A multiple near isogenic line (multi-NIL) RNA-seq approach to identify candidate genes underpinning QTL.
This study demonstrates how identification of genes underpinning disease-resistance QTL based on differential expression and SNPs can be improved by performing transcriptomic analysis on multiple near isogenic lines. Transcriptomic analysis has been widely used to understand the genetic basis of a trait of interest by comparing genotypes with contrasting phenotypes. However, these approaches identify such large sets of differentially expressed genes that it proves difficult to isolate which genes underpin the phenotype of interest. This study tests whether using multiple near isogenic lines (NILs) can improve the resolution of RNA-seq-based approaches to identify genes underpinning disease-resistance QTL. A set of NILs for a major effect Fusarium crown rot-resistance QTL in barley on the 4HL chromosome arm were analysed under Fusarium crown rot using RNA-seq. Differential gene expression and single nucleotide polymorphism detection analyses reduced the number of putative candidates from thousands within individual NIL pairs to only one hundred and two genes, which were differentially expressed or contained SNPs in common across NIL pairs and occurred on 4HL. Our findings support the value of performing RNA-seq analysis using multiple NILs to remove genetic background effects. The enrichment analyses indicated conserved differences in the response to infection between resistant and sensitive isolines suggesting that sensitive isolines are impaired in systemic defence response to Fusarium pseudograminearum.


Evolutionarily conserved genetic interactions between nphp-4 and bbs-5 mutations exacerbate ciliopathy phenotypes.
Primary cilia are sensory and signaling hubs with a protein composition that is distinct from the rest of the cell due to the barrier function of the transition zone (TZ) at the base of the cilium. Protein transport across the TZ is mediated in part by the BBSome, and mutations disrupting TZ and BBSome proteins cause human ciliopathy syndromes. Ciliopathies have phenotypic variability even among patients with identical genetic variants, suggesting a role for modifier loci. To identify potential ciliopathy modifiers, we performed a mutagenesis screen on nphp-4 mutant Caenorhabditis elegans and uncovered a novel allele of bbs-5. Nphp-4;bbs-5 double mutant worms have phenotypes not observed in either individual mutant strain. To test whether this genetic interaction is conserved, we also analyzed zebrafish and mouse mutants. While Nphp4 mutant zebrafish appeared overtly normal, Bbs5 mutants exhibited scoliosis. When combined, Nphp4;Bbs5 double mutant zebrafish did not exhibit synergistic effects, but the lack of a phenotype in Nphp4 mutants makes interpreting these data difficult. In contrast, Nphp4;Bbs5 double mutant mice were not viable and there were fewer mice than expected carrying three mutant alleles. In addition, postnatal loss of Bbs5 in mice using a conditional allele compromised survival when combined with an Nphp4 allele. As cilia are still formed in the double mutant mice, the exacerbated phenotype is likely a consequence of disrupted ciliary signaling. Collectively, these data support an evolutionarily conserved genetic interaction between Bbs5 and Nphp4 alleles that may contribute to the variability in ciliopathy phenotypes.


Bacteriophages Isolated From Turkeys Infecting Diverse Salmonella Serovars.
Salmonella is one of the leading causes of foodborne illnesses worldwide. The rapid emergence of multidrug-resistant Salmonella strains has increased global concern for salmonellosis. Recent studies have shown that bacteriophages (phages) are novel and the most promising antibacterial agents for biocontrol in foods because phages specifically kill target bacteria without affecting other bacteria, do not alter organoleptic properties or nutritional quality of foods, and are safe and environmentally friendly. Due to the vast variation in Salmonella serotypes, large numbers of different and highly virulent Salmonella phages with broad host ranges are needed. This study isolated 14 Salmonella phages from turkey fecal and cecal samples. Six phages (Φ205, Φ206, Φ207, ΦEnt, ΦMont, and Φ13314) were selected for characterization. These phages were from all three families in the Caudovirales order. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that each phage had a unique structural protein profile. Each phage had a distinct host range. Φ207 and ΦEnt are both siphophages. They shared eight hosts, including seven different Salmonella serovars and one Shigella sonnei strain. These two phages showed different restriction banding patterns generated through EcoRI or HindIII digestion, but shared three bands from EcoRI digestion. ΦEnt displayed the broadest and very unusual host range infecting 11 Salmonella strains from nine serovars and three Shigella strains from two species, and thus was further characterized. The one-step growth curve revealed that ΦEnt had a short latent period (10 min) and relatively large burst size (100 PFU/infected cell). ΦEnt and its host showed better thermal stabilities in tryptic soy broth than in saline at 63 or 72°C. In the model food system (cucumber juice or beef broth), ΦEnt infection [regardless of the multiplicity of infections (MOIs) of 1, 10, and 100] resulted in more than 5-log10 reduction in Salmonella concentration within 4 or 5 h. Such high lytic activity combined with its remarkably broad and unusual host range and good thermal stability suggested that ΦEnt is a novel Salmonella phage with great potential to be used as an effective biocontrol agent against diverse Salmonella serovars in foods.


External Drainage of Giant Infantile Choledochal Cyst before Definitive Repair: Is it Worth?
Infantile Choledochal Cysts (IFCC) usually present with jaundice, acholic stool and abdominal lump or abdominal distension. If the surgical intervention is delayed, they rapidly progress to liver fibrosis which is considered to be irreversible if progressed to cirrhosis. We present the data of four cases (aged one month to seven months) of IFCC presented with cholangitis managed in one surgical unit in last two years. In one case, cholangitis was treated with prolonged antibiotic course before definitive repair whereas in rest, external drainage of cyst was done in addition to intravenous antibiotic to treat cholangitis. All the infants had features of cholangitis at time of presentation. Total leucocyte count ranged from 18x1000/UL to 30.6x1000/UL. Total bilirubin level at presentation ranged from 8.2 mg/dl to 18 mg/dl and Prothrombin time (INR) ranged from 1.33 to 1.9. Hepatic fibrosis was observed in all cases but cirrhosis was observed in only one case. There was no mortality but one patient had postoperative complication with prolonged hospital stay. External drainage helps in early recovery from cholangitis and better optimization of liver function. It also delays further progression to liver fibrosis by relieving the biliary outflow obstruction while waiting for definitive repair.


Coarctation induces alterations in basement membranes in the cardiovascular system.
A coarctation hypertensive rat model was used to examine the effects of elevated blood pressure on basement membrane component synthesis by cardiac myocytes and aorta using immunohistochemistry and Northern blot analysis. Carotid arterial pressure increased immediately on coarctation, and left ventricular hypertrophy was maximal within 5 days. In immunohistochemical studies, fibronectin and laminin were increased and the basement membrane chondroitin sulfate proteoglycan decreased in both the subendothelial space and smooth muscle cell basement membranes of the aorta above the clip compared with controls, whereas only fibronectin was elevated in the aorta below the clip. No change in basement membrane staining intensity for the cardiac myocytes was observed. Alterations in steady-state mRNA levels for fibronectin and laminin in the aorta paralleled those observed by immunohistochemical analysis with regard to protein and tissue type affected as well as intensity of the changes. However, changes in mRNA levels (but not protein deposition) for perlecan and type IV collagen were also observed in aortas from hypertensive rats compared with controls. Increases in steady-state mRNA levels for all basement membrane components in the heart and vasculature peaked before maximal cardiac hypertrophy (5 days). These studies indicate that alterations in basement membrane component deposition in the hypertrophied vasculature occur at both transcriptional and translational levels and suggest that the cell attachment glycoproteins fibronectin and laminin may be important factors in the vascular response to elevated transmural pressure.


A novel method for combating dispersion induced power fading in dispersion compensating fiber.
We experimentally investigate the performance of 60 GHz double sideband (DSB) radio over fiber (RoF) links that employ dispersion compensating fiber (DCF). Error free transmission of 3 Gbps signals over 1 m of wireless distance is reported. In order to overcome experimentally observed chromatic dispersion (CD) induced power fading of radio frequency (RF) signal, we propose a method for improvement of RF carrier-to-noise (C/N) ratio through introduction of a degree of RF frequency tunability. Overall results improve important aspects of directly modulated RoF systems and demonstrate the feasibility of high carrier frequency and wide bandwidth RF signals delivery in RoF links including DCF fiber. Error free performance that we obtain for 3 Gbps amplitude shift-keying (ASK) signals enables uncompressed high-definition 1080p video delivery.


Chemical destruction of brain noradrenergic neurons affects splenic cytokine production.
The neurotransmitter noradrenaline (NA) plays a pivotal role in immune regulation. Here we used the selective neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) to investigate the impact of central NA depletion on cytokine production by splenic monocytes/macrophages and T cells. Intraperitoneal administration of DSP-4 in adult rats induced a substantial reduction of noradrenergic neurons in the locus coeruleus and the A5 cell group. The degeneration of brainstem noradrenergic neurons was accompanied by a significant decrease in the production of interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha by lipopolysaccharide-stimulated splenocytes. In addition, upon T cell receptor stimulation with anti-CD3, isolated splenocytes of DSP-4 treated animals produced significantly less interferon (IFN)-gamma but not IL-2 and IL-4. The proportion of monocytes/macrophages and T cells in the spleen remained unaffected by the neurotoxin treatment, however, the percentage of natural killer cells decreased significantly. The findings suggest that a certain level of central noradrenergic tone is required for normal functioning of peripheral immune cells.


Enhanced accessory cell function by alveolar macrophages from patients infected with the human immunodeficiency virus: potential role for depletion of CD4+ cells in the lung.
Mononuclear phagocytes, including alveolar macrophages (AM), can be infected by the human immunodeficiency virus (HIV). Acting as accessory cells (AC), AM could infect CD4 lymphocytes through cell-to-cell contact and by inducing T cell proliferation, which increases lymphocyte susceptibility to infection. Using normal allogeneic T cells as responders, AM from infected individuals demonstrated an enhanced ability to stimulate a Con A and pokeweed mitogen lymphocyte proliferation assay compared with normal AM. Exogenous IL 1 enhanced the stimulation of a mitogen response by normal AM, but not from HIV-positive individuals, suggesting increased levels of this cytokine may explain the observed enhancement. However, increased IL 1 secretion by AM from HIV-infected patients could not be demonstrated, either in a bioassay or antigenically using an ELISA for IL-1 beta. Syncytia formation was observed when AM from asymptomatic HIV-positive individuals were cultured with normal T cells, suggesting viral transmission was occurring. Finally, in individual patients the stimulation of a mitogen response was inversely correlated with the CD4/CD8 ratio and total CD4 count, suggesting that enhanced AC function and CD4 cell depletion may be related in vivo. These findings indicate that enhanced AM accessory cell function is seen in HIV-infected individuals and could be a potential mechanism for CD4 cell depletion in the lung.


