Inhibition of GSK-3 induces differentiation and impaired glucose metabolism in renal cancer.
Glycogen synthase kinase-3 (GSK-3), a constitutively active serine/threonine kinase, is a key regulator of numerous cellular processes ranging from glycogen metabolism to cell-cycle regulation and proliferation. Consistent with its involvement in many pathways, it has also been implicated in the pathogenesis of various human diseases, including type II diabetes, Alzheimer disease, bipolar disorder, inflammation, and cancer. Consequently, it is recognized as an attractive target for the development of new drugs. In the present study, we investigated the effect of both pharmacologic and genetic inhibition of GSK-3 in two different renal cancer cell lines. We have shown potent antiproliferative activity of 9-ING-41, a maleimide-based GSK-3 inhibitor. The antiproliferative activity is most likely caused by G(0)-G(1) and G(2)-M phase arrest as evident from cell-cycle analysis. We have established that inhibition of GSK-3 imparted a differentiated phenotype in renal cancer cells. We have also shown that GSK-3 inhibition induced autophagy, likely as a result of imbalanced energy homeostasis caused by impaired glucose metabolism. In addition, we have demonstrated the antitumor activity of 9-ING-41 in two different subcutaneous xenograft renal cell carcinoma tumor models. To our knowledge, this is the first report describing autophagy induction due to GSK-3 inhibition in renal cancer cells.


Water turbidity by algal blooms causes mating system breakdown in a shallow-water fish, the sand goby Pomatoschistus minutus.
Eutrophication as a result of human activity has resulted in increased algal blooms and turbidity in aquatic environments. We investigated experimentally the effect of algal turbidity on the mating system and sexual selection in the sand goby, Pomatoschistus minutus (Pallas), a marine fish with a resource-defence mating system and paternal care. Owing to male-male competition and female choice, large males can monopolize multiple mates, while some males do not achieve mating at all. We show that the number of eggs laid was the same in both turbid and clear tanks but that mating success was more evenly distributed among males in turbid than in clear water. The opportunity for sexual selection was lower in turbid conditions. In turbid conditions mating success was less skewed towards large males. Our results suggest that increased turbidity can change mating systems and decrease the opportunity for sexual selection as well as selection intensity.


Existential suffering in the palliative care setting: an integrated literature review.
Existential and spiritual concerns in relation to palliative end-of-life care have received increasing attention over the past decade. To review the literature specifically related to existential suffering in palliative care in terms of the significance of existential suffering in end-of-life care, definitions, conceptual frameworks, and interventions. A systematic approach was undertaken with the aim of identifying emerging themes in the literature. Databases using CINAHL (1980-2009), MEDLINE (1970-2009), and PsychINFO (1980-2009) and the search engine of Google Scholar were searched under the key words existential suffering, existential distress, existential pain, palliative and end of life care. The search yielded a total of 156 articles; 32% were peer-reviewed empirical research articles, 28% were peer-reviewed theoretical articles, and 14% were reviews or opinion-based articles. After manually searching bibliographies and related reference lists, 64 articles were considered relevant and are discussed in this review. Overall analysis identifies knowledge of the following: 1) emerging themes related to existential suffering, 2) critical review of those identified themes, 3) current gaps in the research literature, and 4) recommendations for future research. Findings from this comprehensive review reveal that existential suffering and deep personal anguish at the end of life are some of the most debilitating conditions that occur in patients who are dying, and yet the way such suffering is treated in the last days is not well understood. Given the broad range of definitions attributed to existential suffering, palliative care clinicians may need to be mindful of their own choices and consider treatment options from a critical perspective.


A longitudinal study of anxiety, depression and distress as predictors of sexual and urinary quality of life in men with prostate cancer.
To evaluate the prevalence of depression, anxiety and distress among active surveillance (AS) and radical prostatectomy (RP) patients. To evaluate the impact of these symptoms at baseline on urinary and sexual quality of life at follow-up. Patients managed with AS or RP who completed validated questionnaires assessing levels of depression, anxiety, distress and urinary (UF) and sexual function (SF) and bother comprised the final analytic cohort. These measures were completed at baseline, within 1 year, and between 1 and 3 years from baseline. Mixed model repeated measures analysis was used to examine associations between mental health at baseline and sexual and urinary outcomes in a subset of RP patients with complete follow-up. Among 679 men who comprised the study cohort, baseline prevalence of moderate or higher levels of depression or anxiety were low (<5%), while levels of mild depression or anxiety ranged from 3-16% over time. Baseline levels of elevated distress ranged from 8-20%. Among men who provided data at baseline and follow-up, there were no significant differences between AS and RP patients in the proportion of men with elevated levels of depression, anxiety, or distress. Among 177 men who underwent RP and had complete follow-up moderate or higher levels of depression or anxiety appeared to be associated with post-treatment SF and bother, while elevated levels of distress were associated with post-treatment UF. Moderate or higher levels of depression or anxiety were low in men with localised prostate cancer but were associated with sexual outcomes, while elevated distress was associated with urinary outcomes. Greater attention should be paid to mental health symptoms among men with prostate cancer, as these symptoms may be associated with quality of life outcomes.


"No-o-o-o Peeking": Preschoolers' Executive Control, Social Competence, and Classroom Adjustment.
The goals of this study were to evaluate (1) how specific aspects of executive control, briefly assessed, predict social competence and classroom adjustment during preschool; and (2) differences between two aspects of executive control, according to child's age, socioeconomic risk status, and gender. The facets of executive control were defined as cool executive control (CEC; affectively neutral, slow acting, and late developing) and hot executive control (HEC; more emotional, fast acting, and early developing). Two hundred eighty-seven 3- to 5-year-old children from private child care and Head Start centers were directly assessed during executive control tasks, and preschool teachers provided information on their school success. Aspects of executive control varied with age, socioeconomic risk, and gender. Specifically, older children performed better on CEC tasks across three age levels; for HEC tasks, change was seen only between 3-year-olds and 4-year-olds. Children of mothers with less formal education performed less well on CEC than those whose mothers had more education; girls performed better than boys on HEC tasks. Further, facets of executive control were differentially related to later social competence and classroom adjustment. HEC predicted social competence, whereas CEC uniquely predicted classroom adjustment. Implications for everyday practice and specific curricula formulation are discussed.


Biofilm cohesiveness measurement using a novel atomic force microscopy methodology.
Biofilms can be undesirable, as in those covering medical implants, and beneficial, such as when they are used for waste treatment. Because cohesive strength is a primary factor affecting the balance between growth and detachment, its quantification is essential in understanding, predicting, and modeling biofilm development. In this study, we developed a novel atomic force microscopy (AFM) method for reproducibly measuring, in situ, the cohesive energy levels of moist 1-day biofilms. The biofilm was grown from an undefined mixed culture taken from activated sludge. The volume of biofilm displaced and the corresponding frictional energy dissipated were determined as a function of biofilm depth, resulting in the calculation of the cohesive energy. Our results showed that cohesive energy increased with biofilm depth, from 0.10 +/- 0.07 nJ/microm(3) to 2.05 +/- 0.62 nJ/microm(3). This observation was reproducible, with four different biofilms showing the same behavior. Cohesive energy also increased from 0.10 +/- 0.07 nJ/microm(3) to 1.98 +/- 0.34 nJ/microm(3) when calcium (10 mM) was added to the reactor during biofilm cultivation. These results agree with previous reports on calcium increasing the cohesiveness of biofilms. This AFM-based technique can be performed with available off-the-shelf instrumentation. It could therefore be widely used to examine biofilm cohesion under a variety of conditions.


Polypeptide N-acetylgalactosaminyltransferase 6 disrupts mammary acinar morphogenesis through O-glycosylation of fibronectin.
A high expression of short and immature O-glycans is one of the prominent features of breast cancer cells, which would be attributed to the upregulated expression of glycosyltransferases. Therefore, a detailed elucidation of glycosyltransferases and their substrate(s) may improve our understandings for their roles in mammary carcinogenesis. Here we report that overexpression of polypeptide N-acetylgalactosaminyltransferase 6 (GALNT6), a glycosyltransferase involved in the initial step of O-glycosylation, has transformational potentials through disruptive acinar morphogenesis and cellular changes similar to epithelial-to-mesenchymal transition in normal mammary epithelial cell, MCF10A. As one of the critical O-glycan substrates, we identified fibronectin that was O-glycosylated in vivo and thereby stabilized by GALNT6. Because knockdown of fibronectin abrogated the disruptive proliferation caused by introduction of GALNT6 into epithelial cells, our findings suggest that GALNT6-fibronectin pathway should be a critical component for breast cancer development and progression.


Five-Month Follow-up Shows No Improvement in Dermatological Complications in Children With Type 1 Diabetes Using Continuous Glucose Monitoring Systems and Insulin Pumps.
A Danish study showed that 90% of the pediatric patients who participated had some time experienced dermatological complications due to treatment with continuous subcutaneous insulin infusion (CSII). This follow-up study describes dermatological complications due to CSII and/or continuous glucose monitoring (CGM) between the two study periods and includes health-related quality of life (HrQoL) measurements. A total of 138 patients (95%) out of 145 patients from the initial study answered an online questionnaire regarding dermatological complications related to CSII and/or CGM, five months later. A second questionnaire (DISABKIDS) regarding HrQoL was sent out to those 138 of which 111 patients completed it. The patients were aged from 2 to 20 years. Descriptive statistics, χ2 tests, and univariate and multivariate analyses were used to analyze the data. In total, 81% of the 138 patients continued to have dermatological complications at follow-up. Itching was the most frequently reported complication. Patients using Enlite reported more dermatological complications than those using Libre. In total, 79% of the patients who used barrier cream in the initial study still had dermatological complications five months later. Age, gender, Body Mass Index (BMI), or HbA1c levels showed no significant association with dermatological complications. Patients who perceived dermatological complications as a greater problem had lower HrQoL scores. Once dermatological complications start to appear, they become chronic, indicating that the treatments currently available are inadequate. Patients perceiving dermatological complications as a greater problem were associated with lower HrQoL. These findings highlight the need for additional preventive studies.


Inhomogeneous high harmonic generation in krypton clusters.
High order harmonic generation from clusters is a controversial topic: conflicting theories exist, with different explanations for similar experimental observations. From an experimental point of view, separating the contributions from monomers and clusters is challenging. By performing a spectrally and spatially resolved study in a controlled mixture of clusters and monomers, we are able to isolate a region of the spectrum where the emission purely originates from clusters. Surprisingly, the emission from clusters is depolarized, which is the signature of statistical inhomogeneous emission from a low-density source. The harmonic response to laser ellipticity shows that this generation is produced by a new recollisional mechanism, which opens the way to future theoretical studies.


