Formation of radical anions and not deprotonation starts the reaction of gamma-butyrolactone with potassium anions.
A mechanism is proposed for the reaction of gamma-butyrolactone with the potassium anion as a two-electron-transfer reagent. Potassium hydride and potassium 4-potassiobutyrate are formed in this process as intermediates. These compounds deprotonate gamma-butyrolactone. Potassium lactone enolate, potassium butyrate, and hydrogen are the final reaction products.


EI24 alleviates renal interstitial fibrosis through inhibition of epithelial-mesenchymal transition and fibroblast activation.
Etoposide-induced 2.4 (EI24) exerts tumor suppressor activity through participating in cell apoptosis, autophagy, and inflammation. However, its role in renal diseases has not been elucidated. This study showed that the EI24 level decreased gradually in the kidneys of mice with unilateral ureteral obstruction (UUO) and in another fibrosis model induced by diabetic kidney disease. The overexpression of EI24 was used to investigate the possible role both in vivo and in vitro. The overexpression 1 day after UUO through tail vein injection alleviated the progression of renal interstitial fibrosis (RIF). EI24 inhibited epithelial-mesenchymal transition, excessive deposition of the extracellular matrix, and activation of fibroblasts. Furthermore, administration of EI24-overexpressing plasmids restrained the phosphorylation of nuclear factor-κB (NF-κB) and c-Jun kinase (JNK) through regulating the proteasome-dependent degradation of TRAF2, and then, inhibited the expression of downstream inflammation-associated cytokines (interleukin-6, tumor necrosis factor-α, and monocyte chemotactic protein-1) and infiltration of macrophages and neutrophils in mouse kidney after UUO. In conclusion, the data indicated that EI24, a novel anti-fibrosis regulator, was important in the progression of RIF.


Self-diagnosis and self-treatment of malaria in hard-to-reach and mobile populations of the Amazon: results of Malakit, an international multicentric intervention research project.
Illegal gold miners are currently key hosts for malaria in French Guiana (FG), with a risk of emergence of resistance linked to improper use of artemisinin-based combination therapy (ACT). The remoteness of the mines and regulatory issues hinder their access to health care. A quasi-experimental researched project (Malakit) implemented in FG borders with Brazil and Suriname aimed at determining the effectiveness of distributed kits for self-diagnosis and self-treatment to illegal gold miners, after training, at strategic border staging areas. Evaluation relied on questionnaires at inclusion and follow-up visits, and pre/post intervention surveys. The primary outcome was the proportion of persons reporting a use of certified ACT after a positive malaria diagnosis. The secondary outcomes assessed antimalarial adherence, kit use and impact on malaria epidemiology. The proportion of patients reporting a use of certified ACT after a positive diagnosis increased after the intervention (OR 1.8, 95%CI [1.1-3.0]). From April 2018 to March 2020, 3,733 persons participated in the intervention. The kit was used correctly by 71.7% [65.8-77.7] of the 223 persons reporting having used a malakit during the follow-up visits. No serious adverse events related to the misuse of malakit have been reported. The intervention appears to have accelerated the decline in malaria incidence in the region by 42.9%. This innovative international project showed that people with low education can correctly self-manage their malaria symptoms. This strategy could be integrated in the malaria control programs of the countries involved and considered in other regions with residual malaria in remote areas. This project was funded by the European Union, the Global Fund, Brazil MoH, Cayenne Hospital and FG Health Regional Agency. This translation in French was submitted by the authors and we reproduce it as supplied. It has not been peer reviewed. Our editorial processes have only been applied to the original abstract in English, which should serve as reference for this manuscript. The Lancet Group takes a neutral position with respect to territorial claims in published maps and institutional affiliations. Les chercheurs d'or illégaux sont actuellement un réservoir clé du paludisme en Guyane, avec un risque d'émergence de résistance lié à une mauvaise utilisation des combinaisons thérapeutiques à base d'artémisinine (ACT). L'isolement de ces sites miniers clandestins et des contraintes règlementaires entravent leur accès aux soins. Un projet de recherche opérationnelle quasi-expérimental (Malakit) a été mis en œuvre aux frontières de la Guyane avec le Brésil et le Suriname. Il visait à déterminer l'efficacité de la distribution de kits d'autodiagnostic et d'autotraitement à des orpailleurs illégaux, après une formation adaptée, dans des zones stratégiques transfrontalières. L'évaluation s'est appuyée sur des questionnaires lors des visites d'inclusion et de suivi, et sur des enquêtes pré/post intervention. L'indicateur principal était la proportion de personnes déclarant avoir utilisé une ACT certifiée après un diagnostic positif de paludisme. Les indicateurs secondaires reposaient sur l'adhérence aux traitements antipaludiques, l'utilisation des kits et l'impact sur l'épidémiologie du paludisme. La proportion de patients déclarant une utilisation d'ACT certifiée après un diagnostic positif a augmenté après l'intervention (OR 1,8, 95%CI [1,1-3,0]). D'avril 2018 à mars 2020, 3 733 personnes ont participé à l'intervention. Le kit a été utilisé correctement par 71,7% [65,8-77,7] des 223 personnes revues en visites de suivi ayant déclaré avoir utilisé un malakit. Aucun événement indésirable grave lié à une mauvaise utilisation du malakit n'a été signalé. L'intervention semble avoir accéléré la diminution de l'incidence du paludisme dans la région de 42,9%. Ce projet international innovant a montré que les personnes ayant un faible niveau d'éducation peuvent se prendre en charge par eux-mêmes pour des symptômes de paludisme. Cette stratégie pourrait être intégrée dans les programmes de lutte contre le paludisme des pays impliqués et envisagée dans d'autres régions où du paludisme résiduel persiste dans des zones isolées. Ce projet a été financé par l'Union Européenne, le Fonds Mondial, le Ministère de la santé du Brésil, le Centre Hospitalier de Cayenne et l'Agence Régionale de Santé de Guyane.


Prions remodel gene expression in yeast.
Epigenetic mechanisms participate in the regulation of gene transcription in eukaryotes. Two studies in yeast have revealed an additional mechanism for controlling global gene transcription that is based on an inherited self-perpetuating change in the conformation of two different components of key transcriptional regulatory complexes.


Single-cell RNA sequencing reveals the heterogeneity of liver-resident immune cells in human.
The liver plays a critical role in both immune defense and tolerance in the body. The liver-resident immune cells (LrICs) determine the immune properties, but the unique composition and heterogeneity of these cells are incompletely understood. Here, we dissect the diversity of LrICs by a comprehensive transcriptomic profiling using the unbiased single-cell RNA-sequencing (scRNA-seq). A total of 70, 706 of CD45+ immune cells from the paired liver perfusion, spleen and peripheral blood as references were profiled. We identified more than 30 discrete cell populations comprising 13 of T and NK cell, 7 of B cell, 4 of plasma cell, and 8 of myeloid cell subsets in human liver and donor-paired spleen and blood, and characterized their tissue distribution, gene expression and functional modules. Especially, four of CXCR6+ T and NK cell subsets were found to be present preferentially in the liver, where they manifested heterogeneity, distinct function and prominent homeostatic proliferation. We propose a universal category system of T and NK cells based on distinct chemokine receptors, confirmed subsequently by phenotype, transcriptional factors and functionality. We also identified adaptive changes by the spleen and liver-derived monocyte and macrophage populations. Finally, we give a global glimpse on B cell and plasma cell subsets in human spleen and liver. We, therefore, reveal the heterogeneity and functional diversity of LrICs in human. This study presents comprehensively the landscape of LrICs and will enable further study on their roles in various human diseases.


Thirteen novel mutations in the NR0B1 (DAX1) gene as cause of adrenal hypoplasia congenita.
X-linked adrenal hypoplasia congenita (AHC) is a rare developmental disorder associated with primary adrenal insufficiency and combined primary and secondary male hypogonadism. It is caused by deletions or mutations of the NR0B1 (DAX1) gene encoding DAX1, an atypical orphan member of the nuclear receptor superfamily. The continuous molecular genetic analysis of male patients with primary adrenal insufficiency revealed 13 novel mutations within the coding region of the NR0B1 gene which are predicted to inactivate the DAX1 function. These were three nonsense mutations (c.312C>A, p.Cys104X, c.670C>T, p.Gln224X; and c.873G>A, p.Trp291X), five duplications (c.269_270dup, c.421_422dup, c.895_896dup, c.989dup, c.999_1000dup), and five deletions (c.483del, c.745_746del, c.734_740del, c.1092del, and c.1346del). All of the mutations resulted in a premature stop codon destroying the ligand binding domain of the predictive DAX1 protein.


IL-10 delivery by AAV5 vector attenuates inflammation in mice with Pseudomonas pneumonia.
Lung infections with Pseudomonas aeruginosa and other pathogens in cystic fibrosis (CF) cause progressive airway obstruction and tissue damage, the predominant cause of morbidity and mortality in CF. We investigated whether a recombinant adeno-associated virus type 5 (AAV5) vector expressing murine interleukin (IL)-10 (AAV5.Cbeta-mIL-10), a regulatory/anti-inflammatory cytokine, could decrease airway inflammation in IL-10 knockout mice chronically infected with mucoid P. aeruginosa. Mice that received AAV5.Cbeta-mIL10 through intratracheal inoculation produced IL-10 at an average of 25 000 pg/ml in the epithelial lining fluid (ELF) and 12 000 pg/g-lung tissue 6 weeks post-vector delivery, significantly higher levels than in placebo-treated mice. At 3 days post-infection, proinflammatory cytokines (IL-1beta, tumor necrosis factor (TNF)-alpha, macrophage inhibitory protein (MIP)-1alpha and (KC) in the ELF and lung homogenate were decreased (1-9 folds) in the AAV5.Cbeta-mIL10-treated mice accompanied by less pronounced and more localized neutrophil infiltration in lung sections, when compared with placebo-treated mice. These results suggest that AAV5.Cbeta-mIL10 induces IL-10 levels in the lungs mediating a significant anti-inflammatory response and making AAV-IL-10 gene transfer a potentially useful therapy in the treatment of CF lung disease.


Identifying and quantifying genotypes in polyclonal infections due to single species.
Simultaneous infection with multiple pathogens of the same species occurs with HIV, hepatitis C, Epstein-Barr virus, dengue, tuberculosis, and malaria. However, available methods do not distinguish among or quantify pathogen genotypes in individual patients; they also cannot test for novel insertions and deletions in genetically modified organisms. The strategy reported here accomplishes these goals with real-time polymerase chain reaction (PCR) and capillary electrophoresis. Real-time PCR with allotype-specific primers defines the allotypes (strains) present and the intensity of infection (copy number). Capillary electrophoresis defines the number of genotypes within each allotype and the intensity of infection by genotype. This strategy can be used to study the epidemiology of emerging infectious diseases with simultaneous infection by multiple genotypes, as demonstrated here with malaria. It also permits testing for insertions or deletions in genetically modified organisms that may be used for bioterrorism.