Time-periodic oscillation reaction in an organic-solvent dominated electrolyte.
Time-periodic phenomena widely exist in natural life sustaining systems but are rarely reported in highly efficient artificial electrochemical energy-harvesting systems. Herein, we observed for the first time the periodic oscillation reaction in organic-solvent dominated electrolytes that could be also derived from some electrochemical energy harvesting cells. Owing to different reaction activity and acidity in the organic solvent, the oscillation reaction could occur under milder pH conditions and exhibit better durability. Its influence on electric output and related differences from an aqueous B-Z reaction are discussed from a mechanism point of view. Our findings may contribute to smart self-oscillation materials and new strategies for highly efficient long-term energy harvesting.


Serum total cholesterol concentration and 10-year mortality in an 85-year-old population.
Little is known about the association between total cholesterol (TC) and all-cause mortality in the elderly (especially the very elderly). Here we examined the association between TC and all-cause mortality in 207 very elderly (85-year-old) participants. In 2003, we performed a baseline laboratory blood examination, and blood pressure (BP) and body mass index (BMI) measurements, and lifestyle questionnaires were completed by the participants. The participants were followed for the subsequent 10 years. As of 2013, of the 207 participants in 2003, 70 participants had survived, 120 individuals had died, and 17 were lost to follow up. The TC values were divided into high-TC (≥209 mg/dL), intermediate-TC (176-208 mg/dL), and low-TC (≤175 mg/dL) categories. With the Kaplan-Meier method, we found that both the high-TC and intermediate-TC participants survived longer than the low-TC participants. The men with high TC survived longer than those with low TC, but no corresponding difference was found for the women. A multivariate Cox proportional hazards regression model, with adjustment for gender, smoking, alcohol intake, history of stroke or heart disease, serum albumin concentration, BMI, and systolic BP, revealed that the total mortality in the low-TC group was 1.7-fold higher than that in the high-TC group. Mortality, adjusted for the same factors, decreased 0.9% with each 1 mg/dL increase in the serum TC concentration and decreased 0.8% with each 1 mg/dL increase in the serum (low-density lipoprotein) LDL-cholesterol (LDL-C) concentration. Our results indicate an association between lower serum TC concentrations and increased all-cause mortality in a community-dwelling, very elderly population. Mortality decreased with the increases in both TC and LDL-C concentrations, after adjustment for various confounding factors. These findings suggest that low TC and low LDL-C may be independent predictors of high mortality in the very elderly.


MicroRNA‑24‑2 is associated with cell proliferation, invasion, migration and apoptosis in renal cell carcinoma.
Micro (mi)RNAs are involved in multiple cellular processes, and alterations in miRNA expression have been demonstrated to lead to tumorigenesis. Previous microarray analysis revealed that miRNA (miR)‑24 was downregulated in renal cell carcinoma (RCC). Additionally, miR‑24 has been identified as an oncogene and tumor suppressor in various cancers. The present study assessed the expression levels of two stem‑loops of miR‑24, miR‑24‑1 and miR‑24‑2, in RCC tissues and paired healthy tissues by reverse transcription‑quantitative polymerase chain reaction. The results revealed that miR‑24‑2 was upregulated in RCC tissues and ACHN, 786‑O and 769P cell lines compared with healthy tissues and HEK‑293T cells, respectively, whereas miR‑24‑1 was almost absent in RCC and healthy kidney tissues. To investigate the role of miR‑24‑2 in RCC, a synthesized miR‑24‑2 mimic, negative control (NC), inhibitor or inhibitor NC was transfected into 786‑O and ACHN RCC cells, and cell proliferation, mobility and apoptosis assays were performed. The results of the present study revealed that miR‑24‑2 was associated with cell proliferation, migration, invasion and apoptosis, thus demonstrating that miR‑24‑2 may serve a role as an oncogene in RCC. Further studies are required to investigate the signaling pathways of miR‑24‑2, and the potential of miR‑24‑2 as a therapeutic target or biomarker for the early detection of RCC.


Conservation of primary structure in the lipoyl-bearing and dihydrolipoyl dehydrogenase binding domains of mammalian branched-chain alpha-keto acid dehydrogenase complex: molecular cloning of human and bovine transacylase (E2) cDNAs.
The subunit structures and conservation of the dihydrolipoyl transacylase (E2) components of bovine and human branched-chain alpha-keto acid dehydrogenase complexes were investigated by Western blotting, peptide sequencing, and cDNA cloning methods. Rabbit antiserum prepared against the sodium dodecyl sulfate (SDS) denaturated bovine E2 subunit recognized the inner E2 core, and the first hinge region of the E2 chain, but failed to react with the lipoyl-bearing domain as determined by Western blot analysis. The lack of antigenicity in the lipoyl-bearing domain was confirmed with antibodies directed against the native E2 component. A human E2 cDNA (1.6 kb) was isolated from a human liver cDNA library in lambda gt11 with a combination of the above anti-native and anti-SDS-denatured E2 immunoglobulin G's as a probe. The fidelity of the human E2 cDNA was established by nucleotide sequencing which showed the determined peptide sequences of the amino terminus and tryptic fragments of bovine E2. A bovine E2 cDNA (0.7 kb) was also isolated from a bovine liver cDNA library in lambda ZAP with the human E2 cDNA as a probe. Northern blot analysis using the human E2 cDNA probe showed that E2 mRNAs in bovine liver and human kidney mesangial cells are 3.3 and 4.6 kb in size, respectively. Primary structures derived from human and bovine E2 cDNAs show leader sequences including the initiator methionine and the homologous mature peptides consisting of complete lipoyl-bearing and dihydrolipoyl dehydrogenase (E3) binding domains and two hinge regions. In addition, the human E2 cDNA contains a portion of the inner E2 core sequence, a 3'-untranslated region, and a poly(A+) tail. Deduced amino acid sequences of the mammalian E2's were compared with those of Escherichia coli transacetylase and transsuccinylase and bovine kidney transacetylase. The results indicate a high degree of conservation in the sequence flanking the lipoyl-attachment site and in the E3-binding domain. Models are presented to discuss implications for the conserved structure-function relationship in the lipoyl-bearing and E3-binding domains of alpha-keto acid dehydrogenase complexes.


Automated collection of imaging and phenotypic data to centralized and distributed data repositories.
Accurate data collection at the ground level is vital to the integrity of neuroimaging research. Similarly important is the ability to connect and curate data in order to make it meaningful and sharable with other investigators. Collecting data, especially with several different modalities, can be time consuming and expensive. These issues have driven the development of automated collection of neuroimaging and clinical assessment data within COINS (Collaborative Informatics and Neuroimaging Suite). COINS is an end-to-end data management system. It provides a comprehensive platform for data collection, management, secure storage, and flexible data retrieval (Bockholt et al., 2010; Scott et al., 2011). It was initially developed for the investigators at the Mind Research Network (MRN), but is now available to neuroimaging institutions worldwide. Self Assessment (SA) is an application embedded in the Assessment Manager (ASMT) tool in COINS. It is an innovative tool that allows participants to fill out assessments via the web-based Participant Portal. It eliminates the need for paper collection and data entry by allowing participants to submit their assessments directly to COINS. Instruments (surveys) are created through ASMT and include many unique question types and associated SA features that can be implemented to help the flow of assessment administration. SA provides an instrument queuing system with an easy-to-use drag and drop interface for research staff to set up participants' queues. After a queue has been created for the participant, they can access the Participant Portal via the internet to fill out their assessments. This allows them the flexibility to participate from home, a library, on site, etc. The collected data is stored in a PostgresSQL database at MRN. This data is only accessible by users that have explicit permission to access the data through their COINS user accounts and access to MRN network. This allows for high volume data collection and with minimal user access to PHI (protected health information). An added benefit to using COINS is the ability to collect, store and share imaging data and assessment data with no interaction with outside tools or programs. All study data collected (imaging and assessment) is stored and exported with a participant's unique subject identifier so there is no need to keep extra spreadsheets or databases to link and keep track of the data. Data is easily exported from COINS via the Query Builder and study portal tools, which allow fine grained selection of data to be exported into comma separated value file format for easy import into statistical programs. There is a great need for data collection tools that limit human intervention and error while at the same time providing users with intuitive design. COINS aims to be a leader in database solutions for research studies collecting data from several different modalities.


Incidence of childhood linear scleroderma and systemic sclerosis in the UK and Ireland.
Childhood scleroderma encompasses a rare, poorly understood spectrum of conditions. Our aim was to ascertain the incidence of childhood scleroderma in its different forms in the UK and Ireland, and to describe the age, sex, and ethnicity of the cases. The members of 5 specialist medical associations including pediatricians, dermatologists, and rheumatologists were asked to report all cases of abnormal skin thickening suspected to be localized (including linear) scleroderma or systemic sclerosis (SSc) in children <16 years of age first seen between July 2005 and July 2007. We received notification of 185 potential cases, and 94 valid cases were confirmed: 87 (93%) with localized scleroderma and 7 (7%) with SSc. This gave an incidence rate per million children per year of 3.4 (95% confidence interval [95% CI] 2.7-4.1) for localized scleroderma, including an incidence rate of 2.5 (95% CI 1.8-3.1) for linear scleroderma, and 0.27 (95% CI 0.1-0.5) for SSc. Of the 87 localized cases, 62 (71%) had linear disease. Of localized disease cases, 55 (63%) were female, 71 (82%) were classified as white British, and the patients' mean age when first seen in secondary care was 10.4 years. Of the 7 SSc cases, all were female, 6 (86%) were white British, and the mean age when first seen was 12.1 years. The median delay between onset and being first seen was 13.1 months for localized scleroderma and 7.2 months for SSc. These data provide additional estimates of the incidence of this rare disorder and its subforms.