Bombing alone: tracing the motivations and antecedent behaviors of lone-actor terrorists,.
This article analyzes the sociodemographic network characteristics and antecedent behaviors of 119 lone-actor terrorists. This marks a departure from existing analyses by largely focusing upon behavioral aspects of each offender. This article also examines whether lone-actor terrorists differ based on their ideologies or network connectivity. The analysis leads to seven conclusions. There was no uniform profile identified. In the time leading up to most lone-actor terrorist events, other people generally knew about the offender's grievance, extremist ideology, views, and/or intent to engage in violence. A wide range of activities and experiences preceded lone actors' plots or events. Many but not all lone-actor terrorists were socially isolated. Lone-actor terrorists regularly engaged in a detectable and observable range of activities with a wider pressure group, social movement, or terrorist organization. Lone-actor terrorist events were rarely sudden and impulsive. There were distinguishable behavioral differences between subgroups. The implications for policy conclude this article.


The effect of the late 2000s financial crisis on suicides in Spain: an interrupted time-series analysis.
The current financial crisis is having a major impact on European economies, especially that of Spain. Past evidence suggests that adverse macro-economic conditions exacerbate mental illness, but evidence from the current crisis is limited. This study analyses the association between the financial crisis and suicide rates in Spain. An interrupted time-series analysis of national suicides data between 2005 and 2010 was used to establish whether there has been any deviation in the underlying trend in suicide rates associated with the financial crisis. Segmented regression with a seasonally adjusted quasi-Poisson model was used for the analysis. Stratified analyses were performed to establish whether the effect of the crisis on suicides varied by region, sex and age group. The mean monthly suicide rate in Spain during the study period was 0.61 per 100 000 with an underlying trend of a 0.3% decrease per month. We found an 8.0% increase in the suicide rate above this underlying trend since the financial crisis (95% CI: 1.009-1.156; P = 0.03); this was robust to sensitivity analysis. A control analysis showed no change in deaths from accidental falls associated with the crisis. Stratified analyses suggested that the association between the crisis and suicide rates is greatest in the Mediterranean and Northern areas, in males and amongst those of working age. The financial crisis in Spain has been associated with a relative increase in suicides. Males and those of working age may be at particular risk of suicide associated with the crisis and may benefit from targeted interventions.


The floppy mitral valve. Study on pathogenesis.
The pathogenesis of the floppy valve syndrome is not fully solved. An almost invariable histological finding is the great accumulation of mucinous material in the valve leaflets and constitutes the basis of the valvular theory of the syndrome. The presence of a mucinous layer in normal valves-- the zona spongiosa--is not well recognised. To establish the normal range of the extent of this zone, 50 excised mitral valves from patients aged 2 to 89 years and who died as a result of road traffic accidents or non-cardiac causes have been analysed by measuring the thickness of the zone in relation to the valve thickness. A range of 0 to 60 per cent was found and this was not influenced by age. The findings were compared with 50 patients clinically diagnosed as suffering from the floppy valve syndrome. A value of over 60 per cent (range 62 to 94%) was found in 43 patients. The increase in the extent of the mucinous material was considered to be a secondary change in the thickened fibrosa which normally accompanies the floppy valve syndrome. Measurements of zona spongiosa falling within the normal range were found in seven patients. The clinical features, complications, and accompanying conditions have also been analysed. Chordal rupture had occurred in 20 patients, infective endocarditis in three, and calcification was found in four valves. In four patients the aortic valve was also involved and accompanying aortic root dilatation in an additional patient. It is suggested that these patients should not be included in the group of Marfan's forme fruste, nor in the typical floppy mitral valve syndrome. Apart from the valvular theory, the myocardial theory in the pathogenesis of the syndrome has been discussed and the components ensuring normal mitral valve function have been reviewed. It is concluded that an inherent, prominent zona spongiosa predisposes to the floppy valve syndrome, particularly if any one of the components of normal valve function is abnormal.


19F and 1H MRI detection of amyloid beta plaques in vivo.
Formation of senile plaques composed of amyloid beta peptide, a pathological hallmark of Alzheimer disease, in human brains precedes disease onset by many years. Noninvasive detection of such plaques could be critical in presymptomatic diagnosis and could contribute to early preventive treatment strategies. Using amyloid precursor protein (APP) transgenic mice as a model of amyloid beta amyloidosis, we demonstrate here that an intravenously administered (19)F-containing amyloidophilic compound labels brain plaques and allows them to be visualized in living mice by magnetic resonance imaging (MRI) using (19)F and (1)H. Our findings provide a new direction for specific noninvasive amyloid imaging without the danger of exposure to radiation. This approach could be used in longitudinal studies in mouse models of Alzheimer disease to search for biomarkers associated with amyloid beta pathology as well as to track disease course after treatment with candidate medications.


Tissue engineering of ligament and tendon healing.
Ligaments and tendons are bands of dense connective tissue that mediate normal joint movement and stability. Injury to these structures may result in significant joint dysfunction because they either heal by production of inferior matrix or do not heal at all. The process of ligament and tendon healing is complex and the roles of cellular and biochemical mediators continue to be elucidated. The expression of growth factors and growth factor receptors is modulated after injury, and cells from healing tissues are responsive to growth factors. Tissue engineering offers the potential to improve the quality of ligaments and tendons during the healing process. The concept is based on the manipulation of cellular and biochemical mediators to affect protein synthesis and improve tissue remodeling. Recently, novel techniques such as application of growth factors, gene transfer techniques, and cell therapy have shown promise and may become effective biologic therapies in the future. Many groups have been successful in introducing marker and therapeutic genes into ligaments and tendons. Cell therapy involves the introduction of mesenchymal progenitor cells as a pluripotent cell source into the healing environment. The combination of cell therapy with growth factor application via gene transfer offers new avenues to improve ligament and tendon healing.


Teriflunomide-exposed pregnancies in a French cohort of patients with multiple sclerosis.
To describe pregnancies exposed to teriflunomide (TERIF) in women with multiple sclerosis (MS) in France over the period 2014-2016. All 15- to 49-year-old women with MS in the national health insurance database were included. Pregnancies that had started between August 2014 and March 2016 were identified from their outcomes. Three groups according to treatment exposure were compared: TERIF, interferons (IFNs) or glatiramer acetate, and no medication. Among the 44,008 women with MS followed 24.5 months on average, 2,639 pregnancies were identified. There were 1,538 pregnancies (58.3%) that were not exposed to any MS treatment in accordance with the guidelines. A total of 673 pregnancies (25.5%) were exposed to IFN and/or glatiramer acetate, and possible or probable exposure to contra-indicated treatments was observed in 428 pregnancies (16.2%), of whom 47 pregnancies were exposed to TERIF. The annual incidence rate of pregnancies exposed to TERIF was 1.4 per 100 patient-years; i.e., 3 times less than the 2 control groups (5.6 and 4.7, respectively). The median exposure duration to TERIF was 45 days after conception. The outcomes comprised 23 live births, 22 abortions (3 times more than the 2 other groups), and 2 miscarriages. All newborns were healthy at birth. Despite specific TERIF guidelines for pregnancy-related issues and the availability of alternative therapies, some pregnancies exposed to TERIF were identified. Most of the cases were because of the absence of the recommended accelerated elimination procedure and appeared to be mostly unplanned pregnancies that probably reflect a lack of effective contraception.


High-throughput single-molecule studies of protein-DNA interactions.
Fluorescence and force-based single-molecule studies of protein-nucleic acid interactions continue to shed critical insights into many aspects of DNA and RNA processing. As single-molecule assays are inherently low-throughput, obtaining statistically relevant datasets remains a major challenge. Additionally, most fluorescence-based single-molecule particle-tracking assays are limited to observing fluorescent proteins that are in the low-nanomolar range, as spurious background signals predominate at higher fluorophore concentrations. These technical limitations have traditionally limited the types of questions that could be addressed via single-molecule methods. In this review, we describe new approaches for high-throughput and high-concentration single-molecule biochemical studies. We conclude with a discussion of outstanding challenges for the single-molecule biologist and how these challenges can be tackled to further approach the biochemical complexity of the cell.


Detection of tissue kallikrein in the bronchoalveolar lavage fluid of asthmatic subjects.
Kininogenase activity was detected by cleavage of radiolabeled substrate (125I-high molecular weight kininogen [HMWK]) in 22 of 24 bronchoalveolar lavage (BAL) fluid samples from 17 asthmatics who either responded to aerosolized allergen challenge or had symptoms of active asthma. In contrast, six of seven normal controls lacked enzymatic activity. Levels of free immunoreactive kinin found in BAL fluid correlated with the presence of kininogenase activity (P = 0.002). The cleavage pattern of 125I-HMWK by the BAL fluid kininogenase (a dominant 65,000-mol wt fragment), and synthetic inhibitor profile (phe-phe-arg-CH2Cl and phenylmethylsulfonyl fluoride) were compatible with a tissue kallikrein. Peak kininogenase activity eluted at an apparent molecular weight of 20,000-34,000 by HPLC gel filtration. Its antigenic identity was established by immunoblotting with anti-human urinary kallikrein antibody and its activity was inhibited by this antibody. Lysylbradykinin was generated during incubation of fractionated BAL fluid and purified HMWK, the characteristic cleavage product of the tissue kallikreins. We conclude that elevated amounts of tissue kallikrein and kinin are present in the bronchoalveolar spaces of asthmatic subjects. Kinin generation may contribute to the asthmatic response directly through edema formation and smooth muscle contraction and by augmenting release and/or production of preformed (histamine) and secondary mediators such as leukotrienes and platelet-activating factor.


