Costal Chondrocyte-Derived Pellet-Type Autologous Chondrocyte Implantation for Treatment of Articular Cartilage Defect.
Because articular chondrocyte-based autologous chondrocyte implantations (ACIs) have restrictively restored articular cartilage defects, alternative cell sources as a new therapeutic option for cartilage repair have been introduced. To assess whether implantation of a costal chondrocyte-derived pellet-type (CCP) ACI allows safe, functional, and structural restoration of full-thickness cartilage defects in the knee. Case series; Level of evidence, 4. In this first-in-human study, 7 patients with symptomatic, full-thickness cartilage lesions were enrolled. The chondrocytes isolated from the patients' costal cartilage were expanded, followed by 3-dimensional pellet culture to prepare the CCP-ACI. Implantation of the pellets was performed via minimal arthrotomy and secured with a fibrin sealant. Clinical scores, including the International Knee Documentation Committee (IKDC) subjective, Lysholm, and Tegner activity scores, were estimated preoperatively and at 1, 2, and 5 years postoperatively. High-resolution magnetic resonance imaging was also performed to evaluate cartilage repair as well as to calculate the MOCART (magnetic resonance observation of cartilage repair tissue) score. The costal chondrocytes of all patients formed homogeneous-sized pellets, which showed the characteristics of the hyaline cartilaginous tissue with lacunae-occupied chondrocytes surrounded by glycosaminoglycan and type II collagen-rich extracellular matrix. There were no treatment-related serious adverse events during the 5-year follow-up period. Significant improvements were seen in all clinical scores from preoperative baseline to the 5-year follow-up (IKDC subjective score, 34.67 to 75.86; Lysholm score, 34.00 to 85.33; Tegner activity score, 1.17 to 4.67; and MOCART score, 28.33 to 83.33). Two patients had complete defect filling on magnetic resonance imaging evaluation at 1 year. Moreover, at 5 years postoperatively, complete defect filling was observed in 4 patients, and hypertrophy or incomplete defect filling (50%-100%) was observed in 2 patients. The overall results of this clinical study suggest that CCP-ACI can emerge as a promising therapeutic option for articular cartilage repair with good clinical outcomes and structural regeneration and with stable results at midterm follow-up. NCT03517046 ( ClinicalTrials.gov identifier).


Rab7D small GTPase is involved in phago-, trogocytosis and cytoskeletal reorganization in the enteric protozoan Entamoeba histolytica.
Rab small GTPases regulate membrane traffic between distinct cellular compartments of all eukaryotes in a tempo-spatially specific fashion. Rab small GTPases are also involved in the regulation of cytoskeleton and signalling. Membrane traffic and cytoskeletal regulation play pivotal role in the pathogenesis of Entamoeba histolytica, which is a protozoan parasite responsible for human amebiasis. E. histolytica is unique in that its genome encodes over 100 Rab proteins, containing multiple isotypes of conserved members (e.g., Rab7) and Entamoeba-specific subgroups (e.g., RabA, B, and X). Among them, E. histolytica Rab7 is the most diversified group consisting of nine isotypes. While it was previously demonstrated that EhRab7A and EhRab7B are involved in lysosome and phagosome biogenesis, the individual roles of other Rab7 members and their coordination remain elusive. In this study, we characterised the third member of Rab7, Rab7D, to better understand the significance of the multiplicity of Rab7 isotypes in E. histolytica. Overexpression of EhRab7D caused reduction in phagocytosis of erythrocytes, trogocytosis (meaning nibbling or chewing of a portion) of live mammalian cells, and phagosome acidification and maturation. Conversely, transcriptional gene silencing of EhRab7D gene caused opposite phenotypes in phago/trogocytosis and phagosome maturation. Furthermore, EhRab7D gene silencing caused reduction in the attachment to and the motility on the collagen-coated surface. Image analysis showed that EhRab7D was occasionally associated with lysosomes and prephagosomal vacuoles, but not with mature phagosomes and trogosomes. Finally, in silico prediction of structural organisation of EhRab7 isotypes identified unique amino acid changes on the effector binding surface of EhRab7D. Taken together, our data suggest that EhRab7D plays coordinated counteracting roles: a inhibitory role in phago/trogocytosis and lyso/phago/trogosome biogenesis, and an stimulatory role in adherence and motility, presumably via interaction with unique effectors. Finally, we propose the model in which three EhRab7 isotypes are sequentially involved in phago/trogocytosis.


Seeking a better balance between efficiency and interpretability: Comparing the likert response format with the Guttman response format.
The Likert item response format for items is almost ubiquitous in the social sciences and has particular virtues regarding the relative simplicity of item-generation and the efficiency for coding responses. However, in this article, we critique this very common item format, focusing on its affordance for interpretation in terms of internal structure validity evidence. We suggest an alternative, the Guttman response format, which we see as providing a better approach for gathering and interpreting internal structure validity evidence. Using a specific survey-based example, we illustrate how items in this alternative format can be developed, exemplify how such items operate, and explore some comparisons between the results from using the two formats. In conclusion, we recommend usage of the Guttman response format for improving the interpretability of the resulting outcomes. Finally, we also note how this approach may be used in tandem with items that use the Likert response format to help balance efficiency with interpretability. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Utilization of a magnetic field-driven microscopic motion for piezoelectric energy harvesting.
In spite of the recent advances in the development of high performing piezoelectric materials, their applications are typically limited to the direct conversion of mechanical impact energy to electrical energy, potentially risking mechanical failures. In this study, we developed piezoelectric poly(vinylidenefluoride-trifluoroethylene) (P(VDF-TrFE)) nanofibers integrated with SiO2-shelled Fe3O4 magnetic nanoparticles, to utilize magnetic energy to reliably drive the piezoelectric effect. Specifically, we show that the shape of the magnetic nanoparticles exerts a significant effect on the efficiency of the magneto-mechano-electrical energy conversion as magnetic nanorods exhibit approximately 70% enhancement in electric field generation under cyclic magnetic fields as compared to nanospheres. Under an alternating magnetic field of 200 mT, the magnetic nanorod-piezoelectric nanofiber composite generated a peak-to-peak voltage of approximately 30 mVp-p with a superior durability without any performance degradation after over 1 million cycles. This study demonstrates the potential of magnetic-field responsive, piezoelectric-based materials in energy harvesting applications from non-mechanical energy sources.


Human RAD52 stimulates the RAD51-mediated homology search.
Homologous recombination (HR) is a DNA repair mechanism of double-strand breaks and blocked replication forks, involving a process of homology search leading to the formation of synaptic intermediates that are regulated to ensure genome integrity. RAD51 recombinase plays a central role in this mechanism, supported by its RAD52 and BRCA2 partners. If the mediator function of BRCA2 to load RAD51 on RPA-ssDNA is well established, the role of RAD52 in HR is still far from understood. We used transmission electron microscopy combined with biochemistry to characterize the sequential participation of RPA, RAD52, and BRCA2 in the assembly of the RAD51 filament and its activity. Although our results confirm that RAD52 lacks a mediator activity, RAD52 can tightly bind to RPA-coated ssDNA, inhibit the mediator activity of BRCA2, and form shorter RAD51-RAD52 mixed filaments that are more efficient in the formation of synaptic complexes and D-loops, resulting in more frequent multi-invasions as well. We confirm the in situ interaction between RAD51 and RAD52 after double-strand break induction in vivo. This study provides new molecular insights into the formation and regulation of presynaptic and synaptic intermediates by BRCA2 and RAD52 during human HR.


[Significance of urinary concentrations of S-benzyl-N-acetylcysteine (S-BMA) in subjects exposed to toluene].
Toluene is a widely diffuse solvent for oils, resins, rubber and paints, either alone or as a major component in a mixture; in the industrial environment it is currently present at concentrations of ppm. Toluene can be absorbed via the lungs or via the skin. The absorption of toluene via inhalation is related to the exposure level as well as the activities level of workers. Once absorbed into the body, toluene is metabolized in man to benzoic acid, followed by hepatic cytochrome P450 catalyzed glycine conjugation to form hippuric acid. Relatively small amounts appear in urine as o-cresol and p-cresol where they occur as glucoronide and sulfate derivate. Only a minor fraction of inhaled solvent is conjugated with glutathione with the production of S-benzyl-N-acetylcysteine (S-BMA). Several biological indicators have been proposed for evaluating toluene exposure in the workplace. These include urinary hippuric acid, toluene in blood, toluene in breath, o-cresol in urine and toluene in urine. We examined a group of 18 workers occupationally exposed to toluene, determining the concentrations of toluene in ambient air and S-BMA in urine. All urine samples were collected at the end of work shift. The renal excretion of S-BMA showed highly significant correlations with environmental data and with the other established parameters of biological monitoring of toluene. The median ambient air concentration was 15.7 ppm ranging from 2.9 to 70.3 ppm, the median concentration of S-BMA was 16.0 micrograms/g creatinine. S-BMA was detectable in urine samples of a control group of 87 subjects non occupationally exposed to toluene. Most of unexposed subjects showed S-BMA values lower than 10 micrograms/g creatinine both in smokers and in nonsmokers and no significant difference was found in samples (20) collected at three intervals during one day. Our finding further indicates that the metabolite S-BMA could be a marker of occupational toluene exposure.


Challenges in Diagnosis and Therapeutic Approach of Acute on Chronic Liver Failure-A Review of Current Evidence.
Acute-on-chronic liver failure (ACLF) is a syndrome characterized by acute and severe decompensation of chronic liver disease (CLD) correlated with multiple organ failure, poor prognosis, and increased mortality. In 40-50% of ACLF cases, the trigger is not recognized; for many of these patients, bacterial translocation associated with systemic inflammation is thought to be the determining factor; in the other 50% of patients, sepsis, alcohol consumption, and reactivation of chronic viral hepatitis are the most frequently described trigger factors. Other conditions considered precipitating factors are less common, including acute alcoholic hepatitis, major surgery, TIPS insertion, or inadequate paracentesis without albumin substitution. Host response is likely the primary factor predicting ACLF severity and prognosis, the host immune response having a particular significance in this syndrome, together with the inflammatory cascade. The management of ACLF includes both the prevention of the precipitating factors that lead to acute liver decompensation and the support of vital functions, the prevention and management of complications, the estimation of prognosis, and the opportunity for liver transplantation.