A novel ochre mutation in the beta-thalassemia gene of a Thai. Identification by direct cloning of the entire beta-globin gene amplified using polymerase chain reactions.
The beta-globin genes from a Thai patient compound heterozygous for beta-thalassemia and HbE disease were investigated. The 3.0-kilobase fragment containing the entire beta-globin gene was amplified by polymerase chain reaction, using Taq DNA polymerase followed by direct cloning of the amplified product into plasmid DNA. Sequence analysis of the thalassemia gene revealed only one base change, a C-A transversion within codon for an amino acid 35. This new mutation creates a premature terminator, TAA, an ochre codon, and results in a beta 0-thalassemia phenotype. The same result was obtained when this mutation was analyzed using a conventional cloning technique, direct sequencing of the amplified product, and hybridization with allele-specific oligonucleotide probes. No misincorporation was detected in the sequence analysis of the 3.0-kilobase insert of five clones of the amplified products obtained from genomic DNA of a normal individual. This approach is a rapid and accurate method for molecular cloning of the beta-globin gene and also other genes, the partial nucleotide sequences of which are known.


Palliative Care Triggers in the Intensive Care Unit: A Pilot Success Story.
There is growing recognition that electronic medical record triggers in the intensive care unit (ICU) have led to an increase in palliative care consultations. One suburban health care system adopted triggers unique to their culture and setting in a pilot study and saw an increase in palliative consultations in the ICU. Implementing triggers is often a complex and multifaceted process to adopt. This review shares the steps from concept to implementation of establishing palliative prompts in 1 ICU within an integrated health care system.


Does acute stress disorder predict subsequent posttraumatic stress disorder in pediatric burn survivors?
This study examined the prevalence of posttraumatic stress disorder (PTSD) in pediatric burn survivors who had been treated for acute stress disorder (ASD) symptoms during their initial hospitalization and compared them to patients who had been asymptomatic for ASD symptoms. Participants were identified from electronic medical records from 1995 to 2008 and data were collected from 2006 to 2008. Participants were primarily matched on total body surface area burned and gender, and as close as possible on age at time of burn and number of years postburn. Pediatric burn survivors completed a semistructured clinical interview, the Missouri Assessment of Genetics Interview for Children-PTSD section, which is based on criteria from the DSM-IV for evaluating lifetime PTSD. There were 183 participants in the study, and from this sample 85 matched pairs were identified. Most were 5 years postburn. The prevalence of PTSD at the time of follow-up was 8.24% (7 of 85) for the ASD group and 4.71% (4 of 85) for the non-ASD comparison group. No significant differences were found between these groups at P value ≥ .05. A logistic regression analysis was conducted to determine if prior ASD diagnosis, burn size, gender, ethnicity, age at time of study participation, and number of years postburn predicted subsequent PTSD. None of the variables were significant predictors. The prevalence of PTSD was similar in children who had ASD symptoms and those without ASD symptoms. The lifetime prevalence of PTSD was lower than reported in other studies. A possible explanation for this finding is that children received timely pharmacotherapy and psychotherapy during their acute hospitalization.


Adaptation and the cost of complexity.
Adaptation is characterized by the movement of a population toward a many-character optimum, movement that results in an increase in fitness. Here I calculate the rate at which fitness increases during adaptation and describe the curve giving fitness versus time as a population approaches an optimum in Fisher's model of adaptation. The results identify several factors affecting the speed of adaptation. One of the most important is organismal complexity--complex organisms adapt more slowly than simple ones when using mutations of the same phenotypic size. Thus, as Fisher foresaw, organisms pay a kind of cost of complexity. However, the magnitude of this cost is considerably larger than Fisher's analysis suggested. Indeed the rate of adaptation declines at least as fast as n-1, where n is the number of independent characters or dimensions comprising an organism. The present results also suggest that one can define an effective number of dimensions characterizing an adapting species.


Comparison of acrylic and polymethyl methacrylate lenses in a pediatric population.
To compare the intra-patient incidence of posterior capsular opacification (PCO) and their post operative course, in children with bilateral cataract, following implantation of acrylic (Group 1) and polymethyl methacrylate (PMMA) (Group 2) intraocular lenses (IOLs). This was a prospective, interventional intra-patient case series. Twenty-five children with bilateral cataract, 5 years and older, underwent cataract surgery and implantation of an acrylic (AcrySof MA30BA, Alcon, Fortworth, TX) in one eye and PMMA (Indo Am IAB 203, Ocular Vision, Inc.) IOL in the other eye of each patient. They were followed up for an average of 11.1 +/- 9.5 months to assess the incidence of clinically significant PCO and occurrence of postoperative complications. Twenty-three children (46 eyes), were included in this study. Post-operatively, 22(95.6%) patients with acrylic IOLs and 20 (86.9%) patients with PMMA IOLs, either maintained or improved their vision. In the acrylic and PMMA IOL groups respectively, the incidence of clinically significant PCO was 21% (4) and 75% (12) ( P =0.002), with a median onset at 2.9 months and 0.7 months. Other complications included pupillary capture in 2 (8.7%) eyes and uveal prolapse in 1 (4.3%) eye in the acrylic group and increased uveal inflammation in 6 (26.1%) eyes and presumed noninfectious endophthalmitis in 2 (8.7%) eyes in the PMMA group. Incidence of PCO and post operative uveal inflammation is significantly less with acrylic lenses and were safe to use in pediatric eyes.


Heparin-induced thrombocytopenia in the critically ill: interpreting the 4Ts test in a randomized trial.
Thrombocytopenia occurs in 20% to 45% of critically ill medical-surgical patients. The 4Ts heparin-induced thrombocytopenia (HIT) score (with 4 domains: Thrombocytopenia, Timing of thrombocytopenia, Thrombosis and oTher reason[s] for thrombocytopenia) might reliably identify patients at low risk for HIT. Interobserver agreement on 4Ts scoring is uncertain in this setting. To evaluate whether a published clinical prediction rule (the "4Ts score") reliably rules out HIT in "low-risk" intensive care unit (ICU) patients as assessed by research coordinators (who prospectively scored) and 2 adjudicators (who scored retrospectively) during an international heparin thromboprophylaxis trial (PROTECT, NCT00182143). Of 3746 medical-surgical ICU patients in PROTECT, 794 met the enrollment criteria for this HIT substudy. Enrollment was predicated on one of the following occurring in ICU: platelets less than 50×10(9)/L, platelets decreased to 50% of ICU admission value (if admission value<100×10(9)/L), any venous thrombosis, or if HIT was otherwise clinically suspected. Independently, 4Ts scores were completed in real time by research coordinators blinded to study drug and laboratory HIT results, and retrospectively by 2 adjudicators blinded to study drug, laboratory HIT results, and research coordinators' scores; the adjudicators arrived at consensus in all cases. Of the 763 patients, 474 had a central or local laboratory HIT test performed and had 4Ts scoring by adjudicators; 432 were scored by trained research coordinators. Heparin-induced thrombocytopenia was defined by a centrally performed positive serotonin release assay (SRA). Of the 474 patients with central adjudication, 407 (85.9%) had a 4Ts score of 3 or lower, conferring a low pretest probability (PTP) of HIT; of these, 6 (1.5% [95% confidence interval, 0.7%-3.2%) had a positive SRA. Fifty-nine (12.4%) had a moderate PTP (4Ts score of 4-5); of these, 4 (6.8%) had a positive SRA. Eight patients had a high PTP (4Ts score of ≥6); of these, 1 (12.5%) had a positive SRA. Raw agreement between research coordinators and central adjudication on each domain of the 4Ts score and low, intermediate, and high PTP was good. However, chance-corrected agreement was variable between adjudicators (weighted κ values of 0.31-0.93) and between the adjudicator consensus and research coordinators (weighted κ values of 0.13 and 0.78). Post hoc review of the 6 SRA-positive cases with an adjudicated low PTP demonstrated that their scores would have been increased if the adjudicators had had additional information on heparin exposure prior to ICU admission. In general, the fourth domain of 4Ts (oTher causes of thrombocytopenia) generated the most disagreement. Real-time 4Ts scoring by research coordinators at the time of testing for HIT was not consistent with 4Ts scores obtained by central adjudicators. The results of this comprehensive HIT testing highlight the need for further research to improve the assessment of PTP scoring of HIT for critically ill patients.


The analysis of genetic diversity and differentiation of six Chinese cattle populations using microsatellite markers.
A total of 321 individuals from six cattle populations of four species in a bovine subfamily in China were studied using 12 pairs of microsatellite markers. The genetic diversities within and between populations were calculated. The phylogenetic trees were constructed by (delta mu)(2) and DA distances, and the divergence times between populations were estimated by (delta mu)(2). Altogether, 144 microsatellite alleles were detected including 24 private alleles and nine shared alleles. Chinese Holstein had the largest number of private alleles (10), whereas, Bohai black and Buffalo had the smallest number of private alleles (2). Chinese Holstein showed the highest genetic variability. Its observed number of alleles (Na), mean effective number of alleles (MNA), and mean heterozygosity (He) were 7.7500, 4.9722, and 0.7719, respectively, whereas, the Buffalo and Yak showed low genetic variability. In the phylogenetic trees, Luxi and Holstein grouped first, followed by Bohai and Minnan. Yak branched next and buffalo emerged as the most divergent population from other cattle populations. Luxi and Bohai were estimated to have diverged 0.039-0.105 million years ago (MYA), however, buffalo and Holstein diverged 0.501-1.337 MYA. The divergence time of Yak versus Minnan, Holstein and buffalo was 0.136-0.363, 0.273-0.729, and 0.326-0.600 MYA, respectively.


