The secular and the supernatural: madness and psychiatry in the short stories of Muriel Spark.
Edinburgh-born Muriel Spark is one of modern Scotland's greatest writers. Examination of her work reveals that the subjects of madness and psychiatry are recurrent themes in her writing. She herself had a mental breakdown when she was a young woman and she took an interest in the world of psychiatry and psychoanalysis. In her short stories, Spark approaches the subject of madness in a variety of ways: she relates it to the supernatural; to writing fiction; and to religion. She frequently juxtaposes secular and supernatural explanations of mental disturbance. Spark adopts a sceptical and, at times, mocking view of psychiatrists and psychiatric treatment. Both psychoanalysis and pills are seen as problematic.


Validation of the Slovene Version of the Stop-Bang Questionnaire in a Primary Practice Setting.
The aim of our study was to validate the Slovene translation of the STOP-BANG (SBQ) questionnaire for use in the primary practice setting. We recruited 158 randomly selected visitors at four primary practice clinics who came to the practice for any reason. Participants completed the Slovene SBQ and underwent type 3 respiratory polygraphy, which was analysed by an experienced somnologist. The SBQ was previously translated in to Slovene and validated for the sleep clinic. Of 158 participants, 153 had valid recordings. The mean age of the participants was 49.5 years (±13.0 years), and 47.7% were male. OSA was identified in 49.0% of the participants. The questionnaire, with a cutoff of ≥3, demonstrated an area under the curve of 0.823 for any OSA (REI≥5), 0.819 for moderate and severe OSA (REI≥15) and 0.847 for severe OSA (REI≥30). Sensitivity was 65.3%, 81.8%, and 90.0%, and specificity was 87.2%, 73.3% and 65.0% for any, moderate to severe and severe OSA, respectively. The Slovene translation of the SBQ is a reliable instrument for OSA risk stratification in the primary practice setting.


The somatomedin hypothesis: 2001.
Since the original somatomedin hypothesis was conceived, a number of important discoveries have allowed investigators to modify the concept. Originally somatic growth was thought to be controlled by pituitary GH and mediated by circulating insulin-like growth factor-I (IGF-I, somatomedin C) expressed exclusively by the liver. With the discovery that IGF-I is produced by most, if not all, tissues, the role of autocrine/paracrine IGF-I vs. the circulating form has been hotly debated. Recent experiments using transgenic and gene-deletion technologies have attempted to answer these questions. In the liverspecific igf-1 gene-deleted mouse model, postnatal growth and development are normal despite the marked reduction in circulating IGF-I and IGF-binding protein levels; free IGF-I levels are normal. Thus, the normal postnatal growth and development in these animals may be due to normal free IGF-I levels (from as yet unidentified sources), although the role of autocrine/paracrine IGF-I has yet to be determined.


Therapeutic efficacy of MUC1-specific cytotoxic T lymphocytes and CD137 co-stimulation in a spontaneous breast cancer model.
To study immunology in breast tumors, we have utilized a mammary gland adenocarcinoma model in which mice develop spontaneous tumors of the mammary gland which are initiated at puberty and express a human tumor antigen, MUC1. MUC1 (CD227) is over-expressed in 90% of human breast cancers and its glycosylation status and pattern of expression in cancer cells is altered. Humoral and cellular responses to MUC1 have been reported in breast cancer patients and therefore, MUC1 is being evaluated as a target for immune intervention. This mouse model of spontaneous breast cancer allows the evaluation of anti-MUC1 immune responses at all stages of the disease. In this report, we review the model as it pertains to a) the development of the tumor, b) MUC1 expression, and the native immune responses against MUC1 as tumors progress, and c) the immune suppressive microenvironment within the developing tumor. Finally, we report our latest findings describing the therapeutic efficacy of adoptively transferred MUC1-specific cytotoxic T lymphocytes (MUC1-CTL) in these mice and discuss ways to increase their effectiveness by agonistic monoclonal antibody against CD137 T cell costimulatory molecule.


Pesticide applications of copper on perennial crops in California, 1993 to 1998.
Inorganic copper is used as a broad-spectrum fungicide and bacteriocide on a variety of agricultural crops. After application, the copper residue typically accumulates in the upper 15 cm of soil. Data from the California Pesticide Use Reports were used to estimate the augmentation of copper in the soil that resulted from pesticide applications for the six years from 1993 to 1998 on 12 crops that are grown without rotation. The estimated mean mg Cu kg(-1) soil added to the upper 15 cm during the six years was the following: walnut (Juglans regia L.), 28; peach [Prunus persica (L.) Batsch var. persica], 22; nectarine [Prunuspersica (L.) Batsch var. nucipersica (Suckow) C.K. Schneid], 19; cherry (Pseudolmedia oxyphyllaria Donn. Sm.), 18; rice (Orvza sativa L.), 16; apricot (Prunus armeniaca L.), 11; orange [Citrus sinensis (L.) Osbeck) and plum (Prunus domestica L. subsp. domestica ), 9; lemon [Citrus limon (L.) Burm.f.] and almond [Prunus dulcis (Mill.) D.A. Webb], 6; pear (Pyrus communis L.), 4; and grape (Vitis vinifera L.), 3. In addition, for the first five of these crops, we estimated the area that was treated with each level of kg Cu ha(-1). For example, for walnut orchards, we estimated that 12 500 ha, or 17% of the planted area, was treated with a quantity of Cu that would increase the total concentration of Cu in the upper 15 cm of soil by at least 50 mg Cu kg(-1) soil. A comparison of the amount of Cu per unit planted area that was applied in the first and second half of the study indicated that the intensity of copper use is either relatively constant or increasing, depending on the crop. The findings are discussed in relation to the potential effect of continued long-term use of Cu pesticides on soil sustainability.


Nematic liquid crystals in planar and cylindrical hybrid cells: Role of elastic anisotropy on the director deformations.
Nematic samples filling a flat cell or the annular region between two concentric cylinders with hybrid anchoring conditions at the boundaries are investigated by setting up and minimizing their Frank elastic free energy. The coupling with the surfaces is taken to be strong on one side and weak on the other. The equations are numerically solved and the conditions for which the molecular organization inside the cell becomes uniform are analyzed. The classical calculation performed by G. Barbero and R. Barberi [J. Phys. 44, 609 (1983)] is reproduced and investigated from a different point of view, in order to compare the results of planar and cylindrical geometries. The results suggest that the cylindrical cell presents some unusual features deserving a more complete investigation. Although most part of the transitional phenomena are found for K(11)>K(33), a case not common for ordinary (lyotropic and thermotropic) liquid crystals, it is possible to find a completely uniform cell even for K(11)<K(33) in both the geometries considered here.