Adoptive transfer of tumor-reactive transforming growth factor-beta-insensitive CD8+ T cells: eradication of autologous mouse prostate cancer.
Transforming growth factor (TGF)-beta is a potent immunosuppressant. Overproduction of TGF-beta by tumor cells may lead to tumor evasion from the host immune surveillance and tumor progression. The present study was conducted to develop a treatment strategy through adoptive transfer of tumor-reactive TGF-beta-insensitive CD8+ T cells. The mouse TRAMP-C2 prostate cancer cells produced large amounts of TGF-beta1 and were used as an experimental model. C57BL/6 mice were primed with irradiated TRAMP-C2 cells. CD8+ T cells were isolated from the spleen of primed animals, were expanded ex vivo, and were rendered TGF-beta insensitive by infecting with a retrovirus containing dominant-negative TGF-beta type II receptor. Results of in vitro cytotoxic assay revealed that these CD8+ T cells showed a specific and robust tumor-killing activity against TRAMP-C2 cells but were ineffective against an irrelevant tumor line, B16-F10. To determine the in vivo antitumor activity, recipient mice were challenged with a single injection of TRAMP-C2 cells for a period up to 21 days before adoptive transfer of CD8+ T cells was done. Pulmonary metastasis was either eliminated or significantly reduced in the group receiving adoptive transfer of tumor-reactive TGF-beta-insensitive CD8+ T cells. Results of immunofluorescent studies showed that only tumor-reactive TGF-beta-insensitive CD8+ T cells were able to infiltrate into the tumor and mediate apoptosis in tumor cells. Furthermore, transferred tumor-reactive TGF-beta-insensitive CD8+ T cells were able to persist in tumor-bearing hosts but declined in tumor-free animals. These results suggest that adoptive transfer of tumor-reactive TGF-beta-insensitive CD8+ T cells may warrant consideration for cancer therapy.


Wild edible onions - Allium flavum and Allium carinatum - successfully prevent adverse effects of chemotherapeutic drug doxorubicin.
The objective of this study was to evaluate potential of two chemically characterized edible wild onion species, Allium flavum and Allium carinatum, to reduce side effects of cytostatic doxorubicin (Dox). Since Dox application is mainly limited due to its high cardiotoxicity, while there are no approved cardioprotective agents for the prevention of Dox adverse effects, new co-treatments are urgently needed. Here, we showed that methanol extracts expressed high antioxidant activity and synergistically increased Dox anticancer activity against human hepatoma (HepG2) and lung carcinoma (A549) cells, while protected normal human fibroblasts (MRC-5) from Dox cytotoxicity. Analysis of the antioxidative enzymes level (catalase and superoxide dismutases) showed that the catalase level was differently altered in cancer cells compared to normal cells upon applied treatments. In vivo toxicity evaluation in the zebrafish model revealed significantly lower toxicity of extracts compared to Dox, and no teratogenic effects at applied doses. We found that extracts successfully rescued the Dox-treated embryos of life-threating cardiomyopathy, while at the same time reduced developmental toxicity and neutropenia. Further analysis demonstrated that extracts had higher anti-angiogenic activity than sunitinib or auranofin, clinically used antiangiogenic drugs. In addition, angiogenesis was markedly more suppressed in Dox-extract cotreatments than upon single treatments.


[The morphological characteristics of osteoarthrosis].
Osteoarthrosis (OA) is a group of overlapping diseases that have various etiologies, but equal biological, morphological, and clinical outcomes. OA is characterized by degenerative and destructive changes in the articular hyaline cartilage, subchondral bone, spongiosis, synovium, capsule, and articular ligamentous apparatus. The clinical symptoms of OA are closely associated with morphological changes in articular tissue elements, primarily in the articular hyaline cartilage. Six stages that reflect the magnitude of changes in the hyaline cartilage and 4 degrees of the extent of the process along the articular surface are identified.


Propylthiouracil associated antineutrophil cytoplasmic antibodies (ANCA) in patients with childhood onset Graves' disease.
Propylthiouracil (PTU) can induce anti-myeloperoxidase (MPO-ANCA) positive vasculitis. We performed a cross-sectional study to estimate the prevalence of MPO-ANCA in patients with childhood onset Graves' disease (GD) receiving PTU and to assess the relationship between ANCA and clinical manifestations of vasculitis. We studied 60 patients (59 girls and one boy) between 7.3 and 25.0 years of age (mean +/- SD, 14.71 +/- 4.49). GD, diagnosed at the age of 3.0 to 14.5 years (11.3 +/- 2.48), was designated as: newly diagnosed, on PTU therapy, and after PTU discontinuation in 4, 50 and 6 patients, respectively. Manifestations of vasculitis were noted and the patients were tested for MPO-ANCA, antinuclear antibodies, blood urea nitrogen, creatinine and urine analysis. Twenty-six patients (43.3%) reacted positively for MPO-ANCA, 23 were on PTU therapy (0.42 to 6.00, median 3.00 years) and three had discontinued PTU. There were 34 (56.7%) ANCA-negative patients and 27 patients on PTU therapy (0.25 to 5.17, median 1.00 years, p = 0.012). Vasculitis presented in 16 patients (26.7%), all of whom were receiving PTU at the time of the study. The percentage of vasculitis among MPO-ANCA positive patients was 27.6% more than in the negative group, p = 0.017. PTU was discontinued in patients with vasculitis and positive for MPO-ANCA. Our findings show a high prevalence of MPO-ANCA positivity and a significantly higher percentage of vasculitis among these patients, suggesting that patients taking PTU should be closely observed for the appearance of MPO-ANCA and signs of vasculitis, especially patients GD who have been treated for a long time.


Developmental changes in polyamine levels and synthesis in the ovine conceptus.
Polyamines (putrescine, spermidine, and spermine) are essential for placental growth and angiogenesis. However, little is known about changes in polyamine synthesis associated with development of the ovine conceptus (embryo/fetus and associated placental membranes). We hypothesized that rates of placental polyamine synthesis were maximal during the rapid placental growth that occurs in the first half of pregnancy. This hypothesis was tested using ewes between Days 30 and 140 of gestation. Columbia cross-bred ewes were hysterectomized on Days 30, 40, 60, 80, 100, 120, or 140 of gestation (Day 0 = mating; n = 4 ewes/day) to obtain placentomes, intercotyledonary placenta, intercaruncular endometrium, and allantoic as well as amniotic fluids. The tissues were analyzed for ornithine decarboxylase (ODC) and arginase activities; arginine, ornithine, and polyamine concentrations; and polyamine synthesis using radiochemical and chromatographic methods. Maximal ODC and arginase activities and the highest rates of polyamine synthesis were observed in all tissues on Day 40 of gestation. Concentrations of ornithine and polyamines in placentomes and intercaruncular endometrium also peaked on Day 40 of gestation. In ovine allantoic and amniotic fluids, polyamines were most abundant during early (Days 40-60) and late (Days 100-140) gestation, respectively. Amniotic fluid spermine increased progressively with advancing gestation. Results of the present study indicate metabolic coordination among the several integrated pathways that support high rates of polyamine synthesis in the placenta and endometrium during early pregnancy. Our findings may have important implications for both intrauterine growth retardation and fetal origins of diseases in adults.


Proton pump inhibitors and risk of Clostridium difficile infection: a multi-country study using sequence symmetry analysis.
To determine the association between incident proton pump inhibitor (PPI) use and Clostridium difficile infections across multiple countries Method: National data covering the total population in Australia and Korea, the Canadian population over 65 years and a 3 million person random sample data set from Taiwan were assessed, as were data from a worker insurance population and a hospital inpatient/outpatient population in Japan. Sequence symmetry analysis was used to assess the association with oral vancomycin dispensing as the outcome of interest. 54,957 patients were included. Positive associations were observed in Australia; adjusted sequence ratio (ASR) 2.48 (95% CI 1.90, 3.12), Korea ASR 2.15 (95%CI 2.11, 2.19), Canada ASR 1.45 (95% CI 1.16, 1.79), Japan hospital dataset ASR 3.21 (95%CI 2.12, 4.55) and Japan worker insurance dataset ASR 5.40 (95% CI 2.73, 8.75). The pooled result was ASR 2.40 (95%CI 1.88, 3.05) and 3.16 (95%CI 1.95, 5.10) when limited to Japan, Korean and Taiwan. Results did not vary by individual PPI. The temporal analysis showed effects within the first two weeks of PPI initiation. Our study confirms the association between PPI initiation and C. difficile infections across countries in the Asia-Pacific region.


Subjective well-being: gender differences in Holocaust survivors-specific and cross-national effects.
Subjective well-being (SWB) has become an important concept in evaluating older adults' quality of life. The cognitive and emotional evaluations which are used to appraise it differ in structure, characteristics, and effects on life. The purpose of this study was to support hypotheses regarding expected Holocaust survivors-specific effects and cross-cultural differences on three indicators of SWB. We recruited samples of 50 male and female Israeli Holocaust survivors, other older Israelis, and older Canadians (N = 300) for allowing us to distinguish survivors-specific effects from cross-national differences. State anxiety, depressive symptoms, and life-satisfaction were compared across groups of men and women. Where univariate differences were detected, post hoc comparisons were computed to determine which of the groups significantly differed. In general, a higher level of SWB was found among Canadians in comparison to both comparative Israeli groups. Level of depressive symptoms was significantly higher among women survivors than in the other two groups. Both groups of Israeli women had higher scores on anxiety than Canadian Women; less apparent were differences across groups of men. Life-satisfaction did not differ among the groups. Our findings regarding depression support the survivor-specific effect hypothesis for women, and a national effect on anxiety, but not any effect on life-satisfaction. These findings suggest significant differences in impacts of traumatic life events on cognitive versus emotional indicators of SWB. This issue should be further investigated due to its practical implications in use of various measures of SWB with people who experienced traumatic events.


Glycan remodeled erythrocytes facilitate antigenic characterization of recent A/H3N2 influenza viruses.
During circulation in humans and natural selection to escape antibody recognition for decades, A/H3N2 influenza viruses emerged with altered receptor specificities. These viruses lost the ability to agglutinate erythrocytes critical for antigenic characterization and give low yields and acquire adaptive mutations when cultured in eggs and cells, contributing to recent vaccine challenges. Examination of receptor specificities of A/H3N2 viruses reveals that recent viruses compensated for decreased binding of the prototypic human receptor by recognizing α2,6-sialosides on extended LacNAc moieties. Erythrocyte glycomics shows an absence of extended glycans providing a rationale for lack of agglutination by recent A/H3N2 viruses. A glycan remodeling approach installing functional receptors on erythrocytes, allows antigenic characterization of recent A/H3N2 viruses confirming the cocirculation of antigenically different viruses in humans. Computational analysis of HAs in complex with sialosides having extended LacNAc moieties reveals that mutations distal to the RBD reoriented the Y159 side chain resulting in an extended receptor binding site.