Geographic Access to Specialty Mental Health Care Across High- and Low-Income US Communities.
With the future of the Affordable Care Act and Medicaid program unclear, it is critical to examine the geographic availability of specialty mental health treatment resources that serve low-income populations across local communities. To examine the geographic availability of community-based specialty mental health treatment resources and how these resources are distributed by community socioeconomic status. Measures of the availability of specialty mental health treatment resources were derived using national data for 31 836 zip code tabulation areas from 2013 to 2015. Analyses examined the association between community socioeconomic status (assessed by median household income quartiles) and resource availability using logistic regressions. Models controlled for zip code tabulation area-level demographic characteristics and state indicators. Dichotomous indicators for whether a zip code tabulation area had any (1) outpatient mental health treatment facility (more than nine-tenths of which offer payment arrangements for low-income populations), (2) office-based practice of mental health specialist physician(s), (3) office-based practice of nonphysician mental health professionals (eg, therapists), and (4) mental health facility or office-based practice (ie, any community-based resource). Of the 31 836 zip code tabulation areas in the study, more than four-tenths (3382 of 7959 [42.5%]) of communities in the highest income quartile (mean income, $81 207) had any community-based mental health treatment resource vs 23.1% of communities (1841 of 7959) in the lowest income quartile (mean income, $30 534) (adjusted odds ratio, 1.74; 95% CI, 1.50-2.03). When examining the distribution of mental health professionals, 25.3% of the communities (2014 of 7959) in the highest income quartile had a mental health specialist physician practice vs 8.0% (637 of 7959) of those in the lowest income quartile (adjusted odds ratio, 3.04; 95% CI, 2.53-3.66). Similarly, 35.1% of the communities (2792 of 7959) in the highest income quartile had a nonphysician mental health professional practice vs 12.9% (1029 of 7959) of those in the lowest income quartile (adjusted odds ratio, 2.77; 95%, 2.35-3.26). In contrast, outpatient mental health treatment facilities were less likely to be located in the communities in the highest vs lowest income quartiles (12.9% [1025 of 7959] vs 16.5% [1317 of 7959]; adjusted odds ratio, 0.43; 95% CI, 0.37-0.51). More than seven-tenths of the lowest income communities with any resource (71.5% [1317 of 1841]) had an outpatient mental health treatment facility. Mental health treatment facilities are more likely to be located in poorer communities, whereas office-based practices of mental health professionals are more likely to be located in higher-income communities. These findings indicate that mental health treatment facilities constitute the backbone of the specialty mental health treatment infrastructure in low-income communities. Policies are needed to support and expand available resources for this critical infrastructure.


Investigation of sequon engineering for improved O-glycosylation by the human polypeptide N-acetylgalactosaminyl transferase T2 isozyme and two orthologues.
We have been developing bacterial expression systems for human mucin-type O-glycosylation on therapeutic proteins, which is initiated by the addition of α-linked GalNAc to serine or threonine residues by enzymes in the GT-27 family of glycosyltransferases. Substrate preference across different isoforms of this enzyme is influenced by isoform-specific amino acid sequences at the site of glycosylation, which we have exploited to engineer production of Core 1 glycan structures in bacteria on human therapeutic proteins. Using RP-HPLC with a novel phenyl bonded phase to resolve intact protein glycoforms, the effect of sequon mutation on O-glycosylation initiation was examined through in vitro modification of the naturally O-glycosylated human interferon α-2b, and a sequon engineered human growth hormone. As part of the development of our glycan engineering in the bacterial expression system we are surveying various orthologues of critical enzymes to ensure complete glycosylation. Here we present an in vitro enzyme kinetic profile of three related GT-27 orthologues on natural and engineered sequons in recombinant human interferon α2b and human growth hormone where we show a significant change in kinetic properties with the amino acid changes. It was found that optimizing the protein substrate amino acid sequence using Isoform Specific O-Glycosylation Prediction (ISOGlyP, http://isoglyp.utep.edu/index.php) resulted in a measurable increase in kcat/KM, thus improving glycosylation efficiency. We showed that the Drosophila orthologue showed superior activity with our human growth hormone designed sequons compared with the human enzyme.


Simultaneous ODF estimation and tractography in HARDI.
We consider the problem of tracking white matter fibers in high angular resolution diffusion imaging (HARDI) data while simultaneously estimating the local fiber orientation profile. Prior work showed that an unscented Kalman filter (UKF) can be used for this problem, yet existing algorithms employ parametric mixture models to represent water diffusion and to define the state space. To address this restrictive model dependency, we propose to extend the UKF to HARDI data modeled by orientation distribution functions (ODFs), a more generic diffusion model. We consider the spherical harmonic representation of the HARDI signal as the state, enforce nonnegativity of the ODFs, and perform tractography using the directions at which the ODFs attain their peaks. In simulations, our method outperforms filtered two-tensor tractography at different levels of noise by achieving a reduction in mean Chamfer error of 0.05 to 0.27 voxels; it also produced in vivo fiber tracking that is consistent with the neuroanatomy.


Oral mixing ability and cognition in elderly persons with dementia: a cross-sectional study.
Masticatory performance has been positively associated with cognitive ability in both animals and healthy humans. We hypothesised that there would also be a positive correlation between masticatory performance and cognition in older persons suffering from dementia. Older persons suffering from dementia (n = 114) and receiving institutionalised care were studied in a cross-sectional design. The assessments included masticatory performance, which was measured objectively with a two-colour gum mixing ability test, and cognition, which was assessed with a multidomain neuropsychological test battery. Significant relationships were observed between masticatory performance and general cognition and between masticatory performance and verbal fluency. Hierarchical regression analysis revealed that the correlation with general cognition was influenced by the scores for dependency in activities of daily living. The association between verbal fluency and masticatory performance was not significantly affected by secondary variables. An unexpected limitation of this study was the high dropout rate for the mixing ability test. The clinical implications of these findings are profound; care professionals should endeavour to maintain and stimulate mastication in older persons with dementia in an attempt to preserve cognition.


EUS-B for suspected left adrenal metastasis in lung cancer.
Several studies have reported the efficacy of esophageal ultrasound-guided fine needle aspiration (EUS-FNA) for the detection of metastases in the left adrenal gland (LAG) in patients with lung cancer. Currently we have only limited evidence based on small studies on the usefulness of EUS-B [endobronchial ultrasound (EBUS) scope into the esophagus] to provide tissue proof of suspected LAG metastases. The objectives of this study are to investigate feasibility, safety and diagnostic yield of EUS-B-FNA in LAG analysis in patients with proven or suspected lung cancer. In two Danish hospitals, a systematic search in the electronic database for patients who underwent EUS-B-FNA of the LAG for suspected or proven lung cancer was performed retrospectively between January 1st, 2015 and December 31st, 2017. Computed tomography (CT), positron emission tomography-CT, endoscopy, pathology and follow-up data were acquired. One hundred and thirty-five patients were included; the prevalence of biopsy proven LAG malignancy was 30% (40/135). A total of 87% (117/135) of EUS-B-FNA samples were adequate (i.e., containing adrenal or malignant cells). No complications were observed. We present the largest cohort of patients ever reported showing that EUS-B-FNA of the LAG is a safe and feasible procedure and should therefore be used for staging purposes in patients with lung cancer and a suspicious LAG.


Are Toll-like receptors and decoy receptors involved in the immunopathogenesis of systemic lupus erythematosus and lupus-like syndromes?
In this paper we focus our attention on the role of two families of receptors, Toll-like receptors (TLR) and decoy receptors (DcR) involved in the generation of systemic lupus erythematosus (SLE) and lupus-like syndromes in human and mouse models. To date, these molecules were described in several autoimmune disorders such as rheumatoid arthritis, antiphospholipids syndrome, bowel inflammation, and SLE. Here, we summarize the findings of recent investigations on TLR and DcR and their role in the immunopathogenesis of the SLE.


Understanding change in clinical nursing practice.
The present paper employs a poststructural framework to explore the construction of change in clinical nursing practice, as portrayed in nursing literature. In it a critique of two specific bodies of literature is offered: that which deals with the 'abstract' conception of change (but which is heavily influenced by organizational discourses) and that which describes changes in the clinical setting of critical care nursing practice. It is argued that the discursive constitution of change that is revealed in this analysis disadvantages clinical nurses. Because the context in which change occurs is largely unacknowledged, clinical nursing practice, as experienced and understood by nurses, is also marginalized. Change in clinical practice is under-researched, and such inquiry should be undertaken from the perspective of nurses themselves, and requires an exploration of the discursive context of nursing work.


To pool or not to pool, from whether to when: applications of pooling to biospecimens subject to a limit of detection.
Pooling of biological specimens has been utilised as a cost-efficient sampling strategy, but cost is not the unique limiting factor in biomarker development and evaluation. We examine the effect of different sampling strategies of biospecimens for exposure assessment that cannot be detected below a detection threshold (DT). The paper compares use of pooled samples to a randomly selected sample from a cohort in order to evaluate the efficiency of parameter estimates. The proposed approach shows that a pooling design is more efficient than a random sample strategy under certain circumstances. Moreover, because pooling minimises the amount of information lost below the DT, the use of pooled data is preferable (in a context of a parametric estimation) to using all available individual measurements, for certain values of the DT. We propose a combined design, which applies pooled and unpooled biospecimens, in order to capture the strengths of the different sampling strategies and overcome instrument limitations (i.e. DT). Several Monte Carlo simulations and an example based on actual biomarker data illustrate the results of the article.


A phase I trial of two sequence-specific schedules of decitabine and vorinostat in patients with acute myeloid leukemia.
This phase I trial evaluated two schedules of escalating vorinostat in combination with decitabine every 28 days: (i) sequential or (ii) concurrent. There were three dose-limiting toxicities: grade 3 fatigue and generalized muscle weakness on the sequential schedule (n = 1) and grade 3 fatigue on the concurrent schedule (n = 2). The maximum tolerated dose was not reached on both planned schedules. The overall response rate (ORR) was 23% (three complete response [CR], two CR with incomplete incomplete blood count recovery [CRi], one partial response [PR] and two morphological leukemic free state [MLFS]). The ORR for all and previously untreated patients in the sequential arm was 13% (one CRi; one MLFS) and 0% compared to 30% (three CR; one CRi; one PR; one MLFS) and 36% in the concurrent arm (p = 0.26 for both), respectively. Decitabine plus vorinostat was safe and has clinical activity in patients with previously untreated acute myeloid leukemia. Responses appear higher with the concurrent dose schedule. Cumulative toxicities may limit long-term usage on the current dose/schedules.


In situ identification of bacteria in drinking water and adjoining biofilms by hybridization with 16S and 23S rRNA-directed fluorescent oligonucleotide probes.
Free-water-phase and surface-associated microorganisms from drinking water were detected and roughly identified by hybridization with fluorescence-labeled oligonucleotide probes complementary to regions of 16S and 23S rRNA characteristic for the domains Bacteria, Archaea, and Eucarya and the beta and gamma subclasses of Proteobacteria. Samples of glass-attached biofilms and plankton were taken from a Robbins device installed in a water distribution system. More than 70% of the surface-associated cells and less than 40% of the planktonic cells visualized by 4',6-diamidino-2-phenylindole staining bound detectable amounts of rRNA-targeted probes. These findings are an indication for higher average rRNA content and consequently higher physiological activity of the attached microbial cells compared with the free-living cells. All detectable cells hybridized with the bacterial probe, whereas no Archaea and no Eucarya cells could be detected. Simultaneous hybridization with probes specific for the beta and gamma subclasses of Proteobacteria revealed that microcolonies already consisted of mixed populations in early stages with fewer than 50 cells. These observations provide further evidence that the coexistence and interaction of bacteria in drinking water biofilms may be an integral part of their growth and survival strategies.