An approach to determining the nitrofurazone-induced toxic dynamics for ecotoxicity assessment using protozoan periphytons in marine ecosystems.
With several observable responses and sensitivity of protozoans to nitrofurazone (NFZ), the toxic effects of NFZ on protozoans can be an early warning signal of NFZ contamination in the aquatic environment. To evaluate the toxic dynamics induced by NFZ, protozoan samples were collected using microscopy glass slides and exposed to the five concentrations of NFZ: 0, 1, 2, 4, and 8 mg ml-1. Substantial differences in the species composition and toxic-dynamics patterns were observed among all concentrations. Briefly, periphytic euplotids and pleurostomatids were the most prevalent at each concentration level, while dysteriids were less dominant among all treatments. Multivariate analysis revealed significant (P < 0.05) differences in the taxonomic patterns of the test organisms among the five treatments. Furthermore, significant deviation of protozoan communities from the expected taxonomic breadth was observed to occur in a dose-dependent manner. Based on these findings, it is suggested that protozoan periphytons could be used as bioindicators to assess the ecotoxicity of NFZ in the marine environment.


lncRNA CYTOR promotes aberrant glycolysis and mitochondrial respiration via HNRNPC-mediated ZEB1 stabilization in oral squamous cell carcinoma.
Oral squamous cell carcinoma (OSCC), the most common malignancy of the oral and maxillofacial region, severely affects human health. However, current treatments for OSCC commonly show only a ~60% 5-year survival rate of patients with distant metastases, indicating an urgent need for targeted treatments for patients with advanced metastases. Here, we report a survival-related long non-coding RNA, CYTOR, which is highly expressed in the lesions of oral cancer patients. We found that CYTOR can promote both migration and invasion in oral cancer cells as well as the epithelial-mesenchymal transition (EMT). RNA-sequencing of CYTOR-knockdown oral cancer cells revealed that CYTOR can regulate mitochondrial respiration and RNA splicing. Mechanistically, we found that nuclear-localized CYTOR interacts with HNRNPC, resulting in stabilization of ZEB1 mRNAs by inhibiting the nondegradative ubiquitination of HNRNPC. By synthesizing CYTOR-targeting small interfering RNAs (siRNAs) encapsulated in Nanoscale Metal Organic Frameworks (NMOFs), we demonstrate the targeted suppression of CYTOR to inhibit invasion and metastasis of oral cancer cells in a nude mouse model. Cumulatively, this study reveals the potential role of the CYTOR-HNRNPC-ZEB1 axis in regulating mitochondrial metabolism and glycolysis of oral cancer cells, and illustrates the effective use of lncRNA targeting in anti-metastatic cancer therapies.


A dataset of global ocean alkaline phosphatase activity.
Utilisation of dissolved organic phosphorus (DOP) by marine microbes as an alternative phosphorus (P) source when phosphate is scarce can help sustain non-Redfieldian carbon:nitrogen:phosphorus ratios and efficient ocean carbon export. However, global spatial patterns and rates of microbial DOP utilisation are poorly investigated. Alkaline phosphatase (AP) is an important enzyme group that facilitates the remineralisation of DOP to phosphate and thus its activity is a good proxy for DOP-utilisation, particularly in P-stressed regions. We present a Global Alkaline Phosphatase Activity Dataset (GAPAD) with 4083 measurements collected from 79 published manuscripts and one database. Measurements are organised into four groups based on substrate and further subdivided into seven size fractions based on filtration pore size. The dataset is globally distributed and covers major oceanic regions, with most measurements collected in the upper 20 m of low-latitude oceanic regions during summer since 1997. This dataset can help support future studies assessing global ocean P supply from DOP utilisation and provide a useful data reference for both field investigations and modelling activities.


The integrative regulatory network of circRNA and microRNA in keloid scarring.
Circular RNA (circRNA), a novel type of non-coding RNA that consists of a circular loop, has been demonstrated to act as a "sponge" for microRNAs (miRNAs). However, the role of circRNAs in keloid remains unknown. In this study, we investigated circRNA expression profiles in keloid to identify potential diagnostic and therapeutic circRNAs. We performed a circRNA microarray assay to determine circRNA expression in keloid and paired normal skin tissues. Quantitative reverse transcription polymerase chain reaction was used to evaluate the expression levels of candidate circRNAs. The most significantly over-expressed circRNA was used to predict putative miRNA targets and the binding sites of miRNAs with this circRNA. Finally, we constructed a circRNA-miRNA interaction network and carried out gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. We found 52 significantly upregulated and 24 downregulated circRNAs in keloid compared with normal skin tissue. We confirmed that hsa_circ_0057452, hsa_circ_0007482, hsa_circ_0020792, hsa_circ_0057342, and hsa_circ_0043688 were significantly upregulated in keloid tissues. Analysis of the circRNA-miRNA interaction network revealed that circRNAs could interact with miRNAs, including miRNA-29a, miRNA-23a-5p and miRNA-1976. GO and KEGG analyses indicated that these target genes were involved in biological functions and signaling pathways that may play vital roles in the pathogenesis of keloid. This study revealed that circRNAs are potentially implicated in the development of keloid and could serve as novel diagnostic and therapeutic targets.


Deformation Behavior of Cross-Linked Supercrystalline Nanocomposites: An in Situ SAXS/WAXS Study during Uniaxial Compression.
With the ever-expanding functional applications of supercrystalline nanocomposites (a relatively new category of materials consisting of organically functionalized nanoparticles arranged into periodic structures), it becomes necessary to ensure their structural stability and understand their deformation and failure mechanisms. Inducing the cross-linking of the functionalizing organic ligands, for instance, leads to a remarkable enhancement of the nanocomposites' mechanical properties. It is however still unknown how the cross-linked organic phase redistributes applied loads, how the supercrystalline lattice accommodates the imposed deformations, and thus in general what phenomena govern the overall material's mechanical response. This work elucidates these aspects for cross-linked supercrystalline nanocomposites through an in situ small- and wide-angle X-ray scattering study combined with uniaxial pressing. Because of this loading condition, it emerges that the cross-linked ligands effectively carry and distribute loads homogeneously throughout the nanocomposites, while the superlattice deforms via rotation, slip, and local defects generation.


Inhibition of Nrf2-mediated glucose metabolism by brusatol synergistically sensitizes acute myeloid leukemia to Ara-C.
Chemotherapy resistance remains to be the primary barrier to acute myeloid leukemia (AML) treatment failure. Nuclear factor-erythroid 2-related factor 2 (Nrf2) has been well established as a truly pleiotropic transcription factor. Inhibition of Nrf2 function increases the sensitivity of various chemotherapeutics and overcomes chemoresistance effectively. Brusatol (Bru) has been reported to decrease Nrf2 protein expression specifically by ubiquitin degradation of Nrf2. However, it remains elusive whether combination of Brusatol and Cytarabine (Ara-C) elicits a synergistic antitumor effect in AML. Our results demonstrated that combination of Ara-C and Brusatol synergistically exerted remarkable pro-apoptosis effect in HL-60 and THP-1 cells. Mechanistically, synergistic anti-tumor effect of Ara-C/Brusatol in AML cells is mediated by attenuating Nrf2 expression. To our surprise, Nrf2 inhibition by Brusatol causes downregulation of the expression of glycolysis-related proteins and decreased glucose consumption and lactate production, whereas the level of ROS production was unaffected. The activation of Nrf2 by Sulforaphane (SFP) could reverse the chemotherapeutic effect and changes of glycolysis of concomitant of Ara-C with Brusatol in AML cell lines. Additionally, Ara-C/Brusatol co-treatment decreased Glucose-6-phosphate dehydrogenase (G6PD) protein expression and increased the sensitivity of Ara-C. Moreover, the mouse xenograft in vivo experiment confirmed that combining Ara-C with Brusatol exerted stronger antileukemia than Ara-C alone. The efficacy, together with the mechanistic observations, reveals the potential of simultaneously giving these two drugs and provides a rational basis for targeting glucose catabolism in future clinical therapeutic approach.


Pseudomelanocytic nests mimicking atypical melanocytic proliferations: first reported cases in the oral cavity.
The concept of pseudomelanocytic nests has been recently described in the dermatology literature. To our knowledge, this entity has yet to be published in the oral pathology literature. We report 2 cases with features of pseudomelanocytic nests. In both instances, nests of cells suspicious for melanocytes were observed. Interpretation of melan-A was negative. Both cases showed strong and diffuse immunoreactivity of the nested cells to CD68. This immunohistochemical staining pattern is most consistent with a melanophage identity. Pseudomelanocytic nests are a recently described entity that represents a potential diagnostic pitfall. Distinguishing pseudomelanocytic nests from an authentic atypical melanocytic proliferation can be challenging and is important for appropriate patient management. Clinicopathologic correlation with cautious interpretation of immunohistochemistry may be necessary to arrive at the correct diagnosis. These cases represent the first reports of pseudomelanocytic nests in the oral pathology literature.


Chromosome 4q31-34 panic disorder risk locus: association of neuropeptide Y Y5 receptor variants.
There is strong evidence for a genetic contribution to the pathogenesis of panic disorder, with a recent linkage study pointing toward a risk locus on chromosome 4q31-q34 [Kaabi et al., 2006]. Since the neuropeptide Y (NPY) system has been reported to be involved in the pathophysiology of anxiety and in particular panic disorder and the genes coding for NPY Y1, Y2, and Y5 receptors are located in the suggested risk region (4q31-q32), variants in the NPY, NPY Y1, Y2, and Y5 genes were investigated for association with panic disorder in a sample of 230 German patients with panic disorder and matched healthy controls. A synonymous (Gly-426-Gly) NPY Y5 coding variant (rs11946004) as well as haplotypes including rs11946004 and an intronic NPY Y5 variant (rs11724320) were significantly associated with panic disorder (P = 0.027), with the effect originating from the subgroup of female patients (P = 0.030), particularly with concurrent agoraphobia (P = 0.002-0.019). No association was observed for any variants located in the genes coding for NPY, NPY Y1, or Y2. The present results provide preliminary support for an influence of NPY Y5 receptor variants on the etiology of panic disorder in a potentially gender-specific manner further strengthening the evidence for a risk locus on chromosome 4q31-q34 in anxiety disorders. However, in order to allow for conclusive evaluation of the present finding and to exclude a false positive result, further studies in larger, independent, preferably family based samples are warranted.


