Basophilic degeneration of skeletal muscle in hypothyroid myopathy. Histochemical and ultrastructural studies.
Morphologic and histochemical studies of basophilic degeneration (BD) of skeletal muscle were performed on a 64-year-old woman with hypothyroid myopathy. The BD had staining characteristics of polysaccharide and similarity to the deposits described in cardiac muscle in five of six muscle groups studied and was absent in the diaphragm. The frequent occurrence of BD at the myotendinous junction suggested the necessity of including this area in muscle biopsy specimens of suspected hypothyroid myopathy. Review of the literature indicated a need for more detailed examination of muscle specimens of these patients to determine the clinical implication and biochemical nature of the BD. To my knowledge, the demonstration of a close association of BD and leptomeres and the presence of crystalline structures within the BD have not been described previously.


Aurora-PLK1 cascades as key signaling modules in the regulation of mitosis.
Mitosis is controlled by reversible protein phosphorylation involving specific kinases and phosphatases. A handful of major mitotic protein kinases, such as the cyclin B-CDK1 complex, the Aurora kinases, and Polo-like kinase 1 (PLK1), cooperatively regulate distinct mitotic processes. Research has identified proteins and mechanisms that integrate these kinases into signaling cascades that guide essential mitotic events. These findings have important implications for our understanding of the mechanisms of mitotic regulation and may advance the development of novel antimitotic drugs. We review collected evidence that in vertebrates, the Aurora kinases serve as catalytic subunits of distinct complexes formed with the four scaffold proteins Bora, CEP192, INCENP, and TPX2, which we deem "core" Aurora cofactors. These complexes and the Aurora-PLK1 cascades organized by Bora, CEP192, and INCENP control crucial aspects of mitosis and all pathways of spindle assembly. We compare the mechanisms of Aurora activation in relation to the different spindle assembly pathways and draw a functional analogy between the CEP192 complex and the chromosomal passenger complex that may reflect the coevolution of centrosomes, kinetochores, and the actomyosin cleavage apparatus. We also analyze the roles and mechanisms of Aurora-PLK1 signaling in the cell and centrosome cycles and in the DNA damage response.


Immune escape mechanisms and immunotherapy of urothelial bladder cancer.
Urothelial bladder cancer (UBC) is a common malignant tumor of the urogenital system with a high rate of recurrence. Due to the sophisticated and largely unexplored mechanisms of tumorigenesis of UBC, the classical therapeutic approaches including transurethral resection and radical cystectomy combined with chemotherapy have remained unchanged for decades. However, with increasingly in-depth understanding of the microenvironment and the composition of tumor-infiltrating lymphocytes of UBC, novel immunotherapeutic strategies have been developed. Bacillus Calmette-Guerin (BCG) therapy, immune checkpoint blockades, adoptive T cell immunotherapy, dendritic cell (DC) vaccines, etc., have all been intensively investigated as immunotherapies for UBC. This review will discuss the recent progress in immune escape mechanisms and immunotherapy of UBC. Based on a comprehensive search of the PubMed and ClinicalTrials.gov database, this review included the literature reporting the immune escape mechanisms of UBC and clinical trials assessing the effect of immunotherapeutic strategies on tumor or immune cells in UBC patients published in English between 1999 and 2020. Immune surveillance, immune balance, and immune escape are the three major processes that occur during UBC tumorigenesis. First, the role of immunosuppressive cells, immunosuppressive molecules, immunosuppressive signaling molecules, and DCs in tumor microenvironment is introduced elaborately in the immune escape mechanisms of UBC section. In addition, recent progress of immunotherapies including BCG, checkpoint inhibitors, cytokines, adoptive T cell immunotherapy, DCs, and macrophages on UBC patients are summarized in detail. Finally, the need to explore the mechanisms, molecular characteristics and immune landscape during UBC tumorigenesis and development of novel and robust immunotherapies for UBC are also proposed and discussed. At present, BCG and immune checkpoint blockades have been approved by the US Food and Drug Administration for the treatment of UBC patients and have achieved encouraging therapeutic results, expanding the traditional chemotherapy and surgery-based treatment for UBC. Immunotherapy has achieved desirable results in the treatment of UBC, which not only improve the overall survival but also reduce the recurrence rate and the occurrence of treatment-related adverse events of UBC patients. In addition, the indicators to predict the effectiveness and novel therapy strategies, such as combination regimen of checkpoint inhibitor with checkpoint inhibitor or chemotherapy, should be further studied.


Mutations in C8ORF37 cause Bardet Biedl syndrome (BBS21).
Bardet Biedl syndrome (BBS) is a multisystem genetically heterogeneous ciliopathy that most commonly leads to obesity, photoreceptor degeneration, digit anomalies, genito-urinary abnormalities, as well as cognitive impairment with autism, among other features. Sequencing of a DNA sample from a 17-year-old female affected with BBS did not identify any mutation in the known BBS genes. Whole-genome sequencing identified a novel loss-of-function disease-causing homozygous mutation (K102*) in C8ORF37, a gene coding for a cilia protein. The proband was overweight (body mass index 29.1) with a slowly progressive rod-cone dystrophy, a mild learning difficulty, high myopia, three limb post-axial polydactyly, horseshoe kidney, abnormally positioned uterus and elevated liver enzymes. Mutations in C8ORF37 were previously associated with severe autosomal recessive retinal dystrophies (retinitis pigmentosa RP64 and cone-rod dystrophy CORD16) but not BBS. To elucidate the functional role of C8ORF37 in a vertebrate system, we performed gene knockdown in Danio rerio and assessed the cardinal features of BBS and visual function. Knockdown of c8orf37 resulted in impaired visual behavior and BBS-related phenotypes, specifically, defects in the formation of Kupffer's vesicle and delays in retrograde transport. Specificity of these phenotypes to BBS knockdown was shown with rescue experiments. Over-expression of human missense mutations in zebrafish also resulted in impaired visual behavior and BBS-related phenotypes. This is the first functional validation and association of C8ORF37 mutations with the BBS phenotype, which identifies BBS21. The zebrafish studies hereby show that C8ORF37 variants underlie clinically diagnosed BBS-related phenotypes as well as isolated retinal degeneration.


The co-transplantation of bone marrow derived mesenchymal stem cells reduced inflammation in intramuscular islet transplantation.
Although the muscle is one of the preferable transplant sites in islet transplantation, its transplant efficacy is poor. Here we attempted to determine whether an intramuscular co-transplantation of mesenchymal stem cells (MSCs) could improve the outcome. We co-cultured murine islets with MSCs and then analyzed the morphological changes, viability, insulin-releasing function (represented by the stimulation index), and gene expression of the islets. We also transplanted 500 islets intramuscularly with or without 5 × 105 MSCs to diabetic mice and measured their blood glucose level, the glucose changes in an intraperitoneal glucose tolerance test, and the plasma IL-6 level. Inflammation, apoptosis, and neovascularization in the transplantation site were evaluated histologically. The destruction of islets tended to be prevented by co-culture with MSCs. The stimulation index was significantly higher in islets co-cultured with MSCs (1.78 ± 0.59 vs. 7.08 ± 2.53; p = 0.0025). In terms of gene expression, Sult1c2, Gstm1, and Rab37 were significantly upregulated in islets co-cultured with MSCs. Although MSCs were effective in the in vitro assays, they were only partially effective in facilitating intramuscular islet transplantation. Co-transplanted MSCs prevented an early inflammatory reaction from the islets (plasma IL-6; p = 0.0002, neutrophil infiltration; p = 0.016 inflammatory area; p = 0.021), but could not promote neovascularization in the muscle, resulting in the failure of many intramuscular transplanted islets to engraft. In conclusion, co-culturing and co-transplanting MSCs is potentially useful in islet transplantation, especially in terms of anti-inflammation, but further augmentation for an anti-apoptosis effect and neovascularization is necessary.


G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA.
It has been previously shown that the DHX36 gene product, G4R1/RHAU, tightly binds tetramolecular G4-DNA with high affinity and resolves these structures into single strands. Here, we test the ability of G4R1/RHAU to bind and unwind unimolecular G4-DNA. Gel mobility shift assays were used to measure the binding affinity of G4R1/RHAU for unimolecular G4-DNA-formed sequences from the Zic1 gene and the c-Myc promoter. Extremely tight binding produced apparent K(d)'s of 6, 3 and 4 pM for two Zic1 G4-DNAs and a c-Myc G4-DNA, respectively. The low enzyme concentrations required for measuring these K(d)'s limit the precision of their determination to upper boundary estimates. Similar tight binding was not observed in control non-G4 forming DNA sequences or in single-stranded DNA having guanine-rich runs capable of forming tetramolecular G4-DNA. Using a peptide nucleic acid (PNA) trap assay, we show that G4R1/RHAU catalyzes unwinding of unimolecular Zic1 G4-DNA into an unstructured state capable of hybridizing to a complementary PNA. Binding was independent of adenosine triphosphate (ATP), but the PNA trap assay showed that unwinding of G4-DNA was ATP dependent. Competition studies indicated that unimolecular Zic1 and c-Myc G4-DNA structures inhibit G4R1/RHAU-catalyzed resolution of tetramolecular G4-DNA. This report provides evidence that G4R1/RHAU tightly binds and unwinds unimolecular G4-DNA structures.


Spores of Beauveria bassiana and Trichoderma lignorum as a bioinsecticide for the control of Atta cephalotes.
The leafcutter ant (Atta cephalotes) is associated with losses in the agricultural sector, due to its defoliating activity; for its control, biological, mechanical and chemical methods have been developed, the latter associated with adverse effects on human and environmental health. This research validated in the field for the control of the leafcutter ant (A. cephalotes) using a mixture of Beauveria bassiana and Trichoderma lignorum spores. The effectiveness from the combination of spores of B. bassiana and T. lignorum with an initial concentration of 2 × 109 spores/ml, in the following proportions of B. bassiana and T. lignorum, A (1:1), of each fungus. It was evaluated within the university campus, comparing it with two commercial formulations, Mycotrol (B. bassiana) and Mycobac (T. lignorum). Additionally, this formulation was evaluated in 49 nests distributed 16 in 14 locations in Colombia. The formulation application was carried out by direct application, using a pump at a speed of 10 ml/m2. The effectiveness was estimated from the reduction of the flow of ants, evaluating the statistically significant differences using the ANOVA and Tukey-test. Effective control of 90% of the nests was observed in the field phase in 60 days, except in nests with areas > 50 m2 that were located in regions with high rainfall (annual average precipitation above 7000 mm), such as Buenaventura. In this work, it was demonstrated that the combination of B. bassiana and T. lignorum spores represent a viable alternative for the control of the leafcutter ant, in which the effectiveness is related to several factors, including the size of the nest and the rainfall in the area.


