditions (135 mM Cs+), a significant membrane current was recorded in isolated rabbit ventricular myocytes. This current displayed the voltage-dependent onset of and recovery from inactivation that are characteristic to I(Kr). Consistently, the current was selectively inhibited by the specific I(Kr) blockers. The biophysical and pharmacological properties of the Cs+-carried human ether-a-go-go-related gene (hERG) current were very similar to those of the Cs+-carried I(Kr) in ventricular myocytes. The primary sequence of the selectivity filter in hERG was in part responsible for the Cs+ permeability, which was lost when the sequence was changed from GFG to GYG, characteristic of other, Cs+-impermeable K+ channels. Thus the unique high Cs+ permeability in I(Kr) channels provides an effective way to isolate I(Kr) current. Although the biophysical and pharmacological properties of the Cs+-carried I(Kr) are different from those of the K+-carried I(Kr), such an assay enables I(Kr) current to be recorded at a level that is large enough and sufficiently robust to evaluate any I(Kr) alterations in native tissues in response to physiological or pathological changes. It is particularly useful for exploring the role of reduction of I(Kr) in arrhythmias associated with heart failure and long QT syndrome due to the reduced hERG channel membrane expression.


Radiation-associated colon cancer: A case report.
Radiation-associated colon cancer is a rare clinical entity. We herein describe the case of a patient with radiation-associated colon cancer who had undergone low anterior resection for rectal cancer following preoperative radiotherapy. Certain characteristics of radiation-associated colon cancer are highlighted. The patient was a 48-year-old man who had undergone low anterior resection for rectal cancer following preoperative radiotherapy at a total dose of 50 Gy, at the age of 29 years. When the patient presented at the University of Tokyo Hospital, 19 years after the surgery, he complained of severe anal pain and frequent defecation. Colonoscopy revealed two flat tumors in the sigmoid colon, located 10 cm to the oral side of the anastomosis site, which were diagnosed as well-differentiated adenocarcinomas. In addition, colonoscopy identified five flat polyps near the tumors, which were resected endoscopically. Computed tomography and magnetic resonance imaging revealed a mass in the sigmoid colon and no evidence of distant metastasis. Laparoscopic-assisted intersphincteric resection of the rectum and sigmoid colon with diverting ileostomy was performed. There were no specific postoperative complications and the patient was discharged from the hospital on the 20th postoperative day. On pathological examination, the resected rectum and sigmoid colon contained two separate tumors and six flat polyps. The two tumors were diagnosed as well-differentiated adenocarcinomas with invasion of the subserosa and submucosa, respectively. A total of 17 regional lymph nodes without metastasis were resected. The six flat polyps were diagnosed as tubular adenomas. We herein present a case of a radiation-associated colon cancer in a patient who had undergone low anterior resection for rectal cancer following preoperative radiotherapy 19 years prior. Colonoscopic surveillance of radiation-associated colon cancer may be indicated for rectal cancer patients treated with preoperative radiotherapy, particularly for those with long-standing radiation-induced colitis.


Antimicrobial activities of polyhexamethylene guanidine hydrochloride-based disinfectant against fungi isolated from cocoa beans and reference strains of bacteria.
This study was conducted to assess the antibacterial and the antifungal activity of a polyhexamethylene guanidine hydrochloride (PHMGH)-based disinfectant and to determine if it could be used as a disinfectant for the treatment of cocoa beans. The activity of PHMGH was tested in vitro for efficacy against five reference strains of pathogenic bacteria and six strains of fungi isolated from cocoa beans. All the strains tested were sensitive to the disinfectant. The MICs reported were between 0.01 and 1.9 mg/ml and equal to the MBC or minimum fungicidal concentration (MFC) regardless of the strains of those microorganisms. The bacteria were more sensitive to PHMGH than were the fungi. Enterobacter cloacae was the most sensitive bacterium with a MIC and MBC of 0.01 mg/ml, whereas the genus Aspergillus was the least susceptible of the microorganisms tested, with a MIC and MFC from 1.0 to 1.9 mg/ml. The time required for the activity of PHMGH varies from 2 min for Enterobacter cloacae to 12 min for Aspergillus tamarii and generally increases with the MBC or the MFC. Through this in vitro study, the PHMGH has been proved to be bactericidal and fungicidal on the strains studied. Hence, it could probably serve as a fungicidal disinfectant for the treatment of cocoa beans after harvesting.


WWP2 ubiquitin ligase and its isoforms: new biological insight and promising disease targets.
A number of recent papers on the WWP2 E3 ubiquitin ligase and two novel WWP2 isoforms have revealed important biological insight and disease-specific functions, and also impacted on our understanding of ubiquitin ligases in cell cycle regulation, apoptosis and differentiation. Gene knockout studies suggest a developmental role for WWP2 in chondrogenesis via mechanisms involving cartilage-specific transcription factors. Furthermore, WWP2 isoforms have been shown to selectively target oncogenic signaling pathways linked to both the pTEN tumour suppressor and the TGFβ/Smad signaling pathway. Here, it is suggested that WWP2 isoforms have now emerged as central physiological regulators as well as promising new disease targets, and that the challenge ahead is to now develop highly selective WWP2 inhibitors with utility in cartilage disease such as osteoarthritis and as new anticancer strategies.


Synthesis, characterization, and application of chiral ionic liquids and their polymers in micellar electrokinetic chromatography.
Two amino acid-derived (leucinol and N-methylpyrrolidinol) chiral ionic liquids are synthesized and characterized in both monomeric and polymeric forms. Leucinol-based chiral cationic surfactant is a room-temperature ionic liquid, and pyrrolidinol-based chiral cationic surfactant melts at 30-35 degrees C to form an ionic liquid (IL). The monomeric and polymeric ILs are thoroughly characterized to determine critical micelle concentration, aggregation number, polarity, optical rotation, and partial specific volume. Herein, we present the first enantioseparation using chiral IL as a pseudostationary phase in capillary electrophoresis. Chiral separation of two acidic analytes, (+/-)-alpha-bromophenylacetic acid and (+/-)-2-(2-chlorophenoxy)propanoic acid (+/-)-(2-PPA) can be achieved with both monomers and polymers of undecenoxycarbonyl-L-pryrrolidinol bromide (L-UCPB) and undecenoxycarbonyl-L-leucinol bromide (L-UCLB) at 25 mM surfactant concentration using phosphate buffer at pH 7.50. The chiral recognition seems to be facilitated by the extent of interaction of the acidic analytes with the cationic headgroup of chiral selectors. Polysodium N-undecenoxycarbonyl-L-leucine sulfate (poly-L-SUCLS) and polysodium N-undecenoxycarbonyl-L-leucinate (poly-L-SUCL) were compared at high and low pH for the enantioseparation of (+/-)-(2-PPA). At pH 7.5, poly-L-SUCLS, poly-L-SUCL, and (+/-)-(2-PPA) are negatively charged resulting in no enantioseparation. However, chiral separation was observed for (+/-)-(2-PPA) using poly-L-SUCLS at low pH (pH 2.00) at which the analyte is neutral. The comparison of chiral separation of anionic and cationic surfactants demonstrates that the electrostatic interaction between the acidic analyte and cationic micelle plays a profound role in enantioseparation.