CSF1 expression in nongynecological leiomyosarcoma is associated with increased tumor angiogenesis.
Leiomyosarcoma (LMS) is a malignant tumor of smooth muscle cells for which few effective therapies exist. A subset of LMS cases express macrophage colony-stimulating factor (CSF1) and the resultant tumor-associated macrophage (TAM) infiltration predicts poor clinical outcome. Further, TAMs have been shown to increase tumor angiogenesis. Here, we analyzed 149 LMS cases by immunohistochemistry for vascular marker CD34 and show that high microvessel density (MVD) in nongynecological LMS cases significantly predicts poor patient outcome. The majority of high MVD cases were also CSF1-positive, and when combining high MVD with CSF1 expression, an even stronger prognostic correlation with patient outcome was obtained. Gene expression profiling revealed that MVD has a stronger correlation with CSF1 expression than with expression of vascular endothelial growth factor isoforms, which have traditionally been used as markers of angiogenesis and as anti-angiogenic therapeutic targets. Finally, patterns of CSF1 expression and TAM recruitment remained consistent between primary tumors and their metastases, and between primary tumors and those grown as xenografts in mice, highlighting the stability of these features to the biology of LMS tumors. Together, these findings suggest an important role for CSF1 and the resulting TAM infiltration in the pathological neovascularization of LMS tumors and provide a rationale for CSF1-targeted therapies in LMS.


Long-Time Memory and Ternary Logic Gate Using a Multistable Cavity Magnonic System.
Multistability is an extraordinary nonlinear property of dynamical systems and can be explored to implement memory and switches. Here we experimentally realize the tristability in a three-mode cavity magnonic system with Kerr nonlinearity. The three stable states in the tristable region correspond to the stable solutions of the frequency shift of the cavity magnon polariton under specific driving conditions. We find that the system staying in which stable state depends on the history experienced by the system, and this state can be harnessed to store the history information. In our experiment, the memory time can reach as long as 5.11 s. Moreover, we demonstrate the ternary logic gate with good on-off characteristics using this multistable hybrid system. Our new findings pave a way towards cavity magnonics-based information storage and processing.


[Cancer of the prostate: what treatment for what stage?].
The treatment of locally advanced prostatic cancer is the most controversial subject in urological oncology. Localized disease is best treated by radical prostatectomy or radiotherapy. Metastatic cancer is treated by surgical or medical castration. Second line therapeutics will be overviewed. Each treatment decision is based upon the pathological stage and differentiation of the tumour, the life expectancy and comorbidity of the patient.


Isozyme selectivity of the inhibition of rat liver cytochromes P-450 by chloramphenicol in vivo.
The isozyme-selectivity of chloramphenicol as an inhibitor of rat liver cytochromes P-450 has been investigated. Untreated rats and rats treated with the inducers phenobarbital, beta-naphthoflavone, pregnenolone 16 alpha-carbonitrile, and clofibrate have been injected intraperitoneally with chloramphenicol, and inhibition of specific cytochrome P-450 isozymes has been assessed by monitoring the metabolism of warfarin, testosterone, isosafrole, or lauric acid in subsequently prepared hepatic microsomal preparations. Of eight major cytochrome P-450 isozymes which could be monitored in this fashion, three were inhibited by more than 50% by a dose of chloramphenicol of 300 mg/kg, whereas no evidence of inhibition of the remaining isozymes was obtained. P-450PB-C, an isozyme which is present in significant amounts in untreated rats and which is induced approximately 2-fold by phenobarbital, was the most susceptible cytochrome P-450 to inhibition by chloramphenicol both in vivo and in vitro. P-450PB-B, the major phenobarbital-inducible isozyme, and P-450UT-A, a male-specific testosterone 2 alpha- and 16 alpha-hydroxylase, were intermediate in their susceptibility to chloramphenicol. In contrast, the major isozymes induced by beta-naphthoflavone, pregnenolone 16 alpha-carbonitrile, and clofibrate, as well as a constitutive testosterone 7 alpha-hydroxylase, were not inhibited by chloramphenicol.


Interpretability of Cancer Clinical Trial Results Using Restricted Mean Survival Time as an Alternative to the Hazard Ratio.
In a comparative clinical study with progression-free survival (PFS) or overall survival (OS) as the end point, the hazard ratio (HR) is routinely used to design the study and to estimate the treatment effect at the end of the study. The clinical interpretation of the HR may not be straightforward, especially when the underlying model assumption is not valid. A robust procedure for study design and analysis that enables clinically meaningful interpretation of trial results is warranted. To discuss issues of conventional trial design and analysis and to present alternatives to the HR using a recent immunotherapy study as an illustrative example. By comparing 2 groups in a survival analysis, we discuss issues of using the HR and present the restricted mean survival time (RMST) as a summary measure of patients' survival profile over time. We show how to use the difference or ratio in RMST between 2 groups as an alternative for designing and analyzing a clinical study with an immunotherapy study as an illustrative example. Overall survival or PFS. Group contrast measures included HR, RMST difference or ratio, and the event rate difference. For the illustrative example, the HR procedure indicates that nivolumab significantly prolonged patient OS and was numerically better than docetaxel for PFS. However, the median PFS time of docetaxel was significantly better than that of nivolumab. Therefore, it may be difficult to use median OS and/or PFS to interpret of the HR value clinically. On the other hand, using RMST difference, nivolumab was significantly better than docetaxel for both OS and PFS. We also provide details regarding design of a future study with RMST-based measures. The design and analysis of a conventional cancer clinical trial can be improved by adopting a robust statistical procedure that enables clinically meaningful interpretations of the treatment effect. The RMST-based quantitative method may be used as a primary tool for future cancer trials or to help us to better understand the clinical interpretation of the HR even when its model assumption is plausible.


A Conceptual Framework of Social Communication: Clinical Applications to Pediatric Traumatic Brain Injury.
Social communication impairments are common following pediatric traumatic brain injury (TBI) and can lead to social isolation, and poor social outcomes. Social communication has been documented as a persistent area of need in terms of proper assessment and intervention; however, this is not consistently addressed in clinical practice. While there is a body of evidence regarding social communication impairments and pediatric TBI, this area is not yet fully understood and remains underrecognized. To meet this gap, we provide a conceptual framework of social communication from a neurodevelopmental perspective, which can be applied to better understand the social communication impairments associated with pediatric TBI. We propose a general model of social communication with component constructs and consideration of internal factors such as sex and gender. These can inform considerations, clinical applications, and future research in assessment and evidence-based interventions within the domain of social communication.


A Turn-On Fluorescent Probe for Detection of Sub-ppm Levels of a Sulfur Mustard Simulant with High Selectivity.
A new type of fluorescent probe capable of detecting a sulfur mustard (SM) simultant at a concentration of 1.2 μM in solution and 0.5 ppm in the gas phase has been developed. Owing to its molecular structure with a thiocarbonyl component and two piperidyl moieties integrated into the xanthene molecular skeleton, this probe underwent a highly selective nucleophilic reaction with the SM simultant and generated a thiopyronin derivative emitting intensive pink fluorescence. The distinct difference in electronic structure between the probe and thiopyronin derivative generated a marked shift of the absorption band from 445 to 567 nm, which enabled an optimal wavelength propitious for exciting the thiopyronin derivative but adverse to the probe. Such efficient separation of the excitation wavelength and tremendous increase in fluorescence quantum yield, from less than 0.002 to 0.53, upon conversion from the probe to the thiopyronin derivative, jointly led to a distinct contrast in the beaconing fluorescence signal (up to 850-fold) and therefore the unprecedented sensitivity for detecting SM species.


Normalization of flow-mediated dilation to shear stress area under the curve eliminates the impact of variable hyperemic stimulus.
Normalization of brachial artery flow-mediated dilation (FMD) to individual shear stress area under the curve (peak FMD:SSAUC ratio) has recently been proposed as an approach to control for the large inter-subject variability in reactive hyperemia-induced shear stress; however, the adoption of this approach among researchers has been slow. The present study was designed to further examine the efficacy of FMD normalization to shear stress in reducing measurement variability. Five different magnitudes of reactive hyperemia-induced shear stress were applied to 20 healthy, physically active young adults (25.3 +/- 0. 6 yrs; 10 men, 10 women) by manipulating forearm cuff occlusion duration: 1, 2, 3, 4, and 5 min, in a randomized order. A venous blood draw was performed for determination of baseline whole blood viscosity and hematocrit. The magnitude of occlusion-induced forearm ischemia was quantified by dual-wavelength near-infrared spectrometry (NIRS). Brachial artery diameters and velocities were obtained via high-resolution ultrasound. The SSAUC was individually calculated for the duration of time-to-peak dilation. One-way repeated measures ANOVA demonstrated distinct magnitudes of occlusion-induced ischemia (volume and peak), hyperemic shear stress, and peak FMD responses (all p < 0.0001) across forearm occlusion durations. Differences in peak FMD were abolished when normalizing FMD to SSAUC (p = 0.785). Our data confirm that normalization of FMD to SSAUC eliminates the influences of variable shear stress and solidifies the utility of FMD:SSAUC ratio as an index of endothelial function.


Contrast-enhanced Ultrasound in evaluating of angiogenesis and tumor staging of nasopharyngeal carcinoma in nude mice.
To explore the use of Contrast-enhanced Ultrasound (CEUS) in evaluating angiogenesis in a xenograft nasopharyngeal carcinoma (NPC) model in nude mice and the evolution of CEUS parameters according to the growth of NPC. Nude mice were divided into three groups according to experiments conducted at various times from tumor implantation (8 mice/group; group A: 4 weeks from implantation; group B:6 weeks from implantation; group C:8 weeks from implantation). CNE-2 cells were transplanted in 24 nude mice and CEUS evaluations of the tumors were performed at 4, 6 or 8 weeks from implantation. CEUS parametric perfusion images and pathological findings were recorded. R version 3.4.4 software was used to analyze the CEUS parameters and pathological findings. One-way anova analysis indicated statistically significant differences among the three groups with the parameters of peak intensity (PI) (p<0.001), area wash in (AWI) (p<0.001), area wash out (AWO) (p<0.001) and tumor volumes (p<0.001).Pearson correlation coefficient analysis indicated that microvessel density (MVD) was correlated with tumor volume (r = 0.644, p = 0.001), PI (r = 0.904, p<0.0001), AWI (r = 0.547, p = 0.008) and AWO (r = 0.744, P<0.0001). Tumor volume was correlated with MVD (r = 0.644, p = 0.001), PI (r = 0.625, p = 0.002), AWI (r = 0.528, p = 0.012) and AWO (r = 0.784, p<0.001). The percentage of necrosis in histological sections was correlated with the percentage of CEUS unperfused area (r = 0.446,p = 0.038). Spearman rank correlation coefficient analysis indicated that vascular endothelial growth factor (VEGF) was correlated with PI (r = 0.462, P = 0.032). Welch t test indicated PI, AWI and AWO parameters were significantly lower than that of kidneys (p<0.001, p = 0.009, p = 0.005). The CEUS parameters PI, AWI and AWO indirectly reflect the MVD and the tumor volume in our model of subcutaneous transplanted NPC in nude mice, providing precious information on angiogenesis and tumor growth. VEGF may play a role in promoting angiogenesis of NPC.