Genes and pathways for CO2 fixation in the obligate, chemolithoautotrophic acidophile, Acidithiobacillus ferrooxidans, carbon fixation in A. ferrooxidans.
Acidithiobacillus ferrooxidans is chemolithoautotrophic γ-proteobacterium that thrives at extremely low pH (pH 1-2). Although a substantial amount of information is available regarding CO2 uptake and fixation in a variety of facultative autotrophs, less is known about the processes in obligate autotrophs, especially those living in extremely acidic conditions, prompting the present study. Four gene clusters (termed cbb1-4) in the A. ferrooxidans genome are predicted to encode enzymes and structural proteins involved in carbon assimilation via the Calvin-Benson-Bassham (CBB) cycle including form I of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO, EC 220.127.116.11) and the CO2-concentrating carboxysomes. RT-PCR experiments demonstrated that each gene cluster is a single transcriptional unit and thus is an operon. Operon cbb1 is divergently transcribed from a gene, cbbR, encoding the LysR-type transcriptional regulator CbbR that has been shown in many organisms to regulate the expression of RubisCO genes. Sigma70-like -10 and -35 promoter boxes and potential CbbR-binding sites (T-N11-A/TNA-N7TNA) were predicted in the upstream regions of the four operons. Electrophoretic mobility shift assays (EMSAs) confirmed that purified CbbR is able to bind to the upstream regions of the cbb1, cbb2 and cbb3 operons, demonstrating that the predicted CbbR-binding sites are functional in vitro. However, CbbR failed to bind the upstream region of the cbb4 operon that contains cbbP, encoding phosphoribulokinase (EC 188.8.131.52). Thus, other factors not present in the assay may be required for binding or the region lacks a functional CbbR-binding site. The cbb3 operon contains genes predicted to encode anthranilate synthase components I and II, catalyzing the formation of anthranilate and pyruvate from chorismate. This suggests a novel regulatory connection between CO2 fixation and tryptophan biosynthesis. The presence of a form II RubisCO could promote the ability of A. ferrooxidans to fix CO2 at different concentrations of CO2. A. ferrooxidans has features of cbb gene organization for CO2-assimilating functions that are characteristic of obligate chemolithoautotrophs and distinguish this group from facultative autotrophs. The most conspicuous difference is a separate operon for the cbbP gene. It is hypothesized that this organization may provide greater flexibility in the regulation of expression of genes involved in inorganic carbon assimilation.


Health check-ups in general practice: a patient perspective.
One of the strategies identified by the NHS to achieve improvements in the health status of the Scottish population is to implement health promotion programmes through the general medical practitioner service. Grampian Local Health Council undertook a survey of Grampian residents to identify factors which might encourage greater public participation in such programmes. The results which are analysed and discussed in this paper show that factors identified by the public include: a preference for doctors to carry out health checks; the need for public education on the roles of different health professionals; the need for information on health checks and their benefits to be supplied in advance; a choice of appointment times and the offer of a follow-up appointment to discuss test results. These results are of interest to health professionals as they are likely to contribute to higher levels of patient involvement in health promotion programmes.


Monocular spatial distortions induced by marked accommodation.
Contraction of the ciliary muscle during marked accommodation causes the leading edge of the retina to advance as much as 0.5 centimeter. Near the posterior pole of the eye, the upward and downward extensional strains on the retina should be reasonably balanced. In the horizontal meridian an asymmetry is introduced because of the nasal location of the optic nerve head. Observers were asked to bisect the space between two parallel lines while fixating a movable line lying near the midpoint of the two lines. The test was conducted with the target far from and near the subject, in the horizontal and vertical meridians, and was repeated with accommodation paralyzed by a cycloplegic agent. Marked accommodation induced significant spatial distortions in the horizontal meridian. The effect is largely retinal.


Chest Radiographs in Congestive Heart Failure: Visualizing Neural Network Learning.
Purpose To examine Generative Visual Rationales (GVRs) as a tool for visualizing neural network learning of chest radiograph features in congestive heart failure (CHF). Materials and Methods A total of 103 489 frontal chest radiographs in 46 712 patients acquired from January 1, 2007, to December 31, 2016, were divided into a labeled data set (with B-type natriuretic peptide [BNP] result as a marker of CHF) and unlabeled data set (without BNP result). A generative model was trained on the unlabeled data set, and a neural network was trained on the encoded representations of the labeled data set to estimate BNP. The model was used to visualize how a radiograph with high estimated BNP would look without disease (a "healthy" radiograph). An overfitted model was developed for comparison, and 100 GVRs were blindly assessed by two experts for features of CHF. Area under the receiver operating characteristic curve (AUC), κ coefficient, and mixed-effects logistic regression were used for statistical analyses. Results At a cutoff BNP of 100 ng/L as a marker of CHF, the correctly trained model achieved an AUC of 0.82. Assessment of GVRs revealed that the correctly trained model highlighted conventional radiographic features of CHF as reasons for an elevated BNP prediction more frequently than the overfitted model, including cardiomegaly (153 [76.5%] of 200 vs 64 [32%] of 200, respectively; P < .001) and pleural effusions (47 [23.5%] of 200 vs 16 [8%] of 200, respectively; P = .003). Conclusion Features of congestive heart failure on chest radiographs learned by neural networks can be identified using Generative Visual Rationales, enabling detection of bias and overfitted models. © RSNA, 2018 See also the editorial by Ngo in this issue.


The MABIC project: An effectiveness trial for reducing risk factors for eating disorders.
Challenges in the prevention of disordered eating field include moving from efficacy to effectiveness and developing an integrated approach to the prevention of eating and weight-related problems. A previous efficacy trial indicated that a universal disordered eating prevention program, based on the social cognitive model, media literacy educational approach and cognitive dissonance theory, reduced risk factors for disordered eating, but it is unclear whether this program has effects under more real-world conditions. This effectiveness trial tested whether this program has effects when previously trained community providers in an integrated approach to prevention implement the intervention. The research design involved a multi-center non-randomized controlled trial with baseline, post-test and 1-year follow-up measures. The sample included girls in the 8th grade from six schools (n = 152 girls) in a city near Barcelona (intervention group), and from eleven schools (n = 413 girls) in four neighboring towns (control group). The MABIC risk factors of disordered eating were assessed as main outcomes. Girls in the intervention group showed significantly greater reductions in beauty ideal internalization, disordered eating attitudes and weight-related teasing from pretest to 1-year follow-up compared to girls in the control group, suggesting that this program is effective under real-world conditions.


Fidelity of DNA synthesis in a mammalian in vitro replication system.
We have used the simian virus 40 (SV40)-based shuttle vector pZ189 in a forward-mutation assay to determine the fidelity of DNA replication in the in vitro DNA replication system developed by J.J. Li and T.J. Kelly (Proc. Natl. Acad. Sci. USA 81:6973-6977, 1984). We find that very few base substitution errors (approximately 1/180,000 bases incorporated) are made during in vitro replication of the pZ189 vector in a system derived from CV-1 monkey cells. This replication is completely dependent on added SV40 T antigen and presumably reflects synthesis that is initiated at the SV40 replication origin. The observed level of fidelity is far greater than that reported for in vitro replication of DNA by conventionally purified eucaryotic DNA polymerases alpha and beta. Thus, there must be additional cellular factors in the crude in vitro system that serve to enhance the fidelity of DNA replication.


Uptake of tetraphenylporphycene and its photoeffects on actin and cytokeratin elements of HeLa cells.
In the present work we have continued our studies in the photobiological properties of the 2,7,12,17-tetraphenylporphycene (TPPo). In particular, the uptake, the subcellular localization and the photoeffects on two cytoskeletal elements (actin, microfilaments and cytokeratin intermediate filaments) of HeLa cells have been analyzed. The uptake kinetics of TPPo, determined by fluorescence spectroscopy, was initially very rapid, reaching saturation at approximately 6 h of incubation. This porphycene tends to be accumulated mainly in rounded particles distributed throughout the cytoplasm. The morphological comparison of the localization pattern of TPPo and those of acridine orange and rhodamine 123, which are fluorescence markers for lysosomes and mitochondria respectively, allowed us to confirm that this porphycene is mainly accumulated in lysosomal organelles. The results obtained after treatment with TPPo and red light indicated that this compound is very effective in mediating the photodestruction of lysosomes. The photosensitizing effects on the cytoskeletal elements studied depended on both the irradiation time and the elapsed time after treatment. The implications of damage to lysosomes and actin and cytokeratin filaments on the process of cell death is discussed.


The role of positive emotions in positive psychology. The broaden-and-build theory of positive emotions.
In this article, the author describes a new theoretical perspective on positive emotions and situates this new perspective within the emerging field of positive psychology. The broaden-and-build theory posits that experiences of positive emotions broaden people's momentary thought-action repertoires, which in turn serves to build their enduring personal resources, ranging from physical and intellectual resources to social and psychological resources. Preliminary empirical evidence supporting the broaden-and-build theory is reviewed, and open empirical questions that remain to be tested are identified. The theory and findings suggest that the capacity to experience positive emotions may be a fundamental human strength central to the study of human flourishing.


Feasibility of glucagon eyedrops for clinical use in hypoglycemia.
Although sufficient amount of glucagon can be taken up into systemic circulation through ocular route, the concentration of eyedrops needed at 7.5% would be too high and the price of the doses would be too expensive to be desirable. Attempts have been made to reduce the doses of glucagon needed for systemic delivery through ocular route by adding absorption enhancers such as 0.5% BL-9 or 0.5% Brij-78. With these absorption enhancers, the effective concentrations of glucagon eyedrops can be lowered to 0.05% in rabbit eyes and could be lowered to 1.25% in human eyes which would be reasonable to be used in the clinics.