Cerebral outcome after extracorporeal circulation. Comparison between prospective and retrospective evaluations.
The cerebral outcome of 100 consecutive patients who had cardiac valvular replacement was evaluated by comparing the results of prospective neurological examination with retrospective data. The latter showed that the overall prevalence of cerebral abnormalities was 6% (4% among survivors) up to ten days postoperatively and 9% thereafter. This contrasts with the 35% (37% among survivors) obtained by careful neurological investigations that showed five patients with residual signs one year after operation. Electroencephalographic and neuropsychological studies disclosed additional patients who had subclinical involvement. The results question the reportedly ever-falling cerebral complication values claimed particularly in retrospective studies and reflect what is missed when using rough clinical criteria. There is no justification in overlooking slight clinical or even subclinical dysfunction, since the elimination of them is the only acceptable criterion of cerebral safety in cardiac operations.


Real-time kinetic studies of Mycobacterium tuberculosis LexA-DNA interaction.
Transcriptional repressor, LexA, regulates the 'SOS' response, an indispensable bacterial DNA damage repair machinery. Compared with its Escherichia coli ortholog, LexA from Mycobacterium tuberculosis (Mtb) possesses a unique N-terminal extension of additional 24 amino acids in its DNA-binding domain (DBD) and 18 amino acids insertion at its hinge region that connects the DBD to the C-terminal dimerization/autoproteolysis domain. Despite the importance of LexA in 'SOS' regulation, Mtb LexA remains poorly characterized and the functional importance of its additional amino acids remained elusive. In addition, the lack of data on kinetic parameters of Mtb LexA-DNA interaction prompted us to perform kinetic analyses of Mtb LexA and its deletion variants using Bio-layer Interferometry (BLI). Mtb LexA is seen to bind to different 'SOS' boxes, DNA sequences present in the operator regions of damage-inducible genes, with comparable nanomolar affinity. Deletion of 18 amino acids from the linker region is found to affect DNA binding unlike the deletion of the N-terminal stretch of extra 24 amino acids. The conserved RKG motif has been found to be critical for DNA binding. Overall, the present study provides insights into the kinetics of the interaction between Mtb LexA and its target 'SOS' boxes. The kinetic parameters obtained for DNA binding of Mtb LexA would be instrumental to clearly understand the mechanism of 'SOS' regulation and activation in Mtb.


A kinematics-based method for creating deformed patient-derived head and neck CT scans.
CT scans of the head and neck have multiple clinical uses, and simulating deformation of these CT scans allows for predicting patient motion and data augmentation for machine-learning methods. Current methods for creating patient-derived deformed CT scans require multiple scans or use unrealistic head and neck motion. This paper describes the CTHeadDeformation software package which allows for realistic synthetic deformation of head and neck CT scans for small amounts of motion. CTHeadDeformation is a python-based package that uses a kinematics-based approach using anatomical landmarks, and rigid/non-rigid registration to create a realistic patient-derived deformed CT scan. CTHeadDeformation is also designed for simple clinical implementation. The CTHeadDeformation software package was demonstrated on a head and neck CT scan of one patient. The CT scan was deformed in the anterior-posterior, superior-inferior, and left-right directions. Internal organ motion and more complex combination motions were also simulated. The results showed the patient's CT scan was able to be deformed in a way that preserved the shape and location of the anatomy.Clinical Relevance- This method allows for the realistic simulation of head and neck motion in CT scans. Clinical applications including simulating how patient motion affects radiation therapy treatment effectiveness. The CTHeadDeformation software can also be used to train machine-learning networks that are robust to patient motion, or to generate ground truth images for imaging or segmentation grand challenges.


Physico-mechanical and antimicrobial properties of an elastomeric ligature coated with reduced nanographene oxide-nano curcumin subjected to dual-modal photodynamic and photothermal inactivation against Streptococcus mutans biofilms.
White spot lesions (WSLs) are a common side effect of fixed orthodontic treatment. Streptococcus mutans is the primary causative agent of WSLs and dental caries on the teeth during treatment. According to the unique features of reduced graphene oxide-nano curcumin (rGO-nCur), this study aimed to investigate the mechanical properties and antimicrobial potency of rGO-nCur coated orthodontic elastomeric ligatures as a novel coating composite following dual-modal photodynamic inactivation (PDI) and photothermal inactivation (PTI) against S. mutans biofilms. After confirmation of rGO-nCur synthesis and coating elastomeric ligatures with different concentration levels of 1.25, 2.5, 5, 7.5, and 10 % of rGO-nCur, tensile strength, force decay, extension to tensile strength, and contact angle of the coated elastomeric ligatures were measured using universal testing machine and sessile drop method, respectively. To investigate the mechanism through which irradiated rGO-nCur can inhibit the formation of S. mutans biofilms, intracellular reactive oxygen species (ROS) generation, and increase in temperature of rGO-nCur solutions under the 450 and 980 nm laser irradiation, respectively, were measured. The anti-biofilm activity and inhibition of water-insoluble extracellular polysaccharide (EPS) production ability of irradiated rGO-nCur coated elastomeric ligatures using a 450 nm diode laser (195 J/cm2), a 980 nm diode laser (195 J/cm2), and a combination of both (78 J/cm2 of irradiation from each one) (i.e., PDI, PTI, and dual-modal PDI/PTI, respectively) were determined. Also, the expression of virulence genes involved in biofilm formation (comDE, gtfD, and smuT) was assessed by quantitative real-time polymerase chain reaction (RT-qPCR) following the mentioned treatment. One-way ANOVA test and Tukey post-hoc test at a p-value equal to/or less than 0.05 were used to analyze the obtained data. The synthesis of GO nano-sheets in a layered structure with a thickness of 0.76 nm was confirmed by AFM analysis. FESEM showed that the exfoliated sheet of synthesized GO had several micrometers in lateral size. DLS revealed that the mean particle size and density index of synthesized nCur were 57.47 ± 2.14 nm and 10 % respectively. In DLS analysis, rGO-nCur showed more positive surface charge (24 mV) than the nano-sheets of GO. FESEM confirmed the coating of rGO-nCur on elastomeric ligatures. ANOVA revealed that tensile strength of 1.25, 2.5, and 5 % rGO-nCur coated elastomeric ligatures were not decreased statistically significantly (P > 0.05). Mean tensile strength and recorded force of 7.5 and 10 % rGO-nCur coated elastomeric ligatures decreased significantly after 14 days' immersion in the artificial saliva (P < 0.05). On the 28th day of the study, the mean of the tensile strength of elastomeric ligatures coated with 10 % rGO-nCur (13.03 ± 0.10 N) was recorded as 55.90 % of the initial tensile strength (23.31 ± 0.41 N in uncoated elastomeric ligatures), while the mean tensile strength of elastomeric ligatures coated with 7.5 % rGO-nCur (16.01 ± 0.10 N) was measured as 68.94 % of the initial tensile strength (23.22 ± 0.09 N in uncoated elastomeric ligatures). When comparing the coated elastomeric ligatures at 7.5 % and 10 % to the original uncoated elastomeric ligatures at similar time intervals, statistically significant decreases in extension to tensile strength (0.42 to 0.71 mm or 3.02 to 5.05 %; all P < 0.05) were observed. The largest contact angle was measured in elastomeric ligatures coated by 10 % rGO-nCur followed by 7.5 and 5 % rGO-nCur (128 ± 2.19°, 117 ± 2.23°, and 99 ± 1.83°; respectively). The results revealed a rise of 6.4-fold in intracellular ROS and an 11.2 °C increase in the temperature of rGO-nCur solutions following the 450 nm and 980 nm laser irradiation, respectively. The 5 % rGO-nCur coated elastomeric ligature mediated dual-modal PDI/PTI showed the most inhibition of the biofilm formation of S. mutans by 83.62 % (P = 0.00). Significant reductions in water-insoluble EPS were detected in biofilm cultures of S. mutans on 1.25 % rGO-nCur coated elastomeric ligatures following irradiation with dual waves of the 450 nm and 980 nm diode lasers (i.e., dual-modal PDI/PTI; 96.17 %; P = 0.00). The expression levels of comDE, gtfD, and smuT virulence genes were significantly downregulated (7.52-, 13.92-, and 8.23-fold, respectively) in the biofilm cultures of S. mutans on 1.25 % rGO-nCur coated elastomeric ligatures following dual-modal PDI/PTI in comparison with biofilm cultures on non-coated elastomeric ligatures. 5 % rGO-nCur coated elastomeric ligatures following irradiation with dual waves of the 450 and 980 nm diode lasers (dual-modal PDI/PTI), without adverse effects on the physico-mechanical properties of elastomeric ligatures, can be used to inhibit the formation of S. mutans biofilms on the coated elastomeric ligatures around orthodontic brackets.


Projected Increases in Precipitation Are Expected To Reduce Nitrogen Use Efficiency and Alter Optimal Fertilization Timings in Agriculture in the South East of England.
Nitrogen fertilization is vital for productive agriculture and efficient land use. However, globally, approximately 50% of the nitrogen applied is lost to the environment, causing inefficiencies, pollution, and greenhouse gas emissions. Rainfall and its effect on soil moisture are the major components controlling nitrogen losses in agriculture. Thus, changing rainfall patterns could accelerate nitrogen inefficiencies. We used a mechanistic modeling platform to determine how precipitation-optimal nitrogen fertilization timings and resulting crop nitrogen uptake have changed historically (1950-2020) and how they are predicted to change under the RCP8.5 climate scenario (2021-2069) in the South East of England. We found that historically, neither precipitation-optimal fertilization timings nor resulting plant uptake changed significantly. However, there were large year-to-year variations in both. In the 2030s, where it is projected to get wetter, precipitation-optimal fertilization timings are predicted to be later in the season and the resulting plant uptake noticeably lower. After 2040, the precipitation-optimal uptakes are projected to increase with earlier precipitation-optimal timings closer to historical values, corresponding to the projected mean daily rainfall rates decreasing to the historical values in these growing seasons. It seems that the interannual variation in precipitation-optimal uptake is projected to increase. Ultimately, projected changes in precipitation patterns will affect nitrogen uptake and precipitation-optimal fertilization timings. We argue that the use of bespoke fertilization timings in each year can help recuperate the reduced N uptake due to changing precipitation.