Identification of TFIID components required for transcriptional activation by upstream stimulatory factor.
A TATA box-binding initiation factor, TFIID, plays a central role in the transcriptional regulation by activators. Using anti-TFIID tau (a TATA box-binding component of native TFIID) immunoaffinity chromatography, nine polypeptides (230, 110, 85, 62, 58, 42, 28, 22, and 21 kDa) were identified as native Drosophila TFIID components that are tightly associated with TFIID tau. To verify the functional activity of the purified TFIID complex, template DNA and other transcription factors were reconstituted with purified TFIID bound to the antibody-Sepharose matrix. Immobilized TFIID mediated not only basal transcription but transcriptional activation by upstream stimulatory factor (USF). On the other hand, recombinant TFIID tau immobilized on the same antibody-Sepharose matrix could not mediate activation by USF. These results suggest that one or more of these additional polypeptides are required as functional TFIID subunits for activator-dependent transcription in conjunction with TFIID tau. As further evidence of the relevance of the Drosophila TFIID components identified in this analysis, including the previously unrecognized p230 (Dynlacht, B. D., Hoey, T., and Tjian, R. (1991) Cell 66, 563-576), protein blot analysis showed that TFIID tau interacts specifically and exclusively with p230. This suggests that p230 is an integral subunit of TFIID and that it may play a major role in tethering other subunits to TFIID tau.


Establishment of an Academic Tissue Microarray Platform as a Tool for Soft Tissue Sarcoma Research.
Soft tissue sarcoma (STS) is a heterogeneous family of rare mesenchymal tumors, characterized by histopathological and molecular diversity. Tissue microarray (TMA) is a tool that allows performing research in orphan diseases in a more efficient and cost-effective way. TMAs are paraffin blocks consisting of multiple small representative tissue cores from biological samples, for example, from multiple donors, diverse sites of disease, or multiple different diseases. In 2015, we began constructing TMAs using archival tumor material from STS patients. Specimens were well annotated in terms of histopathological diagnosis, treatment, and clinical follow-up of the tissue donors. Each TMA block contains duplicate or triplicate 1.0-1.5 mm tissue cores from representative tumor areas selected by sarcoma pathologists. The construction of TMAs was performed with TMA Grand Master (3DHistech). So far, we have established disease-specific TMAs from 7 STS subtypes: gastrointestinal stromal tumor (72 cases included in the array), alveolar soft part sarcoma (n = 12 + 47), clear cell sarcoma (n = 22 + 32), leiomyosarcoma (n = 55), liposarcoma (n = 42), inflammatory myofibroblastic tumor (n = 12 + 21), and alveolar rhabdomyosarcoma (n = 24). We also constructed a multisarcoma TMA covering a representative number of important histopathological subtypes on arrays for screening purposes, namely, angiosarcoma, dedifferentiated liposarcoma, pleomorphic liposarcoma, and myxoid liposarcoma, leiomyosarcoma, malignant peripheral nerve sheath tumor, myxofibrosarcoma, rhabdomyosarcoma, synovial sarcoma, and undifferentiated pleomorphic sarcoma, with 7-11 individual cases per subtype. We are currently expanding the list of TMAs with additional sarcoma entities, considering the heterogeneity of this family of tumors. Our extensive STS TMA platform is suitable for rapid and cost-effective morphological, immunohistochemical, and molecular characterization of the tumor as well as for the identification of potential novel diagnostic markers and drug targets. It is readily available for collaborative projects with research partners.


Effects of epithelial-to-mesenchymal transition on acute stress response in human peritoneal mesothelial cells.
During peritoneal dialysis, mesothelial cells undergo epithelial-to-mesenchymal transition (EMT), resulting in markedly altered protein expression. This potentially includes heat-shock proteins (HSP), the main effectors of cellular repair. Thus, chronic cellular processes, such as EMT, may influence acute stress responses and thus survival of mesothelial cells following non-lethal injury upon exposure to peritoneal dialysis fluid (PDF). In this study, we investigated the effects of EMT on acute stress responses and cytoresistance in human peritoneal mesothelial cells. In vivo EMT was defined as a fibroblast-like growth pattern in mesothelial cells grown from peritoneal effluents, and in vitro EMT was induced by TGF-beta1 in mesothelial cells grown from omental tissue. Morphologic EMT was validated by western blot analysis of EMT marker proteins (ezrin, alpha-SMA). Expression of HSP and cellular survival was evaluated in a simple in vitro PDF exposure model. In vivo and in vitro EMT resulted in marked effects on phenotypes of mesothelial cells, associated with differential HSP expression. In vivo 'chronic' EMT resulted in lower expression of HSP-27 and HSP-72, whereas in vitro 'acute' EMT was associated with increased HSP-27 and decreased HSP-72 expression. Following PDF exposure, there were no effects of in vivo EMT on the stress induction of HSP, and survival of epithelial versus fibroblast-like phenotypes was comparable. The non-stressful induction of HSP-27 following TGF-beta1 pretreatment resulted in the attenuated stress induction of HSP, and in improved survival in following PDF exposure. Taken together, this study confirms that mesothelial cells are not 'unchanged' or 'static targets' during the clinical course of PD treatment. The cellular processes during EMT play a complex role in acute cellular stress response and cytoresistance of mesothelial cells. Sequential analysis at different stages of EMT will be essential to provide more insights on cytoprotective cellular processes in in vitro and in vivo models of PD.