Farmers' Decision Making on Livestock Trading Practices: Cowshed Culture and Behavioral Triggers Amongst New Zealand Dairy Farmers.
Studies of farmers' failure to implement biosecurity practices frequently frame their behavior as a lack of intention. More recent studies have argued that farmers' behaviors should be conceptualized as emergent from farming experiences rather than a direct consequence of specific intentions. Drawing on the concepts of "cowshed" culture and the "Trigger Change Model," we explore how farmers' livestock purchasing behavior is shaped by farms' natural and physical environments and identify what triggers behavioral change amongst farmers. Using bovine tuberculosis (bTB) in New Zealand as a case example, qualitative research was conducted with 15 New Zealand dairy producers with varying bTB experiences. We show how farmers' livestock purchasing behavior evolve with culture under a given farm environment. However, established cultures may be disrupted by various triggers such as disease outbreaks, introductions of animals with undesired characteristics, and farm relocation. While dealing with economic and socio-emotional impacts posed by triggers, farmers reorganize their culture and trading behaviors, which may involve holistic biosecurity strategies. Nevertheless, we also show that these triggers instigate only small behavioral changes for some farmers, suggesting the role of the trigger is likely to be context-dependent. Using voluntary disease control schemes such as providing disease status of source farms has attracted great interest as a driver of behavioral change. One hopes such schemes are easily integrated into existing farm practices, however, we speculate such an integration is challenging for many farmers due to path-dependency. We therefore argue that these schemes may fail to bring their intended behavioral changes without a greater understanding of how different types of triggers work in different situations. We need a paradigm shift in how we frame farmers' livestock trading practices. Otherwise, we may not able to answer our questions about farm biosecurity if we continue to approaching these questions solely from a biosecurity point of view.


HDAC6 is a microtubule-associated deacetylase.
Reversible acetylation of alpha-tubulin has been implicated in regulating microtubule stability and function. The distribution of acetylated alpha-tubulin is tightly controlled and stereotypic. Acetylated alpha-tubulin is most abundant in stable microtubules but is absent from dynamic cellular structures such as neuronal growth cones and the leading edges of fibroblasts. However, the enzymes responsible for regulating tubulin acetylation and deacetylation are not known. Here we report that a member of the histone deacetylase family, HDAC6, functions as a tubulin deacetylase. HDAC6 is localized exclusively in the cytoplasm, where it associates with microtubules and localizes with the microtubule motor complex containing p150(glued) (ref. 3). In vivo, the overexpression of HDAC6 leads to a global deacetylation of alpha-tubulin, whereas a decrease in HDAC6 increases alpha-tubulin acetylation. In vitro, purified HDAC6 potently deacetylates alpha-tubulin in assembled microtubules. Furthermore, overexpression of HDAC6 promotes chemotactic cell movement, supporting the idea that HDAC6-mediated deacetylation regulates microtubule-dependent cell motility. Our results show that HDAC6 is the tubulin deacetylase, and provide evidence that reversible acetylation regulates important biological processes beyond histone metabolism and gene transcription.


Lesion sites that predict the ability to gesture how an object is used.
We used a two stage procedure to predict which stroke patients would have chronic difficulties gesturing how to use an object when object recognition and hand movements were intact. First, we searched our PLORAS database by behavior and identified 5 patients who had chronic difficulty gesturing object use but no difficulty recognising objects, comprehending words or moving their hands. High definition lesion analyses showed that all 5 patients had damage to the white matter underlying the left ventral supramarginal gyrus, (A) close to the cortex, (B) deep towards the midline and (C) extending into the temporal lobe. In addition, 2 patients had damage to (D) the left posterior middle temporal cortex, and 3 patients had damage to (E) the left dorsal supramarginal gyrus and (F) the left premotor cortex. Second, we searched our database by lesion location for patients who had damage to any part of regions ABCDEF. The incidence of gesturing difficulties was higher in patients with damage to ABCD (7/9), ABCE (7/10) or ABCDE (10/13) than ABCF (7/13), ABC (8/16) or partial damage to ABCF (6/32). Thus behaviour was best predicted by the combination of regions that were damaged (a "network-lesion") rather than on the basis of each region alone or overall lesion size. Our results identify which parts of the temporal and parietal lobes impair the ability to gesture object use and which parts need to be intact to support it after damage. Our methods provide a framework for future studies aiming to predict the consequences of brain damage.


Detection of large numbers of novel sequences in the metatranscriptomes of complex marine microbial communities.
Sequencing the expressed genetic information of an ecosystem (metatranscriptome) can provide information about the response of organisms to varying environmental conditions. Until recently, metatranscriptomics has been limited to microarray technology and random cloning methodologies. The application of high-throughput sequencing technology is now enabling access to both known and previously unknown transcripts in natural communities. We present a study of a complex marine metatranscriptome obtained from random whole-community mRNA using the GS-FLX Pyrosequencing technology. Eight samples, four DNA and four mRNA, were processed from two time points in a controlled coastal ocean mesocosm study (Bergen, Norway) involving an induced phytoplankton bloom producing a total of 323,161,989 base pairs. Our study confirms the finding of the first published metatranscriptomic studies of marine and soil environments that metatranscriptomics targets highly expressed sequences which are frequently novel. Our alternative methodology increases the range of experimental options available for conducting such studies and is characterized by an exceptional enrichment of mRNA (99.92%) versus ribosomal RNA. Analysis of corresponding metagenomes confirms much higher levels of assembly in the metatranscriptomic samples and a far higher yield of large gene families with >100 members, approximately 91% of which were novel. This study provides further evidence that metatranscriptomic studies of natural microbial communities are not only feasible, but when paired with metagenomic data sets, offer an unprecedented opportunity to explore both structure and function of microbial communities--if we can overcome the challenges of elucidating the functions of so many never-seen-before gene families.