S-100 protein and calmodulin levels in cerebrospinal fluid after subarachnoid hemorrhage.
The levels of two calcium-binding proteins, S-100 protein and calmodulin, were measured serially in the cerebrospinal fluid (CSF) of patients after subarachnoid hemorrhage (SAH) and aneurysm surgery. These two proteins have a similar molecular structure and are highly concentrated in the central nervous system (CNS). The levels of S-100 protein found in the earliest postoperative CSF samples correlated with the preoperative SAH grades. High S-100 protein levels in the CSF were found in patients with poor SAH grades. Moreover, the prognosis of the patients correlated with the S-100 protein levels in the CSF samples taken during the immediate postoperative period and with the daily changes of the S-100 protein levels. Severe diffuse cerebral vasospasm was followed by a sharp S-100 protein increase. These results suggest that S-100 protein levels in the CSF provide a useful index of organic damage in the CNS, and furthermore that S-100 protein levels and their changes may have prognostic value for patients after SAH. On the other hand, there was a lack of correlation between the calmodulin levels and the preoperative grade or outcome. It would be inappropriate, however, to speculate from the results of this study that these calcium-binding proteins in the CSF play any causative role in pathological processes such as cerebral vasospasm or brain ischemia after SAH, since changes in the levels of these proteins followed the onset of clinical signs of deterioration.


Privacy Concerns About Health Information Disclosure in Mobile Health: Questionnaire Study Investigating the Moderation Effect of Social Support.
Mobile health (mHealth) provides a new opportunity for disease prediction and patient health self-management. However, privacy problems in mHealth have drawn significant attention to patients' online health information disclosure and to the possibility that privacy concerns may hinder mHealth development. Privacy calculus theory (PCT) has been widely used to understand personal information disclosure behaviors with the basic assumption of a rational and linear decision-making process. However, cognitive behavior processes are complex and mutual. In an attempt to gain a fuller understanding of information disclosure behavior, we further optimize a PCT-based information disclosure model by identifying the mutual relationship between costs (privacy concerns) and benefits. Social support, which has been proven to be a distinct and significant disclosure benefit of mHealth, was chosen as the representative benefit of information disclosure. We examine a structural equation model that incorporates privacy concerns, health information disclosure intention in mHealth, and social support from mHealth, all at the individual level. A validated questionnaire was completed by 253 randomly selected participants. The result indicated that perceived health information sensitivity positively enhances patients' privacy concern (beta path coefficient 0.505, P<.001), and higher privacy concern levels will decrease their health information disclosure intention (beta path coefficient -0.338, P<.001). Various individual characteristics influence perceived health information sensitivity in different ways. One dimension of social support, informational support, negatively moderates the effect of the relationship between perceived health information sensitivity and privacy concerns (beta path coefficient -0.171, P=.092) and the effect of the relationship between privacy concerns and health information disclosure intention (beta path coefficient -0.105, P=.092). However, another dimension, emotional support, has no direct moderation effect on the relationship between privacy concerns and health information disclosure intention. The results indicate that social support can be regarded as a disutility reducer. That is, on the one hand, it reduces patients' privacy concerns; on the other hand, it also reduces the negative impact of privacy concerns on information disclosure intention. Moreover, the moderation effect of social support is partially supported. Informational support, one dimension of social support, is significant (beta path coefficient -0.171, P=.092), while the other dimension, emotional support, is not significant (beta path coefficient -0.137, P=.146), in mHealth. Furthermore, the results are different among patients with different individual characteristics. This study also provides specific theoretical and practical implications to enhance the development of mHealth.


Management of self-harm wounds.
Self-harm may be used by individuals as a means to relieve distress and may be a coping and suicide prevention strategy. This article focuses on risk assessment of patients who may self-harm as well as the treatment of those presenting with self-harm wounds.


Hierarchical population structure and habitat differences in a highly mobile marine species: the Atlantic spotted dolphin.
Recent molecular studies have shown that highly mobile species with continuous distributions can exhibit fine-scale population structure. In this context, we assessed genetic structure within a marine species with high dispersal potential, the Atlantic spotted dolphin (Stenella frontalis). Using 19 microsatellite loci and mitochondrial control region sequences, population structure was investigated in the western North Atlantic, the Gulf of Mexico and the Azores Islands. Analyses of the microsatellite data identified four distinct genetic clusters, which were supported by the control region sequences. The highest level of divergence was seen between two clusters corresponding to previously described morphotypes that inhabit oceanic and shelf waters. The combined morphological and genetic evidence suggests these two lineages are on distinct evolutionary trajectories and could be considered distinct subspecies despite their parapatry. Further analysis of the continental shelf cluster resulted in three groups: animals inhabiting shelf waters in the western North Atlantic, the eastern Gulf of Mexico and the western Gulf of Mexico. Analyses of environmental data indicate the four genetic clusters inhabit distinct habitats in terms of depth and sea surface temperature. Contemporary dispersal rate estimates suggest all of these populations should be considered as distinct management units. Conversely, no significant genetic differentiation was observed between S. frontalis from offshore waters of the western North Atlantic and the Azores, which are separated by approximately 4500 km. Overall, the hierarchical structure observed within the Atlantic spotted dolphin shows that the biogeography of the species is complex because it is not shaped solely by geographic distance.


A retrospective chart review of the patient population accessing augmentative & alternative communication at an urban assistive technology center.
The purpose of this study was to 1) identify and describe the population seeking alternative and augmentative communication (AAC) evaluations at an assistive technology (AT) center in the Midwestern United States and 2) describe the AAC device features or services participants identify as most important at their initial AAC evaluations. Charts of 53 participants seeking AAC interventions at an AT center in the Midwestern United States were retrospectively reviewed. Information from the QUEST 2.0 was used to determine what AT features were identified as most important. The majority of participants seen at the AT center had progressive diseases. Across all of the participants, ease of use and effectiveness were identified as the "most important" aspects of an AAC device satisfaction. These findings highlight the importance of determining who is accessing AAC services across AT centers to determine whether barriers to AAC services exist. Moreover, patients report regarding what variables they deem as most important reflect the fact that excellent service-delivery may not overcome the importance of other variables, such as ease of use that impact AAC use.


The other frontiers of science.
Frontiers of science are usually considered as those areas where the boundares of human knowledge are being pushed most vigorousl into the unknown. These are the intemal frontiers. But no less important are the external frontiers. Those bordering on the federal govermment, on education, on private industry and on intemational affairs and the developing world are among the most critical and demanding. Some of the outstanding problems facing science, and scientists in these extemal interactions, are discussed within the context of our changing national and international priorities.


The Identification of a Quantative Trait Loci-Allele System of Antixenosis against the Common Cutworm (Spodoptera litura Fabricius) at the Seedling Stage in the Chinese Soybean Landrace Population.
Common cutworm (CCW) is an omnivorous insect causing severe yield losses in soybean crops. The seedling-stage mini-tray identification system with the damaged leaf percentage (DLP) as an indicator was used to evaluate antixenosis against CCW in the Chinese soybean landrace population (CSLRP) under three environments. Using the innovative restricted two-stage multi-locus genome-wide association study procedure (RTM-GWAS), 86 DLP QTLs with 243 alleles (2-11/QTL) were identified, including 66 main-effect loci with 203 alleles and 57 QTL-environment interaction loci with 172 alleles. Among the main-effect loci, 12 large-contribution loci (R2 ≥ 1%) explained 25.45% of the phenotypic variation (PV), and 54 small-contribution loci (R2 < 1%) explained 16.55% of the PV. This indicates that the CSLRP can be characterized with a DLP QTL-allele system complex that has not been found before, except for a few individual QTLs without alleles involved. From the DLP QTL-allele matrix, the recombination potentials expressed in the 25th percentile of the DLP of all possible crosses were predicted to be reduced by 41.5% as the maximum improvement and 14.2% as the maximum transgression, indicating great breeding potential in the antixenosis of the CSLRP. From the QTLs, 62 candidate genes were annotated, which were involved in eight biological function categories as a gene network of the DLP. Changing from susceptible to moderate plus resistant varieties in the CSLRP, 26 QTLs had 32 alleles involved, in which 19 genes were annotated from 25 QTL-alleles, including eight increased negative alleles on seven loci and 11 decreased positive alleles on 11 loci, showing the major genetic constitution changes for the antixenosis enhancement at the seedling stage in the CSLRP.


Mid- to Long-Term Outcomes After Resection of Thoracic Dumbbell Tumors Managed by Laminectomy and Unilateral Total Facetectomy Without Instrumented Fusion.
Retrospective multicenter study. To evaluate mid- to long-term surgical outcomes of thoracic dumbbell tumors managed by laminectomy and unilateral total facetectomy without instrumented fusion. A total of 15 patients with thoracic dumbbell tumors who underwent primary resection by laminectomy and unilateral total facetectomy without spinal instrumented fusion between 2000 and 2015 were reviewed. Patient characteristics, surgical outcomes (including spinal alignment and stability), disc degeneration, pain, disability, and health-related quality of life were evaluated. Additionally, to analyze the impact of the affected levels on these outcomes, we divided the patients into 2 groups: a middle thoracic group and a thoracolumbar group. The mean duration of follow-up was 100.5 months (range, 36-190 months). The affected level was T3-T4 or below in all patients. Although the local kyphosis angle (8.1° to 12.7°), thoracic kyphosis angle (25.6° to 33.9°), and coronal Cobb angle (6.6° to 9.5°) significantly increased from preoperative to the final visit (P ≤ .02), no patient demonstrated spinal instability. From magnetic resonance imaging, no patient had a worse grade of disc degeneration in the affected level than those in the adjacent levels. The percentage of patients who presented with an Oswestry disability index ≤ 22% was 80%. Moreover, the surgical region did not adversely affect the outcomes. No patient required additional surgery due to spinal instability or deformity. Unilateral total facetectomy without fusion to resect thoracic dumbbell tumors caused neither spinal deformity nor instability requiring additional surgery at the mid- to long-term follow-up.