Extracorporeal Membrane Oxygenation and Perfluorocarbon in a Therapy Refractory Case of Acute Respiratory Distress Syndrome.
Perfluorocarbons are oxygen-carrying, dense liquids initially intended for the use in partial or total liquid ventilation of patients with severe acute respiratory distress syndrome but which did not show beneficial effects in clinical studies. However, perfluorocarbons may be used for lung lavage in severe alveolar proteinosis. In acute respiratory distress syndrome, oxygenation may be so severely compromised that the use of nonoxygenated perfluorocarbons may not be possible. We report a case of severe, nonresolving acute respiratory distress syndrome treated with extracorporeal membrane oxygenation to secure oxygenation, using perfluorocarbon in a single instillation to aid the clearance of debris and proteinacous edema.


Regulated upon activation, normal T cell expressed and secreted (RANTES) and monocyte chemotactic protein 1 in follicular fluid accumulate differentially in patients with and without endometriosis undergoing in vitro fertilization.
To compare the amount of regulated upon activation, normal T cell expressed and secreted (RANTES) and monocyte chemotactic protein 1 (MCP-1) in follicular fluid (FF) of patients with and without endometriosis and to determine their oocyte fertilization and pregnancy rates. Case-control study. Department of Obstetrics and Gynecology, University Hospital Schleswig-Holstein, Campus Luebeck, Germany. Thirty-two women with endometriosis stages I-IV and 28 women without endometriosis, both groups surgically and histologically confirmed. Diagnostic laparoscopy, IVF-ET. RANTES and MCP-1 levels in follicular fluid, measured by ELISA, and oocyte fertilization and pregnancy rates. Follicular response and days of gonadotropin stimulation were similar between the two groups. The levels of RANTES in FF from patients with endometriosis were significantly higher (460.4 +/- 90.3 pg/mL) compared with concentrations in patients with tubal infertility (243.8 +/- 70.9 pg/mL; P<.05). In contrast, MCP-1 concentrations in FF from women with endometriosis (330.0 +/- 29.2 pg/mL) were lower than in women with tubal infertility (420.5 +/- 46.6 pg/mL; P<.05). Oocyte fertilization rates in the endometriosis group (54%) were significantly lower than those of the tubal infertility group (73%; P<.05), as were the pregnancy rates (19% vs. 35%, respectively; P<.01). Women with endometriosis-associated infertility have a poor IVF outcome. Immune cell recruitment into the ovary might affect follicular function and lead to impaired oocyte quality.


Electron transfer dissociation facilitates the measurement of deuterium incorporation into selectively labeled peptides with single residue resolution.
Mass spectrometry is routinely applied to measure the incorporation of deuterium into proteins and peptides. The exchange of labile, heteroatom-bound hydrogens is mainly used to probe the structural dynamics of proteins in solution, e.g., by hydrogen-exchange mass spectrometry, but also to study the gas-phase structure and fragmentation mechanisms of polypeptide ions. Despite considerable effort in recent years, there is no widely established mass spectrometric method to localize the incorporated deuterium to single amino acid residues, and typically, only the overall deuterium content of peptides or proteins is obtained. The main reason for this is that CID and related techniques induce intramolecular migration of hydrogens ("hydrogen scrambling") upon vibrational excitation of the even-electron precursor ion, thus randomizing the positional distribution of the incorporated deuterium atoms before fragmentation. In contrast, decomposition of radical gas-phase peptide cations upon electron capture dissociation was recently demonstrated to proceed with a very low level of amide hydrogen scrambling. Employing model peptides developed to enable sensitive detection of hydrogen scrambling, we show in the present study that electron transfer dissociation in a 3D-quadrupole ion trap retains the site-specific solution-phase deuterium incorporation pattern and allows for localization of incorporated deuterium with single residue resolution. Furthermore, we exploit this finding to monitor how collisional activation induces proton mobility in a gaseous peptide ion at various levels of vibrational excitation.


Effect of race on asthma management and outcomes in a large, integrated managed care organization.
Morbidity from asthma disproportionately affects black people. Whether this excess morbidity is fully explained by differences in asthma severity, access to care, or socioeconomic status (SES) is unknown. We assessed whether there were racial disparities in asthma management and outcomes in a managed care organization that provides uniform access to health care and then determined to what degree these disparities were explained by differences in SES, asthma severity, and asthma management. We prospectively studied 678 patients from a large, integrated health care delivery system. Patients who had been hospitalized for asthma were interviewed after discharge to ascertain information about asthma history, health status, and SES. Small-area socioeconomic data were ascertained by means of geocoding and linkage to the US Census 2000. Patients were followed up for subsequent emergency department (ED) visits or hospitalizations (median follow-up, 1.9 years). Black race was associated with a higher risk of ED visits (hazard ratio [HR], 1.93; 95% confidence interval [CI], 1.39-2.66) and hospitalizations (HR, 1.89; 95% CI, 1.30-2.76). This finding persisted after adjusting for SES and differences in asthma therapy (adjusted HR for ED visits, 1.73; 95% CI, 1.07-2.81; and adjusted HR for hospitalizations, 2.01; 95% CI, 1.33-3.02). Even in a health care setting that provides uniform access to care, black race was associated with worse asthma outcomes, including a greater risk of ED visits and hospitalizations. This association was not explained by differences in SES, asthma severity, or asthma therapy. These findings suggest that genetic differences may underlie these racial disparities.


[Dynamic pedography for assessing functional ankle joint instability].
Chronic functional instability of the lateral ankle may be difficult to distinguish from mechanical instability when radiological stress test reveal only small ligamentous defects. When a decision for or against surgical reconstruction of the ligaments has to be made it can be helpful to use additional information on joint function. Therefore, a prospective study of 65 patients (mean age 24.1 +/- 4.6 years) with long-standing chronic ankle instability was conducted to demonstrate that the dynamic measurement of plantar pressure distribution can identify patients with functional ankle instability. Plantar pressure patterns were measured during gait by means of a capacitive platform, the EMED-SF system. The impulses at eight points of the foot were calculated intraindividually and compared with those in a group of 100 healthy subjects. On the basis of clinical criteria alone, the two groups of patients were distinguished, 35 with functional instability and 30 with mechanical instability. The patients with functional instability showed significantly increased lateral loading of the unstable foot (P = 0.01), whereas the group with mechanical instability tended to walk more on the medial side of the unstable foot. This finding is explained by a deficit or peroneal strength during the stance phase, based on a proprioceptive deficit after trauma. The new technique provides additional information, which is relevant and sufficiently important to help in making decisions about the individual patient with chronic instability.