Influence of Nutrient Stress on the Relationships between PAM Measurements and Carbon Incorporation in Four Phytoplankton Species.
Two methods of measuring primary production, modulated fluorimetry (PAM) and the traditional carbon incorporation method ((13)C), were compared in four phytoplankton species, two diatoms (Pseudo-nitzschia pungens and Asterionellopsis glacialis), and two dinoflagellates (Heterocapsa sp and Karenia mikimotoï), under N (nitrogen), P (phosphorus) and Si (silicon) limited semi-continuous culture. N and Si-limited cultures showed relatively high quantum efficiency of the PSII (Fv/Fm) values, confirming that Fv/Fm is not a good proxy for nutrient stress in balanced systems, whereas P limitation had a drastic effect on many physiological parameters. In all species, the physiological capacity of phytoplankton cells to acclimate to nutrient limitations led to changes in the cellular biochemical composition and the structure of the photosynthetic apparatus. The observed physiological responses were species and nutrient specific. The values of the chlorophyll-specific absorption cross section (a*) increased with nutrient limitation due to package effect, while the carbon/Chl a ratio was higher under N and P limitations. In diatoms, Si limitation did not affect photosynthesis confirming the uncoupling between Si and carbon metabolisms. In all four species and under all treatments, significant relationships were found between photosynthetic activities, ETR(Chl) (electron transport rate) and P(Chl) (carbon fixation rate) estimated using PAM measurements and (13)C incorporation, showing that the fluorescence technique can reliably be used to estimate carbon fixation by phytoplankton. The relationship between ETR(Chl) and P(Chl) can be described by the shape and the slope of the curve (ΦC.e). Linear relationships were found for dinoflagellates and P. pungens under all treatments. A decrease in ΦC.e was observed under N and P limitation probably due to structural damage to the photosynthetic apparatus. A. glacialis showed a logarithmic relationship in N and P limited conditions, due to the alternative electron flow which takes place to optimise photosynthetic performances under high light and/or nutrient stress.


Manometric findings of esophageal motor disorders in 240 Brazilian patients with non-cardiac chest pain.
Two hundred and forty Brazilian patients with chest pain and normal cardiac evaluation were submitted to computerized esophageal manometry. Endoscopic examination and/or swallow barium studies had excluded obstructive lesions. Motor disorders were found in 63% of patients; non-specific motors disorders and hypotensive lower esophageal sphincter were the most common. The finding of nutcracker esophagus in only 6% of the patients is a quite different rate from what has been previously described in the literature. Esophagitis was observed at endoscopy in 13.4% of the patients, hiatus hernia in 19.7% and peptic gastric or duodenal ulcer in 4.9%. It should be emphasized that after excluding pain as being of cardiac origin an abnormal manometry result points to the esophagus as the probable site of origin of the pain; esophageal investigation is important for establishing proper treatment for these patients.


Update on the Mechanisms of Antibiotic Resistance and the Mobile Resistome in the Emerging Zoonotic Pathogen Streptococcus suis.
Streptococcus suis is a zoonotic pathogen causing important economic losses in swine production. The most commonly used antibiotics in swine industry are tetracyclines, beta-lactams, and macrolides. Resistance to these antibiotics has already been observed worldwide (reaching high rates for macrolides and tetracyclines) as well as resistance to aminoglycosides, fluoroquinolones, amphenicols, and glycopeptides. Most of the resistance mechanisms are encoded by antibiotic resistance genes, and a large part are carried by mobile genetic elements (MGEs) that can be transferred through horizontal gene transfer. This review provides an update of the resistance genes, their combination in multidrug isolates, and their localization on MGEs in S. suis. It also includes an overview of the contribution of biofilm to antimicrobial resistance in this bacterial species. The identification of resistance genes and study of their localization in S. suis as well as the environmental factors that can modulate their dissemination appear essential in order to decipher the role of this bacterium as a reservoir of antibiotic genes for other species.


Origin of intracellular calcium and quantitation of mobilizable calcium in neutrophils stimulated with chemotactic peptide.
The origin and amount of mobilized Ca2+ in chemotactic peptide-stimulated guinea pig neutrophils were examined using biochemical techniques. The total amount of releasable Ca2+ by 20 microM A23187 from the unstimulated intact cells was 0.91 nmol/4 X 10(6) cells, as assessed by change in absorbance of the antipyrylazo III-Ca2+ complex. Two types of internal vesicular Ca2+ pool, mitochondrial and non-mitochondrial pool were identified in the saponin-permeabilized cells. The total amount of releasable Ca2+ was comparable to that accumulated by the non-mitochondrial pool at (1-2) X 10(-7) M of a free Ca2+ concentration. The mitochondrial uncoupler, capable of releasing Ca2+ from the mitochondrial pool, neither modified the basal cytosolic free Ca2+ in quin 2-loaded cells nor caused a Ca2+ efflux from the intact cells. These results suggest that the releasable Ca2+ may be located in the non-mitochondrial pool of unstimulated intact cells, and the mitochondrial pool contains little releasable Ca2+. The addition of fMet-Leu-Phe increased the cytosolic free Ca2+ by two processes: Ca2+ mobilization from internal stores and Ca2+ influx through the surface membrane. The Ca2+ mobilized and effluxed from the intact cells by stimulation with the maximal doses of fMet-Leu-Phe was estimated to be 0.27 nmol/4 X 10(6) cells. Almost equal amounts were released by the maximal doses of inositol 1,4,5-trisphosphate from the non-mitochondrial pool of saponin-treated cells that had accumulated Ca2+ at a free Ca2+ concentration of 1.4 X 10(-7) M. The mechanism related to the Ca2+ influx by fMet-Leu-Phe stimulation was also examined. The addition of nifedipine or phosphatidic acid did not affect the change in the cytosolic free Ca2+ induced by fMet-Leu-Phe, thereby suggesting that the receptor-mediated Ca2+ channel may be involved in the Ca2+ influx.


Inhibitor or promoter? The performance of polysaccharides from Ganoderma lucidum on human tumor cells with different p53 statuses.
Polysaccharides from Ganoderma lucidum (GLPs) have been taken as effective supplements by both healthy people and cancer patients for many years. However, this short survey indicates that instead of inhibiting cancer cell growth, both submerge-cultured intracellular GLP and fruiting body GLP can stimulate the growth of human carcinoma cell lines lacking functional p53, such as HCT-116 p53(-/-), Saos-2, H1299, HL-60, MDA-MB-157. Conversely, the two GLPs inhibit all other assayed cells with functional p53. These results could be an alert since mutational inactivation of the tumor suppressor protein p53 is the most frequent genetic alteration found in human tumors.


Low-molecular-weight heparins and unstable angina--current perspectives.
Thrombosis is responsible for most acute manifestations of coronary artery disease, including unstable angina and non-Q-wave myocardial infarction (MI). Antiplatelet therapy plays a major role in reducing the risk of ischaemic events in such patients. Since thrombin generation is key in the pathogenesis of thrombosis, recent studies have focussed on thrombin inhibition in the management of acute ischaemia. Heparin is the most widely used anticoagulant for acute management of thrombosis and is the treatment of choice in the prevention and treatment of venous thromboembolism. Heparin given in therapeutic doses intravenously has been shown to be more effective than aspirin at reducing the risk of death or MI in patients with unstable angina. Low-molecular-weight heparins (LMWHs) have improved pharmacologic and pharmacokinetic properties over standard heparin that may result in greater efficiency and safety. LMWH may be given in fixed doses subcutaneously without monitoring, resulting in greater clinical utility and cost-effectiveness compared with standard heparin. A number of LMWHs-dalteparin, enoxaparin and nadroparin-have been evaluated in unstable angina. In a small open trial, nadroparin reduced the risk of ischaemic outcomes compared with aspirin alone or a combination of aspirin and standard heparin. Dalteparin has been evaluated in two large clinical trials in the management of unstable angina. The low-Molecular-Weight Heparin (Fragmin) During Instability in Coronary Artery Disease (FRISC) trial showed that dalteparin resulted in a 63% reduction in risk of death or acute MI compared with aspirin alone. The Fragmin in Unstable Coronary Artery Disease (FRIC) trial showed that dalteparin was as effective as intravenous heparin. Enoxaparin resulted in a statistically significant 16% reduction in the combined outcome of death, MI and recurrence of angina in comparison with standard heparin in the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-Wave Coronary Events (ESSENCE) trial. There is accumulating evidence that LMWHs are safe and effective alternatives to standard heparin in unstable coronary artery disease and that they offer practical and therapeutic advantages.


Mesophilic continuous fermentative hydrogen production from acid pretreated de-oiled jatropha waste hydrolysate using immobilized microorganisms.
Mesophilic hydrogen production from acid pretreated hydrolysate (biomass concentration of 100g/L and 2% hydrochloric acid) of de-oiled jatropha waste was carried out in continuous system using immobilized microorganisms at various hydraulic retention times (HRTs) ranging from 48 to 12h. The experimental results of the reusability of immobilized microorganisms showed their stability up to 10 cycles with an average cumulative hydrogen production of 770mL/L. The peak hydrogen production rate and hydrogen yield were 0.9L/L*d and 86mL/greducing sugars added, respectively at 16h HRT, with butyrate as the predominant volatile fatty acid. The microbial community analysis revealed that majority of the PCR-DGGE bands were assigned to genus Clostridium and were perhaps the key drivers of the higher hydrogen production.


Histone deacetylase inhibition increases levels of choline kinase α and phosphocholine facilitating noninvasive imaging in human cancers.
Histone deacetylase (HDAC) inhibitors are currently approved for cutaneous T-cell lymphoma and are in mid-late stage trials for other cancers. The HDAC inhibitors LAQ824 and SAHA increase phosphocholine (PC) levels in human colon cancer cells and tumor xenografts as observed by magnetic resonance spectroscopy (MRS). In this study, we show that belinostat, an HDAC inhibitor with an alternative chemical scaffold, also caused a rise in cellular PC content that was detectable by (1)H and (31)P MRS in prostate and colon carcinoma cells. In addition, (1)H MRS showed an increase in branched chain amino acid and alanine concentrations. (13)C-choline labeling indicated that the rise in PC resulted from increased de novo synthesis and correlated with an induction of choline kinase α expression. Furthermore, metabolic labeling experiments with (13)C-glucose showed that differential glucose routing favored alanine formation at the expense of lactate production. Additional analysis revealed increases in the choline/water and phosphomonoester (including PC)/total phosphate ratios in vivo. Together, our findings provide mechanistic insights into the impact of HDAC inhibition on cancer cell metabolism and highlight PC as a candidate noninvasive imaging biomarker for monitoring the action of HDAC inhibitors.


[Tetrabutyl titanate hydrolysis prepared TiO2 photocatalysis loaded with platinum technology].
Photocatalysts Pt/TiO2 coated on hollow glass beads were prepared by tetrabutyl titanate hydrolysis with Sodium silicate on hollow glass beads at various condition and loaded with platinum varying from 0.2% to 2.4% by weight. Sodium pentachlorophenolate (PCP-Na) solution were used to examined for their photoactivity and characterized by X-ray and BET. The results indicated that the optimization condition to prepare photocatalysts: Water to titanium alkoxides was 100, Sintering temperature was 650 degrees C, Diameter of hollow glass beads was 0.5 mm, TiO2: sodium silicate: hollow glass beads was 10:2.5:20, Platinum content of photocatalysts was about 1.4%-1.6%. When the experiments were carried out in such conditions, the initial concentration of PCP-Na was 100 mg/L, initial pH was 6.5, oxygen flux was 1.6 mL/s, illumination intensity was 30 kW.m-2, catalysts was 2 g/L, illumination time was 2 hours, respectively. Then the rates removals of PCP-Na could reach 92.0%.