Idiopathic versus secondary venous thromboembolism. Findings of the RIETE registry.
The Computerized Registry of Patients with Venous Thromboembolism (RIETE) is a prospective registry that consecutively includes patients diagnosed with venous thromboembolism. We compared the clinical presentation and response to anticoagulant treatment in patients with idiopathic venous thromboembolism (IVT) versus secondary venous thromboembolism (SVT, associated with a risk factor). We analyzed the differences in clinical characteristics, comorbidity, treatment and events during the first 3months after the diagnosis of venous thromboembolism in patients with IVT or SVT and according to their initial clinical presentation. A total of 39,921 patients with IVT (n=18,029; 45.1%) or SVT (n=21,892; 54.9%) were enrolled. The patients with IVT had a greater history of venous thromboembolism than those diagnosed with SVT (p<.001). The initial treatment was similar for the 2 groups, but more inferior vena cava filters were placed in the SVT group (p<.001). In the long term, low-molecular-weight heparin was used more often in the SVT group than in the IVT group. At 90days, bleeding, death and the recurrence of venous thromboembolism were significantly more frequent in the SVT group. The multivariate analysis confirmed that IVT was associated with fewer major (OR, 0.60; 95%CI, 0.50-0.61; p<.001) and fatal (OR, 0.41; 95%CI, 0.29-0.62; p<.001) bleedings, fewer relapses (OR, 0.58; 95%CI, 0.39-0.78; p<.001) and fewer fatal pulmonary embolisms (OR, 0.29; 95%CI, 0.12-0.52; p<.001). These differences were maintained in patients whose venous thromboembolism started with a pulmonary embolism or with deep vein thrombosis. IVT has a better prognosis than SVT at 90days of the diagnosis.


Evaluating DNA damage response through immunofluorescence staining of primordial germ cells in Caenorhabditis elegans L1 larva.
C. elegans L1 larvae have two well-defined primordial germ cells embedded in a niche comprising two somatic gonad precursor cells. Thus, C. elegans provides an ideal model for studying intercellular signaling in response to DNA damage. However, existing staining protocols are focused on worms in later developmental stages and are not optimized for the L1 larvae. Here, we present a revised protocol for assessing the DNA damage response utilizing immunofluorescence staining specifically in C. elegans L1 larva. For complete details on the use and execution of this protocol, please refer to Ou et al. (2019).


Chrysanthemum stunt viroid disturbs the photoperiodic response for flowering of chrysanthemum plants.
Chrysanthemum ( Dendranthema grandiflorum Kitam.) is one of the qualitative short-day flowering plants. Therefore, the flowering of chrysanthemum can usually be controlled by photoperiod. However, it was noted that 'Piato' plants infected by the chrysanthemum stunt viroid (CSVd) flowered autonomously even under long-day conditions. In this study, CSVd-free and CSVd-infected plants were prepared by culturing different-sized dissected shoot apical meristems (SAMs) of 'Piato'. Using these CSVd-free and CSVd-infected plants, we clarified the relationship between CSVd infection and the autonomous flowering of 'Piato'. Under natural short-day conditions, the flowering of plants regenerated from SAMs containing leaf primordia (LPs) was 1 month earlier than plants regenerated from LP-free SAMs. CSVd was detected from these early flowering plants by reverse transcription-polymerase chain reaction. On the other hand, CSVd was not detected in plants regenerated from LP-free SAMs. CSVd-infected and CSVd-free plants were grown under long-day conditions simulated by night-break lighting at 22:00 p.m. to 2:00 a.m. All CSVd-infected plants flowered autonomously even under long-day conditions; on the other hand, CSVd-free chrysanthemum plants maintained their vegetative growth. When the CSVd-free plants were inoculated with CSVd by grafting them to CSVd-infected rootstocks, they flowered autonomously even under night-break lighting. In this study, the results suggest that CSVd may control the qualitative development process, flowering, i.e. CSVd can induce the autonomous flowering of chrysanthemum.


A new validated international questionnaire on health professionals' knowledge of hypertension diagnosis.
Arterial hypertension is the main factor in attributable mortality. It is therefore considered one of the most important public health problems. Health professionals require special training and skills to make a diagnosis. No studies have been found in the literature search that use a validated instrument (questionnaire) to assess health professionals' theoretical and practical knowledge in diagnosing hypertension or measuring blood pressure. To design and validate an instrument for gauging health professionals' theoretical knowledge in measuring blood pressure for the initial diagnosis of hypertension. Design, development, and validation of a questionnaire in three languages (English, Spanish, and Catalan) to assess knowledge based on the Rasch-item response theory model. A questionnaire in three languages was constructed and validated. It consisted of 23 questions on the theoretical knowledge of the initial diagnosis of hypertension and was called the ARC questionnaire. It met all the Rasch-IRT model criteria: item- and person-fit measurement, unidimensionality, local independence, invariance, targeting, and reliability. The ARC questionnaire is a validated tool that enables objective and uniform analyses of knowledge in the initial diagnosis of hypertension among medical and nursing professionals, comparing them over time. It allows for established strategies to be developed to enhance this knowledge.


Predictors of the maximal oxygen consumption in adult patients with type 1 diabetes treated with personal insulin pumps.
Regular physical activity for adults with type 1 diabetes mellitus improves cardiorespiratory fitness (CF) and quality of life. The aim of our study was to evaluate clinical and biochemical features that might be associated with CF in a homogenous group of adults with type 1 diabetes mellitus who are all treated with a personal insulin pump (continuous subcutaneous insulin infusion). We assessed CF in 62 patients (74.2% of whom were men) who fulfilled the eligibility criteria. To determine maximal oxygen consumption, the march-running test on the treadmill was carried out. Two hours before the test, the patients consumed a defined meal covered by a dose of rapid acting insulin analog that was reduced by 25% from their regular dose. Basal insulin infusion was reduced by 50% for an hour. Additionally, the Perceived Stress Scale-10 questionnaire was used to measure the perception of stress. There was no episode of severe hypoglycemia during or after the test. In the final model, independent predictors of maximal oxygen consumption were sex, body fat percentage, lactate at 20 min after CF test and Perceived Stress Scale-10 score. Of interest, neither short-term (continuous glucose monitoring) nor long-term (glycosylated hemoglobin) metabolic control parameters were predictors of CF. In our selected homogenous group of patients with type 1 diabetes mellitus treated with personal insulin pumps, higher CF was associated with a lower percentage of body fat, male sex, higher lactate level after the CF test and the Perceived Stress Scale-10 score. The proposed protocol in our cohort proved to be safe with regard to glycemic control.


An update on Huntington's disease: from the gene to the clinic.
This review highlights the recent advances in Huntington's disease, with a particular focus on development of disease biomarkers for use in therapeutic trials in the premotor phase of the disease, as well as the growing literature regarding pathophysiological mechanisms and their relevance to potential therapeutic targets. There have been continued advances in the development of disease biomarkers, and promising neuroprotection trials are beginning to emerge in the premotor stage of Huntington's disease. Deeper understanding of the pathophysiological mechanisms is being translated into potential therapeutic strategies. The premotor stage of Huntington's disease provides an ideal time to trial disease-modifying therapy, but reliable biomarkers are required for monitoring disease progression, and this remains an area of intense research. Our understanding of the underlying pathophysiological mechanisms continues to expand, and a number of promising therapeutic strategies are emerging, including strategies to silence mutant huntingtin expression.


Expression of matrix metalloproteinases in ovarian endometriomas: immunohistochemical study and enzyme immunoassay.
Like carcinoma, endometriosis has the unique characteristics, of invasion and metastasis, though pathologically, it is a benign tumor. However, the mechanism of destruction of the surrounding tissue in endometriosis is still unclear. In this study, the expression and localization of matrix metalloproteinases (MMP)-1, -2, -3, -7, -9 and tissue inhibitors of metalloproteinases-1 (TIMP-1) were evaluated by immunohistochemistry for 20 cases and the amounts of MMP-1, TIMP-1 and MMP-1/TIMP-1 complex in the fluid of endometrioma, were analyzed by ELISA and western blotting for 20 cases, which were analyzed by immunohistochemical study. MMP-1, -2 and -9 were detected strongly in both stromal and epithelial cells and MMP-7 in the epithelial cells in the menstrual period. MMP-3 was mainly expressed in macrophage containing hemosiderin but the change of expression was not clear. TIMP-1 was intensively detected in both stromal and epithelial cells in the menstrual period but the expression decreased in other stages of the menstrual cycle. ELISA for MMP-1 also showed results similar to immunohistochemistry, suggesting that it was released to the cyst in the menstrual period when it was released to the extracellular space from the cytoplasm. The expression of TIMP-1 was not clearly changed during the menstrual cycle. From these results, it was suggested that the destruction of the surrounding matrix by endometriosis might be caused by various MMPs, which are mainly produced in stromal cells.


Built Environment and Frailty: Neighborhood Perceptions and Associations With Frailty, Experience From the Nagoya Longitudinal Study.
Neighborhood physical characteristics have been consistently associated with the health of older adults. This article investigates links between frailty and perceptions of the neighborhood environment. Using a cross-sectional analysis of 370 community-dwelling older adults from Nagoya, Japan, neighborhood perceptions were assessed using the Neighborhood Environmental Walkability Scale (NEWS) in addition to frailty, using a frailty index. Frailty was associated with the NEWS composite index, land use mix diversity, land use mix access, street connectivity, walking infrastructure, aesthetics, and crime safety, after adjustment for covariates. Older adults with increasing frailty have poorer perceptions of their neighborhoods, which could lead to further constriction of the life-space, less social and physical engagement, and worsening of frailty status.


The acute administration of trimetazidine modified myocardial perfusion and left ventricular function in 31 patients with ischaemic ventricular dysfunction.
Trimetazidine (TMZ) increases the mithocondrial oxidative metabolism and improves Tc-99m sestamibi uptake in myocardial single photon emission tomography (SPECT). The aim of this study was to evaluate whether the acute administration of TMZ improved myocardial perfusion and modified left ventricular ejection fraction (LVEF) in ischaemic left ventricular impairment. Thirty-one patients (23 males, age 66 years) with prior myocardial infarction (>6 months) and echocardiographic LVEF < or = 45% underwent coronary angiography, rest basal myocardial SPECT (after 3-day placebo administration) and rest TMZ myocardial SPECT [after 3-day TMZ administration (60mg/die)]. The left ventricle was analysed in 16 segments. The summed placebo score (SPS) and the summed TMZ score (STS) were calculated with a 5-point scale (from 0 = normal uptake to 4 = absent uptake) by two blinded operators. The GATED Tc-99m SPECT was always provided. After TMZ administration GATED LVEF improved from 26.5+/-9.7% to 29.1+/-11.3% (p = 0.04) and left ventricular end-systolic volume (LVESV) was reduced from 90.2+/-40.7 to 85.6+/-39.2 ml/mq (p = 0.006). Similarly the addition of TMZ to myocardial SPECT significantly reduced the STS compared to SPS (21.5+/-11 vs. 26.6+/-10.5 p = 0.0001). Eleven patients (35.5%) had an echocardiographic LVEF < or = 30%; in these patients who had severe ventricular dysfunction, GATED LVEF and LVESV did not change after TMZ (20.2+/-5.7% vs. 21+/-6.9% p =0.6; 116.7+/-35.3 ml vs. 112.6+/-32.3 ml p = 0.08, respectively). In comparison with placebo, the addition of TMZ to myocardial Tc-99m tetrofosmin SPECT improved myocardial perfusion and LVEF, reducing LVESV. These effects were lost in patients with more severe ventricular dysfunction.