Towards an ethics of authentic practice.
This essay asks how we might best elaborate an ethics of authentic practice. Will we be able to agree on a set of shared terms through which ethical practice will be understood? How will we define ethics and the subject's relation to authoritative structures of power and knowledge? We begin by further clarifying our critique of evidence-based medicine (EBM), reflecting on the intimate relation between theory and practice. We challenge the charge that our position amounts to no more than 'subjectivism' and 'antiauthoritarian' theory. We argue that an ethical practice ought to question the authority of EBM without falling into the trap of dogmatic antiauthoritarianism. In this, we take up the work of Hannah Arendt, who offers terms to help understand our difficult political relation to authority in an authentic ethical practice. We continue with a discussion of Michel Foucault's use of 'free speech' or parrhesia, which he adopts from Ancient Greek philosophy. Foucault demonstrates that authentic ethical practice demands that we 'speak truth to power.' We conclude with a consideration of recent biotechnologies, and suggest that these biomedical practices force us to re-evaluate our theoretical understanding of the ethical subject. We believe that we are at a crucial juncture: we must develop an ethics of authentic practice that will be commensurable with new and emergent biomedical subjectivities.


The effects of detethering on the urodynamics profile in children with a tethered cord.
Tethering of the spinal cord is a pathological fixation of the cord in the vertebral column that can result in neurogenic bladder dysfunction and other neurological problems. It occurs in patients with closed spinal dysraphisms and those in whom postoperative scarring develops following spina bifida closure procedures. The authors of this study sought to determine the effects of detethering on the urodynamic profile of children with a tethered cord. The authors retrospectively reviewed the records of children who underwent surgical release of a tethered cord at a single institution between 2001 and 2003. They identified 17 children (nine girls and eight boys) who had undergone both preoperative and postoperative urodynamic evaluation. Preoperatively, 10 (59%) of the children with a tethered cord had abnormal urodynamic study (UDS) results. Only two (20%) of these patients had urological symptoms. All seven patients with normal preoperative UDS results had normal UDS results after detethering. In addition, in five (50%) of the 10 children with abnormal preoperative UDS results, the postoperative UDS demonstrated improved or normal urodynamics. Because more than half of the children who underwent detethering were found to have abnormal preoperative UDS results, preoperative urodynamic evaluation should be performed in all cases in which detethering is considered. With regard to voiding function, detethering is relatively safe for children with normal preoperative UDS results. In children with abnormal preoperative UDS results, detethering may lead to improvement or even normalization of voiding, especially if the procedure is performed prior to 1 year of age. Finally, children with anorectal anomalies and a tethered cord may represent a subset of patients who are particularly likely to experience urodynamic improvement after detethering.


Pulmonary toxicity syndrome in breast cancer patients undergoing BCNU-containing high-dose chemotherapy and autologous hematopoietic cell transplantation.
We performed a retrospective review to investigate pulmonary toxicity syndrome (PTS) in a cohort of breast cancer patients undergoing BCNU-containing high-dose chemotherapy (HDC). Our aim was to characterize presentation, identify risk factors, determine outcome following therapy, and find any association with differences in survival. We reviewed the data of 152 patients with stage II or III or metastatic breast cancer treated with cyclophosphamide 5625 mg/m2, cisplatin 165 mg/m2, and BCNU 600 mg/m2 followed by autologous peripheral blood hematopoietic cell transplantation. During follow-up, PTS was diagnosed when the following criteria were met: (1) presentation with typical clinical symptoms of PTS, (2) an absolute carbon monoxide diffusion capacity (DLCO) decline of 10% compared with pre-HDC DLCO, and (3) no clinical evidence of active pulmonary infection. Patients were then treated with a course of corticosteroid therapy. The incidence of PTS for all 152 patients was 59%, with a median onset at 45 days (range, 21-149 days) post-HDC. The median absolute DLCO decrement was 26% (range, 10%-73%) at diagnosis of PTS. There was no significant correlation between patient age, stage of breast cancer, pre-HDC chemotherapy regimen, pre-HDC chest wall radiotherapy, tobacco use, prior lung disease, or baseline pulmonary function test results and the development of PTS. We did observe an interesting association between PTS and the development of a noncholestatic elevation of transaminases. Of PTS patients treated with prednisone therapy for a median of 105.5 days (range, 44-300 days), 91% achieved resolution of their PTS without pulmonary sequelae. At 3 years, the overall survival (OS) of stage II or III patients who developed PTS was 84% (95% confidence interval [CI], 73%-95%); of metastatic breast cancer patients with PTS, the OS was 58% (95% CI, 38%-78%). These values were not significantly different from those of patients who did not develop PTS (91% [95% confidence interval [CI], 81%-100%] and 53% [95% CI, 32%-74%], respectively). No significant differences in disease-free or event-free survival were observed between patients with and without PTS. The incidence of PTS in breast cancer patients treated with a BCNU-containing HDC regimen can be remarkably high. Treatment with a course of corticosteroid therapy is successful in the vast majority.


Predicting Functional Independence Measure Scores During Rehabilitation with Wearable Inertial Sensors.
Evaluating patient progress and making discharge decisions regarding inpatient medical rehabilitation rely upon standard clinical assessments administered by trained clinicians. Wearable inertial sensors can offer more objective measures of patient movement and progress. We undertook a study to investigate the contribution of wearable sensor data to predict discharge functional independence measure (FIM) scores for 20 patients at an inpatient rehabilitation facility. The FIM utilizes a 7-point ordinal scale to measure patient independence while performing several activities of daily living, such as walking, grooming, and bathing. Wearable inertial sensor data were collected from ecological ambulatory tasks at two time points mid-stay during inpatient rehabilitation. Machine learning algorithms were trained with sensor-derived features and clinical information obtained from medical records at admission to the inpatient facility. While models trained only with clinical features predicted discharge scores well, we were able to achieve an even higher level of prediction accuracy when also including the wearable sensor-derived features. Correlations as high as 0.97 for leave-one-out cross validation predicting discharge FIM motor scores are reported.


Nonreplicating influenza A virus vaccines confer broad protection against lethal challenge.
New vaccine technologies are being investigated for their ability to elicit broadly cross-protective immunity against a range of influenza viruses. We compared the efficacies of two intranasally delivered nonreplicating influenza virus vaccines (H1 and H5 S-FLU) that are based on the suppression of the hemagglutinin signal sequence, with the corresponding H1N1 and H5N1 cold-adapted (ca) live attenuated influenza virus vaccines in mice and ferrets. Administration of two doses of H1 or H5 S-FLU vaccines protected mice and ferrets from lethal challenge with homologous, heterologous, and heterosubtypic influenza viruses, and two doses of S-FLU and ca vaccines yielded comparable effects. Importantly, when ferrets immunized with one dose of H1 S-FLU or ca vaccine were challenged with the homologous H1N1 virus, the challenge virus failed to transmit to naive ferrets by the airborne route. S-FLU technology can be rapidly applied to any emerging influenza virus, and the promising preclinical data support further evaluation in humans. Influenza viruses continue to represent a global public health threat, and cross-protective vaccines are needed to prevent seasonal and pandemic influenza. Currently licensed influenza vaccines are based on immunity to the hemagglutinin protein that is highly variable. However, T cell responses directed against highly conserved viral proteins contribute to clearance of the virus and confer broadly cross-reactive and protective immune responses against a range of influenza viruses. In this study, two nonreplicating pseudotyped influenza virus vaccines were compared with their corresponding live attenuated influenza virus vaccines, and both elicited robust protection against homologous and heterosubtypic challenge in mice and ferrets, making them promising candidates for further evaluation in humans.


Plasma-derived C1-INH for managing hereditary angioedema in pediatric patients: A systematic review.
Presently, medications approved for children with Hereditary Angioedema (HAE) are extremely limited. This is especially the case for children under 12 years of age. For this reason we reviewed and summarized the data on treatment of children with HAE. Available data indicate that plasma derived C1-inhibitor is a safe, effective treatment option for HAE in pediatric patients, including those below 12 years of age. Other therapies are also appear safe for the under 12 year of age, but less data are available. Importantly, home-based treatment of HAE in this age group appears to be safe and effective and can improve quality of life. These findings support current HAE consensus guidelines which strongly recommend the use of plasma derived C1-inhibitor as a first-line treatment in children and encourage home and self-treatment.


The mitochondrial ATP synthase is a negative regulator of the mitochondrial permeability transition pore.
The mitochondrial permeability transition pore (mPTP) is a channel in the inner mitochondrial membrane whose sustained opening in response to elevated mitochondrial matrix Ca2+ concentrations triggers necrotic cell death. The molecular identity of mPTP is unknown. One proposed candidate is the mitochondrial ATP synthase, whose canonical function is to generate most ATP in multicellular organisms. Here, we present mitochondrial, cellular, and in vivo evidence that, rather than serving as mPTP, the mitochondrial ATP synthase inhibits this pore. Our studies confirm previous work showing persistence of mPTP in HAP1 cell lines lacking an assembled mitochondrial ATP synthase. Unexpectedly, however, we observe that Ca2+-induced pore opening is markedly sensitized by loss of the mitochondrial ATP synthase. Further, mPTP opening in cells lacking the mitochondrial ATP synthase is desensitized by pharmacological inhibition and genetic depletion of the mitochondrial cis-trans prolyl isomerase cyclophilin D as in wild-type cells, indicating that cyclophilin D can modulate mPTP through substrates other than subunits in the assembled mitochondrial ATP synthase. Mitoplast patch clamping studies showed that mPTP channel conductance was unaffected by loss of the mitochondrial ATP synthase but still blocked by cyclophilin D inhibition. Cardiac mitochondria from mice whose heart muscle cells we engineered deficient in the mitochondrial ATP synthase also demonstrate sensitization of Ca2+-induced mPTP opening and desensitization by cyclophilin D inhibition. Further, these mice exhibit strikingly larger myocardial infarctions when challenged with ischemia/reperfusion in vivo. We conclude that the mitochondrial ATP synthase does not function as mPTP and instead negatively regulates this pore.