Dependence of Working Memory on Coordinated Activity Across Brain Areas.
Neural signatures of working memory (WM) have been reported in numerous brain areas, suggesting a distributed neural substrate for memory maintenance. In the current manuscript we provide an updated review of the literature focusing on intracranial neurophysiological recordings during WM in primates. Such signatures of WM include changes in firing rate or local oscillatory power within an area, along with measures of coordinated activity between areas based on synchronization between oscillations. In comparing the ability of various neural signatures in any brain area to predict behavioral performance, we observe that synchrony between areas is more frequently and robustly correlated with WM performance than any of the within-area neural signatures. We further review the evidence for alteration of inter-areal synchrony in brain disorders, consistent with an important role for such synchrony during behavior. Additionally, results of causal studies indicate that manipulating synchrony across areas is especially effective at influencing WM task performance. Each of these lines of research supports the critical role of inter-areal synchrony in WM. Finally, we propose a framework for interactions between prefrontal and sensory areas during WM, incorporating a range of experimental findings and offering an explanation for the observed link between intra-areal measures and WM performance.


Dual-modality optical coherence tomography and fluorescence tethered capsule endomicroscopy.
OCT tethered capsule endomicroscopy (TCE) is an emerging noninvasive diagnostic imaging technology for gastrointestinal (GI) tract disorders. OCT measures tissue reflectivity that provides morphologic image contrast, and thus is incapable of ascertaining molecular information that can be useful for improving diagnostic accuracy. Here, we introduce an extension to OCT TCE that includes a fluorescence (FL) imaging channel for attaining complementary, co-registered molecular contrast. We present the development of an OCT-FL TCE capsule and a portable, plug-and-play OCT-FL imaging system. The technology is validated in phantom experiments and feasibility is demonstrated in a methylene blue (MB)-stained swine esophageal injury model, ex vivo and in vivo.


Multi-omics data reveals aberrant gut microbiota-host glycerophospholipid metabolism in association with neuroinflammation in APP/PS1 mice.
Numerous studies have described the notable impact of gut microbiota on the brain in Alzheimer's disease (AD) via the gut - brain axis. However, the molecular mechanisms underlying the involvement of gut microbiota in the development of AD are limited. This study aimed to explore the potential mechanisms of gut microbiota in AD by integrating multi-omics data. In this study, APP/PS1 and WT mice at nine months of age were used as study mouse model. Cognitive function was assessed using the Morris water maze test. The levels of Aβ plaque and neuroinflammation in the brain were detected using immunofluorescence and PET/CT. In addition, we not only used 16S rRNA gene sequencing and metabolomics to explore the variation characteristics of gut microbiota and serum metabolism abundance, but also combined spatial metabolomics and transcriptomics to explore the change in the brain and identify their potential correlation. APP/PS1 mice showed significant cognitive impairment and amyloid-β deposits in the brain. The abundance of gut microbiota was significantly changed in APP/PS1 mice, including decreased Desulfoviobrio, Enterococcus, Turicibacter, and Ruminococcus and increased Pseudomonas. The integration of serum untargeted metabolomics and brain spatial metabolomics showed that glycerophospholipid metabolism was a common alteration pathway in APP/PS1 mice. Significant proliferation and activation of astrocyte and microglia were observed in APP/PS1 mice, accompanied by alterations in immune pathways. Integration analysis and fecal microbiota transplantation (FMT) intervention revealed potential association of gut microbiota, host glycerophospholipid metabolism, and neuroinflammation levels in APP/PS1 mice.


Multiple environmental controls on phytoplankton growth strategies determine adaptive responses of the N : P ratio.
The controls on the 'Redfield' N : P stoichiometry of marine phytoplankton and hence the N : P ratio of the deep ocean remain incompletely understood. Here, we use a model for phytoplankton ecophysiology and growth, based on functional traits and resource-allocation trade-offs, to show how environmental filtering, biotic interactions, and element cycling in a global ecosystem model determine phytoplankton biogeography, growth strategies and macromolecular composition. Emergent growth strategies capture major observed patterns in marine biomes. Using a new synthesis of experimental RNA and protein measurements to constrain per-ribosome translation rates, we determine a spatially variable lower limit on adaptive rRNA:protein allocation and hence on the relationship between the largest cellular P and N pools. Comparison with the lowest observed phytoplankton N : P ratios and N : P export fluxes in the Southern Ocean suggests that additional contributions from phospholipid and phosphorus storage compounds play a fundamental role in determining the marine biogeochemical cycling of these elements.


[Stabilization of roseofungin by antioxidants].
Stabilization of roseofungin, a pentaene antibiotic, by antioxidants of different chemical structure was studied. The highest inhibitory effect was observed with the use of antioxidants X-1 (a partially hydrated oxyquinoline) and P-1 (a polyaromatic nitrogen containing compound). The results of the study correlated with the data on nystatin and levorin. The stabilizing effect markedly depended on the temperature: with increasing of the temperature the antioxidant effect decreased. The spectrophotometric study revealed that the decrease of the biological activity of the antibiotic in the presence of the antioxidants was more pronounced than the change of its structure.


A Putative Lignin Copper Oxidase from Trichoderma reesei.
The ability of Trichoderma reesei, a fungus widely used for the commercial production of hemicellulases and cellulases, to grow and modify technical soda lignin was investigated. By quantifying fungal genomic DNA, T. reesei showed growth and sporulation in solid and liquid cultures containing lignin alone. The analysis of released soluble lignin and residual insoluble lignin was indicative of enzymatic oxidative conversion of phenolic lignin side chains and the modification of lignin structure by cleaving the β-O-4 linkages. The results also showed that polymerization reactions were taking place. A proteomic analysis conducted to investigate secreted proteins at days 3, 7, and 14 of growth revealed the presence of five auxiliary activity (AA) enzymes in the secretome: AA6, AA9, two AA3 enzymes), and the only copper radical oxidase encoded in the genome of T. reesei. This enzyme was heterologously produced and characterized, and its activity on lignin-derived molecules was investigated. Phylogenetic characterization demonstrated that this enzyme belonged to the AA5_1 family, which includes characterized glyoxal oxidases. However, the enzyme displayed overlapping physicochemical and catalytic properties across the AA5 family. The enzyme was remarkably stable at high pH and oxidized both, alcohols and aldehydes with preference to the alcohol group. It was also active on lignin-derived phenolic molecules as well as simple carbohydrates. HPSEC and LC-MS analyses on the reactions of the produced protein on lignin dimers (SS ββ, SS βO4 and GG β5) uncovered the polymerizing activity of this enzyme, which was accordingly named lignin copper oxidase (TrLOx). Polymers of up 10 units were formed by hydroxy group oxidation and radical formation. The activations of lignin molecules by TrLOx along with the co-secretion of this enzyme with reductases and FAD flavoproteins oxidoreductases during growth on lignin suggest a synergistic mechanism for lignin breakdown.


Applications of a nanocomposite-inspired in-situ broadband ultrasonic sensor to acousto-ultrasonics-based passive and active structural health monitoring.
A novel nanocomposite-inspired in-situ broadband ultrasonic sensor previously developed, with carbon black as the nanofiller and polyvinylidene fluoride as the matrix, was networked for acousto-ultrasonic wave-based passive and active structural health monitoring (SHM). Being lightweight and small, this kind of sensor was proven to be capable of perceiving strain perturbation in virtue of the tunneling effect in the formed nanofiller conductive network when acousto-ultrasonic waves traverse the sensor. Proof-of-concept validation was implemented, to examine the sensor performance in responding to acousto-ultrasonic waves in a broad frequency regime: from acoustic emission (AE) of lower frequencies to guided ultrasonic waves (GUWs) of higher frequencies. Results have demonstrated the high fidelity, ultrafast response and high sensitivity of the sensor to acousto-ultrasonic waves up to 400kHz yet with an ultra-low magnitude (of the order of micro-strain). The sensor is proven to possess sensitivity and accuracy comparable with commercial piezoelectric ultrasonic transducers, whereas with greater flexibility in accommodating curved structural surfaces. Application paradigms of using the sensor for damage evaluation have spotlighted the capability of the sensor in compromising "sensing cost" with "sensing effectiveness" for passive AE- or active GUW-based SHM.


DFAST: a flexible prokaryotic genome annotation pipeline for faster genome publication.
We developed a prokaryotic genome annotation pipeline, DFAST, that also supports genome submission to public sequence databases. DFAST was originally started as an on-line annotation server, and to date, over 7000 jobs have been processed since its first launch in 2016. Here, we present a newly implemented background annotation engine for DFAST, which is also available as a standalone command-line program. The new engine can annotate a typical-sized bacterial genome within 10 min, with rich information such as pseudogenes, translation exceptions and orthologous gene assignment between given reference genomes. In addition, the modular framework of DFAST allows users to customize the annotation workflow easily and will also facilitate extensions for new functions and incorporation of new tools in the future. The software is implemented in Python 3 and runs in both Python 2.7 and 3.4-on Macintosh and Linux systems. It is freely available at https://github.com/nigyta/dfast_core/under the GPLv3 license with external binaries bundled in the software distribution. An on-line version is also available at https://dfast.nig.ac.jp/. email@example.com. Supplementary data are available at Bioinformatics online.


MAD2 depletion triggers premature cellular senescence in human primary fibroblasts by activating a p53 pathway preventing aneuploid cells propagation.
The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures faithful chromosome segregation during mitosis and its failure can result in aneuploidy. Previously, it was suggested that reduction of the MAD2 gene, encoding a major component of the SAC, induced aneuploidy in human tumor cells. However, tumor cell lines contain multiple mutations that might affect or exacerbate the cellular response to Mad2 depletion. Thus, the scenario resulting by Mad2 depletion in primary human cells could be different and more complex that the one depicted so far. We used primary human fibroblasts (IMR90) and epithelial breast cells (MCF10A) to gain further insight on the effects of genomic instability caused by transient Mad2 depletion. To this aim we depleted Mad2 by RNAi to a level shown by Mad2 haplo-insufficient cells and found that induced aneuploidy caused premature cellular senescence in IMR90 cells. IMR90 cells showed typical features of senescent cells, like senescence-associated (SA) beta galactosidase expression, including up-regulation of p53 and p14ARF proteins and of p21(waf1) as well, but not of p16(INK4A) cyclin-dependent kinase (Cdk) inhibitor. In contrast, after MAD2 post-transcriptional silencing MCF10A cells in which the INK4A/ARF locus is deleted, showed both aneuploidy and a small increase of p53 and p21(waf1) proteins, but not premature cellular senescence. Finally, our results provides an explanation of how a p53 controlled pathway, involving initially p21(waf1) and then p14ARF, could minimize the occurrence of genomic alterations derived from chromosome instability induced by low amounts of MAD2 protein.