7-Chloro-4-(Phenylselanyl) Quinoline Is a Novel Multitarget Therapy to Combat Peripheral Neuropathy and Comorbidities Induced by Paclitaxel in Mice.
Paclitaxel-induced peripheral neuropathy (PIPN) is a very common and complex painful condition related to paclitaxel (PTX) exposure, severely impacting patients' quality of life, and contributing to the emergence of clinical signs of anxiety and cognitive loss. At present, no sufficient treatment options are available for PIPN and its exact pathophysiology remains unclear. Based on the therapeutic potential of the 7-chloro-4-(phenylselanyl) quinoline (4-PSQ), we assessed its ability to reverse PIPN and its comorbities induced by PTX. The effect of 4-PSQ was evaluated on pathophysiological processes involved in PIPN, such as oxidative stress (oxidative damage and antioxidant enzymes), neuroinflammation (mRNA expression levels of nuclear factor-kappa B, interleukin-1beta, tumor necrosis factor-alpha, and inducible nitric oxide synthase), and calcium homeostasis (Ca2+ATPase activity) in the spinal cord, cerebral cortex, and hippocampus of mice. Male Swiss mice received PTX (2 mg/kg) or vehicle by intraperitoneal route (days 1, 2, and 3). Oral administration of 4-PSQ (1 mg/kg) or vehicle was performed on days 3 to 14. It was observed that 4-PSQ reduced the mechanical and thermal hypersensitivities induced by PTX. Likewise, 4-PSQ reduced both anxious behavior and cognitive impairment in mice with PIPN. We believe that effects of 4-PSQ may be associated, at least in part, with the modulation of oxidative stress, reduction of neuroinflammation, and normalizing Ca2+ATPase activity in the spinal cord, cerebral cortex, and hippocampus of mice with PIPN. Taken together, the 4-PSQ might be a good prototype for the development of a more effective drug for the treatment of PIPN and its comorbities.


Algorithm for diagnosing hypertension using out-of-office blood pressure measurements.
The authors developed and validated a diagnostic algorithm using the optimal upper and lower cut-off values of office and home BP at which ambulatory BP measurements need to be applied. Patients presenting with high BP (≥140/90 mm Hg) at the outpatient clinic were referred to measure office, home, and ambulatory BP. Office and home BP were divided into hypertension, intermediate (requiring diagnosis using ambulatory BP), and normotension zones. The upper and lower BP cut-off levels of intermediate zone were determined corresponding to a level of 95% specificity and 95% sensitivity for detecting daytime ambulatory hypertension by using the receiver operator characteristic curve. A diagnostic algorithm using three methods, OBP-ABP: office BP measurement and subsequent ambulatory BP measurements if office BP is intermediate zone; OBP-HBP-ABP: office BP, subsequent home BP measurement if office BP is within intermediate zone and subsequent ambulatory BP measurement if home BP is within intermediate zone; and HBP-ABP: home BP measurement and subsequent ambulatory BP measurements if home BP is within intermediate zone, were developed and validated. In the development population (n = 256), the developed algorithm yielded better diagnostic accuracies than 75.8% (95%CI 70.1-80.9) for office BP alone and 76.2% (95%CI 70.5-81.3) for home BP alone as follows: 96.5% (95%CI: 93.4-98.4) for OBP-ABP, 93.4% (95%CI: 89.6-96.1) for OBP-HBP-ABP, and 94.9% (95%CI: 91.5-97.3%) for HBP-ABP. In the validation population (n = 399), the developed algorithm showed similarly improved diagnostic accuracy. The developed algorithm applying ambulatory BP measurement to the intermediate zone of office and home BP improves the diagnostic accuracy for hypertension.


Very early changes in serum creatinine are associated with 30-day mortality after cardiac surgery: A cohort study.
Acute kidney injury predicts adverse outcomes after cardiac surgery. To determine whether ultra-short-term changes (within 120 min) in serum creatinine (SCrea) levels after cardiac surgery predict clinical outcomes (30-day mortality). Observational cohort study. Austrian tertiary referral centre. A total of 7651 patients scheduled to undergo elective cardiac surgery. We analysed SCrea levels measured pre-operatively (baseline) and within 120 min after surgery. We also adjusted the postoperative SCrea levels for fluid balance. Patients were grouped according to the difference between the pre and postoperative SCrea levels (ΔSCreaAdmICU). We performed univariable and multivariable analyses to determine the association between changes in SCrea levels and 30-day mortality. After cardiac surgery, the SCrea level decreased in 5923 patients and increased in 1728 patients. Increased SCrea levels were associated with a 21% increase in 30-day mortality. Even minimal increases in SCrea (0 to <26.5 μmol l) were significantly associated with 30-day mortality [hazard ratio (HR), 1.98; 95% confidence interval (CI), 1.54 to 2.55; P < 0.001]. Adjustments for fluid balance strengthened the above association (increases of 0 to <26.5 μmol l: HR, 1.78; 95% CI, 1.40 to 2.26; P < 0.001; increases of at least 26.5 μmol l: HR, 2.40; 95% CI, 1.68 to 3.42; P < 0.001). Even minimal, ultra-short-term increases in SCrea levels after cardiac surgery are associated with increased 30-day mortality. Adjustment for fluid balance strengthens this association. The change in SCrea between baseline and after admission to the Intensive Care Unit (ΔSCreaAdmICU) can serve as a simple, cheap and widely available marker for very early risk stratification after cardiac surgery.


TikTok, Tide Pods and Tiger King: health implications of trends taking over pediatric populations.
With children and adolescents constantly connected to their devices in an ever-changing technological landscape, new and emerging trends in entertainment media have created potential health implications for pediatric patients. The purpose of this review is to provide pediatricians with explanations of specific trends on streaming services, the popular new mobile application TikTok, recent widespread 'Internet Challenges' that include pediatric participants, and new technologies. These emerging trends can pose potential harms to the health and wellbeing of children and adolescents. Recent reports and news articles have discussed popular technological trends and the adverse effects that can occur. Although technology provides access to information and social connection for youth, it also creates platforms where children can be exposed to disturbing content, peer pressure, cyberbullying, and encouragement to participate in activities that could cause potential physical harm or even death. Pediatricians need to be aware of trends to best understand the implications of these technologies, especially in clinical practice. Pediatricians play an important role in educating parents and patients about the risks of these trends, in facilitating conversations about related topics including self-esteem, cyberbullying and peer pressure to promote healthy internet use, and providing necessary resources.


Isolated isoquercitrin from Green ball apple peel inhibits photoaging in CCD-986Sk fibroblasts cells via modulation of the MMPs signaling.
Ultraviolet B (UVB) irradiation is viewed as the main factor of skin aging, associated with acceleration of elastin, collagen degradation and expression of matrix metalloproteinases (MMPs). Apples are one of the most commonly eaten fruits in the world, and isoquercitrin is the main active ingredient in new bred varieties "Green ball" apple. Therefore, we are studying the functionality of the active ingredient of apple, a natural raw material that does not have toxicity or sensitivity problems. The aim of this study, we scrutinized the effects of isoquercitrin on skin photoaging in UVB-exposed human fibroblasts (CCD-986Sk). To investigate the inhibition effect on photoaging factor regulation, isolated isoquercitrin were treated with UVB, which induces photoaging-related factors in CCD-986Sk fibroblast cells. Pro-inflammatory factors were measured by ELISA, Western blotting and real-time PCR. Isoquercitrin exhibited antioxidant activity and UVB-induced generation of photoaging-related factor inhibition without showing any toxicity. Anti-photoaging effect for protein levels using Isoquercitin was competent, of both the combate MMP-1 and MMP-9. Also, effect of COL1A2 product significantly increase, from up regulating the TIMP-1 mediated pathway in CCD-986Sk cells via the inhibition of MMPs. Isoquercitrin also downregulated the mRNA gene expression of MMPs while upregulating type I procollagen, HAS2 by modulating TIMP-1 and TGF-β in UVB-irradiated CCD-986Sk cells. Collectively, our results show that isoquercitrin might be useful as a functional food while being a good candidate in the development of cosmetic products and medicines for the remedy of UVB-induced skin photoaging.


Autoimmune mechanisms in children with systemic lupus erythematosus.
Although the pathogenesis of SLE remains poorly understood, there is consensus that it involves a combination of genetic, hormonal, and environmental factors. New technologies applied to genomic and gene expression studies have revealed novel gene mutations and cytokine alterations in this disease. Recently, advances in monoclonal antibodies and recombinant DNA technology have resulted in the development of new drugs to arrest disease progression and restore physiologic immune responses without major side effects. Clinical trials to test several of these novel therapies are currently underway.


New chromatographic system for the rapid analysis and preparation of colostrum sialyloligosaccharides.
A new thin-layer chromatographic system on silica gel for the separation of sialyloligosaccharides is described. Calibration of the system with standard milk and colostrum sialyloligosaccharides is presented. The use of the system in monitoring different oligosaccharides is demonstrated for the purification of bovine colostrum sialyllactose isomers and a commercial sialyllactose product, and is discussed with respect to other biological fluids. The large-scale preparation of pure sialyllactose isomers from bovine colostrum is achieved using an improved ion-exchange separation on Dowex 1-X2 (less than 400 mesh) employing isomolar elution at 20 mM for monosialyloligosaccharides and 200 mM for disialyllactose. The purification of four major monosialyltrisaccharides, the 2-3 and 2-6 isomers of N-acetylneuraminyllactose, N-glycolylneuraminyl2-3lactose and N-acetylneuraminyl2-6-N-acetyllactosamine, and the disialyltetrasaccharide di-N-acetylneuraminyllactose is reported. The detection and partial purification of three new minor monosialyloligosaccharides is described.


[Real-life data and Covid-19: The third avenue of reseach].
The analysis of real-life data from hospital information systems could make possible to decide on the efficacy and safety of Covid-19 treatments by avoiding the pitfalls of preliminary studies and randomized clinical trials. The different drugs tested in current clinical trials are already widely prescribed to patients by doctors in hospitals, and can therefore be immediately analysed according to validated methodological standards.