People, organizational, and leadership factors impacting informatics support for clinical and translational research.
In recent years, there have been numerous initiatives undertaken to describe critical information needs related to the collection, management, analysis, and dissemination of data in support of biomedical research (J Investig Med 54:327-333, 2006); (J Am Med Inform Assoc 16:316-327, 2009); (Physiol Genomics 39:131-140, 2009); (J Am Med Inform Assoc 18:354-357, 2011). A common theme spanning such reports has been the importance of understanding and optimizing people, organizational, and leadership factors in order to achieve the promise of efficient and timely research (J Am Med Inform Assoc 15:283-289, 2008). With the emergence of clinical and translational science (CTS) as a national priority in the United States, and the corresponding growth in the scale and scope of CTS research programs, the acuity of such information needs continues to increase (JAMA 289:1278-1287, 2003); (N Engl J Med 353:1621-1623, 2005); (Sci Transl Med 3:90, 2011). At the same time, systematic evaluations of optimal people, organizational, and leadership factors that influence the provision of data, information, and knowledge management technologies and methods are notably lacking. In response to the preceding gap in knowledge, we have conducted both: 1) a structured survey of domain experts at Academic Health Centers (AHCs); and 2) a subsequent thematic analysis of public-domain documentation provided by those same organizations. The results of these approaches were then used to identify critical factors that may influence access to informatics expertise and resources relevant to the CTS domain. A total of 31 domain experts, spanning the Biomedical Informatics (BMI), Computer Science (CS), Information Science (IS), and Information Technology (IT) disciplines participated in a structured surveyprocess. At a high level, respondents identified notable differences in theaccess to BMI, CS, and IT expertise and services depending on the establishment of a formal BMI academic unit and the perceived relationship between BMI, CS, IS, and IT leaders. Subsequent thematic analysis of the aforementioned public domain documents demonstrated a discordance between perceived and reported integration across and between BMI, CS, IS, and IT programs and leaders with relevance to the CTS domain. Differences in people, organization, and leadership factors do influence the effectiveness of CTS programs, particularly with regard to the ability to access and leverage BMI, CS, IS, and IT expertise and resources. Based on this finding, we believe that the development of a better understanding of how optimal BMI, CS, IS, and IT organizational structures and leadership models are designed and implemented is critical to both the advancement of CTS and ultimately, to improvements in the quality, safety, and effectiveness of healthcare.


A maize cystatin suppresses host immunity by inhibiting apoplastic cysteine proteases.
Ustilago maydis is a biotrophic pathogen causing maize (Zea mays) smut disease. Transcriptome profiling of infected maize plants indicated that a gene encoding a putative cystatin (CC9) is induced upon penetration by U. maydis wild type. By contrast, cc9 is not induced after infection with the U. maydis effector mutant Δpep1, which elicits massive plant defenses. Silencing of cc9 resulted in a strongly induced maize defense gene expression and a hypersensitive response to U. maydis wild-type infection. Consequently, fungal colonization was strongly reduced in cc9-silenced plants, while recombinant CC9 prevented salicylic acid (SA)-induced defenses. Protease activity profiling revealed a strong induction of maize Cys proteases in SA-treated leaves, which could be inhibited by addition of CC9. Transgenic maize plants overexpressing cc9-mCherry showed an apoplastic localization of CC9. The transgenic plants showed a block in Cys protease activity and SA-dependent gene expression. Moreover, activated apoplastic Cys proteases induced SA-associated defense gene expression in naïve plants, which could be suppressed by CC9. We show that apoplastic Cys proteases play a pivotal role in maize defense signaling. Moreover, we identified cystatin CC9 as a novel compatibility factor that suppresses Cys protease activity to allow biotrophic interaction of maize with the fungal pathogen U. maydis.


Somatosensory Discrimination in People With Autism Spectrum Disorder: A Scoping Review.
Sensory symptoms in people with autism spectrum disorder (ASD) are commonly reported by researchers. However, an often overlooked sensory aspect of ASD is sensory discrimination in general, and somatosensory discrimination in particular. To examine what has and what has not yet been learned concerning the somatosensory discrimination abilities of people with ASD and to reveal gaps warranting further research. Scoping review of clinical studies published 1995-2017 located through searches of PsycNET, PubMed, ERIC, and Google Scholar. Inclusion criteria were English-language peer-reviewed studies with (1) participants diagnosed with ASD, (2) a specific somatosensory discrimination measure, and (3) a comparison group. No age or intellectual exclusion criteria were established; studies were excluded if they were theoretical or descriptive, did not incorporate a control group, focused only on neurology or genetics, or used simple threshold detection measures or somatosensory measures integrated with other measures. The final search yielded 12 comparative articles discussing tactile and proprioceptive discrimination in people with ASD. Overall, most results showed atypical somatosensory discrimination in people with ASD, especially among young children. The relationship between sensory discrimination abilities and other sensory symptoms and ASD symptoms is briefly discussed. Heterogeneous findings concerning somatosensory discrimination in people with ASD shed light on underlying mechanisms of these disorders and can contribute to improvement of occupational therapy intervention for this population. The occupational therapy evaluation of people with ASD can benefit from addressing somatosensory discrimination and its contribution to other clinical symptoms. This type of assessment can help improve intervention strategies for people with ASD by promoting a focus on the effect of discrimination deficits on daily function.


Involvement of endothelin in renal processes.
This study examined the effect of various doses of endothelin (from 0.2 to 2 nmol/kg body wt) on regional hemodynamics in conscious unrestrained rats. Normal rats were instrumented chronically with femoral artery and vein catheters and pulsed Doppler flow probes simultaneously on the renal and superior mesenteric arteries and the abdominal aorta. Endothelin induced a biphasic response of mean arterial pressure. First, endothelin provoked a sharp hypotension with tachycardia, vasodilation of the hindquarter, and a pronounced decrease in renal and mesenteric blood flows. After this initial response, endothelin induced a dose-dependent increase of mean arterial pressure. Changes in the hindquarter vascular resistance were less pronounced than those in renal and mesenteric vascular resistances. Endothelin (2 nmol/kg) reduced renal flow (-86%) resulting from a vasoconstriction (+1,818%) significantly more pronounced than for the mesenteric vascular bed. In another set of experiments, endothelin (2 nmol/kg) induced an increase in proteinuria, characterized by an increase in excreted albumin and by the appearance of proteins with molecular weights of 20,000-280,000. Renal vascular bed exhibited a pronounced sensitivity to the vasoconstrictive effect of endothelin associated with changes in renal function.


The role of approximate entropy in predicting ventricular defibrillation threshold.
The role of myocardial tissue mass on ventricular defibrillation threshold (DFT) is unclear. We hypothesized that changes in tissue mass modulate DFT by changing ventricular fibrillation (VF) wavefront regularity (entropy). The right ventricles (RV) of seven farm pigs were isolated, superfused and perfused through the right coronary artery with oxygenated Tyrode's solution at 37 degrees C. The epicardial surface was stained with the voltage sensitive dye, di-4-ANEPPS, and activation wavefront numbers (AWN) during VF were determined from the optical maps using a CCD camera (96 x 96 pixels over a 3.5 x 3.5 cm area). The RV mass was progressively reduced by sequential cutting of 1 to 2 g of tissue (approximately 12 cuts in total) distal to the perfusion site. After each cut, VF was reinduced, optical maps obtained, and the 50% probability of successful DFT(50) determined using an up-down algorithm. After each cut, the approximate entropy (ApEn) was also computed using 5 seconds of VF data obtained with a bipolar electrode and a pseudo-electrocardiogram. Tissue mass reduction of up to one third of the RV mass (ie, from 48.4 +/- 4.25 g to 34 +/- 4.7 g) caused little or no change in the DFT, ApEn or AWN. However, further progressive reduction of the RV mass near the critical mass of VF resulted in a significant (P < 0.05) progressive decrease in all three measured parameters. DFT energy was reduced by 27% (1.47 +/- 0.34 J vs. 1.02 +/- 0.14 J). There was a significant (P < 0.01) correlation between the DFT and ApEn, which significantly further increased (P < 0.001) near the critical mass. In a separate series of 6 isolated RVs, the ApEn correlated well with the Kolmogorov-Sinai (K-S) entropy, the standard method of calculating entropy. Tissue mass reduction significantly reduces DFT when the mass reduction increases VF wavefront regularity.


Major complications associated with transcatheter atrial septal occluder implantation: a review of the medical literature and the manufacturer and user facility device experience (MAUDE) database.
To summarize major complications and outcome for patients receiving percutaneous closure of atrial septal communications. The Medline database and the United States Food and Drug Administration manufacturer and user facility device experience databases (MAUDE) were searched for reports related to complications with atrial septal occluding devices. The medical literature documenting complication rates for these devices were reviewed and summarized. The MAUDE database complication reports were compared with those reported in the medical literature using national implant estimates. The MAUDE database correlated in the type of complications most frequently encountered with each device. However, based on estimated total implant numbers, there is a higher incidence of major complications, including death. AGA devices had a 0.3% erosion/perforation rate with a higher morbidity and mortality (29%) than previously reported. NMT devices had a lower incidence of erosion/perforation rate of 0.05%. Embolization rates for the NMT devices were also lower than published European studies, possibly reflecting the US restriction of the device for closure of patent foramen ovale. Thrombus was more frequently encountered on the NMT device. Both AGA and NMT devices have been shown to be safe and effective alternatives to cardiac surgery. The MAUDE database correlated, with a very low overall complication rate, but showed a higher estimated major complication rate than the medical literature. These data demonstrate the difficulty in quantifying rare complications in the premarketing analysis and the obligation providers have to report and evaluate complications through vigilant postmarketing surveillance.


The malnutrition-related increase in early visceralization of Leishmania donovani is associated with a reduced number of lymph node phagocytes and altered conduit system flow.
In a murine model of moderate childhood malnutrition we found that polynutrient deficiency led to a 4-5-fold increase in early visceralization of L. donovani (3 days post-infection) following cutaneous infection and a 16-fold decrease in lymph node barrier function (p<0.04 for all). To begin to understand the mechanistic basis for this malnutrition-related parasite dissemination we analyzed the cellularity, architecture, and function of the skin-draining lymph node. There was no difference in the localization of multiple cell populations in the lymph node of polynutrient deficient (PND) mice, but there was reduced cellularity with fewer CD11c(+)dendritic cells (DCs), fibroblastic reticular cells (FRCs), MOMA-2(+) macrophages, and CD169(+) subcapsular sinus macrophage (p<0.05 for all) compared to the well-nourished (WN) mice. The parasites were equally co-localized with DCs associated with the lymph node conduit network in the WN and PND mice, and were found in the high endothelial venule into which the conduits drain. When a fluorescent low molecular weight (10 kD) dextran was delivered in the skin, there was greater efflux of the marker from the lymph node conduit system to the spleens of PND mice (p<0.04), indicating that flow through the conduit system was altered. There was no evidence of disruption of the conduit or subcapsular sinus architecture, indicating that the movement of parasites into the subcortical conduit region was due to an active process and not from passive movement through a leaking barrier. These results indicate that the impaired capacity of the lymph node to act as a barrier to dissemination of L. donovani infection is associated with a reduced number of lymph node phagocytes, which most likely leads to reduced capture of parasites as they transit through the sinuses and conduit system.