Tuning Plant Promoters Using a Simple Split Luciferase Method to Assess Transcription Factor-DNA Interactions.
Sequence features, including the affinity of binding motifs for their cognate transcription factors, are important contributors to promoter behavior. The ability to predictably recode affinity enables the development of synthetic promoters with varying levels of response to known cellular signals. Here we describe a luminescence-based microplate assay for comparing the interactions of transcription factors with short DNA probes. We then demonstrate how these data can be used to design synthetic plant promoters of varying strengths that respond to the same transcription factor.


Cognitive Impairment in Multiple Sclerosis: Clinical Manifestation, Neuroimaging Correlates, and Treatment.
Cognitive impairment is found in up to 70% of patients with multiple sclerosis (MS). Once thought of as a variant of subcortical dementia with a characteristic set of deficits, we now know that MS-related cognitive impairment can have many faces. This conceptual change in neuropsychology is embedded in a paradigm shift in the neuroscientific understanding of MS over the past 25 years: Partly based on modern neuroimaging techniques, the classical view of MS as an inflammatory demyelinating disease affecting the white matter of the central nervous system has been extended. In particular, many studies have shown that the MS pathology also includes neurodegeneration, and that gray matter structures such as the cerebral cortex can also show focal lesions, atrophy, or both. The authors present an updated summary of the clinical manifestation and neuroimaging correlates of cognitive impairment in MS, and discuss the relatively few treatment options available to date.


Atopy and Sensitization to Allergens Known to Cause Systemic Contact Dermatitis.
Atopic dermatitis (AD) associated with respiratory atopy may represent a form of systemic contact dermatitis (SCD), whereby AD flares after ingestion or inhalation of allergens. The aim of the study was to compare the prevalence of positive patch tests to allergens known to cause SCD in AD patients with and without respiratory atopy. This is a retrospective study of patients with AD patch tested to 23 allergens known to cause SCD. Positive patch tests were compared between AD patients with and without respiratory atopy, stratified by age and wet or dry work occupation. Children and adolescents, but not adults, with AD and respiratory atopy were more likely than age-matched AD patients without respiratory atopy to have positive patch tests to these allergens (odds ratio, 2.33; 95% confidence interval, 1.13-4.79). Moreover, AD patients with respiratory atopy and engaging in wet work, but not dry work, occupations were more likely than AD patients without respiratory atopy to have positive patch tests to allergens known to cause SCD (odds ratio, 1.47; 95% confidence interval, 1.05-2.06). Thus, respiratory atopy and wet work are associated with sensitization to allergens known to cause SCD in patients with AD, and patch testing may be valuable in identifying systemic triggers of dermatitis in these patients.


Radiogenomic-Based Survival Risk Stratification of Tumor Habitat on Gd-T1w MRI Is Associated with Biological Processes in Glioblastoma.
To (i) create a survival risk score using radiomic features from the tumor habitat on routine MRI to predict progression-free survival (PFS) in glioblastoma and (ii) obtain a biological basis for these prognostic radiomic features, by studying their radiogenomic associations with molecular signaling pathways. Two hundred three patients with pretreatment Gd-T1w, T2w, T2w-FLAIR MRI were obtained from 3 cohorts: The Cancer Imaging Archive (TCIA; n = 130), Ivy GAP (n = 32), and Cleveland Clinic (n = 41). Gene-expression profiles of corresponding patients were obtained for TCIA cohort. For every study, following expert segmentation of tumor subcompartments (necrotic core, enhancing tumor, peritumoral edema), 936 3D radiomic features were extracted from each subcompartment across all MRI protocols. Using Cox regression model, radiomic risk score (RRS) was developed for every protocol to predict PFS on the training cohort (n = 130) and evaluated on the holdout cohort (n = 73). Further, Gene Ontology and single-sample gene set enrichment analysis were used to identify specific molecular signaling pathway networks associated with RRS features. Twenty-five radiomic features from the tumor habitat yielded the RRS. A combination of RRS with clinical (age and gender) and molecular features (MGMT and IDH status) resulted in a concordance index of 0.81 (P < 0.0001) on training and 0.84 (P = 0.03) on the test set. Radiogenomic analysis revealed associations of RRS features with signaling pathways for cell differentiation, cell adhesion, and angiogenesis, which contribute to chemoresistance in GBM. Our findings suggest that prognostic radiomic features from routine Gd-T1w MRI may also be significantly associated with key biological processes that affect response to chemotherapy in GBM.


The effect of β-cyclocitral treatment on the carotenoid content of transgenic Marsh grapefruit (Citrus paradisi Macf.) suspension-cultured cells.
This work reports the development of suspension culture system of transgenic Marsh grapefruit (Citrus paradisi Macf., Rutaceae) callus overexpressing bacterial phytoene synthase; and the use of this suspension culture to investigate the effects of β-cyclocitral on carotenoid content and composition. At a β-cyclocitral concentration of 0.5 mM and after ten days cultivation, analysis of the carotenoids showed a significant increase in the content of β-, α-carotene, and phytoene predominantly. The maximal increase in total provitamin A carotenoids content following β-cyclocitral application was ~2-fold higher than the control, reaching 245.8 μg/g DW. The trend for increased transcript levels of biosynthetic genes PSY and ZDS correlated with the enhancement of the content of these carotenes following β-cyclocitral treatment and GC-MS based metabolite profiling showed significant changes of metabolite levels across intermediary metabolism. These findings suggest that β-cyclocitral can act as a chemical elicitor, to enhance the formation of carotenes in citrus suspension-cultured cells (SCC), which could be utilized in studying the regulation of carotenoid biosynthesis and biotechnological application to the renewable production of nutritional carotenoids.


Integrating DNA methylation and gene expression data in a single gene network using the iNETgrate package.
Analyzing different omics data types independently is often too restrictive to allow for detection of subtle, but consistent, variations that are coherently supported based upon different assays. Integrating multi-omics data in one model can increase statistical power. However, designing such a model is challenging because different omics are measured at different levels. We developed the iNETgrate package ( https://bioconductor.org/packages/iNETgrate/ ) that efficiently integrates transcriptome and DNA methylation data in a single gene network. Applying iNETgrate on five independent datasets improved prognostication compared to common clinical gold standards and a patient similarity network approach.


[Use of laryngeal masks: variants of hemodynamic response to various methods of anesthesia].
Choice of induction agent is the decisive factor in utilization of a laryngeal mask (LM) under spontaneous respiration, as insufficient relaxation and unwanted laryngopharyngeal reflexes deteriorate the efficiency of LM functioning during operation and anesthesia. 132 anesthesias with LM were performed, 86 of these for laparoscopic cholecystectomies (LCE) (group 1) and 42 for saphenectomies (group 2). Control group consisted of 60 patients anesthetized for LCE by means of an endotracheal tube (ETT). Combined induction intravenous anesthesia with ketamine, barbiturates, and diprivan was used in group 1 and controls. In group 2 induction anesthesia was based on ketamine and diprivan. Group 1 and control group were matched for demographic characteristics and concomitant diseases. All groups were divided into subgroups, depending on anesthesia. In all groups pipecuronium bromide (0.06 mg/kg) was administered before insertion of EET or LM after injection of induction agents. In total myoplegia, installation of LM was simple, involving no untoward laryngo-pharyngeal reflexes. LM was correctly installed from the first attempt in 98.7% cases. In combined intravenous ketamine-based anesthesia, pressor reaction to LM installation consisted in an increase of heart rate by 5.6% (1K group--control), by 10.2% in 2K group (fractionated ketamine (1.5-2 mg/kg) + diazepam (0.1 mg/kg) and fentanyl (2.6 micrograms/kg); after barbiturates heart rate increased by 6.7% in 1B group (control), after diprivan it did not increase in 1D group (control), while the mean arterial pressure in 2D group (fractionated diprivan (1 mg/kg) + ketamine (25-50 mg), diazepam (0.08 mg/kg) and fentanyl (2.6 micrograms/kg) increased by 10% and heart rate by 6.6%. Reaction to tracheal intubation was pronounced in all types of induction anesthesia. No regurgitation or aspiration was observed in any case. Hence, use of total myoplegia in installation of LM allows the use of various modern anesthetics in optimal doses and notably simplifies the utilization of LM.


[Different insulin treatment regimens in patient with diabetes mellitus type 1: effects on quality of life].
Despite of contemporary diabetes mellitus (DM) treatment, one half of patients do not achieve an optimal metabolic control. Considering great psychological burden of diabetic patients, the purpose of this study was to assess the effect of different insulin treatment regimens, glycemic control and the presence of vascular complications on self-reported well-being and quality of life (QoL) of subjects with type 1 DM. The patients with type 1 DM (n=122) recruited from the outpatient Diabetes Endocrinology Clinic of Zvezdara University Medical Center were divided into 4 groups according to the specific treatment regimen: 26 were on continuous subcutaneous insulin infusion (CSII), 30 on conventional insulin therapy, 33 on multiple daily injections (MDI) with human insulins, and 33 on MDI with insulin analogues. QoL was assesed by self-reported well-being with the following questionnaires: WHO-5 item well being index (WHO-5), 36 item short form (SF-36) survey, and insulin treatment appraisal scale (ITAS). Objective metabolic control was assessed by glycosylated hemoglobin (HbA1c), lipid levels and the presence of vascular complications. Statistical analyses used in this cross-sectional study included: descriptive statistics, student's t-test, chi-sqare test, contingency tables, ANOVA and correlation methods. The patients on CSII had significantly better metabolic control than all other treatment groups, especially when compared to the one on conventional therapy (CSII HbA1c 7.07 +/- 1.48% vs. conventional therapy, HbA1c 10.04 +/- 1.44; p = 0.000). No significant difference in glycemic control was observed between patients on MDI with human insulins and insulin analogues. Good glycemic control significantly influenced the reported QoL. The patients with retinopathy and nephropathy reported significantly lower physical well-being, and the patients with polyneuropathy and cardiovascular complications reported also lower psychological well being. Insulin treatment regiment selection affects not only objective metabolic control, but also QoL.