Improving exchange with consumers within mental health organizations: Recognizing mental ill health experience as a 'sneaky, special degree'.
Stigmatizing views towards consumers may be held even by those working within mental health organizations. Contemporary mental health policies require organizations to work collaboratively with consumers in producing and delivering services. Using social exchange theory, which emphasises mutual exchange to maximise benefits in partnership, the current study explores the perspectives of those working within organizations that have some level of consumer leadership. Interviews were conducted with 14 participants from a range of mental health organizations. Data were transcribed, and analyzed using thematic analytic and discursive psychological techniques. Findings suggest stigma is still prevalent even in organizations that have consumers in leadership positions, and consumers are often perceived as less able to work in mental health organizations than non-consumers. Several discourses challenged such a view - showing how consumers bring value to mental health organizations through their expertise in the mental health system, and their ability to provide safety and support to other consumers. Through a social exchange theory lens, the authors call for organizations to challenge stigma and promote the value that consumers can bring to maximize mutual benefits.


Impact of mailed and automated telephone reminders on receipt of repeat mammograms: a randomized controlled trial.
This study compares the efficacy of three types of reminders in promoting annual repeat mammography screening. RCT. Study recruitment occurred in 2004-2005. Participants were recruited through the North Carolina State Health Plan for Teachers and State Employees. All were aged 40-75 years and had a screening mammogram prior to study enrollment. A total of 3547 women completed baseline telephone interviews. Prior to study recruitment, women were assigned randomly to one of three reminder groups: (1) printed enhanced usual care reminders (EUCRs); (2) automated telephone reminders (ATRs) identical in content to EUCRs; or (3) enhanced letter reminders that included additional information guided by behavioral theory. Interventions were delivered 2-3 months prior to women's mammography due dates. Repeat mammography adherence, defined as having a mammogram no sooner than 10 months and no later than 14 months after the enrollment mammogram. Each intervention produced adherence proportions that ranged from 72% to 76%. Post-intervention adherence rates increased by an absolute 17.8% from baseline. Women assigned to ATRs were significantly more likely to have had mammograms than women assigned to EUCRs (p=0.014). Comparisons of reminder efficacy did not vary across key subgroups. Although all reminders were effective in promoting repeat mammography adherence, ATRs were the most effective and lowest in cost. Health organizations should consider using ATRs to maximize proportions of members who receive mammograms at annual intervals.


Quercetin and ferulic acid aggravate renal carcinoma in long-term diabetic victims.
Many phytoantioxidants have therapeutic drawbacks due to their potent prooxidant bioactivity. It is hypothesized that phytoantioxidants (PAO) are beneficial only to the early-stage diabetes mellitus (DM) and will become ineffective once renopathy occurs. Gallic acid, rutin, EGCG, ferulic acid (FA), and quercetin were tried on the streptozotocin (STZ)-induced DM rat model for a 28 week experimental period. All of these PAO were shown to be ineffective for hypoglycemic action. The incidence of cataract (50%), injured glomerules, and renal cell carcinoma (RCC) was very common, among which the most severely affected involved the quercetin- and the FA-treated groups. The tumorigenicity of ferulic acid is still unclear. However, for quercetin, this can be attributted to (i) the prooxidant effect, (ii) the insulin-secretagogue bioactivity, and (iii) the competitive and noncompetitive inhibition on the O-methyltransferase to enhance the estradiol-induced tumorigenesis. Conclusively, quercetin and FA are able to aggravate, if not induce, nephrocarcinoma. It is time to reevaluate the tumorigenic detrimental effect of PAO, especially those exhibiting prooxidant bioactivity.


The effects of patient education in lithium therapy on quality of life and compliance.
The purpose of this study was to show the effects of education on medication compliance, symptom level and quality of life of outpatients who were being treated with lithium for bipolar disorder. The study was performed comparing a total of 26 patients (14 study and 12 control) who were a patient group in lithium therapy. In the study one group was given a short education program about the disorder and lithium therapy in three sessions. Data were collected from both groups using a medication knowledge form, Brief Symptom Inventory, and WHO Quality of Life Scale before and after the intervention. At the end of 3 months whereas there was no difference seen in the scores of the control group; the study group had an increase in medication knowledge, a decrease in symptom level, an increase in quality of life, and a beginning of more regular medication use. Findings that were obtained show the importance of education about the disorder and medication in increasing the adaptation to society of patients who have bipolar disorder.


Design, production, and validation of the biological and structural performance of an ecologically engineered concrete block mattress: A Nature-Inclusive Design for shoreline and offshore construction.
Over the past decade, the scientific community has studied, experimented, and published a notable body of literature on the ecological enhancement of coastal and marine infrastructure (CMI). The Nature-Inclusive Design (NID) approach refers to methods and technologies that can be integrated into the design and construction of CMI to create a suitable habitat for native species (or communities) whose natural habitat has been degraded or reduced. To examine the compliance of new environmentally sensitive technologies with structural requirements and fiscal restraints, while providing ecosystem and habitat value, this paper presents the findings of a structural-economical-biological analysis of ecologically engineered Articulated Concrete Block Mattresses (ACBMs). To evaluate the structural and biological performance of the Ecological Articulated Concrete Block Mattresses, a pilot project was deployed in April 2017 at Port Everglades, Florida, USA, and evaluated against controls of adjacent artificial structures and smooth-surface concrete blocks and monitored over a period of two years. The elements of ecological enhancement implemented in the fabrication and design of the ecologically enhanced ACBMs were comprised of bio-enhancing concrete additives and science-based designs. Based on the results of this study, these design alterations have increased the richness and diversity of sessile assemblages compared to control blocks and adjacent artificial structures and supported a higher abundance of mobile species. This ecological improvement was achieved within the operational limitations of conventional manufacturing and installation technologies, while complying with strict structural requirements for standard concrete marine construction. The results supported the working hypothesis and demonstrated that modifications of concrete composition, surface texture, and macro-design have the potential to increase the ecological value of concrete-based CMI and promote a more sustainable and adaptive approach to coastal and marine development in an era of climate resilience-building. Integr Environ Assess Manag 2022;18:148-162. © 2021 SETAC.