Fission yeast Nda1 and Nda4, MCM homologs required for DNA replication, are constitutive nuclear proteins.
The nda1+ and nda4+ genes of the fission yeast Schizosaccharomyces pombe encode proteins similar to budding yeast MCM2 and MCM5/CDC46, respectively, which are required for the early stages of DNA replication. The budding yeast Mcm proteins display cell-cycle dependent localization. They are present in the nucleus specifically from late M phase until the beginning of S phase, so that they were suggested to be components of a replication licensing factor, a positive factor for the onset of replication, which is thought to be inactivated after use, thus restricting replication to only once in a cell cycle. In the present study, we raised antibodies against Nda1 or Nda4 and identified 115 kDa and 80 kDa proteins, respectively. Their immunolocalization was examined in wild-type cells and in various cell-cycle mutants. Both Nda1 and Nda4 proteins remained primarily in the nucleus throughout the cell cycle. In mutants arrested in G1, S, and G2 phases, these proteins were also enriched in the nucleus. These results indicate that the dramatic change in subcellular localization as seen in budding yeast is not essential in fission yeast for the functions of Nda1 and Nda4 proteins to be executed. The histidine-tagged nda1+ gene was constructed and integrated into the chromosome to replace the wild-type nda1+ gene. The resulting His-tagged Nda1 protein was adsorbed to the Ni-affinity column, and co-eluted with the untagged Nda4 protein, suggesting that they formed a complex.


Long-range magnetic response of toroidal boron structures: B16 and [Co@B16]-/3- species.
A correlation between the long-range characteristics of the magnetic response of toroidal boron-based structures is given, involving the uncoordinated B16 cluster and the hypercoordinated [Co@B16]-/3- counterparts. It is found that the perfectly symmetrical doubly aromatic systems share common features, involving a continuous shielding region for the orientation-averaged response (isotropic), and a long-ranged shielding cone under a perpendicularly oriented applied field (B). In contrast, the conflicting aromatic structure given by the slightly distorted species, exhibits an enhanced deshielding cone under B, which dominates the isotropic character of the response. In addition, [Mn@B16]- and [Cu@B16]- clusters were evaluated, denoting the role of the coordinated metal atom in such property. This information is valuable to account for a global magnetic response driven by the bonding pattern acting in each respective compound, and for the possible characterization of intermolecular aggregates or extended structures via NMR experiments.


The alternative reality of plant mitochondrial DNA: One ring does not rule them all.
Plant mitochondrial genomes are usually assembled and displayed as circular maps based on the widely-held view across the broad community of life scientists that circular genome-sized molecules are the primary form of plant mitochondrial DNA, despite the understanding by plant mitochondrial researchers that this is an inaccurate and outdated concept. Many plant mitochondrial genomes have one or more pairs of large repeats that can act as sites for inter- or intramolecular recombination, leading to multiple alternative arrangements (isoforms). Most mitochondrial genomes have been assembled using methods unable to capture the complete spectrum of isoforms within a species, leading to an incomplete inference of their structure and recombinational activity. To document and investigate underlying reasons for structural diversity in plant mitochondrial DNA, we used long-read (PacBio) and short-read (Illumina) sequencing data to assemble and compare mitochondrial genomes of domesticated (Lactuca sativa) and wild (L. saligna and L. serriola) lettuce species. We characterized a comprehensive, complex set of isoforms within each species and compared genome structures between species. Physical analysis of L. sativa mtDNA molecules by fluorescence microscopy revealed a variety of linear, branched, and circular structures. The mitochondrial genomes for L. sativa and L. serriola were identical in sequence and arrangement and differed substantially from L. saligna, indicating that the mitochondrial genome structure did not change during domestication. From the isoforms in our data, we infer that recombination occurs at repeats of all sizes at variable frequencies. The differences in genome structure between L. saligna and the two other Lactuca species can be largely explained by rare recombination events that rearranged the structure. Our data demonstrate that representations of plant mitochondrial genomes as simple, circular molecules are not accurate descriptions of their true nature and that in reality plant mitochondrial DNA is a complex, dynamic mixture of forms.


In situ synthesis of iron sulfide embedded porous carbon hollow spheres for sodium ion batteries.
Iron sulfide embedded porous carbon hollow spheres (FeSx@CS) have been successfully synthesized through a facile and environment friendly in situ synthetic method. Using this method, a porous carbon matrix and uniformly dispersed iron sulfide nanoparticles were formed simultaneously and assembled into hollow spheres. The as-synthesized FeSx@CS material showed high reversible capacity, outstanding cycle performance, and excellent rate capability when it was applied as an anode material for sodium ion batteries. It delivered a remarkable charge capacity of 656.75 mA h g-1 in the first cycle and a capacity retention of 97% after 100 cycles. When the current density was increased from 50 mA g-1 to 1000 and 2000 mA g-1, high capacity retention of 76.3% and 62.5% was achieved, respectively.


Neural precursor cell lines promote neurite branching.
Schwann cells and primary progenitor cells improve regeneration across peripheral nerve defects. This study examined the impact of immortalized neural precursor cells on regeneration of rat nerve defects. Across 10-mm gaps, neuromas formed without neural cables with C17.2- or RN33B-transplanted cells, but neural cables formed across 5-mm gaps seeded with RN33B cells. In vitro, dorsal root ganglia neurites elongated across Schwann and RN33B cells; RN33B cells induced neurite branching with shorter total outgrowth. Neural cable formation in vivo was likely determined by the balance of guidance and branch-inducing factors secreted by Schwann and transplanted precursor cells.


The desmin coil 1B mutation K190A impairs nebulin Z-disc assembly and destabilizes actin thin filaments.
Desmin intermediate filaments intimately surround myofibrils in vertebrate muscle forming a mesh-like filament network. Desmin attaches to sarcomeres through its high-affinity association with nebulin, a giant F-actin binding protein that co-extends along the length of actin thin filaments. Here, we further investigated the functional significance of the association of desmin and nebulin in cultured primary myocytes to address the hypothesis that this association is key in integrating myofibrils to the intermediate filament network. Surprisingly, we identified eight peptides along the length of desmin that are capable of binding to C-terminal modules 160-170 in nebulin. In this study, we identified a targeted mutation (K190A) in the desmin coil 1B region that results in its reduced binding with the nebulin C-terminal modules. Using immunofluorescence microscopy and quantitative analysis, we demonstrate that expression of the mutant desmin K190A in primary myocytes results in a significant reduction in assembled endogenous nebulin and desmin at the Z-disc. Non-uniform actin filaments were markedly prevalent in myocytes expressing GFP-tagged desmin K190A, suggesting that the near-crystalline organization of actin filaments in striated muscle depends on a stable interaction between desmin and nebulin. All together, these data are consistent with a model in which Z-disc-associated nebulin interacts with desmin through multiple sites to provide efficient stability to satisfy the dynamic contractile activity of myocytes.


The effect of clamping a tensioned wire: implications for the Ilizarov external fixation system.
This study demonstrates that clamping a tensioned wire can cause a reduction in wire tension. Tension (about 1275 N) was applied to a wire that was subsequently clamped, using cannulated bolts, to the steel half-ring of an Ilizarov external fixator. The tension in the wire was monitored before, during and after clamping. The apparatus was disassembled and the deformations in the wire caused by the clamps were measured. This experiment was repeated 15 times. When the wire was clamped to the frame, the wire tension was reduced by 22 +/- 7 per cent (mean +/- standard deviation, SD). The drop in wire tension was linearly correlated (r = 0.96; p < 0.001) with the deformation caused by the bolts. A finite element (FE) model of the wire was also constructed. The model was pre-stressed (tensioned), and the clamping effect replicated. This analysis showed that clamping the wire could be considered to squeeze the wire outwards (like toothpaste from a tube) and so reduce its tension during fixator assembly. To assess the magnitude of this effect in the clinical situation, the FE model analysis was repeated to replicate clamping a 1.8-mm-diameter wire to a 180-mm-diameter steel Ilizarov ring component. The analysis showed that for these conditions the tension reduced by 8-29 per cent. The results of this study highlight a general engineering problem: how can a tensioned wire be secured to a structure without an appreciable loss of tension? If the performance of the structure depends on the wire tension, this performance will change when the wire is secured.


GWAS of lifetime cannabis use reveals new risk loci, genetic overlap with psychiatric traits, and a causal influence of schizophrenia.
Cannabis use is a heritable trait that has been associated with adverse mental health outcomes. In the largest genome-wide association study (GWAS) for lifetime cannabis use to date (N = 184,765), we identified eight genome-wide significant independent single nucleotide polymorphisms in six regions. All measured genetic variants combined explained 11% of the variance. Gene-based tests revealed 35 significant genes in 16 regions, and S-PrediXcan analyses showed that 21 genes had different expression levels for cannabis users versus nonusers. The strongest finding across the different analyses was CADM2, which has been associated with substance use and risk-taking. Significant genetic correlations were found with 14 of 25 tested substance use and mental health-related traits, including smoking, alcohol use, schizophrenia and risk-taking. Mendelian randomization analysis showed evidence for a causal positive influence of schizophrenia risk on cannabis use. Overall, our study provides new insights into the etiology of cannabis use and its relation with mental health.


Pain perception in fish: indicators and endpoints.
Recent evidence has shown that fish display aversive behavioral and physiological reactions and a suspension of normal behavior in response to noxious stimuli that cause pain in other animals and humans. In addition to these behavioral responses, scientists have identified a peripheral nociceptive system and recorded specific changes in the brain activity of fish during noxious stimulation. As a result of these observations teleost fish are now considered capable of nociception and, in some opinions, pain perception. From both an experimental and an ethical perspective, it is important that scientists be able to assess possible pain and minimize discomfort that may result from invasive or other noxious procedures. If scientists accept that the definition of pain in animals cannot include direct measurement of subjective experience (the standard for humans), then fish fulfill the criteria for animal pain. In this review, recent evidence for pain is discussed in terms of the physiological properties of nociceptors, central responses to noxious stimulation, and changes in behavior and physiology that are indicative of nociception and are responsive to analgesia. To enable the assessment of potential pain, there are descriptions of newly identified robust indicators and species-specific responses that are easily measurable. The article concludes with a discussion of humane endpoints and of the need for alleviation of pain through the use of analgesia and anesthesia.