A study of variables associated with under-attendance at a psychiatric day centre.
This study examines the problems of client under-attendance at a psychiatric day centre. Demographic, psychiatric, psychometric and Semantic Differential measures were used. Several of these measures were significantly associated with under-attendance. Various methods are suggested for the identification of and reduction of the level of under-attendance.


Escape of intracellular Shigella from autophagy.
The degradation of undesirable cellular components or organelles, including invading microbes, by autophagy is crucial for cell survival. Here, Shigella, an invasive bacteria, was found to be able to escape autophagy by secreting IcsB by means of the type III secretion system. Mutant bacteria lacking IcsB were trapped by autophagy during multiplication within the host cells. IcsB did not directly inhibit autophagy. Rather, Shigella VirG, a protein required for intracellular actin-based motility, induced autophagy by binding to the autophagy protein, Atg5. In nonmutant Shigella, this binding is competitively inhibited by IcsB binding to VirG.


Enzymes involved in organellar DNA replication in photosynthetic eukaryotes.
Plastids and mitochondria possess their own genomes. Although the replication mechanisms of these organellar genomes remain unclear in photosynthetic eukaryotes, several organelle-localized enzymes related to genome replication, including DNA polymerase, DNA primase, DNA helicase, DNA topoisomerase, single-stranded DNA maintenance protein, DNA ligase, primer removal enzyme, and several DNA recombination-related enzymes, have been identified. In the reference Eudicot plant Arabidopsis thaliana, the replication-related enzymes of plastids and mitochondria are similar because many of them are dual targeted to both organelles, whereas in the red alga Cyanidioschyzon merolae, plastids and mitochondria contain different replication machinery components. The enzymes involved in organellar genome replication in green plants and red algae were derived from different origins, including proteobacterial, cyanobacterial, and eukaryotic lineages. In the present review, we summarize the available data for enzymes related to organellar genome replication in green plants and red algae. In addition, based on the type and distribution of replication enzymes in photosynthetic eukaryotes, we discuss the transitional history of replication enzymes in the organelles of plants.


Drug treatments in the secondary prevention of ischaemic stroke.
Stroke is an important cause of death and disability. However, about two thirds of cerebrovascular events are initially minor. They carry a high risk of potentially severe recurrent events, but they also offer an opportunity for secondary prevention to avoid such recurrences. As most recurrent events occur within a short time after the initial presentation, secondary prevention has to be started as soon as possible. Dramatic risk reduction can be achieved with well-established drugs if used in a timely manner. A standard secondary preventive regimen will address multiple vascular risk factors and will usually consist of an antiplatelet agent, a lipid lowering drug, and an antihypertensive agent. Depending on the risk factor profile of each patient, this will have to be adjusted individually, for example, taking into account the presence of cardioembolism or of stenotic disease of the brain-supplying arteries. In recent years, the approach to treating these risk factors has evolved. In addition to absolute blood pressure, blood pressure variability has emerged as an important contributing factor to stroke risk, which is affected differently by different antihypertensive agents. New oral anticoagulants reduce the risk of cerebral haemorrhage and the need for regular blood checks. The best antiplatelet regimen for stroke prevention is still uncertain, and treatment of dyslipidaemia may change if trials with cholesteryl ester transfer protein (CETP) inhibitors, which increase levels of HDL-cholesterol, are successful. This article reviews the current evidence for drug treatments in the secondary prevention of ischaemic stroke.


A multi-center study evaluating the correlation between meibomian gland dysfunction and depressive symptoms.
Increased prevalence of depression has been found in patients with meibomian gland dysfunction (MGD); however, specific conditions of patients suffered from MGD and depression remains unclear. Our aim was to investigate the prevalence of depression in patients with MGD and analyze the risk factors of depression in MGD patients. In this multi-center cross-sectional study, we enrolled 830 MGD patients and 114 normal controls. Demographic information was collected by designed questionnaires about lifestyle habits, systemic and ocular medical history. Ophthalmic examinations were performed in a formal order. Depression symptoms were evaluated with a questionnaire survey using a modified self-rating depression scale (M-SDS). The correlations between the M-SDS score and demographic and clinical information were analyzed. The prevalence of hyperlipidemia and obstructive sleep apnea-hypopnea syndrome (OSAHS) was higher in the MGD group. There were 78 cases (9.4%) with depressive symptoms in the MGD group whereas there were 4 cases (3.5%) in controls. Similarly, M-SDS was higher in the MGD group. The characteristics of depression in the MGD group included: crying spells, sleep disturbance and depressed appetite. Some systemic diseases (OSAHS, allergy, skin disease) and the use of contact lenses was correlated with an increased risk of depressive symptoms in MGD patients.


Purple Sweet Potato Leaf Extract Induces Apoptosis and Reduces Inflammatory Adipokine Expression in 3T3-L1 Differentiated Adipocytes.
Background. Purple sweet potato leaves (PSPL) are widely grown and are considered a healthy vegetable in Taiwan. PSPL contain a high content of flavonoids, and the boiling water-extracted PSPL (PSPLE) is believed to prevent metabolic syndrome. However, its efficacy has not yet been verified. Therefore, we investigated the effect of PSPLE on adipocytes. Methods. The differentiated 3T3-L1 cells used in this study were derived from preadipocytes that were differentiated into adipocytes using an adipogenic agent (insulin, dexamethasone, and 3-isobutyl-1-methylxanthine); approximately 90% of the cells were differentiated using this method. Results. Treating the differentiated 3T3-L1 cells with PSPLE caused a dose-dependent decrease in the number of adipocytes rather than preadipocytes. In addition, treatment with PSPLE resulted in apoptosis of the differentiated 3T3-L1 cells as determined by DAPI analysis and flow cytometry. PSPLE also increased the expression of cleaved caspase-3 and poly ADP-ribose polymerase (PARP). Furthermore, PSPLE induced downregulation of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) gene expression in the differentiated 3T3-L1 cells. Conclusions. These results suggest that PSPLE not only induced apoptosis but also downregulated inflammation-associated genes in the differentiated 3T3-L1 cells.


Blackberry consumption protects against e-cigarette-induced vascular oxidative stress in mice.
Electronic cigarettes (e-cigarettes) have gained popularity; however, evidence for their safety with chronic use is lacking. Acute e-cigarette exposure induces systemic oxidative stress in users and contributes to vascular endothelial dysfunction through reduction in nitric oxide (NO). Polyphenols, abundant in blackberries (BL), mitigate cardiovascular damage. We aimed to determine whether BL was protective against e-cigarette-induced cardiopulmonary detriments. Mice were fed a diet supplemented with or without 5% freeze-dried BL (w/w) for 16 weeks. E-cigarette exposure (1 h, 5 days per week) began at week 4. Additionally, human microvascular endothelial cells (HMVECs) were treated with BL polyphenol extract (200 μg mL-1) and e-cigarette condensate (0.5% v/v). Twelve weeks of e-cigarette exposure induced multi-organ oxidative stress. E-cigarette exposure increased expression of pro-oxidant enzymes in the endothelium resulting in increased superoxide production diminishing NO bioavailability. Additionally, e-cigarettes reduced the phosphorylation of endothelial NO synthase, contributing to decreases in NO. Mice supplemented with BL were protected against decreases in NO and BL pre-treatment in vitro reduced superoxide production. However, BL was not able to attenuate oxidative stress responses in the heart or lungs. These studies demonstrate the contribution of e-cigarettes to vascular pathologies through an increase in superoxide-producing enzymes and the ability of BL polyphenols to mitigate these deleterious effects in the vasculature. Further studies should explore the role of polyphenol-rich foods in protecting against cardiopulmonary conditions induced by chronic e-cigarette use and explore their use in the recovery period post-e-cigarette cessation to properly align with current public health messaging.


Current and potential imaging applications of ferumoxytol for magnetic resonance imaging.
Contrast-enhanced magnetic resonance imaging is a commonly used diagnostic tool. Compared with standard gadolinium-based contrast agents, ferumoxytol (Feraheme, AMAG Pharmaceuticals, Waltham, MA), used as an alternative contrast medium, is feasible in patients with impaired renal function. Other attractive imaging features of i.v. ferumoxytol include a prolonged blood pool phase and delayed intracellular uptake. With its unique pharmacologic, metabolic, and imaging properties, ferumoxytol may play a crucial role in future magnetic resonance imaging of the central nervous system, various organs outside the central nervous system, and the cardiovascular system. Preclinical and clinical studies have demonstrated the overall safety and effectiveness of this novel contrast agent, with rarely occurring anaphylactoid reactions. The purpose of this review is to describe the general and organ-specific properties of ferumoxytol, as well as the advantages and potential pitfalls associated with its use in magnetic resonance imaging. To more fully demonstrate the applications of ferumoxytol throughout the body, an imaging atlas was created and is available online as supplementary material.


Pressure effects on the ADH-induced initiation of water flow in toad bladder.
Earlier studies employed colchicine to demonstrate the need for microtubules in the ADH-induced initiation of increased water permeability in toad bladder. We have used colchicine and hydrostatic pressure together to determine whether formed or growing microtubules are required for initiation of the ADH response in Bufo marinus. When ADH and 8,000 psi were administered simultaneously, the ADH-induced increase in water flow was inhibited while under pressure by 107 +/- 7% (n = 6). Application of 8,000 psi for 10 min before ADH administration resulted in an increased initiation of the ADH osmotic response over the non-pressure-treated control (average acceleration, rate of water flow increase during first 3 min after ADH stimulation, 1.25 +/- 0.27 vs. 0.43 +/- 0.12 mg X cm-2 X min-2, n = 6). In addition, the inhibition of the ADH response brought about by colchicine incubation was overcome with pretreatment of the colchicine-incubated bladders with 8,000 psi for 10 min (average 3-min acceleration, 0.18 +/- 0.04 vs. 1.13 +/- 0.06 mg X cm-2 X min-2, respectively). Repeating the experiments with dibutyryl cAMP gave similar results. We interpret these data as suggesting that growing microtubules are required for initiation. The proposed model is as follows. Pressure removes colchicine inhibition by introducing, through disassembly of formed microtubules, more colchicine-free tubulin subunits. These subunits are then available following decompression to reassemble when the tissue is challenged with hormone.