How to measure metacognition.
The ability to recognize one's own successful cognitive processing, in e.g., perceptual or memory tasks, is often referred to as metacognition. How should we quantitatively measure such ability? Here we focus on a class of measures that assess the correspondence between trial-by-trial accuracy and one's own confidence. In general, for healthy subjects endowed with metacognitive sensitivity, when one is confident, one is more likely to be correct. Thus, the degree of association between accuracy and confidence can be taken as a quantitative measure of metacognition. However, many studies use a statistical correlation coefficient (e.g., Pearson's r) or its variant to assess this degree of association, and such measures are susceptible to undesirable influences from factors such as response biases. Here we review other measures based on signal detection theory and receiver operating characteristics (ROC) analysis that are "bias free," and relate these quantities to the calibration and discrimination measures developed in the probability estimation literature. We go on to distinguish between the related concepts of metacognitive bias (a difference in subjective confidence despite basic task performance remaining constant), metacognitive sensitivity (how good one is at distinguishing between one's own correct and incorrect judgments) and metacognitive efficiency (a subject's level of metacognitive sensitivity given a certain level of task performance). Finally, we discuss how these three concepts pose interesting questions for the study of metacognition and conscious awareness.


Racial/Ethnic Discrimination and Suicidal Ideation in Emerging Adults: The Role of Traumatic Stress and Depressive Symptoms Varies by Gender not Race/Ethnicity.
Drawing from Race-Based Traumatic Stress theory, the present study examined whether traumatic stress and depressive symptoms differentially help explain the relation between racial/ethnic discrimination and suicidal ideation across gender and racial/ethnic groups. A racially/ethnically diverse group of emerging adults (N = 1344; Mage = 19.88, SD = 2.25; 72% female; 46% Hispanic) completed a battery of self-report measures. A cross-sectional design was employed with a series of hierarchical linear regression models and bootstrapping procedures to examine the direct and indirect relation between racial/ethnic discrimination and suicidal ideation through traumatic stress and depressive symptoms across gender and race/ethnicity. The findings suggest an indirect relation through depressive symptoms, but not traumatic stress, and a serial indirect relation through traumatic stress to depressive symptoms in young women and young men, the latter of which was stronger in young women. The indirect relations did not vary by racial/ethnic group. Cumulative experiences of racial/ethnic discrimination may impact suicide-related risk via increases in psychiatric symptomology (i.e., traumatic stress and depressive symptoms), particularly in young women. Racial/ethnic discrimination experiences should be accounted for as a potential source of psychological distress in the assessment, diagnosis, and treatment of suicidal thoughts and behavior, especially among young women endorsing traumatic stress and depressive symptoms. Further research is warranted to better understand the gender difference in the relation between racial/ethnic discrimination and suicide-related risk.


Prevalence of hepatitis C antibodies (HCV) in a dialysis population at one center.
Hepatitis C (HC) has been recently diagnosed by determination of specific antibodies that represent the former so-called non-A, non-B hepatitis. We studied the prevalence of plasma HCV antibodies among 61 unselected patients on hemodialysis (HD) and 43 on continuous ambulatory peritoneal dialysis (CAPD). Plasma C-antibodies were determined through the ELISA test system. Transfusion policy was the same in both groups. The prevalence of hepatitis C virus antibodies was significantly higher in hemodialysis patients than among those on CAPD. Time on dialysis, previous blood transfusions, and renal transplantation seem to increase the prevalence of C hepatitis antibodies among hemodialysis patients. The effect of these parameters on CAPD was smaller. Understanding the reasons for these differences may help prevent this disease among dialysis patients.


Visible-Light-Mediated Divergent and Regioselective Vicinal Difunctionalization of Styrenes with Arylazo Sulfones.
Activated by visible light, arylazo sulfones can serve as multifaceted reactants and are employed in diazenylation, sulfonylation, and arylation reactions under (photo)catalyst-free conditions. Such versatile reactivity enabled us to develop an operationally simple, regioselective, and tunable difunctionalization of styrenes with arylazo sulfones to produce α-sulfonyl arylhydrazones and 1,2-alkoxyarylated products in moderate to excellent yields. Furthermore, such difunctionalized products have been exploited as key building blocks for the synthesis of various heterocycles.


[131I and 99mTc-Uptake in focal thyroid autonomies. Development in Germany since the 1980s].
Germany has developed into a country with a nationwide largely sufficient iodine supply due to improved alimentary iodine supply. The reduction of iodine uptake in focal autonomies induced by reduced iodine avidity was evaluated by Gotthardt et al. 2006, showing a significant decline of pertechnetate uptake up to the year 2004. This study was intended to carry this investigation forward to the present day to analyze the course of a conjectural stabilization of iodine uptake values. 283 patients who underwent radioiodine therapy for focal thyroid were analyzed retrospectively. Pertechnetate uptake was measured scintigraphically, thyroid volume sonographically and iodine uptake by iodine uptake test. The uptake percentage in the autonomous volume was correlated with autonomous volume and the resulting values tracked over a time period of seven years. Mean thyroid volume ranged from 24 to 29 ml, autonomous volume from 7.2 to 9.4 ml. Pertechnetate uptake ranged from 0.2 to 0.25%/ml autonomous volume. Iodine uptake values ranged from 3.2 to 4.2%/ml autonomous volume. None of the changes observed were statistically significant (all p>0.05). Prophylactic measures towards improvement of the general public´s iodine supply in Germany had led to a decline of pertechnetate and iodine uptake in the thyroid up until the turn of the millennium. The here presented data show a stabilization of 99mTc-Uptake. Our study could also show that actual iodine uptake has stabilized at a steady level over the preceding seven years.


From rural beginnings to statewide roll-out: Evaluation of facilitator training for a group-based diabetes prevention program.
To evaluate the approach used to train facilitators for a large-scale group-based diabetes prevention program developed from a rural implementation research project. Orientation day was attended by 224 health professionals; 188 submitted the self-learning task; 175 achieved the satisfactory standard for the self-learning task and attended the workshop; 156 completed the pre- and post-training questionnaires. Two pre- and post-training scales were developed to assess knowledge and confidence in group-based diabetes prevention program facilitation. Principal component analysis found four factors for measuring training effectiveness: knowledge of diabetes prevention, knowledge of group facilitation, confidence to facilitate a group to improve health literacy and confidence in diabetes prevention program facilitation. Self-learning task scores, training discontinuation rates and satisfaction scores were also assessed. There was significant improvement in all four knowledge and confidence factors from pre- to post-training (P < 0.001). The self-learning task mean test score was 88.7/100 (SD = 7.7), and mean assignment score was 72.8/100 (SD = 16.1). Satisfaction with training scores were positive and 'previous training' interacted with 'change in knowledge of diabetes prevention program facilitation' but not with change in 'confidence to facilitate.' The training program was effective when analysed by change in facilitator knowledge and confidence and the positive mean satisfaction score. Learning task scores suggest tasks were manageable and the requirement contributed to facilitator self-selection. Improvement in confidence scores in facilitating a group-based diabetes prevention program, irrespective of previous training and experience, show that program-specific skill development activities are necessary in curriculum design.


Combination of fractional carbon dioxide laser with recombinant human collagen in periocular skin rejuvenation.
The most visible sign of facial aging is often seen in the periocular area. However, periocular rejuvenation remains challenging due to the particularity of periocular anatomic locations. We aimed to evaluate the efficacy and safety of the fractional-ablative CO2 laser-facilitated recombinant human collagen permeation in periocular rejuvenation. This 3-month prospective single-blinded and self-controlled trial enrolled 26 patients with periocular aging who underwent the treatments of fractional-ablative CO2 laser along with laser-facilitated recombinant human collagen permeation. Following the treatments, the patients were quantitatively assessed by various periocular skin aging indices before and after the treatment and monitored for any related adverse events. The patients showed significant improvements with the periocular skin aging indices 3 months after the treatments, which were detailed with a 47.3% decrease in lower eyelid skin rhytids, a 41.4% decrease in the lower eyelid skin texture, a 35.0% decrease in the static crow's feet, a 29.3% decrease in the amount of upper eyelid laxity, and a 20.2% increase in the MRD1 as compared with baseline (p < 0.05). Moreover, total skin thickness under ultrasound was increased in both upper and lower eyelids (5.6% and 3.3%, p < 0.05, respectively). Moreover, six patients (23.1%, 6/26) had erythema for 2 weeks, and two (2/26, 7.7%) had mild hyperpigmentation for 3 months. Fractional-ablative CO2 laser combined with laser-facilitated recombinant human collagen permeation can be a safe and effective treatment for periocular rejuvenation.


The nexus of long noncoding RNAs, splicing factors, alternative splicing and their modulations.
The process of alternative splicing (AS) is widely deregulated in a variety of cancers. Splicing is dependent upon splicing factors. Recently, several long noncoding RNAs (lncRNAs) have been shown to regulate AS by directly/indirectly interacting with splicing factors. This review focuses on the regulation of AS by lncRNAs through their interaction with splicing factors. AS mis-regulation caused by either mutation in splicing factors or deregulated expression of splicing factors and lncRNAs has been shown to be involved in cancer development and progression, making aberrant splicing, splicing factors and lncRNA suitable targets for cancer therapy. This review also addresses some of the current approaches used to target AS, splicing factors and lncRNAs. Finally, we discuss research challenges, some of the unanswered questions in the field and provide recommendations to advance understanding of the nexus of lncRNAs, AS and splicing factors in cancer.


High throughput and wide field of view EUV microscope for blur-free one-shot imaging of living organisms.
We present and demonstrate the use of an extreme ultraviolet (EUV) microscope that was developed in-house. Images are acquired using Bragg reflection multilayer optics and a laser-produced plasma light source. The upper-limit spatial resolution of the EUV microscope is 130 nm with a 10 ns exposure time and 250 x 250 microm(2) field of view. Resolution is superior to that of visible microscopes with the same size of field of view, and the exposure time is short enough to observe fine structures in-vivo. Observation of the cerebral cortex of a mouse is demonstrated.