Chronic neutropenia following gold therapy.
We describe the first case of chronic neutropenia of 17 years' duration following gold therapy in a 53-year-old woman given a 1-g course of gold therapy in 1965 for treatment of seropositive rheumatoid arthritis. Although she had a good response to the gold therapy, her originally normal leukocyte count fell to 1.2 x 10(9)/L. Over the subsequent 17 years, she required multiple hospitalizations for recurrent skin, mouth, and respiratory tract infections. Serial leukocyte counts failed to show a cyclical nature to the chronic neutropenia. Normal results of a technetium Tc 99m spleen scan and lack of increased bone marrow leukocyte precursors rendered a diagnosis of Felty's syndrome unlikely. A bone marrow biopsy specimen revealed an isolated reduction in the number of myeloid precursors, which is consistent with gold-induced bone marrow toxicity. This patient's relative freedom from serious life-threatening infections remains enigmatic, but is undoubtedly related to her ability to augment another phagocytic cell line, and the remarkable phagocytic activity of her monocytes appears to have well compensated for her neutropenia. This activity was most likely responsible for her long-term survival.


Genomic events associated with progression of pleural malignant mesothelioma.
Pleural malignant mesothelioma (MM) is an aggressive cancer with a very long latency and a very short median survival. Little is known about the genetic events that trigger MM and their relation to poor outcome. The goal of our study was to characterize major genomic gains and losses associated with MM origin and progression and assess their clinical significance. We performed Representative Oligonucleotide Microarray Analysis (ROMA) on DNA isolated from tumors of 22 patients who recurred at variable interval with the disease after surgery. The total number of copy number alterations (CNA) and frequent imbalances for patients with short time (<12 months from surgery) and long time to recurrence were recorded and mapped using the Analysis of Copy Errors algorithm. We report a profound increase in CNA in the short-time recurrence group with most chromosomes affected, which can be explained by chromosomal instability associated with MM. Deletions in chromosomes 22q12.2, 19q13.32 and 17p13.1 appeared to be the most frequent events (55-74%) shared between MM patients followed by deletions in 1p, 9p, 9q, 4p, 3p and gains in 5p, 18q, 8q and 17q (23-55%). Deletions in 9p21.3 encompassing CDKN2A/ARF and CDKN2B were characterized as specific for the short-term recurrence group. Analysis of the minimal common areas of frequent gains and losses identified candidate genes that may be involved in different stages of MM: OSM (22q12.2), FUS1 and PL6 (3p21.3), DNAJA1 (9p21.1) and CDH2 (18q11.2-q12.3). Imbalances seen by ROMA were confirmed by Affymetrix genome analysis in a subset of samples.


Codon substitution in evolution and the "saturation" of synonymous changes.
A mathematical model for codon substitution is presented, taking into account unequal mutation rates among different nucleotides and purifying selection. This model is constructed by using a 61 X 61 transition probability matrix for the 61 nonterminating codons. Under this model, a computer simulation is conducted to study the numbers of silent (synonymous) and amino acid-altering (nonsynonymous) nucleotide substitutions when the underlying mutation rates among the four kinds of nucleotides are not equal. It is assumed that the substitution rates are constant over evolutionary time, the codon frequencies being in equilibrium, and, thus, the numbers of synonymous and nonsynonymous substitutions both increase linearly with evolutionary time. It is shown that, when the mutation rates are not equal, the estimate of synonymous substitutions obtained by F. Perler, A. Efstratiadis, P. Lomedico, W. Gilbert, R. Kolodner and J. Dodgson's "Percent Corrected Divergence" method increases nonlinearly, although the true number of synonymous substitutions increases linearly. It is, therefore, possible that the "saturation" of synonymous substitutions observed by Perler et al. is due to the inefficiency of their method to detect all synonymous substitutions.


Pomegranate as a cosmeceutical source: pomegranate fractions promote proliferation and procollagen synthesis and inhibit matrix metalloproteinase-1 production in human skin cells.
Pomegranate (Punica granatum) is an ancient fruit with exceptionally rich ethnomedical applications. The peel (pericarp) is well regarded for its astringent properties; the seeds for conferring invulnerability in combat and stimulating beauty and fertility. Here, aqueous fractions prepared from the fruit's peel and fermented juice and lipophilic fractions prepared from pomegranate seeds were examined for effects on human epidermal keratinocyte and human dermal fibroblast function. Pomegranate seed oil, but not aqueous extracts of fermented juice, peel or seed cake, was shown to stimulate keratinocyte proliferation in monolayer culture. In parallel, a mild thickening of the epidermis (without the loss of ordered differentiation) was observed in skin organ culture. The same pomegranate seed oil that stimulated keratinocyte proliferation was without effect on fibroblast function. In contrast, pomegranate peel extract (and to a lesser extent, both the fermented juice and seed cake extracts) stimulated type I procollagen synthesis and inhibited matrix metalloproteinase-1 (MMP-1; interstitial collagenase) production by dermal fibroblasts, but had no growth-supporting effect on keratinocytes. These results suggest heuristic potential of pomegranate fractions for facilitating skin repair in a polar manner, namely aqueous extracts (especially of pomegranate peel) promoting regeneration of dermis, and pomegranate seed oil promoting regeneration of epidermis.


Vulnerability of drinking water supplies to engineered nanoparticles.
The production and use of engineered nanoparticles (ENPs) inevitably leads to their release into aquatic environments, with the quantities involved expected to increase significantly in the future. Concerns therefore arise over the possibility that ENPs might pose a threat to drinking water supplies. Investigations into the vulnerability of drinking water supplies to ENPs are hampered by the absence of suitable analytical methods that are capable of detecting and quantifiying ENPs in complex aqueous matrices. Analytical data concerning the presence of ENPs in drinking water supplies is therefore scarce. The eventual fate of ENPs in the natural environment and in processes that are important for drinking water production are currently being investigated through laboratory based-experiments and modelling. Although the information obtained from these studies may not, as yet, be sufficient to allow comprehensive assessment of the complete life-cycle of ENPs, it does provide a valuable starting point for predicting the significance of ENPs to drinking water supplies. This review therefore addresses the vulnerability of drinking water supplies to ENPs. The risk of ENPs entering drinking water is discussed and predicted for drinking water produced from groundwater and from surface water. Our evaluation is based on reviewing published data concerning ENP production amounts and release patterns, the occurrence and behavior of ENPs in aquatic systems relevant for drinking water supply and ENP removability in drinking water purification processes. Quantitative predictions are made based on realistic high-input case scenarios. The results of our synthesis of current knowledge suggest that the risk probability of ENPs being present in surface water resources is generally limited, but that particular local conditions may increase the probability of raw water contamination by ENPs. Drinking water extracted from porous media aquifers are not generally considered to be prone to ENP contamination. In karstic aquifers, however, there is an increased probability that if any ENPs enter the groundwater system they will reach the extraction point of a drinking water treatment plant (DWTP). The ability to remove ENPs during water treatment depends on the specific design of the treatment process. In conventional DWTPs with no flocculation step a proportion of ENPs, if present in the raw water, may reach the final drinking water. The use of ultrafiltration techniques improves drinking water safety with respect to ENP contamination.


Obesity-induced gut microbial metabolite promotes liver cancer through senescence secretome.
Obesity has become more prevalent in most developed countries over the past few decades, and is increasingly recognized as a major risk factor for several common types of cancer. As the worldwide obesity epidemic has shown no signs of abating, better understanding of the mechanisms underlying obesity-associated cancer is urgently needed. Although several events were proposed to be involved in obesity-associated cancer, the exact molecular mechanisms that integrate these events have remained largely unclear. Here we show that senescence-associated secretory phenotype (SASP) has crucial roles in promoting obesity-associated hepatocellular carcinoma (HCC) development in mice. Dietary or genetic obesity induces alterations of gut microbiota, thereby increasing the levels of deoxycholic acid (DCA), a gut bacterial metabolite known to cause DNA damage. The enterohepatic circulation of DCA provokes SASP phenotype in hepatic stellate cells (HSCs), which in turn secretes various inflammatory and tumour-promoting factors in the liver, thus facilitating HCC development in mice after exposure to chemical carcinogen. Notably, blocking DCA production or reducing gut bacteria efficiently prevents HCC development in obese mice. Similar results were also observed in mice lacking an SASP inducer or depleted of senescent HSCs, indicating that the DCA-SASP axis in HSCs has key roles in obesity-associated HCC development. Moreover, signs of SASP were also observed in the HSCs in the area of HCC arising in patients with non-alcoholic steatohepatitis, indicating that a similar pathway may contribute to at least certain aspects of obesity-associated HCC development in humans as well. These findings provide valuable new insights into the development of obesity-associated cancer and open up new possibilities for its control.


No unjamming transition in a Voronoi model of biological tissue.
Vertex models are a popular approach to simulating the mechanical and dynamical properties of dense biological tissues, describing the tissue as a network of polygons. Recently a class of two-dimensional vertex models was shown to exhibit a disordered rigidity transition controlled by the preferred cellular geometry, which was subsequently echoed by experimental findings. An attractive variant of these models uses a Voronoi tessellation to describe the cells, which reduces the number of degrees of freedom as compared the original vertex model. The Voronoi model was also endowed with a non-equilibrium model of cellular motility, leading to rich, glassy behavior. This glassy behavior was suggested to be inextricably linked to an underlying jamming transition. We test this conjecture, exploring the low-effective-temperature limit of the 2D Voronoi model by studying cell trajectories from detailed dynamical simulations in combination with rigidity measurements of energy-minimized disordered cell configurations. We find that the zero-temperature limit of this model has no unjamming transition. We show that this absence of an unjamming transition is intimately linked to the marginality of the model, i.e. the fact that the constraints imposed on cell areas and perimeters precisely balance the number of degrees of freedom in the model. Our work suggests that constraint counting arguments are useful to understand rigidity in a broad class of models of dense biological tissues.