Preclinical development of a recombinant toxin containing circularly permuted interleukin 4 and truncated Pseudomonas exotoxin for therapy of malignant astrocytoma.
Effective treatment is lacking for malignant glioblastoma/astrocytoma. We have identified interleukin-4 receptors (IL-4R) on human malignant astrocytoma. We demonstrate that 16 of 21 surgical samples of high-grade astrocytoma and glioblastoma but not normal brain tissues expressed IL-4R as assessed by reverse transcriptase PCR. We further demonstrate that human malignant astrocytoma cell lines express high-affinity IL-4R. Using a chimeric protein composed of circularly permuted IL-4 and a truncated form of Pseudomonas exotoxin A, we observed that this toxin IL4(38-37)-PE38KDEL) is highly cytotoxic to IL-4R-bearing glioblastoma cells. Compared with a previously reported IL4-PE chimeric protein (IL-PE4E), IL4(38-37)-PE38KDEL bound with higher affinity and was 3-30-fold more cytotoxic to glioblastoma cell lines. Upon intrathecal administration in monkeys, high cerebrospinal fluid IL4(38-37)-PE38KDEL levels were achieved using 2- and 6-microg/kg doses without any central nervous system or other abnormalities. IL4(38-37)-PE38KDEL levels were not detectable in the serum of any monkey studied. When IL4(38-37)-PE38KDEL was injected into the right frontal cortex of rats, localized necrosis was observed at 1000-ng/ml doses but not at < or = 100-ng/ml doses. We conclude that by localized administration, nontoxic levels of IL4(38-37)-PE38KDEL can be achieved, which may have significant cytotoxic activity against malignant astrocytoma.


Association of a missense mutation in the bovine leptin gene with carcass fat content and leptin mRNA levels.
Previously, we have shown that alleles of the BM1500 microsatellite, located 3.6 kb downstream of the leptin gene in cattle, were associated with carcass fat measures in a population of 154 unrelated beef bulls. Subsequently, a cytosine (C) to thymine (T) transition that encoded an amino acid change of an arginine to a cysteine was identified in exon 2 of the leptin gene. A PCR-RFLP was designed and allele frequencies in four beef breeds were correlated with levels of carcass fat. The T allele was associated with fatter carcasses and the C allele with leaner carcasses. The frequencies of the SNP alleles among breeds indicated that British breeds have a higher frequency of the T allele whereas the continental breeds have a higher occurrence of the C allele. A ribonuclease protection assay was developed to quantify leptin mRNA in a separate group of animals selected by genotype. Animals homozygous for thymine expressed higher levels of leptin mRNA. This may suggest that the T allele, which adds an extra cysteine to the protein, imparts a partial loss of biological function and hence could be the causative mutation.


The effect of ultrasound on lipase-catalyzed regioselective acylation of mangiferin in non-aqueous solvents.
A simple and efficient method for regioselective acylation of mangiferin catalyzed by lipase under ultrasound irradiation is reported. Compared with the conventional methods, its main advantages are shorter reaction time and higher yields. The optimum conditions were screened out. Under the optimal conditions (lipase: PCL, acyl donor: vinyl acetate; reaction solvent: DMSO, reaction temperature: 45 degrees C, ultrasonic power: 200 W; substrate ratio: acyl donor/mangiferin 6/1, enzyme loading: 6 mg/ml), the regioselective acylation yield was up to 84%.


The influence of dust originated in different coal mines on pneumoconiotic changes in white rats.
In experiments on white rats, three different types of dust of equal dispersion, coming from coal mines, were examined. The dusts exhibited different content of free silica and certain metals, such as: nickel, chromium and vanadium. The animals intratracheally administered with 50 mg of the mentioned dusts had hydroxyproline determined. After six months of experiment, histopathological examination of lungs was carried out. It was found that different SiO2 content (5 and 11%) with the same content of trace metals and different content of trace metals with the same SiO2 content (5%) does not affect hydroxyproline content in lungs. A significant increase in hydroxyproline level in rats' lungs was found in a group that obtained dust of the highest SiO2 content (11%) with more trace metals. Findings of histopathological examinations confirmed the data obtained from hydroxyproline levels measurements. It seems that compounds of certain metals, such as: nickel, chromium, vanadium, at a specific SiO2 level in the dust may increase its fibrogenic properties in lungs.


Human epidermal growth factor receptor 2 assessment in a case-control study: comparison of fluorescence in situ hybridization and quantitative reverse transcription polymerase chain reaction performed by central laboratories.
The optimal method to assess human epidermal growth factor receptor 2 (HER2) status remains highly controversial. Before reporting patient HER2 results, American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines mandate that laboratories demonstrate ≥ 95% concordance to another approved laboratory or methodology. Here, we compare central laboratory HER2 assessed by fluorescence in situ hybridization (FISH) and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using Oncotype DX in lymph node-negative, chemotherapy-untreated patients from a large Kaiser Permanente case-control study. Breast cancer specimens from the Kaiser-Genomic Health study were examined. Central FISH assessment of HER2 amplification and polysomy 17 was conducted by PhenoPath Laboratories (ratios > 2.2, 1.8 to 2.2, and < 1.8 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). HER2 expression by RT-PCR was conducted using Oncotype DX by Genomic Health (normalized expression units ≥ 11.5, 10.7 to < 11.5, and < 10.7 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). Concordance analyses followed ASCO/CAP guidelines. HER2 concordance by central FISH and central RT-PCR was 97% (95% CI, 96% to 99%). Twelve percent (67 of 568 patients) and 11% (60 of 568 patients) of patients were HER2 positive by RT-PCR and FISH, respectively. HER2-positive patients had increased odds of dying from breast cancer compared with HER2-negative patients. Polysomy 17 was demonstrated in 12.5% of all patients and 33% of FISH-positive patients. Nineteen of 20 FISH-positive patients with polysomy 17 were also RT-PCR HER2 positive. Although not statistically significantly different, HER2-positive/polysomy 17 patients tended to have the worst prognosis, followed by HER2-positive/eusomic, HER2-negative/polysomy 17, and HER2-negative/eusomic patients. There is a high degree of concordance between central FISH and quantitative RT-PCR using Oncotype DX for HER2 status, and the assay warrants additional study in a trastuzumab-treated population.


Congenital erosive and vesicular dermatosis with reticulated supple scarring: unifying clinical features.
Congenital erosive and vesicular dermatosis (CEVD) healing with reticulated supple scarring, a condition usually observed in premature neonates, presents at birth with vesicles and erosions. Lesions typically heal within a few months, leaving behind scarring with a distinctive supple and reticulated texture. We sought to merge existing literature with new cases to further define CEVD. We analyzed 19 previous reports of CEVD and added 9 additional patients; we identified unifying characteristics of this cohort. In 28 total cases, notable features included: preterm birth (79%), nail abnormalities (46%), hyperthermia/hypohidrosis (46%), a history of maternal chorioamnionitis (43%), alopecia (43%), neurodevelopmental and ophthalmologic abnormalities (36% each), tongue atrophy (29%), or a combination of these. Patients with CEVD may be prone to postnatal herpetic superinfections. Previously unreported findings included: erosive lichen planus, digital tip gangrene, and hydronephrosis. The small patient sampling makes it difficult to define diagnostic criteria. As certain findings are associated with prematurity, it is unclear to what extent these features result from CEVD, premature birth, or another intrauterine pathology. Although rare, CEVD should be considered in the differential diagnosis of neonatal vesicles/erosions in the context of a negative infectious workup. This review strengthens the spectrum of CEVD features, thus facilitating its recognition by clinicians.


Directed connectivity of brain default networks in resting state using GCA and motif.
Nowadays, there is a lot of interest in assessing functional interactions between key brain regions. In this paper, Granger causality analysis (GCA) and motif structure are adopted to study directed connectivity of brain default mode networks (DMNs) in resting state. Firstly, the time series of functional magnetic resonance imaging (fMRI) data in resting state were extracted, and the causal relationship values of the nodes representing related brain regions are analyzed in time domain to construct a default network. Then, the network structures were searched from the default networks of controls and patients to determine the fixed connection mode in the networks. The important degree of motif structures in directed connectivity of default networks was judged according to p-value and Z-score. Both node degree and average distance were used to analyze the effect degree an information transfer rate of brain regions in motifs and default networks, and efficiency of the network. Finally, activity and functional connectivity strength of the default brain regions are researched according to the change of energy distributions between the normals and the patients' brain regions. Experimental results demonstrate that, both normal subjects and stroke patients have some corresponding fixed connection mode of three nodes, and the efficiency and power spectrum of the patient's default network is somewhat lower than that of the normal person. In particular, the Right Posterior Cingulate Gyrus (PCG.R) has a larger change in functional connectivity and its activity. The research results verify the feasibility of the application of GCA and motif structure to study the functional connectivity of default networks in resting state.


Effects of phosphorylation on Drp1 activation by its receptors, actin, and cardiolipin.
Drp1 is a dynamin family GTPase that is required for mitochondrial and peroxisomal division, in which it oligomerizes into a ring and constricts the underlying membrane in a GTP hydrolysis-dependent manner. Oligomerization increases Drp1 GTPase activity through interactions between neighboring GTPase domains. In cells, Drp1 is regulated by several factors including Drp1 receptors, actin filaments, cardiolipin, and phosphorylation at two sites: S579 and S600. Phosphorylation of S579 is widely regarded as activating, while S600 phosphorylation is commonly considered inhibiting. However, the direct effects of phosphorylation on Drp1 GTPase activity have not been investigated in detail. In this study, we compare the effects of S579 and S600 phosphorylation on purified Drp1, using phospho-mimetic mutants and in vitro phosphorylation. The oligomerization state of both phospho-mimetic mutants is shifted toward smaller oligomers. Both phospho-mimetic mutations maintain basal GTPase activity, but eliminate GTPase stimulation by actin and decrease GTPase stimulation by cardiolipin, Mff, and MiD49. Phosphorylation of S579 by Erk2 produces similar effects. When mixed with wild-type Drp1, both S579D and S600D phospho-mimetic mutants reduce the actin-stimulated GTPase activity of Drp1-WT. Conversely, a Drp1 mutant that lacks GTPase activity, the K38A mutant, stimulates Drp1-WT GTPase activity under both basal and actin-stimulated conditions, similar to previous results for dynamin-1. These results suggest that the effect of S579 phosphorylation is not to activate Drp1 directly, and likely requires additional factors for stimulation of mitochondrial fission in cells. In addition, our results suggest that nearest neighbor interactions within the Drp1 oligomer affect catalytic activity.