LAST Double Check - A Comprehensive Pre-Regional Checklist for the Busy Institution.
Wrong site peripheral nerve blocks are included in the National Quality Forum and Joint Commission's category of "never event." Multiple attempts have been made by various groups in an effort to eliminate these events. Prior attempts to eliminate these never events include the Regional Block Preprocedural Checklist provided by the American Society of Regional Anesthesia (ASRA) taskforce. Following a series of errors involving anticoagulation prior to regional anesthesia, our department saw a need for a more comprehensive checklist. An expert panel developed the LAST Double Check Checklist with the aim of identifying and eliminating errors associated with regional anesthesia delivery. This checklist was implemented over the course of two 30 d trial periods. Feedback was collected and any delays associated with implementation were recorded. There were no reported procedures performed on patients taking anticoagulation or reported case delays during the two 30 d trials. A total of 350 regional anesthetics were performed during both trials. During the first week of implementation, a patient was identified as having received enoxaparin, despite the electronic medical record showing the medication as held. The planned regional anesthetic was not performed given increased risk of bleeding. Feedback collected during the trial periods was incorporated into the final draft and implementation of the LAST Double Check for use in all locations where regional anesthesia is performed. There have been no post-implementation events reported (11-mo period, greater than 1,000 regional anesthetics performed). The LAST Double Check is a more comprehensive checklist with the aim of preventing errors associated with wrong site blocks, anticoagulation administration, and care team coordination. This checklist covers areas of the patient history that are routinely reviewed prior to regional anesthesia administration and did not contribute to delay in arrival to the operating room.


Motor Proficiency, Manual Dexterity, and Visual Perception in School-age Children With ADHD: Contribution of Different Comorbidities.
Attention-deficit/hyperactivity disorder (ADHD) is often comorbid with a number of other psychiatric conditions. The goal of this study was to investigate whether motor skills, manual dexterity, and visual perception differ in the presence of ADHD and comorbid psychiatric conditions in school-age boys by comparing them to a group of children with typical development (TD). We evaluated 81 boys 6 to 10 years of age in 5 groups: ADHD only (n=18); ADHD+specific learning disorders (ADHD+SLDs) (n=17); ADHD+oppositional defiant disorder (n=16); ADHD+anxiety disorders (n=15); and TD (n=15). The participants with ADHD only and those with ADHD+comorbidity had significantly lower scores than the participants in the TD group in every area we evaluated. The lowest scores were observed in the ADHD+SLD group in all motor skills domains except for the fine motor precision field. Furthermore, the group with ADHD+SLD had statistically significantly lower scores than the 3 other groups with ADHD+comorbidity in 3 areas: balance, upper extremity coordination, and speed and agility. Motor problems in patients with ADHD and comorbid conditions may not be a focus of attention in clinics. However, especially in the case of comorbid conditions accompanying ADHD, it is important to consider problems in motor skills, manual dexterity, and visual perception in evaluation and treatment to increase these children's quality of life.


Validation of parametric methods for [¹¹C]PE2I positron emission tomography.
The radioligand [(11)C]PE2I is highly selective for dopamine transporter (DAT) and can be used in vivo for investigation of changes in DAT concentration, progression of disease and validation of treatment using positron emission tomography (PET). DAT is an important protein for regulation of central dopamine concentration and DAT deficiency has been associated with several neurodegenerative and neuropsychiatric disorders. Accurate parametric images are a prerequisite for clinical application of [(11)C]PE2I. The purpose of this study was to evaluate different methods for producing [(11)C]PE2I parametric images, showing binding potential (BPND) and relative delivery (R1) at the voxel level, using clinical data as well as simulations. Investigations were made in twelve subjects either with social anxiety disorder (n=6) or parkinsonian syndrome (n=6), each receiving an 80 min dynamic PET scan. All subjects underwent a T1-weighted MRI scan which was co-registered to the PET images and used for definition of regions of interest using a probabilistic template (PVElab). Two basis function implementations (receptor parametric mapping: RPM, RPM2) of the simplified reference tissue model (SRTM) and three multilinear reference tissue models (MRTMo, MRTM and MRTM2) were used for computation of parametric BPND and R1 images. In addition, reference Logan and standard uptake value ratio (SUVr) were investigated. Evaluations of BPND and R1 images were performed using linear regression to compare the parametric methods to region-based analyses with SRTM and cerebellar gray matter as reference region. Accuracy and precision of each method were assessed by simulations. Correlation and slope of linear regression between parametric and region-based BPND and R1 values in both striatum and extra-striatal regions were optimal for RPM (R(2)=0.99 for both BPND and R1; slopes 0.99 and 0.98 for BPND and R1, respectively, in striatum). In addition, accuracy and precision were best for RPM and RPM2. The basis function methods provided more robust estimations of the parameters compared to the other models and performed best in simulations. RPM, a basis function implementation of SRTM, is the preferred method for voxel level analysis of [(11)C]PE2I PET studies.


Functional analysis of an unusual type IV pilus in the Gram-positive Streptococcus sanguinis.
Type IV pili (Tfp), which have been studied extensively in a few Gram-negative species, are the paradigm of a group of widespread and functionally versatile nano-machines. Here, we performed the most detailed molecular characterisation of Tfp in a Gram-positive bacterium. We demonstrate that the naturally competent Streptococcus sanguinis produces retractable Tfp, which like their Gram-negative counterparts can generate hundreds of piconewton of tensile force and promote intense surface-associated motility. Tfp power 'train-like' directional motion parallel to the long axis of chains of cells, leading to spreading zones around bacteria grown on plates. However, S. sanguinis Tfp are not involved in DNA uptake, which is mediated by a related but distinct nano-machine, and are unusual because they are composed of two pilins in comparable amounts, rather than one as normally seen. Whole genome sequencing identified a locus encoding all the genes involved in Tfp biology in S. sanguinis. A systematic mutational analysis revealed that Tfp biogenesis in S. sanguinis relies on a more basic machinery (only 10 components) than in Gram-negative species and that a small subset of four proteins dispensable for pilus biogenesis are essential for motility. Intriguingly, one of the piliated mutants that does not exhibit spreading retains microscopic motility but moves sideways, which suggests that the corresponding protein controls motion directionality. Besides establishing S. sanguinis as a useful new model for studying Tfp biology, these findings have important implications for our understanding of these widespread filamentous nano-machines.


CC-5013 (Celgene).
Celgene, in collaboration with the National Cancer Institute, is developing CC-5013, the lead compound in a series of thalidomide derivatives that inhibit TNFalpha overproduction, for the potential treatment of hematological and solid tumor cancers and inflammatory diseases.