An introductory characterization of a combat-casualty-care relevant swine model of closed head injury resulting from exposure to explosive blast.
Explosive blast has been extensively used as a tactical weapon in Operation Iraqi Freedom (OIF) and more recently in Operation Enduring Freedom(OEF). The polytraumatic nature of blast injuries is evidence of their effectiveness,and brain injury is a frequent and debilitating form of this trauma. In-theater clinical observations of brain-injured casualties have shown that edema, intracranial hemorrhage, and vasospasm are the most salient pathophysiological characteristics of blast injury to the brain. Unfortunately, little is known about exactly how an explosion produces these sequelae as well as others that are less well documented. Consequently, the principal objective of the current report is to present a swine model of explosive blast injury to the brain. This model was developed during Phase I of the DARPA (Defense Advanced Research Projects Agency) PREVENT (Preventing Violent Explosive Neurotrauma) blast research program. A second objective is to present data that illustrate the capabilities of this model to study the proximal biomechanical causes and the resulting pathophysiological, biochemical,neuropathological, and neurological consequences of explosive blast injury to the swine brain. In the concluding section of this article, the advantages and limitations of the model are considered, explosive and air-overpressure models are compared, and the physical properties of an explosion are identified that potentially contributed to the in-theater closed head injuries resulting from explosions of improvised explosive devices (IEDs).


One-year outcome after decompressive surgery for massive nondominant hemispheric infarction.
Massive cerebral infarction is often accompanied by early death secondary to transtentorial herniation. We have tested the hypothesis that decompressive hemicraniectomy for massive nondominant cerebral infarction is lifesaving in a series of 14 patients presenting with right hemispheric infarction and clinical signs of uncal herniation and impending death. We have further analyzed, in prospective follow-up examinations, the levels of physical, psychiatric, and social disabilities in these patients. The methods used included retrospective analysis to determine rates of immediate mortality and morbidity after surgical intervention. Prospective follow-up data were obtained to determine the level of recovery in surviving patients after 1 year. Standardized measures of outcome to assess physical, psychiatric, and social recovery included the Barthel Index, Zung Depression Scale, and Reintegration to Normal Living Index. With decompressive hemicraniectomy, we were able to prevent death secondary to transtentorial herniation in all cases; 11 patients experienced long-term survival after the procedure, and three deaths were related to non-neurological causes. We observed that 8 of the 11 surviving patients were at home, were functioning with minimal to moderate assistance, and had Barthel scores greater than 60. The remaining three patients were severely disabled. Seven of the 11 survivors were able to walk at 1 year after undergoing the procedure. Depression and failure to reintegrate socially were experienced by most patients. This series confirms the lifesaving nature of hemicraniectomy in patients deteriorating because of cerebral edema after infarction. In patients younger than 50 years, recovery to a state of near-independence is possible.


A review of the specificity of exercises designed for conditioning the lumbar extensors.
To review the specificity of exercises designed to condition the lumbar extensor musculature (ie, lumbar erector spinae and multifidus). A review of studies examining effects of exercises designed to condition the lumbar extensors was conducted. Included were studies that examined the acute activation and chronic adaptation of the lumbar extensor musculature in response to benches and roman chair trunk extensions, free weights exercises (ie, deadlifts, squats, good-mornings, etc), floor and stability ball exercise (ie, trunk extensions, bridging, four-point kneeling, etc) and resistance machines (ie, those with and without pelvic restraints). Evidence suggests that the reviewed exercises designed to condition the lumbar extensors all may result in significant activation of this musculature during their performance. However, examination of training studies shows that for benches and roman chair trunk extensions, free weights exercises, floor and stability ball exercise and resistance machines without appropriate pelvic restraints, evidence suggests that they may be less effective for inducing chronic adaptations in the lumbar extensors as a result of their performance. Contrastingly, resistance machines that employ appropriate pelvic restraint to isolate lumbar extension are better evidenced to confer specific adaptations to the lumbar extensors. Numerous exercise approaches have been designed with the intention of conditioning the lumbar extensors. Those examined appear to activate the lumbar extensors; however, the specificity of many of these exercises for producing chronic adaptations may be questionable, potentially due to the compound nature of them allowing involvement of other musculature such as the hip extensors. Many of the reviewed exercises offer potential to condition the lumbar extensors, however, isolation of lumbar extension through appropriate pelvic restraint appears important for optimising specific adaptations in the lumbar extensors.


Cryptic adaptive radiation in tropical forest trees in New Caledonia.
The causes of the species richness of tropical trees are poorly understood, in particular the roles of ecological factors such as soil composition. The nickel(Ni)-hyperaccumulating tree genus Geissois (Cunoniaceae) from the South-west Pacific was chosen as a model of diversification on different substrates. Here, we investigated the leaf element compositions, spatial distributions and phylogeny of all species of Geissois occurring on New Caledonia. We found that New Caledonian Geissois descended from a single colonist and diversified relatively quickly into 13 species. Species on ultramafic and nonultramafic substrates showed contrasting patterns of leaf element composition and range overlap. Those on nonultramafic substrates were largely sympatric but had distinct leaf element compositions. By contrast, species on ultramafic substrates showed similar leaf element composition, but occurred in many cases exclusively in allopatry. Further, earlier work showed that at least three out of these seven species use different molecules to bind Ni. Geissois qualifies as a cryptic adaptive radiation, and may be the first such example in a lineage of tropical forest trees. Variation in biochemical strategies for coping with both typical and adverse soil conditions may help to explain the diversification and coexistence of tropical forest trees on similar soil types.


EfrEF and the Transcription Regulator ChlR Are Required for Chlorhexidine Stress Response in Enterococcus faecalis V583.
Enterococcus faecalis is an opportunistic pathogen and leading cause of health care-associated infections. Daily chlorhexidine gluconate (CHG) bathing of patients is generally regarded as an effective strategy to reduce the occurrence of health care-associated infections. It is likely that E. faecalis is frequently exposed to inhibitory and subinhibitory concentrations of CHG in clinical settings. The goal of this study was to investigate how the vancomycin-resistant strain E. faecalis V583 transcriptionally responds to and tolerates stress from CHG. We used transcriptome (microarray) analysis to identify genes upregulated by E. faecalis V583 in response to CHG. The genes efrE (EF2226) and efrF (EF2227), encoding a heterodimeric ABC transport system, were the most highly upregulated genes. efrEF expression was induced by CHG at concentrations several 2-fold dilutions below the MIC. Deletion of efrEF increased E. faecalis V583 susceptibility to CHG. We found that ChlR, a MerR-like regulator encoded by a sequence upstream of efrEF, mediated the CHG-dependent upregulation of efrEF, and deletion of chlR also increased chlorhexidine susceptibility. Overall, our study gives insight into E. faecalis stress responses to a commonly used antiseptic.


[New pathways and current concepts in intensive care medicine--a summary of important articles of the last month].
One of the most outstanding critical care concepts of the last month is the definition of 5 therapy strategies by the Society of Critical Care for treatment of patients in septic shock. These strategies are: Low tidal volume of 6 ml/kg(-1) body weight in patients with ARDS, early goal-directed therapy, activated protein C therapy, moderate-dose corticosteroids, and tight control of blood sugar. Following further review of the main critical care medicine related journals for strategies to reduce mortality, the non-invasive ventilation is worthwhile mentioning. Moreover, this review summarizes articles on the topics of: Low-dose dopamine does not prevent acute renal insufficiency, increase of mortality is not related to albumin infusion, subglottic suctioning prevents early onset of ventilator associated pneumonia, prone position in ARDS patients has no influence on mortality, the safest site for central venous catheterization, and pulmonary-artery catheter directed therapy has no influence on mortality of high-risk surgical patients.


Comprehensive diagnosis of PCDD/F emission from three hazardous waste incinerators.
Comprehensive diagnosis of polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/F) emissions was systematically conducted on three hazardous waste incinerators (HWIs). Results indicated that PCDD/F mainly existed in the solid phase before the bag filter. This was especially true for higher chlorinated dioxin and furan congeners (hexa-, hepta- and octa-). The aged bag filters tended to increase the gas-phase PCDD/F. Emissions also increased due to PCDD/F desorption from circulated scrubbing solution and plastic packing media used in the wet scrubber. The PCDD/F concentrations were elevated during the start-up process, reaching up to 5.4 times higher than those measured during the normal operating period. The ratios of PCDFs/PCDDs revealed that the surface-catalysed de novo synthesis was the dominant pathway of PCDD/F formation. Installation of more efficient fabric filters, intermittent replacement of circulated scrubbing solution will result in reduced PCDD/F emission. Additionally, 2,3,4,7,8-PeCDF correlated well with the international toxic equivalent quantity (I-TEQ) value, which suggests that 2,3,4,7,8-PeCDF could act as an I-TEQ indicator.


Adrenal hemorrhage: a 25-year experience at the Mayo Clinic.
To characterize the clinical course of adrenal hemorrhage (AH) by using a systematic review of the presentation, associated conditions, and outcomes in patients with AH seen at our institution between 1972 and 1997 (a 25-year period). A computer search of recorded dismissal diagnoses identified 204 patients with a diagnosis of AH, but only 141 fulfilled our study criteria. Their records were analyzed systematically by presentation, bilateral or unilateral hemorrhage, corticosteroid treatment, and survival. AH is a heterogeneous entity that occurs in the postoperative period, in the antiphospholipid-antibody syndrome, in heparin-associated thrombocytopenia, or in the setting of severe physical stress and multiorgan failure. Standard laboratory evaluation is not helpful in establishing the diagnosis. Of the 141 cases of AH, 78 were bilateral, and 63 were unilateral. Corticosteroid treatment in situations of severe stress or sepsis had little effect on outcome (9% vs. 6% survival with and without corticosteroid treatment, respectively). This is in sharp contrast to AH occurring postoperatively (100% vs. 17% survival with or without treatment, respectively) or in the antiphospholipid-antibody syndrome (73% vs. 0% survival, respectively). A high index of suspicion is required to make a timely diagnosis of AH. Fever and hypotension in the appropriate clinical setting necessitate further investigation. Although the diagnosis of AH is infrequently made while the patient is alive, appropriate imaging techniques are useful for establishing a timely diagnosis. In severe physical stress or sepsis, AH may be a marker of severe, preterminal physiologic stress and poor outcome.