Single-Nucleotide Polymorphisms Promote Dysregulation Activation by Essential Gene Mediated Bio-Molecular Interaction in Breast Cancer.
Breast cancer (BRCA) is a malignant tumor with a high mortality rate and poor prognosis in patients. However, understanding the molecular mechanism of breast cancer is still a challenge. In this study, we constructed co-expression networks by weighted gene co-expression network analysis (WGCNA). Gene-expression profiles and clinical data were integrated to detect breast cancer survival modules and the leading genes related to prognostic risk. Finally, we introduced machine learning algorithms to build a predictive model aiming to discover potential key biomarkers. A total of 42 prognostic modules for breast cancer were identified. The nomogram analysis showed that 42 modules had good risk assessment performance. Compared to clinical characteristics, the risk values carried by genes in these modules could be used to classify the high-risk and low-risk groups of patients. Further, we found that 16 genes with significant differential expressions and obvious bridging effects might be considered biological markers related to breast cancer. Single-nucleotide polymorphisms on the CYP24A1 transcript induced RNA structural heterogeneity, which affects the molecular regulation of BRCA. In addition, we found for the first time that ABHD11-AS1 was significantly highly expressed in breast cancer. We integrated clinical prognosis information, RNA sequencing data, and drug targets to construct a breast cancer-related risk module. Through bridging effect measurement and machine learning modeling, we evaluated the risk values of the genes in the modules and identified potential biomarkers for breast cancer. The protocol provides new insight into deciphering the molecular mechanism and theoretical basis of BRCA.


Low concentrations of LL-37 alter IL-8 production by keratinocytes and bronchial epithelial cells in response to proinflammatory stimuli.
The immunomodulatory cationic host defence peptide LL-37 plays an important role in epithelial innate immunity; at higher concentrations (20-50 microg mL(-1)) associated with inflammation, LL-37 elicits the production of cytokines and chemokines. It was demonstrated here that lower, physiologically relevant LL-37 concentrations (2-3 microg mL(-1)) altered epithelial cell responses to proinflammatory stimuli. In combination with interleukin-1beta (IL-1beta) and the Toll-like receptor-5 (TLR5) agonist flagellin, these low concentrations of LL-37 synergistically increased IL-8 production by both proliferating and differentiated keratinocytes and by bronchial epithelial cells. In combination with the TLR2/1 agonist PAM3CSK4, LL-37 synergistically induced transcription and the release of both IL-8 and IL-6 from primary bronchial epithelial cells; the IL-8 response was demonstrated to be regulated by epidermal growth factor receptor signalling. Treatment of bronchial epithelial cells with LL-37 and the TLR3 agonist polyI:C resulted in synergistic increases in IL-8 release and cytotoxicity. These data indicate that low concentrations of LL-37 may alter epithelial responses to infecting microorganisms in vivo.


Risk of hospitalizations/emergency department visits and treatment costs associated with initial maintenance therapy using fluticasone propionate 500 microg/salmeterol 50 microg compared with ipratropium for chronic obstructive pulmonary disease in older adults.
Cost-effective treatments for chronic obstructive pulmonary disease (COPD) are needed to reduce the burden on the Medicare system. The objectives of this retrospective study were to compare the effects of fluticasone propionate 500 microg/salmeterol 50 microg (FSC 500/50) with those of ipratropium (IPR) in older adult patients with COPD on the following factors: (1) the risk of all-cause and COPD-related hospitalization and emergency department (ED) visits; and (2) all-cause and COPD-related treatment costs. This retrospective, observational cohort study assessed commercially insured patients aged > or =65 years (Medicare eligible) with COPD-related medical costs (International Classification of Diseases, Ninth Revision, Clinical Modification codes 490.xx, 491.xx, 492.xx, or 496.xx), 18 months of continuous enrollment (12 months' pre-study and 6 months' poststudy), and > or =1 prescription claim (ie, the index event) for FSC 500/50 or IPR between January 2, 2001, and August 12, 2003. Exacerbation events (hospitalizations/ED visits) were assessed using Cox proportional hazards regression models controlling for baseline factors and preindex events. Treatment costs were estimated using multivariate generalized linear models to adjust for baseline characteristics and preindex utilization and costs. A propensity-matched comparison was conducted as a sensitivity analysis. A total of 1051 patients (540 women, 511 men) were identified: 952 in the IPR group (mean age, 74.26 years) and 99 in the FSC 500/50 group (mean age, 72.38 years). Treatment with FSC 500/50 was not associated with a reduction in the risk of all-cause hospitalization or ED visit compared with IPR (adjusted hazards ratio [HR], 0.913 [95% CI, 0.673-1.238]); however, FSC 500/50 was associated with a significantly lower risk (45%) of a COPD-related event (adjusted HR, 0.547 [95% CI, 0.301-0.995]). Although treatment with FSC 500/50 was associated with significantly lower annual mean all-cause medical costs compared with IPR ($18,642 vs $25,556, P < 0.05), the use of FSC 500/50 was also associated with significantly higher annual all-cause pharmacy costs ($2813 vs $2244, P < 0.05). Compared with the IPR cohort, the FSC 500/50 cohort had lower mean annual COPD-related medical costs (-$464 [P = NS]) and higher COPD-related pharmacy costs ($260 [P < 0.01]). However, increased pharmacy costs may be compensated for by decreased medical costs in both all-cause and COPD-related services. Results from the sensitivity analysis with the propensity score-matched sample of 194 patients were similar. Compared with the IPR cohort, the FSC 500/50 cohort was 45% less likely to have a COPD-related exacerbation event and had similar medical costs. FSC 500/50 was a more effective initial maintenance therapy than IPR for this Medicare population, and, despite the $260 increase in COPD-related pharmacy costs, there was no significant difference in COPD-related medical costs.


Feasibility of a randomized trial of extended lymphadenectomy for pancreatic cancer.
The required sample size of a prospective randomized trial comparing standard pancreaticoduodenectomy with pancreaticoduodenectomy plus extended lymphadenectomy for pancreatic adenocarcinoma is prohibitively large, making such a trial infeasible. Retrospective cohort study. Comprehensive cancer center. We identified 158 patients who underwent pancreaticoduodenectomy for pancreatic adenocarcinoma with separate pathologic analysis of second-echelon lymph nodes, defined as lymph nodes along the proximal hepatic artery and/or the great vessels. To estimate the sample size required for a randomized trial, we devised a biostatistical model with the following assumptions: extended lymphadenectomy can benefit only patients who (1) actually have disease removed from second-echelon nodes, (2) have microscopically negative (R0) primary tumor resection margins, and (3) do not have visceral metastatic (M0) disease. Seventy-six patients (48.1%) had negative first- and second-echelon lymph nodes, 65 (41.1%) had positive first-echelon and negative second-echelon lymph nodes, and 17 (10.8%) had positive first- and second-echelon lymph nodes. Patients with positive second-echelon lymph nodes had an R0 resection rate of 47.1%. At a median follow-up of 65.1 months, 4 patients with positive second-echelon lymph nodes were alive, but 3 had recurrent disease. This implies that only 1 patient (5.9%) with positive second-echelon lymph nodes may have had true M0 disease. Therefore, only 0.3% of patients (10.8% with positive second-echelon lymph nodes x 47.1% with R0 resection x 5.9% with M0 disease) may achieve a survival benefit from extended lymphadenectomy. A randomized trial of standard pancreaticoduodenectomy vs pancreaticoduodenectomy with extended lymphadenectomy would require 202 000 patients in each study arm to detect such a small difference. Definitive evaluation of the potential benefits of extended lymphadenectomy would require a prohibitively large sample size. Adequately powered randomized trials to address the potential benefit of extended lymphadenectomy are infeasible.


Targeting GPR30 with G-1: a new therapeutic target for castration-resistant prostate cancer.
Castration-resistant prostate cancer (CRPC) is an advanced-stage prostate cancer (PC) associated with high mortality. We reported that G-1, a selective agonist of G protein-coupled receptor 30 (GPR30), inhibited PC cell growth by inducing G2 cell cycle arrest and arrested PC-3 xenograft growth. However, the therapeutic actions of G-1 and their relationships with androgen in vivo are unclear. Using the LNCaP xenograft to model PC growth during the androgen-sensitive (AS) versus the castration-resistant (CR) phase, we found that G-1 inhibited growth of CR but not AS tumors with no observable toxicity to the host. Substantial necrosis (approximately 65%) accompanied by marked intratumoral infiltration of neutrophils was observed only in CR tumors. Global transcriptome profiling of human genes identified 99 differentially expressed genes with 'interplay between innate and adaptive immune responses' as the top pathway. Quantitative PCR confirmed upregulation of neutrophil-related chemokines and inflammation-mediated cytokines only in the G-1-treated CR tumors. Expression of murine neutrophil-related cytokines also was elevated in these tumors. GPR30 (GPER1) expression was significantly higher in CR tumors than in AS tumors. In cell-based experiments, androgen repressed GPR30 expression, a response reversible by anti-androgen or siRNA-induced androgen receptor silencing. Finally, in clinical specimens, 80% of CRPC metastases (n=123) expressed a high level of GPR30, whereas only 54% of the primary PCs (n=232) showed high GPR30 expression. Together, these results provide the first evidence, to our knowledge, that GPR30 is an androgen-repressed target and G-1 mediates the anti-tumor effect via neutrophil-infiltration-associated necrosis in CRPC. Additional studies are warranted to firmly establish GPR30 as a therapeutic target in CRPC.