Ten-year experience with endobronchial ultrasound-guided transbronchial needle aspiration: single center results in mediastinal diagnostic and staging.
Endobronchial ultrasonography with transbronchial needle aspiration (EBUS-TBNA) is recognized as an accurate and minimal invasive procedure for diagnosis and staging of lung cancer and lymph nodal malignancies. EBUS is recommended as the first choice procedure for mediastinal staging in lung cancer in international guidelines. A retrospective evaluation was performed on single center experience with EBUS-TBNA in our thoracic surgery department in a 10-year time frame. Main indication for the procedure was suspected non-lymphomatous malignancy in intrathoracic lymph-nodes on computed tomography (CT) or positron emission tomography (PET) scan. All procedures were performed under conscious sedation in a day-hospital setting. All the aspirated specimens were obtained with a 22-gauge needle and were fixed in 10% formalin and paraffin embedded. Sections of 3 micron in thickness were cut and hematoxylin-eosin stained. From October 2005 to August 2016, 496 patients were submitted to EBUS-TBNA. Number of nodal stations punctured was 592 with a mean of 2.25 punctures per patient. Diagnosis of malignancy was obtained in 291 patients (58.6%). In 25 cases a nodal metastasis from an extrathoracic primary tumor was diagnosed. Sensitivity, specificity and diagnostic accuracy were 95%, 100% and 96% respectively. Negative predictive value was 90% and positive predictive value (PPV) was 100%. When molecular tests were requested, mutational analysis was successfully performed on cell block derived material in 55 out of 56 cases (98.2%), and fluorescence in situ hybridization (FISH) analysis in 26 out of 27 cases (96.2%). EBUS-TBNA in our setting was an accurate and safe tool to diagnose non-lymphomatous nodal malignancies. Interestingly, in our series EBUS-TBNA has demonstrated to yield sufficient tissue for molecular analysis.


An artificial 'haemophilic' plasma for one-stage factor-VIII assay.
An 'artificial' plasma for one-stage factor-VIII assays is made by incubating human plasma with EDTA, to destroy factor VIII, and afterwards removing the anticoagulant by dialysis. Bovine factor V is then added to a given level. In the assay, contact activation is controlled by adding contact product. It was confirmed that factor-VIII activity was destroyed and that the EDTA was freely dialysable. The fibrinogen in the treated plasma clotted normally with thrombin. Likely variation in the factor-V activity was found not to be critical. The concentration of fibrinogen and other factors was adequate. Variation between batches was small. The artificial plasma yielded assay results closely comparable to haemophilic plasma in samples with factor-VIII activities in the range 0.01--20.0 iu/ml; the mean results in the artificial system were estimated to be 0.997 x those in haemophilic plasma, with a 95% confidence interval of 0.901--1.103. Biological variability in individual assays was smaller in the artificial system than when haemophilic plasma was used. Instability at the bench was more often detected in the artificial system than in haemophilic plasma assays, but the effect was eliminated from the results by obtaining duplicated readings in a balanced order.


A farnesyltransferase inhibitor activates lysosomes and reduces tau pathology in mice with tauopathy.
Tau inclusions are a shared feature of many neurodegenerative diseases, among them frontotemporal dementia caused by tau mutations. Treatment approaches for these conditions include targeting posttranslational modifications of tau proteins, maintaining a steady-state amount of tau, and preventing its tendency to aggregate. We discovered a new regulatory pathway for tau degradation that operates through the farnesylated protein, Rhes, a GTPase in the Ras family. Here, we show that treatment with the farnesyltransferase inhibitor lonafarnib reduced Rhes and decreased brain atrophy, tau inclusions, tau sumoylation, and tau ubiquitination in the rTg4510 mouse model of tauopathy. In addition, lonafarnib treatment attenuated behavioral abnormalities in rTg4510 mice and reduced microgliosis in mouse brain. Direct reduction of Rhes in the rTg4510 mouse by siRNA reproduced the results observed with lonafarnib treatment. The mechanism of lonafarnib action mediated by Rhes to reduce tau pathology was shown to operate through activation of lysosomes. We finally showed in mouse brain and in human induced pluripotent stem cell-derived neurons a normal developmental increase in Rhes that was initially suppressed by tau mutations. The known safety of lonafarnib revealed in human clinical trials for cancer suggests that this drug could be repurposed for treating tauopathies.


Molecular dynamics simulations of the Cx26 hemichannel: evaluation of structural models with Brownian dynamics.
The recently published crystal structure of the Cx26 gap junction channel provides a unique opportunity for elucidation of the structure of the conductive connexin pore and the molecular determinants of its ion permeation properties (conductance, current-voltage [I-V] relations, and charge selectivity). However, the crystal structure was incomplete, most notably lacking the coordinates of the N-terminal methionine residue, which resides within the pore, and also lacking two cytosolic domains. To allow computational studies for comparison with the known channel properties, we completed the structure. Grand canonical Monte Carlo Brownian dynamics (GCMC/BD) simulations of the completed and the published Cx26 hemichannel crystal structure indicate that the pore is too narrow to permit significant ion flux. The GCMC/BD simulations predict marked inward current rectification and almost perfect anion selectivity, both inconsistent with known channel properties. The completed structure was refined by all-atom molecular dynamics (MD) simulations (220 ns total) in an explicit solvent and POPC membrane system. These MD simulations produced an equilibrated structure with a larger minimal pore diameter, which decreased the height of the permeation barrier formed by the N terminus. GCMC/BD simulations of the MD-equilibrated structure yielded more appropriate single-channel conductance and less anion/cation selectivity. However, the simulations much more closely matched experimentally determined I-V relations when the charge effects of specific co- and posttranslational modifications of Cx26 previously identified by mass spectrometry were incorporated. We conclude that the average equilibrated structure obtained after MD simulations more closely represents the open Cx26 hemichannel structure than does the crystal structure, and that co- and posttranslational modifications of Cx26 hemichannels are likely to play an important physiological role by defining the conductance and ion selectivity of Cx26 channels. Furthermore, the simulations and data suggest that experimentally observed heterogeneity in Cx26 I-V relations can be accounted for by variation in co- and posttranslational modifications.