Tc-99m DTPA aerosol lung clearance test in systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) can affect every organ. Involvement of the lung in systemic lupus erythematosus may be a significant cause of morbidity and mortality. To evaluate alveolar epithelial damage in SLE, we studied lung epithelial permeability by measuring the clearance of inhaled Tc-99m DTPA aerosol. Twenty-three SLE patients without clinical pulmonary manifestations were studied. Of the 23 patients, 16 had normal clearance rates of Tc-99m DTPA (T1/2 > 60mins) and normal chest X-ray findings. A mild increase in the clearance rate was found in 5 cases (T1/2 between 40-60 min) and a significant increase in clearance rate in the remaining 2 (T1/2 < 40mins). Totally, 30% (7/23) of SLE patients have abnormal lung permeability. Of the 7 patients with abnormal clearance of Tc-99m DTPA, 3 had abnormal findings in chest X-rays and four had normal chest X-rays. Our study suggests that the clearance of Tc-99m DTPA aerosol may be a useful complementary study to assess pulmonary involvement in SLE.


[Analysis of implant position after envelope technique in "white" cataract surgery].
Capsulopunture in cataract surgery doesn't allow insertion of intraocular lens in the capsular bag. Envelope technique is a easy way for this insertion. The goal of our study was to analyse the position of the intraocular lens in the capsular bag, after performing envelope technique in eyes suffering from "white" cataract. We realised on 25 eyes with "white" cataract, envelope technique. GALAND method (one superior linear capsulotomy) was realised in 15 cases and double linear capsulotomy (superior and inferior), was realised in 10 cases. The mean age of our patients was 55 years with two extremes (17 and 70 years). The mean visual acuity was 14/20. After verification, in 91.636% eyes, intraocular lenses were in the capsular bag. In 2 cases, we found a decentration. Envelope technique can lessen risks of capsular zip. It protects endothelium cells and is helpful for the insertion of the intraocular lens in the capsular bag. Envelope technique is well adapted to the surgery of "White" Cataract.


Tuning the electronic and phosphorescence properties of blue-emitting iridium(III) complexes through different cyclometalated ligand substituents: a theoretical investigation.
The geometric and electronic structures, phosphorescence properties and the organic light-emitting diode (OLED) performance of a series of Ir(III) complexes based on bis[(4,6-di-fluorophenyl)-pyridinato-N,C2']picolinate (FIrpic) were investigated by using density functional theory/time-dependent density functional theory (DFT/TD-DFT), including Ir(III)bis[2-(2,4-difluorophenyl)-4-(tert-butyl)pyridinato-N,C2']picolinate (1a), Ir(III)bis[2-(2,4-difluorophenyl)-4-(n-heptyl)pyridinato-N,C2']picolinate (2a), Ir(III)bis[2-(2,4-difluorophenyl)-4-(3-ethylheptyl)pyridinato-N,C2']picolinate (3a), Ir(III)bis[2-(2,4-difluorophenyl)-4-(2,4,6-trimethylphenyl)pyridinato-N,C2']picolinate (5a), and Ir(III)bis[2-(2,4-difluoro-3-(2,4,6-trimethylphenyl)phenyl)-pyridinato-N,C2'] picolinate (5b). To explore the influence of the substituted positions on the optical and electronic properties of the Ir(III) complexes, seven other new complexes were designed by introducing the substituted groups on the difluorophenyl rings or pyridine rings. After introducing the phenyl substituted groups on the pyridine or difluorophenyl rings of cyclometalated ligands, the HOMO-LUMO energy gap is decreased. Thus, the absorption spectra of 4a and 4b undergo a red-shifting, especially for 4a, and have a stronger absorption strength that will be beneficial to improving their quantum yields, which is proved by the further evaluation of the radiative (k(r)) and non-radiative (k(nr)) rate constants. The combinations of a larger (3)MLCT-(3)MC energy gap, smaller ΔE(S1-T1), and higher contribution of (3)MLCT in the emission process result in the higher quantum yields for complexes 4a and 4b. Besides, the designed complexes 4a and 4b are considered to be potential candidates as blue-emitting materials with better equilibrium between the hole transport (λ(hole)) and electron transport (λ(electron)).


Two new Haemoproteus species (Haemosporida: Haemoproteidae) from columbiform birds.
Here we describe Haemoproteus (Haemoproteus) multivolutinus n. sp. from a tambourine dove (Turtur timpanistria) of Uganda and Haemoproteus (Haemoproteus) paramultipigmentatus n. sp. (Haemosporida, Haemoproteidae) from the Socorro common ground dove (Columbina passerina socorroensis) of Socorro Island, Mexico. These parasites are described based on the morphology of their blood stages and segments of the mitochondrial cytochrome b gene that can be used for molecular identification and diagnosis of these species. Gametocytes of H. multivolutinus possess rod-like pigment granules and are evenly packed with volutin, which masks pigment granules and darkly stains both macro- and microgametocytes in the early stages of their development. Based on these 2 characters, H. multivolutinus can be readily distinguished from other species of hemoproteids parasitizing columbiform (Columbiformes) birds. Haemoproteus paramultipigmentatus resembles Haemoproteus multipigmentatus; it can be distinguished from the latter parasite primarily due to the broadly ovoid shape of its young gametocytes and significantly fewer pigment granules in its fully developed gametocytes. We provide illustrations of blood stages of the new species, and phylogenetic analyses identify DNA lineages closely related to these parasites. Cytochrome b lineages of Haemoproteus multivolutinus and H. paramultipigmentatus cluster with hippoboscid-transmitted lineages of hemoproteids; thus these parasites likely belong to the subgenus Haemoproteus. We emphasize the importance of using cytochrome b sequences in conjunction with thorough microscopic descriptions to facilitate future identification of these and other avian hemosporidian species.


Analysis of the Bacillus subtilis tyrS gene: conservation of a regulatory sequence in multiple tRNA synthetase genes.
The Bacillus subtilis tyrS gene, which encodes tyrosyl-tRNA synthetase (TyrTS), was isolated, and its nucleotide sequence was determined. The cloned gene was shown to complement an Escherichia coli tyrS (Ts) mutant. The predicted amino acid sequence exhibited 70% identity to that of Bacillus stearothermophilus TyrTS and 55% identity to that of E. coli TyrTS, while identity to a second cryptic B. subtilis TyrTS gene, designated tyrZ, was only 27%. Primer extension analysis indicated that tyrS transcription initiated at a vegetative promoter sequence located 300 nucleotides upstream of the AUG start codon. The mRNA leader region was found to contain an inverted repeat sequence resembling a transcriptional terminator. Expression of a transcriptional tyrS-lacZ fusion was found to be induced by starvation for tyrosine in a tyrosine auxotroph (tyrA1). Transcription initiation was unaffected by tyrosine starvation. Deletion of the terminator region in a tyrS-lacZ fusion resulted in high-level constitutive expression. Immediately preceding the putative terminator was sequence element found to be conserved in the upstream region of a number of Bacillus tRNA synthetase genes as well as in the ilv-leu biosynthetic operon; mutation of this element in tyrS resulted in low-level uninducible expression. The conservation of this sequence element suggests that aminoacyl-tRNA synthetase genes and the ilv-leu operon may be regulated by a common mechanism in Bacillus spp.


Topology of RbsC, the membrane component of the Escherichia coli ribose transporter.
The topology of RbsC, the membrane component of the ribose transporter in Escherichia coli, has been determined by using 34 single-cysteine mutants and a modified fluorescence labeling technique designated multiplex labeling. This technique gives topology, expression, and localization information for a membrane protein from a single batch of bacterial cells. The results indicate that RbsC contains 10 transmembrane-spanning helices, with the N and C termini being in the cytosol. This topology matches predictions from the latest prediction programs and the topology of the similar, recently crystallized membrane protein BtuC.


Mutants of staphylococcal toxic shock syndrome toxin 1: mitogenicity and recognition by a neutralizing monoclonal antibody.
Toxic shock syndrome toxin 1 (TSST-1), a 22-kilodalton protein made by strains of Staphylococcus aureus harboring the chromosomal toxin gene, may elicit toxic shock syndrome in humans. In vitro, TSST-1 induces T cells to proliferate and macrophages to secrete interleukin-1. To conduct a structure-function analysis, point mutations on the TSST-1 gene were generated by site-directed mutagenesis to identify amino acids critical for activity of the toxin. Specific tyrosine and histidine residues were replaced by alanines. Wild-type and mutant TSST-1 gene constructs were expressed in Escherichia coli, and the products were tested for their mitogenic potential and reactivity with a TSST-1 neutralizing monoclonal antibody (MAb 8-5-7). Four of the mutants were similar to the wild type; i.e., the mutant toxins stimulated murine T cells and reacted with MAb 8-5-7 equally as well as the wild type. Two mutants exhibited a decrease in mitogenic activity, but one of these retained the capacity to bind with MAb 8-5-7 while the other was no longer recognized by the same antibody. One double mutant demonstrated minimal mitogenic activity and did not react in enzyme-linked immunosorbent and immunoblot assays with MAb 8-5-7. The data show that specific residues near the carboxy terminus of TSST-1 are essential for mitogenic activity and in forming the epitope recognized by neutralizing MAb 8-5-7.


Analysis of internet-based patient education materials related to pituitary tumors.
The Internet has become a primary and ubiquitous information source for patient education material (PEM); however, the information provided may not be appropriate for the average patient to comprehend. Various national healthcare organizations have recommended that PEM be written at or below the sixth-grade level. The purpose of this study was to assess the readability of pituitary tumor-related PEMs available on the Internet. Fifty-one PEMs on pituitary tumors were downloaded from professional society and clinical practice websites. Analysis of readability was performed using 4 different readability indices: Flesch-Kincaid Grade Level (FKGL), Flesch Reading Ease Score (FRES), Simple Measure of Gobbledygook (SMOG), and Gunning Frequency Measure of Gobbledygook (Gunning FOG). Scores from the FKGL, SMOG, and Gunning FOG scales correspond to reading grade levels. Therefore, a higher number corresponds to higher difficulty and lower readability. The average grade level of the PEMs according to the readability indices were as follows: FKGL = 11.71 (11th to 12th grades), SMOG = 14.56 (college level), and Gunning FOG = 14.86 (college level). For the FRES, higher scores imply easier readability. The average FRES was 40.19 (fairly difficult-between 10th and 11th grades). These findings suggest that online pituitary tumor-related material may be too difficult for comprehension by the majority of the targeted patient population. Keeping the reading level of PEMs at or below the sixth grade may improve understanding of this disease and its management for pituitary tumor patients.