Periploca forrestii Schltr. ameliorate liver injury caused by fluorosis in rat.
To investigate the impact of the ethanoic fractions of Periploca forrestii Schltr. (P. forrestii) in ameliorating the liver injury caused by fluoride ingestion and to explore the potential mechanisms. Initially, an in vitro fluorosis cell model was constructed using the human normal liver cell line (L-02) induced by fluoride. Cell viability was assessed using the CCK-8 assay kit. The lactate dehydrogenase (LDH) assay kit was utilized to measure LDH content in the cell supernatant, while the malonic dialdehyde (MDA) assay kit was employed to determine MDA levels within the cells. Subsequently, a fluorosis rat model was established, and LDH content in the cell supernatant was measured using the LDH assay kit. Various parameters, including MDA, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and reactive oxygen species (ROS) content within the cells, were detected using appropriate assay kits. Additionally, cell apoptosis rate was determined using the Annexin V-FITC/PI cell apoptosis assay kit. The protein expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, Cleaved Caspase-3, Caspase-9, and Cleaved Caspase-9 were analyzed through Western blotting. Compared to the model group, the ethanolic fraction D of P.forrestii (Fr.D) increased cell viability (P < 0.01) and decreased LDH and MDA levels (P < 0.01). In the high-dose Fr.D treatment group of fluoride-poisoned rats, serum ALT, AST, LDH and MDA levels significantly decreased (P < 0.01). Results from rat primary cells exhibited that the Fr.D administration group exhibited significantly higher cell survival rates than the fluoride group (P < 0.01). Similarly, primary rat cells treated with Fr.D showed enhanced cell viability (P < 0.05) and reduced apoptosis rate, LDH, MDA, SOD, GSH-Px, CAT, and ROS levels (P < 0.05) compared to the model group. Western blot analysis indicated that the Fr.D treatment group elevated the Bcl-2/Bax protein expression ratio and reduced Caspase-3 and Caspase-9 activation levels (P < 0.01) compared to the model group. The results suggest that components within the Fr.D from Periploca forrestii may alleviate fluoride-induced liver injury by potentially counteracting oxidative stress and cell apoptosis.


Cell selectivity and anti-inflammatory activity of a Leu/Lys-rich alpha-helical model antimicrobial peptide and its diastereomeric peptides.
To investigate the effect of the number and distribution of d-amino acids introduced into non-cell-selective alpha-helical antimicrobial peptides on the cell selectivity, protease stability and anti-inflammatory activity, we synthesized an 18-meric Leu/Lys-rich alpha-helical model peptide (K(9)L(8)W) and d-amino acid-containing diastereomeric peptides. Increasing in cell selectivity of the peptides was increased in parallel with increasing in the number of d-amino acids introduced. Despite having the same number of d-amino acids, D(9)-K(9)L(8)W-1 had better cell selectivity than D(9)-K(9)L(8)W-2, indicating that a dispersed distribution of d-amino acids in diastereomeric peptides is more effective for cell selectivity than their segregated distribution. D(3)-K(9)L(8)W-2, D(6)-K(9)L(8)W, D(9)-K(9)L(8)W-1 and D(9)-K(9)L(8)W-2 showed complete resistance to tryptic digestion. Furthermore, K(9)L(8)W and all of its diastereomeric peptides significantly inhibited nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) mRNA expression and tumor necrosis factor-alpha (TNF-alpha) release in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells at a lower concentration than bactericidal concentration. The order of anti-inflammatory activity for the peptides was K(9)L(8)W approximately D(3)-K(9)L(8)W-1 approximately D(3)-K(9)L(8)W-2 approximately D(6)-K(9)L(8)W approximately D(9)-K(9)L(8)W-2>D(4)-K(9)L(8)W>D(9)-K(9)L(8)W-1. Increasing in hydrophobicity or alpha-helicity of the peptides was more closely correlated with increasing in hemolytic activity and anti-inflammatory activity than antimicrobial and LPS-disaggregation activities. Collectively, we successfully developed several d-amino acid-containing antimicrobial peptides (D(4)-K(9)L(8)W, D(6)-K(9)L(8)W and D(9)-K(9)L(8)W-1) with good cell selectivity, protease stability and potent anti-inflammatory activity. These antimicrobial peptides could serve as templates for the development of peptide antibiotics for the treatment of sepsis, as well as microbial infection.


Cell-Free DNA Maps Tissue Injury and Correlates with Disease Severity in Lung Transplant Candidates.
Plasma cell-free DNA levels correlate with disease severity in many conditions. Pre-transplant cell-free DNA may risk stratify lung transplant candidates for post-transplant complications. Evaluate if pre-transplant cell-free DNA levels and tissue sources identify patients at high risk of primary graft dysfunction and other pre- and post-transplant outcomes. This multicenter, prospective cohort study recruited 186 lung transplant candidates. Pre-transplant plasma samples were collected to measure cell-free DNA. Bisulfite sequencing was performed to identify the tissue sources of cell-free DNA. Multivariable regression models determined the association between cell-free DNA levels and the primary outcome of primary graft dysfunction and other transplant outcomes including Lung Allocation Score, chronic lung allograft dysfunction and death. Transplant candidates had 2-fold greater cell-free DNA levels than healthy controls (median [IQR]: 23.7 ng/mL [15.1-35.6] vs 12.9 ng/mL [9.9-18.4], p < 0.0001), primarily originating from inflammatory innate immune cells. Cell-free DNA levels and tissue sources differed by native lung disease category and correlated with the Lung Allocation Score (p<0.001). High pre-transplant cell-free DNA increased the risk of primary graft dysfunction (OR 1.60, 95% CI [1.09 - 2.46], p = 0.0220), and death (HR 1.43, 95% CI [1.07 - 1.92], p = 0.0171) but not chronic lung allograft dysfunction (HR 1.37, 95% CI [0.97 - 1.94], p = 0.0767). Lung transplant candidates demonstrate a heightened degree of tissue injury with elevated cell-free DNA, primarily originating from innate immune cells. Pre-transplant plasma cell-free DNA levels predict post-transplant complications.


Rapid modulation of tumor necrosis factor membrane receptors by activators of protein kinase C.
Tumor necrosis factor membrane receptors are rapidly down-regulated upon treatment of activated T lymphocytes with various activators of protein kinase C. Loss of binding-capacity was half maximal after 2 min. incubation in 10 ng/ml of phorbol 12-myristate 13-acetate. A similar modulation could be induced with either the calcium ionophore A 23187 or the protein kinase C activator 1-oleyl-2-acetyl glycerol, whereas 1,2-diolein and dibutyryl cAMP were ineffective. Protein kinase C inhibitor H7 antagonizes the phorbol ester-induced TNF receptor modulation. These data suggest an important role of protein kinase C in the control of TNF responsiveness by regulation of TNF binding-capacity possibly via direct phosphorylation of specific receptor proteins.


Synergistic effect of hydrothermal co-carbonization of sewage sludge with fruit and agricultural wastes on hydrochar fuel quality and combustion behavior.
In order to improve fuel quality of sewage sludge, fruit and agricultural wastes have been selected for hydrothermal co-carbonization. After hydrothermal co-carbonization, organics retention was facilitated, while O/C and H/C atomic ratios of hydrochars were substantially upgraded. Particularly, hydrochar from hydrothermal co-carbonization of sewage sludge with peanut shells at mass ratio of 1:3 (denoted as "SS:PS = 1:3") showed the highest fuel ratio of 0.79 and its carbon content was increased to 50.0% with significantly decreased O/C and H/C atomic ratios. Furthermore, higher heating value of hydrochars from hydrothermal co-carbonization was increased by nearly 2.65-fold and reached 21.72 MJ/kg. Moreover, the most favorable aromatization occurred when sewage sludge and peanut shells blending ratio was 3:1 or 1:1, whereas hydrothermal co-carbonization induced more CO and OH than COOH in hydrochars due to synergistic decarboxylation. A relatively higher value of point of zero charge for hydrochars from hydrothermal co-carbonization implied improved hydrophobicity. Combustion kinetics results indicated that hydrothermal co-carbonization balanced activation energies of hydrochars in devolatilization/combustion stage and char combustion process, rendering a more stable and lasting combustion profile. Hydrochars "SS:PS = 1:3" demonstrated desirable combustion performance. Therefore, hydrothermal co-carbonization can realize sustainable utilization of organic solid wastes towards superior hydrochar solid biofuels.


The spleen's role in transplantation immunology.
While graft rejection has been studied for many years, recent investigations have focused on the generation and maintenance of transplantation tolerance. This review examines the role of secondary lymphoid organs, especially the spleen, characterizes the maintenance mechanism that is key to the development of tolerance and explores the implications of new strategies for inducing tolerance in transplantation.


De novo engineering and metabolic flux analysis of inosine biosynthesis in Bacillus subtilis.
Wild-type B. subtilis strain W168 was de novo engineered for inosine biosynthesis. Inactivation of deoD and purA led to 0.15 ± 0.04 and 6.44 ± 0.39 g inosine/l yields, respectively. The deoD purA double mutant accumulated 7.6 ± 0.34 g inosine/l, with a 4.7% (w/w) conversion ratio from glucose to inosine. Comparative metabolic flux analysis revealed that the fluxes from inosine to hypoxanthine and from inosine monophosphate to adenosine monophosphate in the double mutant decreased to 14.0 and 0.61% of those in the wild-type strain. The major role of purA was demonstrated when inactivation of deoD and purA were found to contribute additively to inosine accumulation. This work is expected to contribute to the improvement of the fermentative production of purine nucleosides in the microbial industry.


Nonoperative management of pancreatic injuries in pediatric patients.
Nonoperative management of minor pancreatic injury is the generally accepted approach. However, the management of major pancreatic injury remains controversial in pediatric patients. The aim of the present study was to determine the safety and efficacy of nonoperative management of pancreatic injury in pediatric patients. Between 2003 and 2009, 31 patients, 28 male and 3 female, with pancreatic injury due to blunt abdominal trauma were treated in our clinic. All patients were evaluated by ultrasonography, computed tomography (CT), and evaluation of serum amylase levels. Patients with ongoing hemodynamic instability after resuscitation or signs of bowel perforation underwent immediate laparotomy, and the remaining patients were conservatively treated. Conservative treatment consisted of nasogastric tube replacement, total parenteral nutrition, monitoring of amylase levels, and serial clinical examination. The most common mechanism of injury was a fall (35.4%). Ten patients (32.2%) had associated extra-abdominal injuries, and 18 patients (58.1%) had associated abdominal injuries. The spleen was the most common site of intra-abdominal injury that was associated with pancreatic trauma. Initial amylase levels were normal in 5 patients, whose CT scans revealed pancreatic injury. Twenty-five patients (80.6%) were conservatively treated. Six patients (19.4%) required surgical intervention because of a hollow viscus or diaphragmatic injury and hemodynamic instability. A pseudocyst developed in 11 of the 25 patients who were nonoperatively treated; 6 patients required intervention for the pseudocyst (percutaneous drainage and cystogastrostomy). No patient succumbed to injury. The majority of the pancreatic injuries in pediatric patients can be successfully treated conservatively, unless there is hemodynamic instability and a hollow viscus injury. The most common complication is a pseudocyst.


Polymorphism and predictability at the alpha-glycerophosphate dehydrogenase locus in Colias butterflies: gradients in allele frequency within single populations.
Heterozygosity at the alpha-glycerophosphate dehydrogenase locus of five species of Colias butterflies is widespread in montane populations; alpine and lowland populations are not heterozygous. Within a single demographically characterized population of C. meadii where the population extends from alpine down into montane habitats, a marked cline in allele frequency is seen. Such within-population clines suggest the involvement of strong selection across the marked ecological interface. Thermal factors are the most likely causative agents, but associative overdominance is not excluded.