Transcranial light-emitting diode therapy for neuropsychological improvement after traumatic brain injury: a new perspective for diffuse axonal lesion management.
The cost of traumatic brain injury (TBI) for public health policies is undeniable today. Even patients who suffer from mild TBI may persist with cognitive symptoms weeks after the accident. Most of them show no lesion in computed tomography or conventional magnetic resonance imaging, but microstructural white matter abnormalities (diffuse axonal lesion) can be found in diffusion tensor imaging. Different brain networks work together to form an important part of the cognition process, and they can be affected by TBI. The default mode network (DMN) plays an important central role in normal brain activities, presenting greater relative deactivation during more cognitively demanding tasks. After deactivation, it allows a distinct network to activate. This network (the central executive network) acts mainly during tasks involving executive functions. The salience network is another network necessary for normal executive function, and its activation leads to deactivation of the DMN. The use of red or near-infrared (NIR) light to stimulate or regenerate tissue is known as photobiomodulation. It was discovered that NIR (wavelength 800-900 nm) and red (wavelength 600 nm) light-emitting diodes (LEDs) are able to penetrate through scalp and skull and have the potential to improve the subnormal, cellular activity of compromised brain tissue. Based on this, different experimental and clinical studies were done to test LED therapy for TBI, and promising results were found. It leads us to consider developing different approaches to maximize the positive effects of this therapy and improve the quality of life of TBI patients.


Obesity and age-related alterations in the gene expression of zinc-transporter proteins in the human brain.
The incidence of Alzheimer's disease (AD) is increasing. Major risk factors for AD are advancing age and diabetes. Lately, obesity has been associated with an increased risk of dementia. Obese and diabetic individuals are prone to decreased circulating levels of zinc, reducing the amount of zinc available for crucial intracellular processes. In the brain, zinc co-localizes with glutamate in synaptic vesicles, and modulates NMDA receptor activity. Intracellular zinc is involved in apoptosis and fluctuations in cytoplasmic Zn(2+) affect modulation of intracellular signaling. The ZNT and ZIP proteins participate in intracellular zinc homeostasis. Altered expression of zinc-regulatory proteins has been described in AD patients. Using microarray data from human frontal cortex (BrainCloud), this study investigates expression of the SCLA30A (ZNT) and SCLA39A (ZIP) families of genes in a Caucasian and African-American sample of 145 neurologically and psychiatrically normal individuals. Expression of ZNT3 and ZNT4 were significantly reduced with increasing age, whereas expression of ZIP1, ZIP9 and ZIP13 were significantly increased. Increasing body mass index (BMI) correlated with a significant reduction in ZNT1 expression similar to what is seen in the early stages of AD. Increasing BMI also correlated with reduced expression of ZNT6. In conclusion, we found that the expression of genes that regulate intracellular zinc homeostasis in the human frontal cortex is altered with increasing age and affected by increasing BMI. With the increasing rates of obesity throughout the world, these findings warrant continuous scrutiny of the long-term consequences of obesity on brain function and the development of neurodegenerative diseases.


Genome-wide association studies of quantitatively measured skin, hair, and eye pigmentation in four European populations.
Pigmentation of the skin, hair, and eyes varies both within and between human populations. Identifying the genes and alleles underlying this variation has been the goal of many candidate gene and several genome-wide association studies (GWAS). Most GWAS for pigmentary traits to date have been based on subjective phenotypes using categorical scales. But skin, hair, and eye pigmentation vary continuously. Here, we seek to characterize quantitative variation in these traits objectively and accurately and to determine their genetic basis. Objective and quantitative measures of skin, hair, and eye color were made using reflectance or digital spectroscopy in Europeans from Ireland, Poland, Italy, and Portugal. A GWAS was conducted for the three quantitative pigmentation phenotypes in 176 women across 313,763 SNP loci, and replication of the most significant associations was attempted in a sample of 294 European men and women from the same countries. We find that the pigmentation phenotypes are highly stratified along axes of European genetic differentiation. The country of sampling explains approximately 35% of the variation in skin pigmentation, 31% of the variation in hair pigmentation, and 40% of the variation in eye pigmentation. All three quantitative phenotypes are correlated with each other. In our two-stage association study, we reproduce the association of rs1667394 at the OCA2/HERC2 locus with eye color but we do not identify new genetic determinants of skin and hair pigmentation supporting the lack of major genes affecting skin and hair color variation within Europe and suggesting that not only careful phenotyping but also larger cohorts are required to understand the genetic architecture of these complex quantitative traits. Interestingly, we also see that in each of these four populations, men are more lightly pigmented in the unexposed skin of the inner arm than women, a fact that is underappreciated and may vary across the world.


Ageing and visual spatiotemporal processing.
Ageing affects many visual functions. Here, we investigated the effects of ageing on vernier acuity and backward masking using the shine-through paradigm. We divided healthy older adults (>60 years) into two groups depending on whether vernier duration was comparable to younger adults (Older Adults 1) or not (Older Adults 2). Backward masking was deteriorated for Older Adults 2 but not for Older Adults 1. In addition, by using complex masking gratings, we found deficits in spatial and temporal vision in Older Adults 2, which cannot be explained by deteriorated visual acuity, pointing to cortical rather than retinal causes. Our results highlight the importance of taking into account individual differences in visual ageing research. In addition, our results have important implications for schizophrenia. Schizophrenia has been suggested to be a form of early brain ageing. Linking our current masking results in ageing to previous masking results in schizophrenia shows that schizophrenia is not a form of early ageing, at least not in the visual domain.


Prevalence of frailty and disability: findings from the English Longitudinal Study of Ageing.
to examine the prevalence of frailty and disability in people aged 60 and over and the proportion of those with disabilities who receive help or use assistive devices. participants were 5,450 people aged 60 and over from the English Longitudinal Study of Ageing. Frailty was defined according to the Fried criteria. Participants were asked about difficulties with mobility or other everyday activities. Those with difficulties were asked whether they received help or used assistive devices. the overall weighted prevalence of frailty was 14%. Prevalence rose with increasing age, from 6.5% in those aged 60-69 years to 65% in those aged 90 or over. Frailty occurred more frequently in women than in men (16 versus 12%). Mobility difficulties were very common: 93% of frail individuals had such difficulties versus 58% of the non-frail individuals. Among frail individuals, difficulties in performing activities or instrumental activities of daily living were reported by 57 or 64%, respectively, versus 13 or 15%, respectively, among the non-frail individuals. Among those with difficulties with mobility or other daily activities, 71% of frail individuals and 31% of non-frail individuals said that they received help. Of those with difficulties, 63% of frail individuals and 20% of non-frail individuals used a walking stick, but the use of other assistive devices was uncommon. frailty becomes increasingly common in older age groups and is associated with a sizeable burden as regards difficulties with mobility and other everyday activities.