[Barriers to tuberculosis awareness and screening: a qualitative study in a French department].
The incidence of tuberculosis in Seine-Saint-Denis is considerably higher than the national average. The north-west part of the department is particularly concerned. Screening teams encounter difficulties communicating on the disease, reaching the populations concerned and obtaining their adhesion to the screening proposal. The objective of this study was to identify and elucidate the obstacles to tuberculosis prevention and screening. A qualitative study was conducted based on observation of screening actions in Seine-Saint-Denis and semi-directive interviews with health professionals in charge of screening, community representatives and associations, and individuals from the population concerned. Obstacles to tuberculosis awareness and screening appear to be linked to the way in which screening is organized and implemented, and communication difficulties in relation to this disease. Three major obstacles were identified: the gap between the little attention paid to this disease and the sanitary crisis it can trigger in the event of an epidemic; the unsuitability of communication tools for the target population and their lack of attractiveness; maladjustment of screening actions to the local context. The study highlights the individual and social marginalization of the tuberculosis issue and the mechanisms producing this marginalization. It encourages the development of more interactive communication tools and modes, and increased awareness of the professionals involved in screening about field realities.


Prediction of Future Epilepsy in Neonates With Hypoxic-Ischemic Encephalopathy Who Received Selective Head Cooling.
Epilepsy outcomes after therapeutic hypothermia for neonates with hypoxic-ischemic encephalopathy are understudied. The authors used multivariable logistic regression to predict epilepsy in neonates after selective head cooling. Sensitivity analyses used magnetic resonance imaging (MRI) and electroencephalogram (EEG) interpretations by different clinicians. Fifty neonates had 2-year follow-up. Nine developed epilepsy. Predictors included pH ≤6.8 on day of birth (adjusted odds ratio [OR] 19 [95% confidence interval (CI) 1-371]), burst suppression on EEG on day 4 (8.2 [1.3-59]), and MRI deep gray matter injury (OR 33 [2.4-460]). These factors stratify neonates into low (0-1 factors; 3% [0%-14%] risk), medium (2 factors; 56% [21%-86%] risk), and high-risk groups (3 factors; 100% [29%-100%] risk) for epilepsy. The stratification was robust to varying clinical interpretations of the MRI and EEG. Neonates with hypoxic-ischemic encephalopathy who undergo selective head cooling appear at risk of epilepsy if they have 2 to 3 identified factors. If validated, this rule may help counsel families and identify children for close clinical follow-up.


Regulation of heparan sulphate metabolism by adenosine 3':5'-cyclic monophosphate in hepatocytes in culture.
Freshly isolated rat hepatocytes maintained as monolayers in a serum-free medium synthesize sulphated glycosaminoglycans, most of which behave as heparan sulphate and are mainly distributed into intracellular compartments. Cyclic AMP, dibutyryl cyclic AMP, glucagon, noradrenaline, prostaglandin E(1), and theophylline, all drugs and hormones known to increase intracellular cyclic AMP concentrations, decreased the incorporation of (35)SO(4) (2-) into heparan sulphate of intra-, extra- and peri-cellular pools. The inhibition mediated by dibutyryl cyclic AMP was dose-dependent and observed as early as 2h after exposure to the drug. In the presence of 1mm-dibutyryl cyclic AMP, incorporation of (35)SO(4) (2-) or [(14)C]glucosamine into heparan sulphate was decreased to 40-50%, suggesting that dibutyryl cyclic AMP interfered with the synthesis of heparan sulphate. This was further supported by pulse-chase experiments, where dibutyryl cyclic AMP had no effect on the degradation of sulphated glycosaminoglycans. Heparan sulphates synthesized and secreted into the extracellular pool in the presence of dibutyryl cyclic AMP were smaller in size, whereas the degree of sulphation and molecular size of the heparan sulphate chains released by beta-elimination from these proteoglycans were not different from control values. In the presence of 1mm-cycloheximide, (35)SO(4) (2-) incorporation was decreased to 5%. Addition of p-nitrophenyl beta-d-xyloside, an artificial acceptor of glycosaminoglycan chain synthesis, enhanced this incorporation to 18%. Dibutyryl cyclic AMP did not have any inhibitory effect on the synthesis of chains initiated on p-nitrophenyl beta-d-xylosides. Incorporation of [(3)H]serine into heparan sulphate was not affected by dibutyryl cyclic AMP, whereas the degree of substitution of serine residues with heparan sulphate chains was less in heparan sulphate synthesized in the presence of dibutyryl cyclic AMP, suggesting that cyclic AMP exerts its effect on the metabolism of sulphated glycosaminoglycans by affecting the transfer of xylose on to the protein core.


Characterization of TEM1/endosialin in human and murine brain tumors.
TEM1/endosialin is an emerging microvascular marker of tumor angiogenesis. We characterized the expression pattern of TEM1/endosialin in astrocytic and metastatic brain tumors and investigated its role as a therapeutic target in human endothelial cells and mouse xenograft models. In situ hybridization (ISH), immunohistochemistry (IH) and immunofluorescence (IF) were used to localize TEM1/endosialin expression in grade II-IV astrocytomas and metastatic brain tumors on tissue microarrays. Changes in TEM1/endosialin expression in response to pro-angiogenic conditions were assessed in human endothelial cells grown in vitro. Intracranial U87MG glioblastoma (GBM) xenografts were analyzed in nude TEM1/endosialin knockout (KO) and wildtype (WT) mice. TEM1/endosialin was upregulated in primary and metastatic human brain tumors, where it localized primarily to the tumor vasculature and a subset of tumor stromal cells. Analysis of 275 arrayed grade II-IV astrocytomas demonstrated TEM1/endosialin expression in 79% of tumors. Robust TEM1/endosialin expression occurred in 31% of glioblastomas (grade IV astroctyomas). TEM1/endosialin expression was inversely correlated with patient age. TEM1/endosialin showed limited co-localization with CD31, alphaSMA and fibronectin in clinical specimens. In vitro, TEM1/endosialin was upregulated in human endothelial cells cultured in matrigel. Vascular Tem1/endosialin was induced in intracranial U87MG GBM xenografts grown in mice. Tem1/endosialin KO vs WT mice demonstrated equivalent survival and tumor growth when implanted with intracranial GBM xenografts, although Tem1/endosialin KO tumors were significantly more vascular than the WT counterparts. TEM1/endosialin was induced in the vasculature of high-grade brain tumors where its expression was inversely correlated with patient age. Although lack of TEM1/endosialin did not suppress growth of intracranial GBM xenografts, it did increase tumor vascularity. The cellular localization of TEM1/endosialin and its expression profile in primary and metastatic brain tumors support efforts to therapeutically target this protein, potentially via antibody mediated drug delivery strategies.


Neurological features of COVID-19 and their treatment: a review.
Severe acute respiratory syndrome-correlated new coronavirus (SARS-Cov-2) infection may result in neurological signs and symptoms through different mechanisms. Although direct infection of the central nervous system is uncertain or very rare and the para-infectious complications (e.g. inflammatory neuropathies) are rare, delirium and septic encephalopathy are common in severely ill patients. Smell dysfunction and headache are very common in mild cases, especially in younger people and females. Muscle pain is common in both mild and severe cases, and in the most compromised patients, it is accompanied by increased creatine kinase levels and by a likely true myopathic damage. Ischemic stroke has been reported as a possible complication of the hypercoagulability associated with severe SARS-Cov-2 infection, but further studies are needed. Most of the neurological manifestations may occur early in the illness. Therefore, during the pandemic period, neurologists need to be involved, alert, and prepared. Neurological practice will not be the same until a vaccine is available.


Evidence for increased postprandial distal nephron calcium delivery in hypercalciuric stone-forming patients.
A main mechanism of idiopathic hypercalciuria (IH) in calcium stone-forming patients (IHSF) is postprandial reduction of renal tubule calcium reabsorption that cannot be explained by selective reduction of serum parathyroid hormone levels; the nephron site(s) responsible are not as yet defined. Using fourteen 1-h measurements of the clearances of sodium, calcium, and endogenous lithium during a three-meal day in the University of Chicago General Clinical Research Center, we found reduced postprandial proximal tubule reabsorption of sodium and calcium in IHSF vs. normal subjects. The increased distal sodium delivery is matched by increased distal reabsorption so that urine sodium excretions do not differ, but distal calcium reabsorption does not increase enough to match increased calcium delivery, so hypercalciuria results. In fact, urine calcium excretion and overall renal fractional calcium reabsorption both are high in IHSF vs. normal when adjusted for distal calcium delivery, strongly suggesting a distal as well as proximal reduction of calcium reabsorption. The combination of reduced proximal tubule and distal nephron calcium reabsorption in IHSF is a new finding and indicates that IH involves a complex, presumably genetic, variation of nephron function. The increased calcium delivery into the later nephron may play a role in stone formation via deposition of papillary interstitial apatite plaque.


Low fertility may be a significant determinant of ovarian cancer worldwide: an ecological analysis of cross- sectional data from 182 countries.
Ageing, socioeconomic level, obesity, fertility, relaxed natural selection and urbanization have been postulated as the risk factors of ovarian cancer (OC56). We sought to identify which factor plays the most significant role in predicting OC56 incidence rate worldwide. Bivariate correlation analysis was performed to assess the relationships between country-specific estimates of ageing (measured by life expectancy), GDP PPP (Purchasing power parity), obesity prevalence, fertility (indexed by the crude birth rate), opportunity for natural selection (Ibs) and urbanization. Partial correlation was used to compare contribution of different variables. Fisher A-to-Z was used to compare the correlation coefficients. Multiple linear regression (Enter and Stepwise) was conducted to identify significant determinants of OC56 incidence. ANOVA with post hoc Bonferroni analysis was performed to compare differences between the means of OC56 incidence rate and residuals of OC56 standardised on fertility and GDP respectively between the six WHO regions. Bivariate analyses revealed that OC56 was significantly and strongly correlated to ageing, GDP, obesity, low fertility, Ibs and urbanization. However, partial correlation analysis identified that fertility and ageing were the only variables that had a significant correlation to OC56 incidence when the other five variables were kept statistically constant. Fisher A-to-Z revealed that OC56 had a significantly stronger correlation to low fertility than to ageing. Stepwise linear regression analysis only identified fertility as the significant predictor of OC56. ANOVA showed that, between the six WHO regions, multiple mean differences of OC56 incidence were significant, but all disappeared when the contributing effect of fertility on OC56 incidence rate was removed. Low fertility may be the most significant determining predictor of OC56 incidence worldwide.