Membranous glomerulonephritis and hepatitis B surface antigen in children.
Of 33 children with membranous nephropathy screened for HBs Ag, 14 were found to be HBs Ag carriers, whereas HBs Ag was detected in 3 of 170 and 4 of 100 children with glomerular and nonglomerular kidney diseases, respectively. HBs Ag was often associated with acute hepatitis at onset (five patients) or with elevated transminases values. This high incidence and the prevalence of an unusual subtype (ayw2) suggest a relationship between HBs Ag and the glomerular lesions. Using immunofluorescence, however, HBs Ag could not be detected within the deposits, so that the nature of the relationship cannot be considered as established. The clinical outcome (50% remission), the plasma complement component disturbances, and findings by immunofluorescence did not differ from those observed in children with MGN without detectable HBs Ag.


Thyroglossal duct carcinoma.
Primary carcinoma of the thyroglossal duct is rare. This discussion reports two cases and reviews the 50 previously reported in the literature. The criteria for diagnosis include evidence of a thyroglossal duct remnant and a normal thyroid gland. The differentiation from cystic metastases to lymph nodes is pointed out. The histologic types parallel those of carcinoma of the thyroid gland, papillary carcinoma being the most common and having a generally favorable prognosis. The clinical presentation of these tumors is similar to that with benign cysts and thus is of limited value in the diagnosis.


The Impact of Sleeve Gastrectomy on Gastroesophageal Reflux Disease in Patients with Morbid Obesity.
The impact of laparoscopic sleeve gastrectomy (LSG) on gastroesophageal reflux disease (GERD) has not been widely quantified, and the data in the literature remain controversial. Candidates for LSG underwent barium swallow, esophageal manometry, ambulatory 24-h esophageal pH monitoring (APM), and gastric emptying scintigraphy before and after surgery (1 and 18 months). Symptoms were evaluated using a gastroesophageal reflux disease questionnaire (GERDq). Esophagogastroduodenoscopy was performed preoperatively in all patients and at 18 months postoperatively in patients who had suffered from preoperative esophagitis. Fifty-two patients were included in the study (64.4% women and 34.6% men) with a median age of 46 years (25-63 years) and BMI of 45.0 ± 5.6 kg/m2. The follow-up rates at 1 and 18 months were 82.7% and 80.8%. At 18 months, the percentage of weight loss (%TWL) was 33.6 ± 10.4% and the percentage of excess BMI loss (%EBMIL) was 77.6 ± 25%. Postoperatively, a significant increase in accelerated gastric emptying and impaired esophageal body motility occurred at 1 and 18 months. A significant worsening of all the values obtained at both 1 and 18 months postoperatively becomes evident when comparing the results of the APM. After surgery, 76.4% of patients had developed "de novo" GERD at 1 month and 41% at 18 months. No improvement was found in patients with symptomatic GERD. Based on the results of this study, LSG led to a considerable rate of postoperative "de novo" GERD. In addition, no improvement was found in patients with symptomatic GERD.


The prostaglandin E2 receptor EP4 is expressed by human platelets and potently inhibits platelet aggregation and thrombus formation.
Low concentrations of prostaglandin (PG) E(2) enhance platelet aggregation, whereas high concentrations inhibit it. The effects of PGE(2) are mediated through 4 G protein-coupled receptors, termed E-type prostaglindin (EP) receptor EP1, EP2, EP3, and EP4. The platelet-stimulating effect of PGE(2) has been suggested to involve EP3 receptors. Here we analyzed the receptor usage relating to the inhibitory effect of PGE(2). Using flow cytometry, we found that human platelets expressed EP4 receptor protein. A selective EP4 agonist (ONO AE1-329) potently inhibited the platelet aggregation as induced by ADP or collagen. This effect could be completely reversed by an EP4 antagonist, but not by PGI(2), PGD(2), and thromboxane receptor antagonists or cyclooxygenase inhibition. Moreover, an EP4 antagonist enhanced the PGE(2)-induced stimulation of platelet aggregation, indicating a physiological antiaggregatory activity of EP4 receptors. The inhibitory effect of the EP4 agonist was accompanied by attenuated Ca(2+) flux, inhibition of glycoprotein IIb/IIIa, and downregulation of P-selectin. Most importantly, adhesion of platelets to fibrinogen under flow and in vitro thrombus formation were effectively prevented by the EP4 agonist. In this respect, the EP4 agonist synergized with acetylsalicylic acid. These results are suggestive of EP4 receptor activation as a novel antithrombotic strategy.


Percutaneous balloon versus surgical closed commissurotomy for mitral stenosis. A prospective, randomized trial.
We performed a prospective, randomized trial comparing percutaneous balloon commissurotomy with surgical closed commissurotomy in 40 patients with severe rheumatic mitral stenosis. Data were analyzed by investigators who were masked to treatment assignment or phase of study. Patients randomized to balloon (n = 20) or surgical (n = 20) commissurotomy had severe mitral stenosis without significant baseline differences (left atrial pressure, 26.1 +/- 4.2 versus 27.6 +/- 6.2 mm Hg; mitral valve gradient, 18.0 +/- 4.2 versus 19.7 +/- 6.3 mm Hg; mitral valve area, 1.0 +/- 0.2 versus 1.0 +/- 0.4 cm2, respectively). At 1-week follow-up after balloon commissurotomy, pulmonary wedge pressure was 14.3 +/- 7.2 mm Hg; mitral valve gradient was 9.6 +/- 5.1 mm Hg; and mitral valve area was 1.6 +/- 0.6 cm2 (all p less than 0.0001). At 1-week follow-up after surgical closed commissurotomy, wedge pressure was 13.7 +/- 5.4 mm Hg; mitral valve gradient was 9.4 +/- 4.2 mm Hg (both p less than 0.0001); and mitral valve area was 1.6 +/- 0.7 cm2 (p less than 0.003). At 8-month follow-up, improvement occurred in both groups: Mitral valve area was 1.6 +/- 0.6 cm2 in the balloon commissurotomy group (p less than 0.002) and was 1.8 +/- 0.6 cm2 in the surgical closed commissurotomy group (p less than 0.0001). There was no difference between the groups at 1-week or 8-month follow-up (all p greater than 0.4). One case of severe mitral regurgitation occurred in each group; complications were otherwise related to transseptal catheterization. There was no death, stroke, or myocardial infarction. Cost analysis revealed that balloon commissurotomy may substantially exceed the cost of surgical commissurotomy in developing countries, whereas it may represent a significant savings in industrialized nations. We conclude that percutaneous balloon commissurotomy and surgical closed commissurotomy result in comparable hemodynamic improvement that is sustained through 8 months of follow-up.


Facile synthesis of high-quality graphene nanoribbons.
Graphene nanoribbons have attracted attention because of their novel electronic and spin transport properties, and also because nanoribbons less than 10 nm wide have a bandgap that can be used to make field-effect transistors. However, producing nanoribbons of very high quality, or in high volumes, remains a challenge. Here, we show that pristine few-layer nanoribbons can be produced by unzipping mildly gas-phase oxidized multiwalled carbon nanotubes using mechanical sonication in an organic solvent. The nanoribbons are of very high quality, with smooth edges (as seen by high-resolution transmission electron microscopy), low ratios of disorder to graphitic Raman bands, and the highest electrical conductance and mobility reported so far (up to 5e(2)/h and 1,500 cm(2) V(-1) s(-1) for ribbons 10-20 nm in width). Furthermore, at low temperatures, the nanoribbons show phase-coherent transport and Fabry-Perot interference, suggesting minimal defects and edge roughness. The yield of nanoribbons is approximately 2% of the starting raw nanotube soot material, significantly higher than previous methods capable of producing high-quality narrow nanoribbons. The relatively high-yield synthesis of pristine graphene nanoribbons will make these materials easily accessible for a wide range of fundamental and practical applications.


Thrombectomy is a cost-saving procedure up to 24 h after onset.
The treatment of ischemic stroke due to large-vessel occlusion has been revolutionized by mechanical thrombectomy (MT), as multiple trials have consistently shown improved functional outcomes compared to standard medical management both in the early and late time windows after symptom onset. However, MT is an interventional procedure that is more costly than best supportive care (BSC). We set out to study the cost-utility and budget impact of MT + BSC versus BSC alone for large-vessel occlusion using a combined decision tree and Markov model. The analysis was conducted from a Belgian payer perspective over a lifetime horizon, and health states were defined by the modified Rankin Scale (mRS). The treatment effect of MT + BSC combined clinical outcomes from all published early and late treatment window studies showing improved mRS after 90 days. Resource use and utilities were informed by an observational Belgian study of 569 stroke patients. Long-term mRS transitions were sourced from the Oxford Vascular study. MT + BSC generated 1.31 additional quality-adjusted life years and resulted in cost savings of €10,216 per patient over lifetime. Deterministic sensitivity analyses demonstrated dominance of MT over a wide range of parameter inputs. In a Belgian setting, adding MT to BSC within an early time window for 1575 eligible stroke patients every year produced cost savings between €6.3 million (year 1) and €14.6 million (year 5), or a total cost saving of €56.2 million over 5 years. Mechanical thrombectomy is a highly cost-effective treatment for ischemic stroke patients, providing quality-adjusted survival at lower health care cost, both when given in an early time window, as well as in a late time window.


Electrophysiological differences between oxytocin and vasopressin neurones recorded from female rats in vitro.
1. Intracellular recordings in vitro from immunochemically identified oxytocin (OT) and vasopressin (VP) neurones in the supraoptic nucleus (SON) of virgin or lactating female rats revealed no differences between neurone types in membrane potential (Vm), input resistance and current-voltage relationships (I-V), when taken at resting membrane potentials. 2. When OT (94%), but not VP, neurones (93%) were current clamped at depolarized voltages (above -50 mV), small hyperpolarizing pulses revealed a time- and voltage-dependent outward rectification that was present above -75 mV and that decreased in amplitude as Vm approached the equilibrium potential for potassium (EK). The rectification was more pronounced when the neurones were held at a more depolarized membrane potential, and was larger the longer the neurone was held depolarized, reaching a maximum at 0.6-0.9 s. 3. A rebound depolarization followed the offset of hyperpolarizing pulses that were associated with the rectification. The peak amplitude of the rebound showed a time and a voltage dependence. It followed a bell-shaped curve as the hyperpolarizing commands were made larger, attaining a peak at -65 +/- 1.5 mV. The rebound amplitude increased with pulse duration, achieving a half-maximal amplitude at 0.5 +/- 0.1 s. 4. The expression of the sustained outward rectification and the rebound in OT neurones was similar in virgin and lactating female rats. 5. These results indicate the presence of significant differences in the intrinsic membrane properties, probably K+ currents, between OT and VP neurones in both lactating and virgin female rats.