GATA3 is redundant for maintenance and self-renewal of hematopoietic stem cells.
GATA3 has been identified as a master regulator of T helper cells, as well as being important for early thymic progenitors and T-cell commitment. However, Gata3 expression initiates already at the hematopoietic stem cell (HSC) level, implicating a potential role also in the regulation of HSCs. Herein we used a conditional Gata3 knockout strategy in which Gata3 expression was completely deleted from the earliest stage of embryonic hematopoietic development after emergence of HSCs from hemogenic endothelium. Through a detailed analysis of HSCs at the phenotypic and functional level, we demonstrate that steady-state levels of HSCs are normal in Gata3(fl/fl)Vav-Cre(tg/+) mice. Moreover, through long-term primary and secondary transplantation experiments, we also unequivocally demonstrate that Gata3 has a redundant role in post-transplantation HSC self-renewal.


The diversity of the fecal bacterial community and its relationship with the concentration of volatile fatty acids in the feces during subacute rumen acidosis in dairy cows.
Sub-acute ruminal acidosis (SARA) is a well-recognized digestive disorder found in particular in well-managed dairy herds. SARA can result in increased flow of fermentable substrates to the hindgut, which can increase the production of volatile fatty acids, alter the structure of the microbial community, and have a negative effect on animal health and productivity. However, little is known about changes in the structure of the microbial community and its relationship with fatty acids during SARA. Four cannulated primiparous (60 to 90 day in milk) Holstein dairy cows were assigned to two diets in a 2 × 2 crossover experimental design. The diets contained (on a dry matter basis): 40% (control diet, COD) and 70% (SARA induction diet, SAID) concentrate feeds. Samples of ruminal fluid and feces were collected on day 12, 15, 17 and 21 of the treatment period, and the pH was measured in the ruminal and fecal samples; the fecal microbiota was determined by pyrosequencing analysis of the V1-V3 region of amplified 16S ribosomal RNA (16S rRNA). SAID decreased ruminal and fecal pH and increased the propionate, butyrate and total volatile fatty acid (TVFA) concentration in feces when compared with the COD. A barcoded DNA pyrosequencing method was used to generate 2116 16S operational taxonomic units (OTUs). A total of 11 phyla were observed, distributed amongst all cattle on both diets; however, only 5 phyla were observed in all animals regardless of dietary treatment, and considerable animal to animal variation was revealed. The average abundance and its range of the 5 phyla were as follows: Firmicutes (63.7%, 29.1-84.1%), Proteobacteria (18.3%, 3.4-46.9%), Actinobacteria (6.8%, 0.4-39.9%), Bacteroidetes (7.6%, 2.2-17.7%) and Tenericutes (1.6%, 0.3-3%). Feeding the SAID resulted in significant shifts in the structure of the fecal microbial community when compared with the traditional COD. Among the 2116 OTUs detected in the present study, 88 OTUs were affected significantly by diet; and the proportion of these OTUs was 20.6% and 17.4% among the total number of sequences, respectively. Among the OTUs affected, the predominant species, including OTU2140 (G: Turicibacter), OTU1695 (G: Stenotrophomonas) and OTU8143 (F: Lachnospiraceae), were increased, while the abundance of OTU1266 (S: Solibacillus silvestris) and OTU2022 (G: Lysinibacillus) was reduced in the SAID group compared with the COD. Further, our results indicated that the fecal volatile fatty acid (VFA) concentrations were significantly related to presence of some certain species of Bacteroidetes and Firmicutes in the feces. This is, to our knowledge, the first study that has used barcoded DNA pyrosequencing to survey the fecal microbiome of dairy cattle during SARA. Our results suggest that particular bacteria and their metabolites in the feces appear to contribute to differences in host health between those given SAID and traditional COD feeding. A better understanding of these microbial populations will allow for improved nutrient management and increased animal growth performance.


Comparative Genomics Reveals Evidence of Genome Reduction and High Extracellular Protein Degradation Potential in Kangiella.
The genus Kangiella has recently been proposed within the family Kangiellaceae, belonging to order Oceanospirillales. Here, we report the complete genome sequence of a novel strain, Kangiella profundi FT102, which is the only Kangiella species isolated from a deep sea sediment sample. Furthermore, gaps in the publicly available genome scaffold of K. aquimarina DSM 16071 (NCBI Reference Sequence: NZ_ARFE00000000.1) were also filled using polymerase chain reaction (PCR) and Sanger sequencing. A comparative genomic analysis of five Kangiella and 18 non-Kangiella strains revealed insights into their metabolic potential. It was shown that low genomic redundancy and Kangiella-lineage-specific gene loss are the key reasons behind the genome reduction in Kangiella compared to that in any other free-living Oceanospirillales strain. The occurrence of relatively diverse and more frequent extracellular protease-coding genes along with the incomplete carbohydrate metabolic pathways in the genome suggests that Kangiella has high extracellular protein degradation potential. Growth of Kangiella strains has been observed using amino acids as the only carbon and nitrogen source and tends to increase with additional tryptone. Here, we propose that extracellular protein degradation and amino acid utilization are significant and prominent features of Kangiella. Our study provides more insight into the genomic traits and proteolytic metabolic capabilities of Kangiella.