Supply-side and demand-side policies for biosimilars: an overview in 10 European member states.
This study aimed to provide an overview of biosimilar policies in 10 EU MSs. Methods: Ten EU MS pharmaceutical markets (Belgium, France, Germany, Greece, Hungary, Italy, Poland, Spain, Sweden, and the UK) were selected. A comprehensive literature review was performed to identify supply-side and demand-side policies in place in the selected countries. Results: Supply-side policies for biosimilars commonly include price linkage, price re-evaluation, and tendering; the use of internal or external reference pricing varies between countries; health technology assessment is conducted in six countries. Regarding demand-side policies, pharmaceutical prescription budgets or quotas and monitoring of prescriptions (with potential financial incentives or penalties) are in place in eight and in seven countries respectively. Switching is generally allowed, but is solely the physician's responsibility. Automatic substitution is not recommended, or even forbidden, in most EU MSs. Prescription conditions or guidelines that apply to biosimilars are established in nearly all surveyed EU MSs. Conclusions: Important heterogeneity in policies on biosimilars was seen between (and even within) selected countries, which may partly explain variations in biosimilar uptake. Supply-side policies targeting price have been reported to limit biosimilar penetration in the long term, despite short-term savings, while demand-side policies are considered to positively impact uptake.


Treatment for lymphedema following head and neck cancer therapy: A systematic review.
To perform the first systematic review evaluating all established treatment modalities of head and neck lymphedema resulting from head and neck cancer therapy. Since craniofacial lymphedema treatment represents unique challenges not addressed by extremity lymphedema therapies, a systematic review and evaluation of treatment modalities specific to this area is needed to guide clinical management and further research. Four electronic databases were searches from inception to September 2018. These included Scopus (Embase), PubMed (Medline), Clinicaltrials.gov, and Cochrane Databases. A search string was developed, and all databases queried for keywords on three subjects: head and neck cancer, lymphedema, and therapy. Results were uploaded to an EndNote database where relevant items were identified by hand-searching all titles and abstracts. Subsequently results were combined, duplicates removed, and full papers screened according to eligibility criteria. Of a total 492 search results, twenty-six items met eligibility criteria for this review. These included fourteen cohort studies, seven case reports, two randomized controlled trials, two systematic reviews, and one narrative review totaling 1018 study subjects. The manual lymph drainage group had the largest number of studies and participants, with fewer studies investigating selenium, liposuction, and lymphaticovenular anastomosis. Evidence for the efficacy of all types of lymphedema therapy is limited by paucity of large randomized controlled trials. While manual lymph drainage is best studied, liposuction and surgical treatments have also been effective in a small number of patients.


Phenotypic switching and mating type switching of Candida glabrata at sites of colonization.
Candida glabrata switches spontaneously at high frequency among the following four graded phenotypes discriminated on agar containing 1 mM CuSO(4): white, light brown, dark brown (DB), and very dark brown. C. glabrata also contains three mating type loci with a configuration similar to that of the Saccharomyces cerevisiae mating type cassette system, suggesting it may also undergo cassette switching at the expression locus MTL1. To analyze both reversible, high-frequency phenotypic switching and mating type switching at sites of colonization, primary samples from the oral cavities and vaginal canals of three patients suffering from C. glabrata vaginitis were clonally plated on agar containing CuSO(4). It was demonstrated that (i) in each vaginitis patient, there was only one colonizing strain; (ii) an individual could have vaginal colonization without oral colonization; (iii) phenotypic switching occurred at sites of colonization; (iv) the DB phenotype predominated at the site of infection in all three patients; (v) genetically unrelated strains switched in similar, but not identical, fashions and caused vaginal infection; (vi) different switch phenotypes of the same strain could simultaneously dominate different body locations in the same host; (vii) pathogenesis could be caused by cells in different mating type classes; and (viii) mating type switching demonstrated at both the genetic and transcription levels occurred in one host.


Comparing Resident Outcomes in Cataract Surgery at Different Levels of Experience.
To evaluate outcomes of resident-performed cataract surgeries in different training levels in a retrospective case series. A total of 730 surgeries performed by residents were evaluated into three groups: surgeries performed during residents' first semester of training in phacoemulsification (Level 1 - L1), surgeries performed during the second semester (Level 2 - L2), and surgeries performed during the third semester (Level 3 - L3). The primary outcome was the incidence of intraoperative complications in each group. Secondary outcomes were the comparisons between initial and final corrected distance visual acuity (CDVA), intraocular pressure (IOP), endothelial cell density (ECD), and central corneal thickness (CCT) in each group. Descriptive statistical analyses were employed in the presentation of the results using central tendency and variance measurements. The rate of complications within six weeks of follow-up was 24 out of 102 eyes (23.53%) in the L1 group, 63 out of 301 eyes (20.93%) in the L2 group, and 37 out of 327 (11.31%) in the L3 group (p=0.001). Posterior capsule rupture (PCR) was the most frequent intercurrence observed in all three semesters: it occurred in 12.7% of the surgeries in the first semester (13/102), 16.9% of surgeries in the second semester (51/301), and 9.5% of surgeries in the third semester (31/327). There was no significant difference in CDVA (p=0.298), ECD (p=0.067), IOP (p=0.217), or CCT (p=0.807) between the groups. When measured by rates of complications and by the aforementioned parameters, surgical competency was found to improve as surgical experience and frequency increased. Therefore, this study identified some patterns of skill development that can be applied to teaching strategies and better assist surgeons in training.


Stimulant Use in the Management of Mild Traumatic Brain Injury: A Qualitative Literature Review.
Objective: Mild traumatic brain injury (mTBI) often presents with cognitive complaints including difficulty with attention and concentration. As these symptoms resemble those of ADHD, stimulants may be a potential treatment for mTBI. This review evaluates the literature on the use of stimulants for the treatment of mTBI. Method: A systematic evaluation of the literature using six databases: Ovidmedline, Pubmed, psychINFO, CINAH, Embase, and Cochrane. Broad search terms were used and studies were included that evaluate the use of stimulant and stimulant-like medications in the mTBI population. Data extracted included stimulant type and dosing, symptoms targeted, outcomes, safety and tolerability, and if the study population had ADHD. Results: Nine studies were identified that met the inclusion criteria. Immediate release methylphenidate and amantadine were used for treatment. Methylphenidate had some impact on attention, fatigue, and depression. However, due to the limited number of studies and heterogeneity of study populations, symptoms targeted, and outcome measures used, meaningful conclusions regarding the effect of stimulants in mTBI could not be made. No study evaluated for the presence of ADHD within the study population, despite stimulants being the mainstay treatment for ADHD. Conclusion: PProspective studies on the use of stimulants in mTBI, that evaluate participants for a diagnosis of ADHD, are needed.


Optimizing gene delivery vectors for the treatment of heart disease.
Cardiac gene therapy is approaching reality, with clinical trials entering Phase II/III. Even so, challenges exist to improve the efficacy of even the most successful therapies. The merits of different gene therapy vectors are weighed to assess the current feasibility of each in specific cardiac applications. Major obstacles are discussed, along with recent advances in vector development to overcome or circumvent those difficulties. This review focuses primarily on gene delivery via naked DNA, adenovirus, lentivirus, and adeno-associated virus (AAV) vectors. Gene therapy via adenovirus and AAV vectors has developed into a promising option for the treatment of heart disease. The merits of gene therapy compared with emerging stem cell and microRNA-based treatments are discussed.


Excess of de novo deleterious mutations in genes associated with glutamatergic systems in nonsyndromic intellectual disability.
Little is known about the genetics of nonsyndromic intellectual disability (NSID). We hypothesized that de novo mutations (DNMs) in synaptic genes explain an important fraction of sporadic NSID cases. In order to investigate this possibility, we sequenced 197 genes encoding glutamate receptors and a large subset of their known interacting proteins in 95 sporadic cases of NSID. We found 11 DNMs, including ten potentially deleterious mutations (three nonsense, two splicing, one frameshift, four missense) and one neutral mutation (silent) in eight different genes. Calculation of point-substitution DNM rates per functional and neutral site showed significant excess of functional DNMs compared to neutral ones. De novo truncating and/or splicing mutations in SYNGAP1, STXBP1, and SHANK3 were found in six patients and are likely to be pathogenic. De novo missense mutations were found in KIF1A, GRIN1, CACNG2, and EPB41L1. Functional studies showed that all these missense mutations affect protein function in cell culture systems, suggesting that they may be pathogenic. Sequencing these four genes in 50 additional sporadic cases of NSID identified a second DNM in GRIN1 (c.1679_1681dup/p.Ser560dup). This mutation also affects protein function, consistent with structural predictions. None of these mutations or any other DNMs were identified in these genes in 285 healthy controls. This study highlights the importance of the glutamate receptor complexes in NSID and further supports the role of DNMs in this disorder.


Site-directed mutagenesis of Azotobacter vinelandii ferredoxin I. Changes in [4Fe-4S] cluster reduction potential and reactivity.
We have used site-directed mutagenesis to obtain two variants of Azotobacter vinelandii ferredoxin I (AvFdI), whose x-ray structures are now available. In the C20A protein, a ligand to the [4Fe-4S] cluster was removed whereas in the C24A mutant a free cysteine next to that cluster was removed. Like native FdI, both mutants contain one [4Fe-4S] cluster and one [3Fe-4S] cluster. The structure of C24A is very similar to that of native FdI, while the structure of C20A is rearranged in the region of the [4Fe-4S] cluster to allow it to use the free Cys-24 as a replacement ligand. Here we compare the properties of the native, C20A, and C24A proteins. Although all three proteins are O2 stable in vitro, the C20A protein is much less stable toward proteolysis than the other two in vivo. Spectroscopic results show that all three proteins exhibit the same general redox behavior during O2-oxidation and dithionite reduction. Electrochemical data show that the [3Fe-4S] clusters in all three proteins have the same pH-dependent reduction potentials (-425 mV versus SHE, pH 7.8), whereas the [4Fe-4S] cluster potentials vary over a approximately 150 mV range from -600 mV (C24A) to -647 mV (native) to -746 mV (C20A). Despite this variation in potential both the C20A and C24A proteins appear to be functional in vivo. Native FdI reacts with three equivalents of Fe(CN)3-(6) to form a paramagnetic species previously proposed to be a cysteinyl-disulfide radical. Neither the C20A nor the C24A variant undergoes this reaction, strongly suggesting that it involves the free Cys-24.