Association between human herpesviruses and multiple sclerosis: A systematic review and meta-analysis.
The aim of this study was to investigate the prevalence and potential association between infection with different herpes viruses and multiple sclerosis (MS). A systematic literature search was performed by finding relevant cross-sectional and case-control studies from a large online database. Heterogeneity, Odds ratio (OR), and corresponding 95% Confidence interval (CI) were applied to all studies by meta-analysis and forest plots. The analysis was performed using Stata Software v.14. One hundred and thirty-four articles (289 datasets) were included in the meta-analysis, 128 (245 datasets) of which were case/control and the rest were cross-sectional. The pooled prevalence of all human herpes viruses among MS patients was 50% (95% CI: 45-55%; I2 = 96.91%). In subgroup analysis, the pooled prevalence of Herpes simplex virus (HSV), Varicella-zoster virus (VZV), Epstein-Barr virus (EBV), Cytomegalovirus (CMV), Human herpes virus 6 (HHV-6), Human herpes virus 7 (HHV-7), and Human herpes virus 8 (HHV-8) was 32%, 52%, 74%, 41%, 39% 28%, and 28%, respectively. An association was found between infection with human herpes viruses and MS [summary OR 2.07 (95% CI (1.80-2.37); I2 = 80%)]. The results of the present study showed that EBV, VZV, and HHV-6 infection are associated with multiple sclerosis and can be considered as potential risk factors for MS. Although the exact molecular mechanism of the role of herpes viruses in the development of MS is still unknown, it seems that molecular mimicry, the release of autoreactive antibodies, and inflammation in the CNS following viral infection can be important factors in the induction of MS.


Phosphoinositides and host-pathogen interactions.
Phosphoinositides control key cellular processes including vesicular trafficking and actin polymerization. Intracellular bacterial pathogens manipulate phosphoinositide metabolism in order to promote their uptake by target cells and to direct in some cases the biogenesis of their replication compartments. In this chapter, we review the molecular strategies that major pathogens including Listeria, Mycobacterium, Shigella, Salmonella, Legionella and Yersinia use to hijack phosphoinositides during infection. This article is part of a Special Issue entitled Phosphoinositides.


Synthesis and characterization of CdSe nanorods using a novel microemulsion method at moderate temperature.
CdSe nanoparticles have been successfully synthesized using a novel microemulsion method at moderate temperature. It is found that with a combination of the surfactant AOT and hydrazine hydrate, it is possible to control the morphology of the nanoparticles. The hydrazine hydrate acts as both a reducing agent and a templating agent that favors the formation of a rodlike structure. The composition, morphology and optical properties of the CdSe nanoparticles were investigated using powder X-ray diffraction (XRD), transmission electron microscopy (TEM), ultraviolet-visible (UV-vis) absorption spectroscopy, photoluminescence (PL) spectroscopy, energy dispersive X-ray spectroscopy (EDX) and Fourier transform infrared (FT-IR) spectroscopy. The nucleation and growth mechanism for this system is also proposed based on a time-dependent study. This synthesis route provides a moderate temperature (100 degrees C) method for synthesizing rodlike CdSe, hence reducing the possibility of oxidation of this chalcogenide compound.


An improved PID switching control strategy for type 1 diabetes.
In order for an "artificial pancreas" to become a reality for ambulatory use, a practical closed-loop control strategy must be developed and critically evaluated. In this paper, an improved PID control strategy for blood glucose control is proposed and evaluated in silico using a physiologic model of Hovorka et al. The key features of the proposed control strategy are: (i) a switching strategy for initiating PID control after a meal and insulin bolus; (ii) a novel time-varying setpoint trajectory, (iii) noise and derivative filters to reduce sensitivity to sensor noise, and (iv) a systematic controller tuning strategy. Simulation results demonstrate that the proposed control strategy compares favorably to alternatives for realistic conditions that include meal challenges, incorrect carbohydrate meal estimates, changes in insulin sensitivity, and measurement noise.


Etretinate in severe psoriasis. Results of double-blind study and maintenance therapy in pustular psoriasis.
A randomized double-blind study was performed in forty patients with severe psoriasis. Etretinate, in contrast to placebo, clearly reduced the psoriatic area and severity index score, and this difference was statistically significant from week 2. The results in psoriasis vulgaris were satisfactory in only three of ten patients; however moderate improvement was noticed in four of ten patients. In pustular psoriasis of von Zumbusch type, the drug produced moderate improvement to complete clearing in all four of the treated patients and was regarded as the first-choice therapy in all of them. In erythrodermic psoriasis satisfactory results were obtained in four of six patients. In three of the five patients with psoriatic arthropathy there was a marked improvement of arthralgia. Laboratory investigations did not show significant abnormalities with the exception of an increase of alanine transaminase (ALT; SGPT) in about 20% of the patients. Ten patients with pustular psoriasis (four of them included in the double-blind trial) on long-term, i.e., maintenance, therapy remained free of lesions up to this writing for 3 years. Triglyceride levels were not elevated.


Interactions of the hepatitis C virus protease inhibitor faldaprevir with cytochrome P450 enzymes: in vitro and in vivo correlation.
The potential inhibition of the major human cytochrome P450 (CYP) enzymes by faldaprevir was evaluated both in vitro and in clinical studies (healthy volunteers and hepatitis C virus [HCV] genotype 1-infected patients). In vitro studies indicated that faldaprevir inhibited CYP2B6, CYP2C9, and CYP3A, and was a weak-to-moderate inactivator of CYP3A4. Faldaprevir 240 mg twice daily in healthy volunteers demonstrated moderate inhibition of hepatic and intestinal CYP3A (oral midazolam: 2.96-fold increase in AUC(0-24 h)), weak inhibition of hepatic CYP3A (intravenous midazolam: 1.56-fold increase in AUC(0-24 h)), weak inhibition of CYP2C9 ([S]-warfarin: 1.29-fold increase in AUC(0-120 h)), and had no relevant effects on CYP1A2, CYP2B6, or CYP2D6. Faldaprevir 120 mg once daily in HCV-infected patients demonstrated weak inhibition of hepatic and intestinal CYP3A (oral midazolam: 1.52-fold increase in AUC(0-∞)), and had no relevant effects on CYP2C9 or CYP1A2. In vitro drug-drug interaction predictions based on inhibitor concentration ([I])/inhibition constant (Ki) ratios tended to overestimate clinical effects and a net-effect model provided a more accurate approach. These studies suggest that faldaprevir shows a dose-dependent inhibition of CYP3A and CYP2C9, and does not induce CYP isoforms.


Noncoding RNA regulates the immune microenvironment in recurrent spontaneous abortion (RSA): new insights into immune mechanisms.
Recurrent spontaneous abortion (RSA) has various causes, including chromosomal abnormalities, prethrombotic state, and abnormal uterine anatomical factors. However, the pathogenesis of RSA is still unclear. Surprisingly, non-coding RNA can stably express at the maternal-fetal interface and regulate immune cells' proliferation, apoptosis, invasion, metastasis, and angiogenesis. Accumulating evidence suggests that the competing endogenous RNA (ceRNA) regulatory network between non-coding RNAs complicates RSA's pathological process and maybe a new starting point for exploring RSA. In this review, we mainly discuss the regulatory network and potential significance of non-coding RNA in the immune microenvironment of RSA patients. In addition, the cellular interactions of non-coding RNA transported through vesicles were introduced from aspects of trophoblast function and immune regulation. Finally, we analyze previous studies and further discuss that the stable expression of non-coding RNA may be used as a biomarker of some disease states and a prediction target of RSA.


A prospective study of hepatitis during antituberculous treatment in Taiwanese patients and a review of the literature.
The present study prospectively investigated the incidence of and factors associated with hepatitis during antituberculous treatment in patients with tuberculosis and various underlying diseases. The results were compared with those of previously published studies. Patients treated with antituberculous agents were enrolled from July 1, 2000 to July 31, 2001, in the divisions of chest and infectious diseases at National Taiwan University Hospital and followed until November 30, 2001. Hepatitis was defined as an aminotransferase level>5 times the upper limit of normal (ULN), or >3 times ULN in the presence of symptoms of hepatitis, or total bilirubin level>3 mg/dL. Studies reporting the incidence of hepatitis during antituberculous treatment were reviewed for comparison. Among 261 patients, median age was 58 years (range, 17-90 years), 17.7% had abnormal baseline liver function tests and 18.4% had concurrent hepatotoxic drug use. Fifteen patients (5.7%) had hepatitis B virus infection, 17 (6.5%) had hepatitis C virus infection, 14 (5.4%) had liver cirrhosis, and 15 (5.7%) had human immunodeficiency virus infection. Hepatitis occurred in 42 patients (16.1%), with 60% of the events in the first 2 months of treatment. Such an incidence was comparable to that in other Asian countries (5.3-18.2%) and slightly higher than that in Western countries (2.4-19%). In multivariate analysis, abnormal liver function tests at baseline and liver cirrhosis were independent factors for development of hepatitis. Elevation of liver function tests was not uncommon during antituberculous treatment, especially in the first 2 months. Patients with abnormal liver function tests at baseline or liver cirrhosis should be closely monitored.


Designing photoswitchable peptides using the AsLOV2 domain.
Photocontrol of functional peptides is a powerful tool for spatial and temporal control of cell signaling events. We show that the genetically encoded light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2) can be used to reversibly photomodulate the affinity of peptides for their binding partners. Sequence analysis and molecular modeling were used to embed two peptides into the Jα helix of the AsLOV2 domain while maintaining AsLOV2 structure in the dark but allowing for binding to effector proteins when the Jα helix unfolds in the light. Caged versions of the ipaA and SsrA peptides, LOV-ipaA and LOV-SsrA, bind their targets with 49- and 8-fold enhanced affinity in the light, respectively. These switches can be used as general tools for light-dependent colocalization, which we demonstrate with photo-activable gene transcription in yeast.


A new Gateway vector and expression protocol for fast and efficient recombinant protein expression in Mycobacterium smegmatis.
A major obstacle associated with recombinant protein over-expression in Escherichia coli is the production of insoluble inclusion bodies, a problem particularly pronounced with Mycobacterium tuberculosis proteins. One strategy to overcome the formation of inclusion bodies is to use an expression host that is more closely related to the organism from which the proteins are derived. Here we describe methods for efficiently identifying M. tuberculosis proteins that express in soluble form in Mycobacterium smegmatis. We have adapted the M. smegmatis expression vector pYUB1049 to the Gateway cloning system by the addition of att recombination recognition sequences. The resulting vector, designated pDESTsmg, is compatible with our in-house Gateway methods for E. coli expression. A target can be subcloned into pDESTsmg by a simple LR reaction using an entry clone generated for E. coli expression, removing the need to design new primers and re-clone target DNA. Proteins are expressed by culturing the M. smegmatis strain mc(2)4517 in autoinduction media supplemented with Tween 80. The media used are the same as those used for expression of proteins in E. coli, simplifying and reducing the cost of the switch to an alternative host. The methods have been applied to a set of M. tuberculosis proteins that form inclusion bodies when expressed in E. coli. We found that five of eight of these previously insoluble proteins become soluble when expressed in M. smegmatis, demonstrating that this is an efficient salvage strategy.