S100B profiles and cognitive function at high altitude.
Exposure to high altitude can lead to acute mountain sickness (AMS) and high altitude cerebral edema (HACE). In this study we investigated the effect of high altitude on neurocognitive function and S100B release. Increased S100B release has been hypothesized to signify a loss of integrity in the blood-brain barrier (BBB). Seven healthy volunteers trekked to Capanna Regina Margherita (4554 m above sea level) in the Monte Rosa massif. During ascent and descent, five test events were undertaken; participants underwent neurocognitive testing, Lake Louise scoring (LLS), and blood sampling to measure levels of S100B. The blood tests revealed that S100B levels increased 42% to 122% from baseline, and mean LLS increased from 0.57 to 2.57. A significant correlation was observed between both S100B levels and LLS and S100B and some neurocognitive scores. The study indicates that S100B can be released by a mild hypoxia during AMS. Moreover, an observed correlation between S100B and a lower score on neurocognitive tests suggests that the pathogenetic mechanisms may be linked. The study indicates that a decline in cognitive function is associated with symptoms of AMS.


Tardive Dystonia Related with Aripiprazole.
Tardive dystonia is characterized by sustained, generally slow involuntary twisting movements. It is estimated to occur at a frequency of 1% to 4% among patients who are taking an antipsychotic agent. Unlike the first generation antipsychotics, the second generation antipsychotics are less likely to cause neuroleptic-induced movement disorder. For aripiprazole, only a few cases have been reported for tardive dystonia. We present a young male, who developed a severe tardive dystonia after taking aripiprazole for 5 years. The patient was admitted to for the treatment of both hisdystonic and psychotic symptoms. Olanzapine was administered instead of aripiprazole and while his psychotic symptoms improved, the dystonic symptoms were continued. Therefore, olanzapine was switched to clozapine while augmenting with benzodiazepine, anti-cholinergic, and ginko biloba to control his tardive dystonia. After 2 weeks of treatment, the dystonic movement decreased remarkably.


General internal medicine and geriatrics: building a foundation to improve the training of general internists in the care of older adults.
Internists--"doctors for adults"--provide most of the medical care given to older Americans, especially those with serious chronic disease. Nonetheless, the United States lacks an adequate physician workforce with mastery in caring for older persons and with expertise in building knowledge about how best to provide this care. This supplement aims to strengthen the physician workforce by fostering incremental and sustained improvements in the training of internal medicine residents in the care of older adults and in the development of geriatrics-oriented general internal medicine faculty. It identifies 3 major barriers to these improvements: lack of adequately trained teachers and mentors, the belief that explicit training in geriatrics has little to offer the generalist, and inadequate funding. Three strategies offer particular promise in overcoming these barriers: engaging directors of internal medicine residency programs, funding centers to promote collaboration between teaching and research programs in general internal medicine and geriatrics, and providing substantial incremental funding on the national level to pay for the time required to care for frail older patients and to teach and do research about this care. The barriers and strategies identified in this supplement may also inform efforts to enhance the skills of practicing physicians and improve training and faculty development in family medicine and other disciplines.


The binding of Xanthophylls to the bulk light-harvesting complex of photosystem II of higher plants. A specific requirement for carotenoids with a 3-hydroxy-beta-end group.
The pigment composition of the light-harvesting complexes (LHCs) of higher plants is highly conserved. The bulk complex (LHCIIb) binds three xanthophyll molecules in combination with chlorophyll (Chl) a and b. The structural requirements for binding xanthophylls to LHCIIb have been examined using an in vitro reconstitution procedure. Reassembly of the monomeric recombinant LHCIIb was performed using a wide range of native and nonnative xanthophylls, and a specific requirement for the presence of a hydroxy group at C-3 on a single beta-end group was identified. The presence of additional substituents (e.g. at C-4) did not interfere with xanthophyll binding, but they could not, on their own, support reassembly. cis isomers of zeaxanthin, violaxanthin, and lutein were not bound, whereas all-trans-neoxanthin and different chiral forms of lutein and zeaxanthin were incorporated into the complex. The C-3 and C-3' diols lactucaxanthin (a carotenoid native to many plant LHCs) and eschscholtzxanthin (a retro-carotenoid) both behaved very differently from lutein and zeaxanthin in that they would not support complex reassembly when used alone. Lactucaxanthin could, however, be bound when lutein was also present, and it showed a high affinity for xanthophyll binding site N1. In the presence of lutein, lactucaxanthin was readily bound to at least one lutein-binding site, suggesting that the ability to bind to the complex and initiate protein folding may be dependent on different structural features of the carotenoid molecule. The importance of carotenoid end group structure and ring-to-chain conformation around the C-6-C-7 torsion angle of the carotenoid molecule in binding and complex reassembly is discussed.


Vaccine platform recombinant measles virus.
The classic development of vaccines is lengthy, tedious, and may not necessarily be successful as demonstrated by the case of HIV. This is especially a problem for emerging pathogens that are newly introduced into the human population and carry the inherent risk of pandemic spread in a naïve population. For such situations, a considerable number of different platform technologies are under development. These are also under development for pathogens, where directly derived vaccines are regarded as too complicated or even dangerous due to the induction of inefficient or unwanted immune responses causing considerable side-effects as for dengue virus. Among platform technologies are plasmid-based DNA vaccines, RNA replicons, single-round infectious vector particles, or replicating vaccine-based vectors encoding (a) critical antigen(s) of the target pathogens. Among the latter, recombinant measles viruses derived from vaccine strains have been tested. Measles vaccines are among the most effective and safest life-attenuated vaccines known. Therefore, the development of Schwarz-, Moraten-, or AIK-C-strain derived recombinant vaccines against a wide range of mostly viral, but also bacterial pathogens was quite straightforward. These vaccines generally induce powerful humoral and cellular immune responses in appropriate animal models, i.e., transgenic mice or non-human primates. Also in the recent first clinical phase I trial, the results have been quite encouraging. The trial indicated the expected safety and efficacy also in human patients, interestingly independent from the level of prevalent anti-measles immunity before the trial. Thereby, recombinant measles vaccines expressing additional antigens are a promising platform for future vaccines.