Enhancing the weight training experience: a comparison of limb kinematics and EMG activity on three machines.
The aim of the study was to compare the kinematics and the timing and amount of electromyographic (EMG) activity during the lat-pull down exercise performed on machines that afforded one, two, or three degrees of freedom for the movement. Seven healthy men (age 29.4 +/- 5.6 years) participated in the study. The exercise was performed with a 30% 1-RM load. Three types of machines with varying degrees of freedom were used: Type 1, the conventional device that restricted the movement to a frontal plane; Type 2, the addition of forearm supination-pronation; Type 3, the addition of forearm supination-pronation and horizontal extension-flexion about the shoulder. All exercises involved a technique known as beginning movement load (BML) training in which light loads are lifted with a relaxation-lengthening-shortening sequence of muscle activation. The Type-3 machine showed: (1) the greatest vertical displacement of the wrist (p < 0.05); (2) the greatest abduction-adduction displacement about the shoulder (p < 0.01); (3) the least flexion-extension displacement about the elbow joint (p < 0.01); (4) a peak vertical velocity for the shoulder that preceded (p < 0.01) those for the elbow and then wrist during the pull-down phase; (5) a progressive proximal-to-distal sequence of EMG activation involving the serratus anterior, posterior deltoid, latissimus dorsi, and triceps brachii muscles; (6) a reversal of the roles for biceps and triceps brachii during the pull-down phase. These results suggest that BML exercises with greater degrees of freedom can enhance the association between training actions and functional activities.


Advances in cell-based therapy for structural heart disease.
Congestive heart failure and coronary artery disease are the leading causes of morbidity and mortality in the United States despite substantial therapeutic advances in the last half century. Only very recently have studies arisen that support possibility of regenerating tissue of damaged human organs including the heart. In this regard, there is growing pre-clinical and clinical evidence demonstrating the safety and efficacy of cell-based myocardial regeneration using a variety of cell lines. Although the data on the exact mechanism of action and the fate of the administered cells is controversial, there is consistent evidence for improved cardiac function and myocardial regeneration using different cell types. This extraordinarily exciting scientific advance has forced cardiovascular scientists to re-evaluate the long-held paradigm of cardiac myocyte terminal differentiation and life-long longevity of the cardiac myocytes that comprise the heart. Whereas, these new ideas originated with attempts to perform cellular transplantation using exogenous stem or precursor cells, mechanistic insights have rapidly evolved to the realization that adult organs harbor stem cells with significant plasticity, capable of repopulating their respective organ. Indeed these cells may be harnessed as a therapeutic agent or may represent the target of regenerative therapeutic strategies.


Early Syphilis Among Men Who Have Sex with Men in the US Pacific Northwest, 2008-2013: Clinical Management and Implications for Prevention.
Substantial increases in syphilis during 2008-2013 were reported in the US Pacific Northwest state of Oregon, especially among men who have sex with men (MSM). The authors aimed to characterize the ongoing epidemic and identify possible gaps in clinical management of early syphilis (primary, secondary, and latent syphilis ≤1 year) among MSM in Multnomah County, Oregon to inform public health efforts. Administrative databases were used to examine trends in case characteristics during 2008-2013. Medical records were abstracted for cases occurring in 2013 to assess diagnosis, treatment, and screening practices. Early syphilis among MSM increased from 21 cases in 2008 to 229 in 2013. The majority of cases occurred in HIV-infected patients (range: 55.6%-69.2%) diagnosed with secondary syphilis (range: 36.2%-52.4%). In 2013, 119 (51.9%) cases were diagnosed in public sector medical settings and 110 (48.0%) in private sector settings. Over 80% of HIV-infected patients with syphilis were in HIV care. Although treatment was adequate and timely among all providers, management differed by provider type. Among HIV-infected patients, a larger proportion diagnosed by public HIV providers than private providers were tested for syphilis at least once in the previous 12 months (89.6% vs. 40.0%; p < 0.001). The characteristics of MSM diagnosed with early syphilis in Multnomah County remained largely unchanged during 2008-2013. Syphilis control measures were well established, but early syphilis among MSM continued to increase. The results suggest a need to improve syphilis screening among private clinics, but few gaps in clinical management were identified.


Quality measures in gastrointestinal endoscopy: the current state.
The purpose of this review is to summarize the current state of endoscopic quality measurement and use of measures in enhancing the value of endoscopic services. Initially, quality measurement of endoscopic procedures was claims based or included small unit or practice-specific efforts. Now we have a mature national registry and large electronic medical or procedural records that are designed to yield valuable data relevant to quality measurement. With the advent of better measures, we are beginning to understand that initial process and surrogate outcome measures (adenoma detection rate) can be improved to provide a better reflection of endoscopic quality. Importantly, however, even measures currently in use relate to important patient outcomes such as missed colon cancers. At a federal level, older cumbersome pay-for-performance initiatives have been combined into a new overarching program named the quality payment program within the centers for medicare and medicaid services. This program is an additional step toward furthering the progress from volume-to-value-based reimbursement. The legislation mandating the movement toward outcomes-linked (value) reimbursement is the medicare access and children's health insurance program reauthorization act, which was passed with overwhelming bipartisan support and will not be walked back by alterations of the affordable care act. Increasing portions of medicare reimbursement (and likely commercial to follow) will be linked to quality metrics, so familiarity with the underlying process and rationale will be important for all proceduralists.


Early pathogenesis during infectious bursal disease in susceptible chickens is associated with changes in B cell genomic methylation and loss of genome integrity.
We propose a model by which an increase in the genomic modification, 5-hydroxymethylcytosine (5hmC), contributes to B cell death within the chicken bursa of Fabricus (BF) infected with infectious bursal disease virus (IBDV). Our findings indicate that, following an IBDV infection, Rhode Island Red (RIR) chickens have fewer surviving B cells and higher levels of 5hmC in the BF than the more resistant 15l line of birds. Elevated genomic 5hmC levels within the RIR BF are associated with markers of immune responses: infiltrating T cells and increased expression of CD40L, FasL and iNOS. Such changes correlate with genomic fragmentation and the presence of IBDV capsid protein, VP2. To explore the effects of CD40L, the immature B cell line, DT40, was exposed to recombinant chicken CD40L that resulted in changes in nuclear 5hmC distribution. Collectively, our observations suggest that T cell infiltration exacerbates early immunopathology within the BF during an IBDV infection contributing to B cell genomic instability and death to facilitate viral egress and immunosuppression.