Effects of metal-contaminated forest soils from the Canadian shield to terrestrial organisms.
The effects of elevated metal concentrations in forest soils on terrestrial organisms were investigated by determining the toxicity of six site soils from northern Ontario and Quebec, Canada, using a battery of terrestrial toxicity tests. Soils were collected from three sites on each of two transects established downwind of nickel (Sudbury, ON, Canada) and copper (Rouyn-Noranda, PQ, Canada) smelting operations. Site soils were diluted to determine if toxicity estimates for the most-contaminated site soils could be quantified as a percent of site soil. Rouyn-Noranda soils were toxic following acute exposure (14 d) to plants, but not to invertebrates (7 d for collembola and 14 d for earthworms). However, Rouyn-Noranda soils were toxic to all species following chronic exposure (21, 35, and 63 d for plants, collembola, and earthworms, respectively). The toxicity of the Rouyn-Noranda site soils did not correspond to the gradient of metal concentrations in soil. Metal-contaminated Sudbury soils were toxic to plants but not to invertebrates, following acute exposure. Chronic exposure to Sudbury soils caused adverse effects to plant growth and invertebrate survival and reproduction. The toxicity of Sudbury soils corresponded to the metal concentration gradient, with one exception: The reference soil collected in October was toxic to collembola following acute and chronic exposure. This study evaluated the applicability of the new Environment Canada terrestrial toxicity test methods, developed using agricultural soils, to forest soils and also provided useful data to assess the ecological risk associated with mixtures of metals in soil.


The "Immunoscore" in rectal cancer: could we search quality beyond quantity of life?
Because of the function and anatomical environment of the rectum, therapeutic strategies for local advanced rectal cancer (LARC) must deal with two challenging stressors that are a high-risk of local and distal recurrences and a high-risk of poor quality of life (QoL). Over the last three decades, advances in screening tests, therapies, and combined-modality treatment options and strategies have improved the prognosis of patients with LARC. However, owing to the heterogeneous nature of LARC and genetic status, the patient may not respond to a specific therapy and may be at increased risk of side-effects without the life-prolonging benefit. Indeed, each therapy can cause its own side-effects, which may worsen by a combination of treatments resulting in long-term poor QoL. In LARC, QoL has become even more essential with the increasing incidence of rectal cancer in young individuals. Herein, we analyzed the value of the Immunoscore-Biopsy (performed on tumor biopsy at diagnosis) in predicting outcomes, alone or in association with clinical and imaging data, for each therapy used in LARC.


Improved adjuvant endocrine therapy for premenopausal women with endocrine responsive disease.
Results from two randomised global trials (SOFT & TEXT) designed to newly define the most effective components of adjuvant endocrine therapy for premenopausal women with endocrine responsive disease, showed that for some, those with high risk of relapse, the use of the aromatase inhibitor exemestane together with ovarian function suppression with GnRH analogue (triptorelin) yielded the most favourable treatment outcome compared with tamoxifen. For women with low risk of relapse, treatment with tamoxifen was similar to ovarian function suppression together with either exemestane or tamoxifen. For women with intermediate risk of relapse, ovarian function suppression added to tamoxifen was not inferior to exemestane, while it resulted in superior outcomes compared to tamoxifen alone. Now, these trials provide critical information for the adjuvant treatment of premenopausal women with endocrine responsive breast cancer and are important for the development of future trials for further improvement of adjuvant endocrine therapies for the younger population.


Transcriptional regulation of the murine Elf3 gene in embryonal carcinoma cells and their differentiated counterparts: requirement for a novel upstream regulatory region.
The transcription factor Elf3, which is one of over 25 Ets family members, is expressed in a wide variety of carcinomas and has been shown to promote the transcription of many genes implicated in cancer. To understand how the Elf3 gene is regulated at the transcriptional level, we probed its 5'-flanking region, and we report here the identification of both proximal and distal regions that regulate murine Elf3 promoter activity. In addition to mapping the transcription start site of the Elf3 gene, the work described in this study identifies four cis-regulatory elements in the proximal promoter region of the gene. These include a cis-regulatory element previously designated ESE, a kappaB site, a POU motif, and a CCAAT box. In addition, we demonstrate that a novel 94 bp region 2 kb upstream of the transcription start site significantly elevates Elf3 promoter activity in F9-differentiated cells, but not in the parental F9 embryonal carcinoma (EC) cells. This region appears to be largely responsible for the increase in Elf3 promoter activity that accompanies the differentiation of embryonal carcinoma cells.


Immunomodulatory Effect of Withania somnifera (Ashwagandha) Extract-A Randomized, Double-Blind, Placebo Controlled Trial with an Open Label Extension on Healthy Participants.
The immunomodulatory effect of Withania somnifera (WS) extract was tested in healthy adults. In this randomized placebo-controlled double-blinded study, subjects were allocated either 60 mg WS extract or placebo. It consists of a blinded 30-day period and an open-label extension study of another 30 days with crossover of only placebo to test. After the 30-day blinded study period, the WS test group reported significant increase (p < 0.05) in Ig's (IgA, IgM, IgG, IgG2, IgG3 and IgG4), Cytokines (IFN-γ, IL4), TBNK (CD45+, CD3+, CD4+, CD8+, CD19+, NK cells) whereas in the placebo group TBNK cells showed significant decrease (p < 0.05) and Ig's and cytokines showed no change (p > 0.05). In the extension period on day 60, the subjects on placebo who were crossed over to the WS test group showed significant increase (p < 0.05) in Ig's, cytokines and TBNK cells and the subjects who continued on the WS group showed a further significant improvement (p < 0.05) in Ig's, cytokines and TBNK cells. There were no adverse events reported in the study. WS extract significantly improved the immune profile of healthy subjects by modulating the innate and adaptive immune systems. Boosting the immune system of people at risk of infection and during widespread infections can be targeted with WS extract.


Glucocorticoids induce a 29 000 Mr protein in DDT1 MF-2 smooth muscle cells but not in the DDT1 MF-2 GR glucocorticoid resistant variant.
We have demonstrated that glucocorticoids induce in DDT1 MF-2 cells by a glucocorticoid mediated mechanism the synthesis of a methionine-cysteine rich protein of 29 000 Mr (p29). Induction of p29 is not observed in DDT1 MF-2 GR glucocorticoid resistant variants which have only 7% of glucocorticoid receptor site per cell compared to wild type cells. Increased synthesis of p29 is specific to glucocorticoids since neither androgens, estrogens, progesterone nor the glucocorticoid antagonist dexamethasone mesylate are effective inducers. Stimulation of p29 synthesis in wild type cells is observed at 10(-10) M triamcinolone acetonide, reaching a maximum at a concentration of 1 X 10(-8) M. The induction of p29 is not a function of glucocorticoid arrest of DDT1 MF-2 cells since DDT1 MF-2 cells promoted to re-enter the cell cycle by 50 ng/ml platelet derived growth factor (PDGF) continue synthesis of p29. Finally, increased levels of p29 translation products are observed in cell free translation assays carried out utilizing poly A+ RNA transcripts isolated from glucocorticoid treated cells. These data suggest that the glucocorticoid stimulation of p29 synthesis is a transcriptional and/or RNA processing event controlled by glucocorticoid receptor complexes.


Trypanosoma cruzi screening in Texas blood donors, 2008-2012.
Chagas disease is an important emerging disease in Texas that results in cardiomyopathy in about 30% of those infected with the parasite Trypanosoma cruzi. Between the years 2008 and 2012, about 1/6500 blood donors were T. cruzi antibody-confirmed positive. We found older persons and minority populations, particularly Hispanic, at highest risk for screening positive for T. cruzi antibodies during routine blood donation. Zip code analysis determined that T. cruzi is associated with poverty. Chagas disease has a significant disease burden and is a cause of substantial economic losses in Texas.


Refined Cryo-EM Structure of the T4 Tail Tube: Exploring the Lowest Dose Limit.
The bacteriophage T4 contractile tail (containing a tube and sheath) was the first biological assembly reconstructed in three dimensions by electron microscopy at a resolution of ∼35 Å in 1968. A single-particle reconstruction of the T4 baseplate was able to generate a 4.1 Å resolution map for the first two rings of the tube using the overall baseplate for alignment. We have now reconstructed the T4 tail tube at a resolution of 3.4 Å, more than a 1,000-fold increase in information content for the tube from 1968. We have used legacy software (Spider) to show that we can do better than the typical 2/3 Nyquist frequency. A reasonable map can be generated with only 1.5 electrons/Å2 using the higher dose images for alignment, but increasing the dose results in a better map, consistent with other reports that electron dose does not represent the main limitation on resolution in cryo-electron microscopy.


Primary Hyperparathyroidism and Hyperthyroidism in a Patient with Myotonic Dystrophy: A Case Report and Review of the Literature.
Various endocrine manifestations are commonly described in myotonic dystrophy (MD), including primary hypogonadism, diabetes mellitus, and thyroid and parathyroid dysfunction. We describe a 46-year-old woman with a family history of MD with her son. She was diagnosed with cardiac arrhythmia and required the implantation of a pacemaker. She was noted to have a bilateral cataract. She complained of muscle weakness, diffuse myalgia, and palpitation. The electromyography (EMG) showed myotonic discharges. Laboratory tests showed high serum calcium 2.83 mmol/L, serum phosphate 1.2 mmol/L, parathormone 362.5 pg/mL, thyroid stimulating hormone TSH 0.02 mIU/L (normal range: 0.34-5.6 mIU/L), FT4 21.17 ng/mL, and negative anti-thyroperoxidase antibodies. Cervical ultrasound revealed a multinodular goiter. The 99mTc-MIBI scintigraphy localized a lower right parathyroid adenoma. The clinical data, the family history of MD, EMG data, and endocrine disturbances were strongly suggestive of MD associated with hyperthyroidism and primary hyperparathyroidism.


Divergence and isolation of cryptic sympatric taxa within the annual legume Amphicarpaea bracteata.
The amphicarpic annual legume Amphicarpaea bracteata is unusual in producing aerial and subterranean cleistogamous flowers that always self-fertilize and, less commonly, aerial chasmogamous flowers that outcross. Although both morphologic and genetic variants are known in this highly selfing species, debate continues over whether this variation is continuous, reflecting the segregation of standing genetic variation, or discontinuous, reflecting distinct taxa that rarely intercross. We characterized SNP variation in 128 individuals in southern Wisconsin to assess within- and among-population variation at 3928 SNPs. We also assessed genotype and leaf morphology in an additional 76 individuals to connect phenotypic variation with genetic variation. Genetic variation maps onto three strongly divergent and highly inbred genetic groups showing little relation to site location. Each group has a distinct phenotype, but the divergence of these groups differs from the varietal divisions previously identified based on morphological characters. Like previous authors, we argue that the taxonomy of this species should be revised. Despite extensive sympatry, estimates of among-group migration rates are low, and hybrid individuals were at low frequency (<2%) in our dataset. Restricted gene flow likely results from high selfing rates and partial reproductive incompatibility as evidenced by the U-shaped distribution of pairwise F ST values reflecting "islands" of genomic divergence. These islands may be associated with hybrid incompatibility loci that arose in allopatry. The coexistence of lineages within sites may reflect density-dependent attack by species-specific strains of pathogenic fungi and/or root-nodulating bacteria specializing on distinct genotypes.