Peregrine rogue waves induced by the interaction between a continuous wave and a soliton.
Based on the soliton solution on a continuous wave background for an integrable Hirota equation, the reduction mechanism and the characteristics of the Peregrine rogue wave in the propagation of femtosecond pulses of optical fiber are discussed. The results show that there exist two processes of the formation of the Peregrine rogue wave: one is the localized process of the continuous wave background, and the other is the reduction process of the periodization of the bright soliton. The characteristics of the Peregrine rogue wave are exhibited by strong temporal and spatial localization. Also, various initial excitations of the Peregrine rogue wave are performed and the results show that the Peregrine rogue wave can be excited by a small localized (single peak) perturbation pulse of the continuous wave background, even for the nonintegrable case. The numerical simulations show that the Peregrine rogue wave is unstable. Finally, through a realistic example, the influence of the self-frequency shift to the dynamics of the Peregrine rogue wave is discussed. The results show that in the absence of the self-frequency shift, the Peregrine rogue wave can split into several subpulses; however, when the self-frequency shift is considered, the Peregrine rogue wave no longer splits and exhibits mainly a peak changing and an increasing evolution property of the field amplitude.


Multivariate analysis as a method to understand variability in a complex excipient, and its contribution to formulation performance.
A key part of the Risk Assessment of excipients is to understand how raw material variability could (or does) contribute to differences in performance of the drug product. Here we demonstrate an approach which achieves the necessary understanding for a complex, functional, excipient. Multivariate analysis (MVA) of the certificates of analysis of an ethylcellulose aqueous dispersion (Surelease) formulation revealed low overall variability of the properties of the systems. Review of the scores plot to highlight batches manufactured using the same ethylcellulose raw material in the formulation, indicated that these batches tend to be more closely related than other randomly selected batches. This variability could result in potential differences in the quality of drug product lots made from these batches. Manufacture of a model drug product from Surelease batches coated using different lots of starting material revealed small differences in the release of a model drug, which could be detected by certain model dependent dissolution modeling techniques, but they were not observed when using model-independent techniques. This illustrates that the techniques are suitable for detecting and understanding excipient variability, but that, in this case, the product was still robust.


Rectal Lipoma in an Elderly Male: A Case Report.
Lipomas of the gastrointestinal tract are uncommon, benign non-epithelial tumors detected incidentally during surgery or endoscopy. Rectal lipomas are extremely rare. Patients may be asymptomatic or present with rectal bleeding, constipation, tenesmus and signs of intestinal obstruction. Preoperative diagnosis is challenging. Management consists of simple observation, endoscopic or laparoscopic removal, and open surgery. We present a case of an elderly male admitted to the Department of Surgery of a general hospital in Crete, complaining of a protruding rectal mass during defecation. CT raised the diagnostic suspicion. The mass was removed by trans-anal excision. Histopathology of the resected specimen confirmed the diagnosis. The patient had an uneventful postoperative course and was discharged home at the second postoperative day.


Measurement temperature plays a pivotal role in the distribution of erythrocyte deformability after LPS.
Reductions in red blood cell membrane deformability (RBC(D)) may perturb microcirculatory blood flow and impair tissue O(2)-availability. We investigated the effect of assay temperature on the distribution of RBC(D) in endotoxin (LPS) incubated and control RBCs. Fresh blood from healthy rats was incubated with and without the presence of LPS for 6 hrs. An index of red blood cell membrane deformability, delta, was measured via the micropipette aspiration technique at 25 degrees C and 37 degrees C at 0, 2 and 6 hrs of incubation. The ATP content of RBC was measured by the luciferin-luciferase technique. At 25 degrees C, LPS caused a significant decrease in mean delta after 2 and 6 hours incubation compared to controls (-10.0%, p=0.03 and -24.0%, p=0.03, respectively) characterized by a left shift in the distribution (skewness: -1.4). However, at 37 degrees C a significant decrease in delta was only detected after 6 hrs of LPS incubation (-13.8%, p=0.01, compared to -5.1%, p=0.7 at 2 hours) and lacked the left shifted distribution (skewness: 0.2). No significant difference in ATP content of RBCs was observed between groups. We have shown that LPS incubation results in a significant decrease in RBC(D) and that room temperature measurement of physical membrane properties may exaggerate the differences between normal and perturbed RBCs.


Omega-3 fatty acids in bipolar patients with a low omega-3 index and reduced heart rate variability: the "BIPO-3" trial.
Research suggests that a low omega-3 index may contribute to the low heart rate variability and the increased risk of cardiovascular morbidity and mortality in bipolar disorders. However, so far, no intervention trial with EPA and DHA has been conducted in bipolar patients attempting to increase their heart rate variability. 119 patients with bipolar disorder according to DSM-IV were screened, with 55 euthymic bipolar patients-owing to inclusion criteria (e.g. low omega-3 index (< 6%), SDNN < 60 ms.)-being enrolled in a randomized, double-blind, 12-week parallel study design with omega-3 fatty acids (4 capsules of 530 mg EPA, 150 mg DHA) or corn oil as a placebo, in addition to usual treatment. Heart rate variability as well as the omega-3 index were measured at baseline and at the endpoint of the study. A total of 42 patients (omega-3: n = 23, corn oil: n = 19) successfully completed the study after 12 weeks. There was a significant increase in the omega-3 index (value at endpoint minus value at baseline) in the omega-3 group compared to the corn oil group (p < 0.0001). However, there was no significant difference in the change of the SDNN (value at endpoint minus value at baseline) between the treatment groups (p = 0.22). In addition, no correlation between changes in SDNN and change in the omega-3 index could be detected in the omega-3 group (correlation coefficient = 0.02, p = 0.94) or the corn oil group (correlation coefficient = - 0.11, p = 0.91). Similarly, no significant differences between corn oil and omega-3 group regarding the change of LF (p = 0.19), HF (p = 0.34) and LF/HF ratio (p = 0.84) could be demonstrated. In our randomized, controlled intervention trial in euthymic bipolar patients with a low omega-3 index and reduced heart rate variability no significant effect of omega-3 fatty acids on SDNN or frequency-domain measures HF, LF and LF/HF ratio could be detected. Possible reasons include, among others, the effect of psychotropic medication present in our trial and/or the genetics of bipolar disorder itself. Further research is needed to test these hypotheses. Trial registration ClinicalTrials.gov, NCT00891826. Registered 01 May 2009-Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT00891826.