Explaining the Muslim employment gap in Western Europe: individual-level effects and ethno-religious penalties.
It is well-documented that Muslims experience economic disadvantages in Western European labor markets. However, few studies comprehensively test individual-level explanations for the Muslim employment gap. Using data from the European Social Survey, this research note briefly examines the role of individual-level differences between Muslims and non-Muslims in mediating employment differences. Results reveal that human capital, migration background, religiosity, cultural values, and perceptions of discrimination jointly account for about 40% of the employment variance between Muslims and non-Muslims. Model specifications for first- and second-generation Muslim immigrants reveal a similar pattern, with migration background and perceived discrimination being of key relevance in mediating employment difference. While individual-level effects are indeed relevant, unexplained variance suggests that symbolic boundaries against Islam may still translate into tangible ethno-religious penalties.


Rehabilitation practice patterns for patients with heart failure: the South American perspective.
Heart failure (HF) is an important public health issue in South America. Economic impacts are substantial. Chagas heart disease is a prevalent HF etiology; it is caused by the protozoan Trypanosoma cruzi. Cardiac rehabilitation (CR) is an integral component of HF care. The benefits of CR in HF patients need to be assessed. The effectiveness and safety of CR delivery, such as home-based interventions, should be explored. Strategies to improve adherence in CR are imperative. We describe past and current CR trends for HF patients and discuss the future of this important intervention.


Comparison of MB-check and Löwenstein-Jensen media for recovery of mycobacteria.
We have compared the rate of recovery of mycobacteria with the MB-Check culture system (liquid phase) and the Löwenstein-Jensen (LJ) medium in 2,907 clinical specimens obtained from 830 patients submitted for mycobacterial culture during 1-year period. Direct smear examination was carried out by auramine-rhodamine staining. All primary isolates from the culture media were confirmed by Ziehl-Neelsen staining and identified by acridinium-ester-labeled DNA probes specific for Mycobaterium tuberculosis complex. A total of 214 isolates were of the M. tuberculosis complex (88 patients) and 54 of "potentially pathogenic environmental mycobacteria" (45 patients). A total of 117 (54.7%) samples were smear-positive and the remaining 97 (45.3%) were smear-negative. There was a significant difference in the percentage of positive cultures obtained by the MB-Check method (99.1%) as compared with the LJ medium (73.8%) (P < 0.05). This difference, however, occurred almost exclusively at the expense of the 97 smear-negative samples (positive cultures 97.95% by the MB-Check method vs. 42.3% by the LJ culture, P < 0.05). The number of patients diagnosed of tuberculosis by the MB-Check was significantly higher as compared with LJ medium (88 [100%] vs. 77 [87.5%], P < 0.05). In 11 (12.5%) patients, the diagnosis was only established by the MB-Check system. In smear-positive samples, the mean (+/-SD) detection time for M. tuberculosis complex was 14.8 +/- 8 days with MB-Check and 19.9 +/- 7 days with LJ medium. The corresponding figures in smear-negative samples were 22.8 +/- 3 days and 27.8 +/- 6 days, respectively. DNA probes directly applied to MB-Check liquid medium showed a sensitivity of 98.8% and specificity of 100%. These results indicate that the MB-Check system is more efficient for the recovery of mycobacteria than LJ medium.


Long-term follow-up after percutaneous treatment of the unprotected left main stenosis in high risk patients not suitable for bypass surgery.
Percutaneous coronary intervention is recommended in patients with unprotected left main stenosis non suitable for coronary artery bypass graft. Long-term follow-up of those patients remains uncertain. All patients with de novo unprotected left main stenosis treated with stent implantation were consecutively enrolled. Percutaneous coronary intervention was indicated according to the standards of care, taking into account clinical and anatomical conditions unfavorable for coronary artery bypass graft. The primary end point was the occurrence of major adverse cardiac events, a composite of death, nonfatal acute myocardial infarction, or target lesion revascularization. Of 226 consecutive patients included, 202 (89.4%) were treated with drug-eluting stents. Mean age was 72.1 years, 41.1% had renal dysfunction, and mean Syntax score and EuroSCORE were 28.9 and 7.4, respectively. Angiographic and procedural success was achieved in 99.6% and 92.9% of patients. At 3 years, the rates of major adverse cardiac events, death, nonfatal acute myocardial infarction and target lesion revascularization were 36.2%, 25.2%, 8.4%, 8.0%, respectively. Target lesion revascularization was more frequently observed when ≥ 2 stents were implanted rather than a single stent (18.5% vs 5.8%, P=.03); and with bare metal stents rather than drug-eluting stents (13.0% vs 7.9%, P=.24). Definite stent thrombosis was observed in 2 patients (0.9%) and probable stent thrombosis in 7 (3.1%). Female sex, impaired left ventricular function, and use of bare metal stents were significantly related with all-cause mortality. High-risk patients with unprotected left main stenosis treated with percutaneous coronary intervention presented with a high rate of major adverse cardiac events at long-term follow-up. Female sex, impaired left ventricular function, and use of bare metal stents were predictors of poor prognosis.


Light signaling and the phytohormonal regulation of shoot growth.
Shoot growth of dicot plants is rigorously controlled by the interactions of environmental cues with several groups of phytohormones. The signaling effects of light on shoot growth are of special interest, as both light irradiance and light quality change rapidly throughout the day, causing profound changes in stem elongation and leaf area growth. Among the several dicot species examined, we have focused on sunflower (Helianthus annuus L.) because its shoots are robust and their growth is highly plastic. Sunflower shoots thus constitute an ideal tissue for assessing responses to both light irradiance and light quality signals. Herein, we discuss the possible roles of gibberellins, auxin, ethylene, cytokinins and brassinosteroids in mediating the stem elongation and leaf area growth that is induced by shade light. To do this we uncoupled the plant's responses to changes in the red to far-red [R/FR] light ratio from its responses to changes in irradiance of photosynthetically active radiation [PAR]. Reducing each of R/FR light ratio and PAR irradiance results in increased sunflower stem elongation. However, the plant's response for leaf area growth differs considerably, with a low R/FR ratio generally promoting leaf area growth, whereas low irradiance PAR inhibits it. The increased stem elongation that occurs in response to lowering R/FR ratio and PAR irradiance is accomplished at the expense of leaf area growth. In effect, the low PAR irradiance signal overrides the low R/FR ratio signal in shade light's control of leaf growth and development. Three hormone groups, gibberellins, auxin and ethylene are directly involved in regulating these light-mediated shoot growth changes. Gibberellins and auxin function as growth promoters, with auxin likely acting as an up-regulator of gibberellin biosynthesis. Ethylene functions as a growth-inhibitor and probably interacts with gibberellins in regulating both stem and leaf growth of the sunflower shoot.


The regulation of COUP-TFII gene expression by Ets-1 is enhanced by the steroid receptor co-activators.
Recent phenotypic analysis of orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) [NR2F2] knockout mice shows that COUP-TFII is involved in the angiogenic process in the developing embryos. Since Ets-1 expression is also correlated with angiogenesis, and both Ets-1 and COUP-TFII mRNA are present in mesenchymal cells, we have sought to determine whether Ets-1 is a potential regulator of COUP-TFII gene expression. For this purpose, we performed transient transfection experiments using a luciferase reporter construct containing the mouse COUP-TFII promoter. We found that the COUP-TFII promoter activity is indeed regulated by Ets-1. We have identified two identical inverted potential ETS-binding sites located 47 nucleotides downstream of the start site. Mutation of both sites reduced the ability of Ets-1 to enhance the COUP-TFII promoter activity. Furthermore, other members of the ETS family such as Ets-2 or ETV1 are also potent regulators of the COUP-TFII promoter. Finally, the induction of the COUP-TFII gene is strongly enhanced by the expression of steroid receptor co-activator factors through a direct interaction with Ets-1. These results indicate that COUP-TFII is a potential downstream target of Ets-1 and it may partially mediate the Ets-1 function in angiogenesis.


A Fluorescence Sensing Method with Reduced DNA Typing and Low-Cost Instrumentation for Detection of Sample Tampering Cases in Urinalysis.
This work presents a method to unequivocally detect urine sample tampering in cases where integrity of the sample needs to be verified prior to urinalysis. The technique involves the detection of distinct patterns of a triplex short tandem repeats system in DNA extracted from human urine. The analysis is realized with single-dye fluorescence detection and using a regular smartphone camera. The experimental results had demonstrated the efficacy of the analytical approach to obtaining distinct profiles of amplicons in urine from different sample providers. Reproducibility tests with fresh and stored urine have revealed a maximum variation in the profiles within an interval of 5 to 9%. Cases of urine sample tampering via mixture were simulated in the study, and the experiments have identified patterns of mixed genotypes from dual mixtures of urine samples. Moreover, sample adulteration by mixing a non-human fluid with urine in a volume ratio over 25% can be detected. The low cost of the approach is accompanied by the compatibility of the technique to use with different DNA sample preparation protocols and PCR instrumentation. Furthermore, the possibility of realizing the method in an integrated microchip system open great perspectives to conducting sample integrity tests at the site of urine sample reception and/or at resource-limited settings.


The relationship between culture, attitude, social networks and quality of life in midlife Australian and Taiwanese citizens.
The objectives of this study were to specifically investigate the differences in culture, attitudes and social networks between Australian and Taiwanese men and women and identify the factors that predict midlife men and women's quality of life in both countries. A stratified random sample strategy based on probability proportional sampling (PPS) was conducted to investigate 278 Australian and 398 Taiwanese midlife men and women's quality of life. Multiple regression modelling and classification and regression trees (CARTs) were performed to examine the potential differences on culture, attitude, social networks, social demographic factors and religion/spirituality in midlife men and women's quality of life in both Australia and Taiwan. The results of this study suggest that culture involves multiple functions and interacts with attitudes, social networks and individual factors to influence a person's quality of life. Significant relationships were found between the interaction between cultural circumstances and a person's internal and external factors. The research found that good social support networks and a healthy optimistic disposition may significantly enhance midlife men and women's quality of life. The study indicated that there is a significant relationship between culture, attitude, social networks and quality of life in midlife Australian and Taiwanese men and women. People who had higher levels of horizontal individualism and collectivism, positive attitudes and better social support had better psychological, social, physical and environmental health, while it emerged that vertical individualists with competitive characteristics would experience a lower quality of life. This study has highlighted areas where opportunities exist to further reflect upon contemporary social health policies for Australian and Taiwanese societies and also within the global perspective, in order to provide enhanced quality care for growing midlife populations.