Serotonin exerts a direct modulatory role on bladder afferent firing in mice.
Functional disorders (i.e. interstitial cystitis/painful bladder syndrome and irritable bowel syndrome) are associated with hyperexcitability of afferent nerves innervating the urinary tract and the bowel, respectively. Various non-5-HT3 receptor mRNA transcripts are expressed in mouse urothelium and exert functional responses to 5-HT. Whilst 5-HT3 receptors were not detected in mouse urothelium, 5-HT3 receptors expressed on bladder sensory neurons plays a role in bladder afferent excitability both under normal conditions and in a mouse model of chronic visceral hypersensitivity. These data suggest that the role 5-HT3 receptors play in bladder afferent signalling warrants further study as a potential therapeutic target for functional bladder disorders. Serotonin (5-HT) is an excitatory mediator that in the gastrointestinal (GI) tract plays a physiological role in gut-brain signalling and is dysregulated in functional GI disorders such as irritable bowel syndrome (IBS). Patients suffering from IBS frequently suffer from urological symptoms characteristic of interstitial cystitis/painful bladder syndrome, which manifests due to cross-sensitization of shared innervation pathways between the bladder and colon. However, a direct modulatory role of 5-HT in bladder afferent signalling and its role in colon-bladder neuronal crosstalk remain elusive. The aim of this study was to investigate the action of 5-HT on bladder afferent signalling in normal mice and mice with chronic visceral hypersensitivity (CVH) following trinitrobenzenesulfonic acid-induced colitis. Bladder afferent activity was recorded directly using ex vivo afferent nerve recordings. Expression of 14 5-HT receptor subtypes, the serotonin transporter (SERT) and 5-HT-producing enzymes was determined in the urothelium using RT-PCR. Retrograde labelling of bladder-projecting dorsal root ganglion neurons was used to investigate expression of 5-HT3 receptors using single cell RT-PCR, while sensory neuronal and urothelial responses to 5-HT were determined by live cell calcium imaging. 5-HT elicited bladder afferent firing predominantly via 5-HT3 receptors expressed on afferent terminals. CVH animals showed a downregulation of SERT mRNA expression in urothelium, suggesting increased 5-HT bioavailability. Granisetron, a 5-HT3 antagonist, reversed bladder afferent hypersensitivity in CVH mice. These data suggest 5-HT exerts a direct effect on bladder afferents to enhance signalling. 5-HT3 antagonists could therefore be a potential therapeutic target to treat functional bladder and bowel disorders.


Effect of intake on fasting heat production, respiratory quotient and plasma metabolites measured using the washed rumen technique.
The objective was to investigate the effect of intake before fasting on concentrations of metabolites and hormones, respiratory quotient (RQ) and fasting heat production (HP) using the washed rumen technique and to compare these values with those from the fed state. Six Holstein steers (360±22 kg) were maintained at 21°C and fed three different energy intakes within a replicated 3×3 Latin square design with 21-day periods. Steers were fed alfalfa cubes to provide 1.0, 1.5 and 2.0×NEm during 19 days of each experimental period. Steers were placed in individual metabolism stalls fitted with indirect calorimetry head-boxes on day 20 of each experimental period (FED steers) and fed their normal meal. On day 21 of each period the reticulorumen was emptied, washed and refilled with ruminal buffer (NaCl=96; NaHCO3=24; KHCO3=30; K2HPO4=2; CaCl2=1.5; MgCl2=1.5 mmol/kg of buffer) aerated with 75% N2 and 25% CO2 before introduction to the rumen (steers were not fed; WASHED steers). Each gas exchange was measured over 24 h. HP for 1.0, 1.5 and 2.0×NEm were 479, 597 and 714 kJ/daykg0.75 (s.e.m. =16), respectively. The plateau RQ was 0.756, 0.824 and 0.860 for the 1.0, 1.5 and 2.0×NEm intakes for the FED steers, respectively. After rumen washing, fasting HP was 331, 359 and 400 kJ/daykg0.75 (s.e.m.=13) for 1.0, 1.5, and 2.0×NEm intakes before fasting, respectively. The RQ for WASHED rumen steers was 0.717, 0.710 and 0.719, respectively. Cortisol and β-hydroxybutyrate concentrations in WASHED rumen steers did not exceed threshold levels for severe energy deficit and stress as can be induced from prolonged fasting. This study demonstrates that a fasting state can be emulated using the washed rumen technique, minimizing the time required as opposed to traditional fasting methodologies, without causing a severe energy deficit and stress.


Exogenous coproporphyrin III production by Corynebacterium aurimucosum and Microbacterium oxydans in erythrasma lesions.
Erythrasma is a superficial skin disease caused by Gram-positive Corynebacterium species. Coral-red fluorescence under Wood's light, strongly suggestive of erythrasma, can be attributed to the presence of porphyrins. Fractionated porphyrin analysis in erythrasma lesions is yet to be reported. We attempted to investigate erythrasma lesions by isolating the responsible bacteria and determining their exogenous porphyrin production by HPLC analysis. We observed a 78-year-old woman with erythrasma who had a well-demarcated slightly scaling patch on her left foot, between the fourth and fifth toes. Two kinds of colonies on 5 % sheep blood agar were obtained from this lesion. Analysis of the 16S rRNA sequence revealed the colonies to be Corynebacterium aurimucosum and Microbacterium oxydans. HPLC analysis demonstrated that coproporphyrin III (Copro III) levels were clearly elevated, although the amounts of protoporphyrin were diminished. These results indicate that the fluorescent substance was Copro III. This study supports the view that excess Copro III synthesis by C. aurimucosum and M. oxydans leads to accumulation of porphyrin in cutaneous tissue, which emits a coral-red fluorescence when exposed to Wood's light.


Isolation of new actinomycete strains for the screening of new bioactive compounds.
In order to facilitate the discovery of novel bioactive compounds from microorganisms, various techniques for isolation of new actinomycete strains have been attempted. Studies of the vertical distribution of actinomycetes in soil, isolation of actinomycetes from desert soils or fallen leaves, selective isolation of Kitasatospora strains using novobiocin or Actinoplanes strains using the chemotactic method, and the use of gellan gum as a solidifying agent were carried out. We discovered 9 novel bioactive compounds from actinomycete strains isolated under unusual conditions, and proposed two new genera, five new species and one new subspecies.


[Cloning and bioinformatic analysis of bone morphological protein 7 (BMP7) in rabbit].
On the basis of known partial coding DNA sequence (GenBank accession No. AF413111), we tried to clone the full mRNA of rabbit BMP7 gene by sub-cloning and RACE methods. Then after, bioinformatic analysis on the acquired sequences were conducted. The sequencing results showed that the all cloned sequences could be assembled into a 1 654 bp long DNA fragment, which contained the near full coding cDNA for the propetide, full coding cDNA for the mature protein, and full length of 3'UTR of rabbit BMP7. The newly cloned sequences extended the 5'and 3'ends of known partial DNA sequence by 395 bp and 628 bp, respectively. Sequence comparisons revealed that the full length of coding cDNA of rabbit BMP7 was 91.89% and 89.32% identical to that of human and mouse, while the deduced amino acids was 96.51% and 96.01% identical, respectively. The 3'UTR of rabbit BMP7 was 446 bp in length, 57.38% and 45.57% identical to that of human or mouse, respectively. In addition, there were two closely arranged AATAAA sites in the 3'UTR of rabbit BMP7 gene, as well as in human BMP7. The deduced mature protein of rabbit BMP7 had the seven conservative cysteines and a TGF-b family signature, which were the characters of all BMPs. Our results suggested that the main part of rabbit BMP7 had been successively cloned. The alternation of polyadenization sites in the 3'UTR of rabbit BMP7 maybe related to the posttranscriptional regulation of the gene.


Stimulation of arterial smooth muscle cell proliferation by remnant lipoprotein particles isolated by immuno-affinity chromatography with anti-apo A-I and anti-apo B-100.
Postprandial lipidemia, characterized by high plasma triglyceride-rich lipoprotein remnants, is associated with atherosclerosis. It has also been known that proliferation of vascular smooth muscle cells is crucial for the development of atherosclerosis. In this study, we investigated the direct effect of remnant lipoprotein particles, which consist of chylomicron remnants and very low density lipoprotein remnants, on vascular smooth muscle cell proliferation. Blood was collected from six patients with postprandial lipidemia two hours after their usual meal. Remnant lipoprotein particles were isolated from plasma by immuno-affinity chromatography containing two monoclonal antibodies, anti-apo A-I (H-12) and anti-apo B-100 (JI-H). Remnant lipoprotein particles, as well as betaVLDL, significantly stimulated the proliferation of porcine coronary artery smooth muscle cells in a concentration-dependent manner, whereas very low density lipoprotein (d < 1.006) was virtually ineffective. These observations are consistent with recent reports that triglyceride-rich lipoprotein remnants, which are rich in apo E as well, are atherogenic.


Small subunit ribosomal RNA sequences unite alternate actinosporean and myxosporean stages of Myxobolus cerebralis the causative agent of whirling disease in salmonid fish.
The alternating myxosporean and actinosporean stages of the myxozoan parasite Myxobolus cerebralis (Hofer 1903) from its salmonid fish and aquatic oligochaete hosts, respectively, were compared for sequence homology of the small subunit (18S) ribosomal RNA genes. A 99.8% similarity between the sequences of these two stages was substantially greater than that of M. cerebralis compared to two other Myxobolus sp. from salmonid fish. Our results are the first molecular evidence confirming the alternating stages, initially described by Wolf and Markiw [25] for the life cycle of M. cerebralis but found in two different taxonomic classes (Myxosporea and Actinosporea) are indeed forms of the same organism. Sequencing of rRNA genes of the actinosporean stage followed by development of specific primers for DNA amplification of the myxosporean stage, as in our study, should be applied to solve other myxozoan life cycles. Additionally, these approaches will in the future provide useful diagnostic reagents for the detection and study of this important group of fish pathogens.


Clinical Performance of the Luminex NxTAG CoV Extended Panel for SARS-CoV-2 Detection in Nasopharyngeal Specimens from COVID-19 Patients in Hong Kong.
In December 2019, the coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) was first reported in the Hubei province of China and later spread all over the world. There was an urgent need of a high-throughput molecular test for screening the COVID-19 patients in the community. The Luminex NxTAG CoV extended panel is a high-throughput FDA emergency use-authorized molecular diagnostic assay for SARS-CoV-2 detection. This system targets three genes (ORF1ab, N, and E genes) of SARS-CoV-2, the ORF1ab region of SARS-CoV, and the ORF5 region of MERS-CoV. In this study, we evaluated the diagnostic performance of this system with nasopharyngeal swab specimens of 214 suspected COVID-19 patients in Hong Kong. The results were compared with our routine COVID-19 reverse transcription-PCR (RT-PCR) protocol with a LightMix SarbecoV E-gene kit and an in-house RdRp/Hel RT-PCR assay. The NxTAG CoV extended panel demonstrated 97.8% sensitivity and 100% specificity to SARS-CoV-2 in nasopharyngeal specimens. On low-viral load specimens, the sensitivity of the NxTAG panel could still maintain at 85.71%. Strong agreement was observed between the NxTAG panel and the routine COVID-19 RT-PCR protocol (kappa value = 0.98). Overall, the E gene target of the NxTAG panel demonstrated the highest sensitivity among the three SARS-CoV-2 targets, while the N gene targets demonstrated the least. In conclusion, the NxTAG CoV extended panel is simple to use, and it has high diagnostic sensitivity and specificity to SARS-CoV-2 in nasopharyngeal specimens. We recommend this diagnostic system for high-throughput COVID-19 screening in the community.