Providing Incentive for Emergency Physician X-Waiver Training: An Evaluation of Program Success and Postintervention Buprenorphine Prescribing.
Emergency department (ED) initiation of buprenorphine for patients with opioid use disorder increases treatment engagement but remains an uncommon practice. One important barrier to ED-initiated buprenorphine is the additional training requirement (X waiver). Our objective is to evaluate the influence of a financial incentive program on emergency physician completion of X-waiver training. Secondary objectives are to evaluate the program's effect on buprenorphine prescribing and to explore physician attitudes toward the incentive. We conducted a prospective, observational cohort study set in 3 urban academic EDs before and after implementation of a financial incentive program providing $750 for completion of X-waiver training. We describe program participation as well as rates of buprenorphine prescribing per opioid use disorder-related encounter before and after the intervention period, using electronic health record data. We also completed a postintervention physician survey assessing attitudes about the incentive program. Overall, 89% of eligible emergency physicians (56/63) completed the X-waiver training during the 6-week incentive period. In the 5 months after the incentive, buprenorphine prescribing per opioid use disorder-related encounter increased from 0.5% to 16% (Δ 15%; 95% confidence interval 10.6% to 19.9%), with substantial variability across sites (range 8% to 22% of opioid use disorder-related encounters). In a postintervention survey, 67% of participating physicians indicated that they would have completed the training for a lower amount. A financial incentive paying approximately half the clinical rate was effective in promoting emergency physician X-waiver training. The effect on ED-based buprenorphine prescribing was positive but variable across sites, and likely dependent on the availability of additional supports.


Afferent contributions to digit force coupling and force level variation during performance of non-lift pinch.
Afferent contributions to the coordination of thumb and index finger forces during non-lift pinch were studied using an anesthetization case study design. Two subjects, one performing with and without digital anesthetization and one with intact sensation, produced dynamic pinch forces against a stable object, with and without visual feedback. Error corrections were less frequent post-anesthetization, and the cross correlation between digit forces was lower when sensation was removed. However, this decrease in cross correlation between digit forces seemed to reflect a loss in the magnitude of tightly coupled error corrections when sensation was removed, rather than more frequent deviations of force magnitude between the digit forces. Force-time output without visual feedback lacked these error corrections, and the correlation between digit forces remained high, irrespective of sensory status. Additionally, with vision occluded, the time rate of force change did not vary in a gradual manner as would be expected from a neural representation of a sinusoidal target, but was instead marked by sudden abrupt reversals of force rate of change, invariant of somatosensory status. The coupling of digit forces and rates of force change during non-lift pinch appear to be controlled primarily with feedforward mechanisms, where the lack of proprioceptive feedback does not seem to disrupt this coupling.


Factors contributing to malnutrition in patients with Parkinson's disease.
Our objective in this study was to evaluate the nutritional status and to identify clinical, psychosocial, and nutritional factors contributing to malnutrition in Korean patients with Parkinson's disease. We used a descriptive, cross-sectional study design. Of 102 enrolled patients, 26 (25.5%) were malnourished and 27 (26.5%) were at risk of malnutrition based on Mini-Nutritional Assessment scores. Malnutrition was related to activity of daily living score, Hoehn and Yahr stage, duration of levodopa therapy, Beck Depression Inventory and Spielberger's Anxiety Inventory scores, body weight, body weight at onset of Parkinson's disease, and body mass index. On multiple logistic regression analysis, anxiety score, duration of levodopa therapy, body weight at onset of Parkinson's disease, and loss of body weight were significant factors predicting malnutrition in Parkinson's disease patients. Therefore, nutritional assessment, including psychological evaluation, is required for Parkinson's disease patients to facilitate interdisciplinary nutritional intervention for malnourished patients.


Online workbenches for neural network connections.
The nervous system is the most complex object we know of. It is a spatially distributed, functionally differentiated network formed by axonal connections between defined neuron populations and effector cells. Computer science provides exciting new tools for archiving, analyzing, synthesizing, and modeling on the Web vast amounts of frequently conflicting and incomplete qualitative and quantitative data about the organization and molecular mechanisms of neural networks. To optimize conceptual advances in systems neuroscience, it is important for the research and publishing communities to embrace three exercises: using defined nomenclatures; populating databases; and providing feedback to developers about improved design, performance, and functionality of knowledge management systems and associated visualization tools.


Microphysiological Models of Lung Epithelium-Alveolar Macrophage Co-Cultures to Study Chronic Lung Disease.
The interactions between immune cells and epithelial cells influence the progression of many respiratory diseases, such as chronic obstructive pulmonary disease (COPD). In vitro models allow for the examination of cells in controlled environments. However, these models lack the complex 3D architecture and vast multicellular interactions between the lung resident cells and infiltrating immune cells that can mediate cellular response to insults. In this study, three complementary microphysiological systems are presented to delineate the effects of cigarette smoke and respiratory disease on the lung epithelium. First, the Transwell system allows the co-culture of pulmonary immune and epithelial cells to evaluate cellular and monolayer phenotypic changes in response to cigarette smoke exposure. Next, the human and mouse precision-cut lung slices system provides a physiologically relevant model to study the effects of chronic insults like cigarette smoke with the dissection of specific interaction of immune cell subtypes within the structurally complex tissue environment. Finally, the lung-on-a-chip model provides an adaptable system for live imaging of polarized epithelial tissues that mimic the in vivo environment of the airways. Using a combination of these models, a complementary approach is provided to better address the intricate mechanisms of lung disease.


Toxinotype V Clostridium difficile in humans and food animals.
Clostridium difficile is a recognized pathogen in neonatal pigs and may contribute to enteritis in calves. Toxinotype V strains have been rare causes of human C. difficile-associated disease (CDAD). We examined toxinotype V in human disease, the genetic relationship of animal and human toxinotype V strains, and in vitro toxin production of these strains. From 2001 through 2006, 8 (1.3%) of 620 patient isolates were identified as toxinotype V; before 2001, 7 (<0.02%) of approximately 6,000 isolates were identified as toxinotype V. Six (46.2%) of 13 case-patients for whom information was available had community-associated CDAD. Molecular characterization showed a high degree of similarity between human and animal toxinotype V isolates; all contained a 39-bp tcdC deletion and most produced binary toxin. Further study is needed to understand the epidemiology of CDAD caused by toxinotype V C. difficile, including the potential of foodborne transmission to humans.