Psychological therapies for the treatment of depression in chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) has been recognised as a global health concern, and one of the leading causes of morbidity and mortality worldwide. Projections of the World Health Organization (WHO) indicate that prevalence rates of COPD continue to increase, and by 2030, it will become the world's third leading cause of death. Depression is a major comorbidity amongst patients with COPD, with an estimate prevalence of up to 80% in severe stages of COPD. Prevalence studies show that patients who have COPD are four times as likely to develop depression compared to those without COPD. Regrettably, they rarely receive appropriate treatment for COPD-related depression. Available findings from trials indicate that untreated depression is associated with worse compliance with medical treatment, poor quality of life, increased mortality rates, increased hospital admissions and readmissions, prolonged length of hospital stay, and subsequently, increased costs to the healthcare system. Given the burden and high prevalence of untreated depression, it is important to evaluate and update existing experimental evidence using rigorous methodology, and to identify effective psychological therapies for patients with COPD-related depression. To assess the effectiveness of psychological therapies for the treatment of depression in patients with chronic obstructive pulmonary disease. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (2018, Issue 11), and Ovid MEDLINE, Embase and PsycINFO from June 2016 to 26 November 2018. Previously these databases were searched via the Cochrane Airways and Common Mental Disorders Groups' Specialised Trials Registers (all years to June 2016). We searched ClinicalTrials.gov, the ISRCTN registry, and the World Health Organization International Clinical Trials Registry Platform (ICTRP) to 26 November 2018 to identify unpublished or ongoing trials. Additionally, the grey literature databases and the reference lists of studies initially identified for full-text screening were also searched. Eligible for inclusion were randomised controlled trials that compared the use of psychological therapies with either no intervention, education, or combined with a co-intervention and compared with the same co-intervention in a population of patients with COPD whose depressive symptoms were measured before or at baseline assessment. Two review authors independently assessed the titles and abstracts identified by the search to determine which studies satisfied the inclusion criteria. We assessed two primary outcomes: depressive symptoms and adverse events; and the following secondary outcomes: quality of life, dyspnoea, forced expiratory volume in one second (FEV1), exercise tolerance, hospital length of stay or readmission rate, and cost-effectiveness. Potentially eligible full-text articles were also independently assessed by two review authors. A PRISMA flow diagram was prepared to demonstrate the decision process in detail. We used the Cochrane 'Risk of bias' evaluation tool to examine the risk of bias, and assessed the quality of evidence using the GRADE framework. All outcomes were continuous, therefore, we calculated the pooled standardised mean difference (SMD) or mean difference (MD) with a corresponding 95% confidence interval (CI). We used a random-effects model to calculate treatment effects. The findings are based on 13 randomised controlled trials (RCTs), with a total of 1500 participants. In some of the included studies, the investigators did not recruit participants with clinically confirmed depression but applied screening criteria after randomisation. Hence, across the studies, baseline scores for depressive symptoms varied from no symptoms to severe depression. The severity of COPD across the studies was moderate to severe.Primary outcomesThere was a small effect showing the effectiveness of psychological therapies in improving depressive symptoms when compared to no intervention (SMD 0.19, 95% CI 0.05 to 0.33; P = 0.009; 6 studies, 764 participants), or to education (SMD 0.23, 95% CI 0.06 to 0.41; P = 0.010; 3 studies, 507 participants).Two studies compared psychological therapies plus a co-intervention versus the co-intervention alone (i.e. pulmonary rehabilitation (PR)). The results suggest that a psychological therapy combined with a PR programme can reduce depressive symptoms more than a PR programme alone (SMD 0.37, 95% CI -0.00 to 0.74; P = 0.05; 2 studies, 112 participants).We rated the quality of evidence as very low. Owing to the nature of psychological therapies, blinding of participants, personnel, and outcome assessment was a concern.None of the included studies measured adverse events.Secondary outcomesQuality of life was measured in four studies in the comparison with no intervention, and in three studies in the comparison with education. We found inconclusive results for improving quality of life. However, when we pooled data from two studies using the same measure, the result suggested that psychological therapy improved quality of life better than no intervention. One study measured hospital admission rates and cost-effectiveness and showed significant reductions in the intervention group compared to the education group. We rated the quality of evidence as very low for the secondary outcomes. The findings from this review indicate that psychological therapies (using a CBT-based approach) may be effective for treating COPD-related depression, but the evidence is limited. Depressive symptoms improved more in the intervention groups compared to: 1) no intervention (attention placebo or standard care), 2) educational interventions, and 3) a co-intervention (pulmonary rehabilitation). However, the effect sizes were small and quality of the evidence very low due to clinical heterogeneity and risk of bias. This means that more experimental studies with larger numbers of participants are needed, to confirm the potential beneficial effects of therapies with a CBT approach for COPD-related depression.New trials should also address the gap in knowledge related to limited data on adverse effects, and the secondary outcomes of quality of life, dyspnoea, forced expiratory volume in one second (FEV1), exercise tolerance, hospital length of stay and frequency of readmissions, and cost-effectiveness. Also, new research studies need to adhere to robust methodology to produce higher quality evidence.


Nanoradian angle sensor and in situ self-calibration.
We describe a high-precision angle sensor and a new calibration system for calibrating the linearity error of an angle sensor under in situ conditions without the need for accurate reference instruments. To achieve accuracy in the nanoradian (nrad) order, we elongated the critical-angle prism to increase the sensitivity, and we added a thermal controller to the angle sensor. This served to suppress thermal drift of the light source to improve stability. The proposed angle sensor demonstrated a stability of 3 nrad over 1 min, and the maximum repeatability error of the calibration curve determined by the in situ self-calibration method was approximately 35 nrad with a measurement range of 500 murad.


Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway.
Chimeric genes were created by fusing DNA sequences encoding the ectodomain of the influenza virus hemagglutinin (HA) to DNA coding for the transmembrane and cytoplasmic domains of either the G glycoprotein of vesicular stomatitis virus or the gC glycoprotein of Herpes simplex virus 1. CV-1 cells infected with SV40 vectors carrying the recombinant genes expressed large amounts of the chimeric proteins, HAG or HAgC on their surfaces. Although the ectodomains of HAG and HAgC differed in their immunological properties from that of HA, the chimeras displayed the biological functions characteristic of the wild-type protein. Both HAG and HAgC bound erythrocytes as efficiently as HA did and, after brief exposure to an acidic environment, induced the fusion of erythrocyte and CV-1 cell membranes. However, the behavior of HAG and HAgC at the cell surface differed from that of HA in several important respects. HAG and HAgC were observed to collect in coated pits whereas wild-type HA was excluded from those structures. In the presence of chloroquine, which inhibits the exit of receptors from endosomes, HAG and HAgC accumulated in intracellular vesicles. By contrast, chloroquine had no effect on the location of wild-type HA. HAG and HAgC labeled at the cell surface exhibited a temperature-dependent acquisition of resistance to extracellular protease at a rate similar to the rates of internalization observed for many cell surface receptors. HA acquired resistance to protease at a rate at least 20-fold slower. We conclude that HAG and HAgC are efficiently routed into the endocytic pathway and HA is not. However, like HA, HAG was degraded slowly, raising the possibility that HAG recycles to the plasma membrane.


Uncovering the basis of protein-protein interaction specificity with a combinatorially complete library.
Protein-protein interaction specificity is often encoded at the primary sequence level. However, the contributions of individual residues to specificity are usually poorly understood and often obscured by mutational robustness, sequence degeneracy, and epistasis. Using bacterial toxin-antitoxin systems as a model, we screened a combinatorially complete library of antitoxin variants at three key positions against two toxins. This library enabled us to measure the effect of individual substitutions on specificity in hundreds of genetic backgrounds. These distributions allow inferences about the general nature of interface residues in promoting specificity. We find that positive and negative contributions to specificity are neither inherently coupled nor mutually exclusive. Further, a wild-type antitoxin appears optimized for specificity as no substitutions improve discrimination between cognate and non-cognate partners. By comparing crystal structures of paralogous complexes, we provide a rationale for our observations. Collectively, this work provides a generalizable approach to understanding the logic of molecular recognition.


Thalidomide for the treatment of uremic pruritus: a crossover randomized double-blind trial.
Our observation that thalidomide administration to a dialysis patient with leprosy alleviated his pruritus led us to conduct this short-term study to assess the efficacy of the drug in this regard. From 210 hemodialysis patients, 29 cases of refractory uremic pruritus were entered into the study. Patients were instructed to score their symptoms from 0 to 3, three times a day and assigned to receive thalidomide or placebo at bed time for 7 days. After a washout period of 7 days, drugs were crossed over. Response was defined as a reduction of at least 50% in the pruritus scoring. Eighteen patients finished the study. In the first phase, 55% of patients responded showing a mean reduction in their pruritus scoring of 78% (p < 0.05 vs. placebo); no response to placebo was observed. A similar proportion of patients responded to thalidomide in the second phase with a mean reduction in their pruritus scoring of 81%. In conclusion, thalidomide can be a precious tool in the handling of uremic pruritus unresponsive to available therapy.


Aging of Transition Metal Dichalcogenide Monolayers.
Two-dimensional sheets of transition metal dichalcogenides are an emerging class of atomically thin semiconductors that are considered to be "air-stable", similar to graphene. Here we report that, contrary to current understanding, chemical vapor deposited transition metal dichalcogenide monolayers exhibit poor long-term stability in air. After room-temperature exposure to the environment for several months, monolayers of molybdenum disulfide and tungsten disulfide undergo dramatic aging effects including extensive cracking, changes in morphology, and severe quenching of the direct gap photoluminescence. X-ray photoelectron and Auger electron spectroscopy reveal that this effect is related to gradual oxidation along the grain boundaries and the adsorption of organic contaminants. These results highlight important challenges associated with the utilization of transition metal dichalcogenide monolayers in electronic and optoelectronic devices. We also demonstrate a potential solution to this problem, featuring encapsulation of the monolayer sheet by a 10-20 nm thick optically transparent polymer (parylene C). This strategy is shown to successfully prevent the degradation of the monolayer material under accelerated aging (i.e., high-temperature, oxygen-rich) conditions.