Molecular characterization of three chloramphenicol acetyltransferases isolated from Haemophilus influenzae.
Three plasmid-mediated chloramphenicol acetyltransferases isolated from different Haemophilus influenzae strains were purified and characterized. All three enzymes had properties in common with the gram-negative family of chloramphenicol acetyltransferase. The Haemophilus enzymes and the enteric type II enzyme were sensitive to 5,5'-dithiobis(2-nitrobenzoic acid), gave the same elution patterns from a highly substituted resin containing a bound chloramphenicol base, and had similar reactions to antisera. All four differed from each other in subunit molecular weight, enzyme activity, and partial protein digestion patterns. The data suggest that the three Haemophilus enzymes belong to the less common type II group and are related, but is not identical, to each other and to the enteric type II enzyme.


Radiotherapy timing in the treatment of limited-stage small cell lung cancer: the impact of thoracic and brain irradiation on survival.
Small cell lung cancer is an aggressive histologic subtype of lung cancer in which the role of chemotherapy and radiotherapy has been well established in limited-stage disease. We retrospectively reviewed a series of limited-stage small cell lung cancers treated with chemotherapy and thoracic and brain radiotherapy. A total of 124 patients affected by limited-stage small cell lung cancer has been treated over 10 years in our Institute. Fifty-three patients (42.8%) had concomitant radio-chemotherapy treatment and 71 patients (57.2%) a sequential treatment. Eighty-eight patients (70.9%) underwent an association of a platinum-derived drug (cisplatinum or carboplatinum) and etoposide. Prophylactic cranial irradiation was planned in all patients with histologically proven complete response to primary radio-chemotherapy. With a mean follow-up of 2.2 years, complete response was obtained in 50.8% of cases. We found a significant difference between different radio-chemotherapy association approaches (P = 0.007): percentages of overall survival were respectively 10.0%, 12.9% and 5.6% in early, late concomitant and sequential radio-chemotherapy timing. Cranial prophylaxis did not seem to influence overall survival (P = 0.21) or disease-free survival for local relapse (P = 0.34). Concomitant radio-chemotherapy is the best approach according to our experience. Our results show a benefit of prophylactic cranial irradiation in distant metastasis-free survival.


Workplace-Based Assessments Using Pediatric Critical Care Entrustable Professional Activities.
Workplace-based assessment (WBA) is critical to graduating competent physicians. Developing assessment tools that combine the needs of faculty, trainees, and governing bodies is challenging but imperative. Entrustable professional activities (EPAs) are emerging as a clinically oriented framework for trainee assessment. We sought to develop an EPA-based WBA tool for pediatric critical care medicine (PCCM) fellows. The goals of the tool were to promote learning through benchmarking and tracking entrustment. A single PCCM EPA was iteratively subdivided into observable practice activities (OPAs) based on national and local data. Using a mixed-methods approach following van der Vleuten's conceptual model for assessment tool utility and Messick's unified validity framework, we sought validity evidence for acceptability, content, internal structure, relation to other variables, response process, and consequences. Evidence was gathered after 1 year of use. Items for assessment were based on correlation between the number of times each item was assessed and the frequency professional activity occurred. Phi-coefficient reliability was 0.65. Narrative comments demonstrated all factors influencing trust, identified by current literature, were cited when determining level of entrustment granted. Mean entrustment levels increased significantly between fellow training years (P = .001). Compliance for once- and twice-weekly tool completion was 50% and 100%, respectively. Average time spent completing the assessment was less than 5 minutes. Using an EPA-OPA framework, we demonstrated utility and validity evidence supporting the tool's outcomes. In addition, narrative comments about entrustment decisions provide important insights for the training program to improve individual fellow advancement toward autonomy.


Influence of various intermittent fasting regimens on body weight and glycemic control in streptozotocin-induced diabetic rats.
This study aims to compare the effects of various intermittent fasting (IF) regimens, i.e., time-restricted fasting (TRF), alternate day fasting (ADF), and periodic fasting (PF) on body weight, glycemic control and associated metabolic parameters in streptozotocin-induced diabetic rats. Sixty male Sprague-Dawley rats (aged 3 months) were randomly assigned to the normal control (NC), diabetic control (DC), TRF, ADF, and PF groups. Type 2 diabetes was induced in all groups, except for the NC group, by intramuscular administration of streptozotocin (55 mg/kg). The IF interventions were administered for 6 weeks. The rats in all the groups, except for the NC group, exhibited significant weight loss (31.4%, 46.4%, 31.0%, and 33.9% in the DC, TRF, ADF, and PF groups, respectively). The fasting blood glucose levels decreased to varying degrees, with the PF group showing the most significant decrease (77.0%), followed by the ADF (55.0%) and TRF (32.2%) groups. The plasma insulin levels were significantly lower in the experimental groups than in the NC group, but no significant effects were observed on the lipid profile. The study findings indicate that while the IF protocols led to body weight loss, they exhibited varying effects on glycemic control and other metabolic parameters.


Medical Students' Attitude and Perception Towards Basic Medical Science Subjects at Wollo University, Northeast Ethiopia.
The knowledge of basic medical science could help to remember a fact and be used to understand causal mechanisms of disease process that improve the accuracy of diagnostic formulations. To assess the perception and attitude of medical students towards basic medical science subjects along with retention rate and clinical relevance in Wollo University. Institutional-based cross-sectional study design was employed in Wollo University from September, 2020 to October, 2020. A pre-tested and structured self-administered questionnaire was used for data collection. All opinions were rated using a positive-point Likert scale, which ranges from "strongly disagree" to "strongly agree." The data were entered and analyzed using SPSS version 20. A total of 153 students participated with a 100% response rate. Among them, 45 (29.4%), 38 (24.8%), 39 (25.5%), and 31 (20.3%) were PC-II, C-I, C-II, and Intern students, respectively. Ninety-six (62.7%) of medical students were very much interested in basic medical science subjects. But, 113 (73.9%) of them did not have plans to join the subjects as a future career. The main reasons in more than half 85 (55.7%) of the respondents were less financial growth followed by less chance of promotion 31 (20.3%). There were 66.7% of the students who considered anatomy as clinically relevant, whereas 53.6% and 47.1% considered physiology and biochemistry, respectively, to be clinically relevant. The number of students who could recall anatomy and physiology during relevant clinical discussions was 102 (66.7%) and 85 (55.6%), respectively. This percentage was relatively less for biochemistry (26.8%). In conclusion, medical students have a positive attitude towards basic medical science subjects. However, they are hesitant to join the field because it offers them less financial growth and few chances of promotion. Moreover, anatomy and physiology were highly relevant subjects during clinical practice. But, the retention rate of basic medical science knowledge during their clinical year was low.


Energy expenditure of adult male rhesus monkeys during the first 30 mo of dietary restriction.
Energy expenditure, activity, and body composition were measured in 30 adult male rhesus monkeys used in a study having the long-term goal of determining the effects of moderate dietary restriction (DR) on aging. All animals were fed a defined diet, with the restricted animals maintained at approximately 70% of the caloric intakes of the controls. After 12 mo of DR, body fat mass of restricted monkeys was 33% less than that of controls (P = 0.004), whereas lean body mass differences were not present until after 24 mo. At the 24- and 30-mo assessments, nighttime energy expenditure was significantly reduced (P < 0.01) in the restricted compared with control monkeys after adjustment for lean body mass differences, whereas morning, afternoon, and total energy expenditure were not significantly different (P > 0.05). No significant differences (P > 0.05) in activity were noticed between treatment groups at any time point. DR resulted in a prolonged decrease in resting energy expenditure, which could contribute to the possible life-extending action of this treatment.


Micro-mesoporous materials obtained by zeolite recrystallization: synthesis, characterization and catalytic applications.
The review covers the recent developments in the field of novel micro-mesoporous materials obtained by zeolite recrystallization. The materials are classified into three distinctly different groups depending on the degree of recrystallization: (i) coated mesoporous zeolites (RZEO-1); (ii) micro-mesoporous nanocomposites (RZEO-2); and (iii) mesoporous materials with zeolitic fragments in the walls (RZEO-3). The first part of the review is focused on the analysis of the synthetic strategies leading to different types of recrystallized materials. In the second part, a comprehensive view on their structure, texture and porosity in connection with acidic and diffusion properties is given. The last part is devoted to the catalytic applications of recrystallized materials. The advantages and disadvantages with respect to pure micro- and mesoporous molecular sieves and other hierarchical zeolites are critically analyzed and the future opportunities and perspectives are discussed.


Calcium sensitization involved in dexmedetomidine-induced contraction of isolated rat aorta.
Dexmedetomidine, a full agonist of the α2B-adrenoceptor that is mainly involved in vascular smooth muscle contraction, is primarily used for analgesia and sedation in intensive care units. High-dose dexmedetomidine produces hypertension in children and adults. The goal of this in vitro study was to investigate the role of the calcium (Ca(2+)) sensitization mechanism involving Rho-kinase, protein kinase C (PKC), and phosphoinositide 3-kinase (PI3-K) in mediating contraction of isolated rat aortic smooth muscle in response to dexmedetomidine. The effect of dexmedetomidine on the intracellular Ca(2+) level ([Ca(2+)]i) and tension was measured simultaneously. Dexmedetomidine concentration-response curves were generated in the presence or absence of the following antagonists: rauwolscine, Y 27632, LY 294002, GF 109203X, and verapamil. Dexmedetomidine-induced phosphorylation of PKC and membrane translocation of Rho-kinase were detected with Western blotting. Rauwolscine, Y 27632, GF 109203X, LY 294002, and verapamil attenuated dexmedetomidine-induced contraction. The slope of the [Ca(2+)]i-tension curve for dexmedetomidine was higher than that for KCl. Dexmedetomidine induced phosphorylation of PKC and membrane translocation of Rho-kinase. These results suggest that dexmedetomidine-induced contraction involves a Ca(2+) sensitization mechanism mediated by Rho-kinase, PKC, and PI3-K that is secondary to α2-adrenoceptor stimulation in rat aortic smooth muscle.


Behavioral variability and autism spectrum disorder.
Restricted and repetitive behavior is a diagnostic characteristic of autism spectrum disorder (ASD). To the extent that the behavior of individuals with ASD can be conceptualized as problems of invariance, our understanding of environmental variables that influence restricted and repetitive behavior may be informed by the basic and applied literature on response variability. The purposes of this paper are (a) to describe how restricted and repetitive behavior can be conceptualized as problems of invariance, (b) to consider the implications of a lack of varied responding for individuals with ASD, (c) to review relevant basic and applied research on response variability, (d) to present methods to address invariant responding for individuals with ASD, and (e) to suggest areas for future research.