Bacterium-Like Particles Displaying the Rift Valley Fever Virus Gn Head Protein Induces Efficacious Immune Responses in Immunized Mice.
Rift Valley fever virus (RVFV), a mosquito-borne zoonotic phlebovirus, causes serious disease in humans and ruminants. According to the World Health Organization, Rift Valley fever is classified as a priority disease, and as such, vaccine development is of high priority due to the lack of licensed vaccines. In this study, a bacterium-like particle vaccine (BLP), RVFV-BLPs, is constructed. A novel display system is described, which is based on non-living and non-genetically modified Gram-positive bacterial cells, designated as Gram-positive enhancer matrix (GEM). The RVFV Gn head protein was displayed on the surface of GEM by co-expression with the peptidoglycan-binding domain (protein anchor) at the C-terminus. We determined that the RVFV Gn head-PA fusion protein was successfully displayed on the GEM. Mice immunized with RVFV-BLPs produced humoral and cellular immunity. Interestingly, comparing the production of RVFV Gn head-specific IgG and its subtype by vaccinating with different antigen doses of the RVFV-BLPs determined that the RVFV-BLPs (50 μg) group showed a greater effect than the other two groups. More importantly, antibodies produced by mice immunized with RVFV-BLPs (50 μg) exhibited potent neutralizing activity against RVFV pseudovirus. RVFV-BLPs (50 μg) also could induce IFN-γ and IL-4 in immunized mice; these mice generated memory cells among the proliferating T cell population after immunization with RVFV-BLPs with effector memory T cells as the major population, which means that RVFV-BLPs is an effective vaccine to establish a long-lived population of memory T cells. The findings suggest that the novel RVFV-BLPs subunit vaccine has the potential to be considered a safe and effective candidate vaccine against RVFV infection.


LincIN, a novel NF90-binding long non-coding RNA, is overexpressed in advanced breast tumors and involved in metastasis.
Recent genome-wide profiling by sequencing and distinctive chromatin signatures has identified thousands of long non-coding RNA (lncRNA) species (>200 nt). LncRNAs have emerged as important regulators of gene expression, involving in both developmental and pathological processes. While altered expression of lncRNAs has been observed in breast cancer development, their roles in breast cancer progression and metastasis are still poorly understood. To identify novel breast cancer-associated lncRNA candidates, we employed a high-density SNP array-based approach to uncover intergenic lncRNA genes that are aberrantly expressed in breast cancer. We first evaluated the potential value as a breast cancer prognostic biomarker for one breast cancer-associated lncRNA, LincIN, using a breast cancer cohort retrieved from The Cancer Genome Atlas (TCGA) Data Portal. Then we characterized the role of LincIN in breast cancer progression and metastasis by in vitro invasion assay and a mouse tail vein injection metastasis model. To study the action of LincIN, we identified LincIN-interacting protein partner(s) by RNA pull-down experiments followed with protein identification by mass spectrometry. High levels of LincIN expression are frequently observed in tumors compared to adjacent normal tissues, and are strongly associated with aggressive breast cancer. Importantly, analysis of TCGA data further suggest that high expression of LincIN is associated with poor overall survival in patients with breast cancer (P = 0.044 and P = 0.011 after adjustment for age). The functional experiments demonstrate that knockdown of LincIN inhibits tumor cell migration and invasion in vitro, which is supported by the results of transcriptome analysis in the LincIN-knockdown cells. Furthermore, knockdown of LincIN diminishes lung metastasis in a mouse tail vein injection model. We also identified a LincIN-binding protein, NF90, through which overexpression of LincIN may repress p21 protein expression by inhibiting its translation, and upregulation of p21 by LincIN knockdown may be associated with less aggressive metastasis phenotypes. Our studies provide clear evidence to support LincIN as a new regulator of tumor progression-metastasis at both transcriptional and translational levels and as a promising prognostic biomarker for breast cancer.


The Silence of PSMC6 Inhibits Cell Growth and Metastasis in Lung Adenocarcinoma.
The proteasome has been validated as an anticancer drug target, while the role of a subunit of proteasome, PSMC6, in lung adenocarcinoma (LUAD) has not been fully unveiled. In this study, we observed that both the RNA and protein of PSMC6 were highly upregulated in LUAD compared with the adjacent normal tissues. Moreover, a high PSMC6 expression was associated with poor prognosis. In accordance with this finding, PSMC6 was associated with poor tumor differentiation. Furthermore, the silence of PSMC6 by small interference RNAs (siRNAs) could significantly inhibit cell growth, migration, and invasion in lung cancer cell lines, suggesting that PSMC6 might serve as a promising therapeutic target in LUAD. To further explore the molecular mechanism of PSMC6 in LUAD, we observed that the proteasome subunits, such as PSMD10, PSMD6, PSMD9, PSMD13, PSMB3, PSMB1, PSMA4, PSMC1, PSMC2, PSMD7, and PSMD14, were highly correlated with PSMC6 expression. Based on the gene set enrichment analysis, we observed that these proteasome subunits were involved in the degradation of AXIN protein. The correlation analysis revealed that the positively correlated genes with PSMC6 were highly enriched in WNT signaling-related pathways, demonstrating that the PSMC6 overexpression may activate WNT signaling via degrading the AXIN protein, thereby promoting tumor progression. In summary, we systematically evaluated the differential expression levels and prognostic values of PSMC6 and predicted its biological function in LUAD, which suggested that PSMC6 might act as a promising therapeutic target in LUAD.


Ultrathin Graphdiyne-Wrapped Iron Carbonate Hydroxide Nanosheets toward Efficient Water Splitting.
We employed a two-step strategy for preparing ultrathin graphdiyne-wrapped iron carbonate hydroxide nanosheets on nickel foam (FeCH@GDY/NF) as the efficient catalysts toward the electrical splitting water. The introduction of naturally porous GDY nanolayers on FeCH surface endows the pristine catalyst with structural advantages for boosting catalytic performances. Benefited from the protection of robust GDY nanolayers with intimate contact between GDY and FeCH, the combined material exhibits high long-term durability of 10 000 cycles for oxygen-evolution reaction (OER) and 9000 cycles for hydrogen evolution reaction (HER) in 1.0 M KOH. Such excellent bifunctional OER/HER performance makes FeCH@GDY/NF quite qualified for alkaline two-electrode electrolyzer. Remarkably, such electrocatalyst can drive 10 and 100 mA cm-2 at 1.49 and 1.53 V, respectively. These results demonstrate the decisive role of GDY in the improvement of electrocatalytic performances, and open up new opportunities for designing cost-effective, efficient, and stable electrocatalysts for sustainable oxygen/hydrogen generation.