A review of the oral health-related quality of life (OHRQL) model for dental hygiene: Eighteen years later.
The purpose of this paper is to review available evidence to determine the impact of the oral health-related quality of life (OHRQL) Model for Dental Hygiene on education, research, and practice. A search was conducted in PubMed using the search terms oral health-related quality of life, OHRQL, and dental hygiene and limited to publications between 1998 and 2016. Google Scholar was also used to identify citations for the two original OHRQL model papers to capture the literature widely read by practicing dental hygienists but not indexed in PubMed. A total of 40 publications met the predetermined search strategy. Full publications were obtained and reviewed to determine how this research is being used in education, research, and practice. The OHRQL is being utilized to a very limited degree in education, research, and practice. The literature shows how the OHRQL is actively being used in the school where the model originated in addition to a dental hygiene programme in Japan. Studies have been conducted on a variety of research instruments, including the OHRQL, designed to capture OHRQL outcomes. Japanese researchers have used the OHRQL in studies designed to determine the impact of oral healthcare delivery on patients' oral health-related quality of life. Eighteen years of the literature confirms that the OHRQL is being used minimally in education, research, and practice. Greater emphasis is needed around the collection and assessment of oral health-related quality of life measures for providing patient-centred care.


Matrix metalloproteinase-2 production and its binding to the matrix are increased in abdominal aortic aneurysms.
Degradation of the elastic media is a hallmark of abdominal aortic aneurysms (AAAs). We examined the expression of 2 elastolytic matrix metalloproteinases (MMPs), MMP-2 and MMP-9, in AAA aortic tissues compared with those from atherosclerotic occlusive disease (AOD) and nondiseased control tissues. Quantitative competitive reverse transcription-polymerase chain reaction and gelatin zymography showed increased MMP-9 mRNA and protein in both AAA and AOD tissues compared with those in control tissue, but there was no significant difference between AAA and AOD. In contrast, MMP-2 mRNA and protein levels were significantly higher in AAA than in AOD or control tissues. Sequential extraction of the MMPs from the aortic tissue with a physiological salt solution, 2% dimethylsulfoxide (DMSO), and 10 mol/L urea showed that large amounts of MMP-2 and MMP-9 were bound to the matrix. The most conspicuous finding was that the levels of MMP-2 were significantly elevated in the DMSO fraction in AAA tissues compared with AOD and control tissues. In addition, a large portion of MMP-2 found in the DMSO and urea fractions was in the active 62-kDa form, indicating that the precursor of MMP-2 in AAA is largely activated locally and binds to the tissue matrix tightly. By immunolocalization, MMP-9 was found to be primarily produced by macrophages and MMP-2 by mesenchymal cells. The production of MMP-2 was prominent when mesenchymal cells were surrounded by inflammatory cells, suggesting paracrine modulation of MMP-2 expression in AAAs. These observations emphasize that MMP-2 participates in the progression of AAAs by degrading aortic tissue matrix components.


Consequences of gestational diabetes in an urban hospital in Viet Nam: a prospective cohort study.
Gestational diabetes mellitus (GDM) is increasing and is a risk for type 2 diabetes. Evidence supporting screening comes mostly from high-income countries. We aimed to determine prevalence and outcomes in urban Viet Nam. We compared the proposed International Association of the Diabetes and Pregnancy Study Groups (IADPSG) criterion, requiring one positive value on the 75-g glucose tolerance test, to the 2010 American Diabetes Association (ADA) criterion, requiring two positive values. We conducted a prospective cohort study in Ho Chi Minh City, Viet Nam. Study participants were 2,772 women undergoing routine prenatal care who underwent a 75-g glucose tolerance test and interview around 28 (range 24-32) wk. GDM diagnosed by the ADA criterion was treated by local protocol. Women with GDM by the IADPSG criterion but not the ADA criterion were termed "borderline" and received standard care. 2,702 women (97.5% of cohort) were followed until discharge after delivery. GDM was diagnosed in 164 participants (6.1%) by the ADA criterion, 550 (20.3%) by the IADPSG criterion. Mean body mass index was 20.45 kg/m(2) in women with out GDM, 21.10 in women with borderline GDM, and 21.81 in women with GDM, p<0.001. Women with GDM and borderline GDM were more likely to deliver preterm, with adjusted odds ratios (aORs) of 1.49 (95% CI 1.16-1.91) and 1.52 (1.03-2.24), respectively. They were more likely to have clinical neonatal hypoglycaemia, aORs of 4.94 (3.41-7.14) and 3.34 (1.41-7.89), respectively. For large for gestational age, the aORs were 1.16 (0.93-1.45) and 1.31 (0.96-1.79), respectively. There was no significant difference in large for gestational age, death, severe birth trauma, or maternal morbidity between the groups. Women with GDM underwent more labour inductions, aOR 1.51 (1.08-2.11). Choice of criterion greatly affects GDM prevalence in Viet Nam. Women with GDM by the IADPSG criterion were at risk of preterm delivery and neonatal hypoglycaemia, although this criterion resulted in 20% of pregnant women being positive for GDM. The ability to cope with such a large number of cases and prevent associated adverse outcomes needs to be demonstrated before recommending widespread screening. Please see later in the article for the Editors' Summary.


Yoga-based exercise improves health-related quality of life and mental well-being in older people: a systematic review of randomised controlled trials.
health-related quality of life (HRQOL) and mental well-being are associated with healthy ageing. Physical activity positively impacts both HRQOL and mental well-being. Yoga is a physical activity that can be modified to suits the needs of older people and is growing in popularity. We conducted a systematic review with meta-analysis to determine the impact of yoga-based exercise on HRQOL and mental well-being in people aged 60+. searches were conducted for relevant trials in the following electronic databases; MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, CINAHL, Allied and Complementary Medicine Database, PsycINFO and the Physiotherapy Evidence Database (PEDro) from inception to January 2017. Trials that evaluated the effect of physical yoga on HRQOL and/or on mental well-being in people aged 60+ years were included. Data on HRQOL and mental well-being were extracted. Standardised mean differences and 95% confidence intervals (CI) were calculated using random effects models. Methodological quality of trials was assessed using the PEDro scale. twelve trials of high methodological quality (mean PEDro score 6.1), totalling 752 participants, were identified and provided data for the meta-analysis. Yoga produced a medium effect on HRQOL (Hedges' g = 0.51, 95% CI 0.25-0.76, 12 trials) and a small effect on mental well-being (Hedges' g = 0.38, 95% CI 0.15-0.62, 12 trials). yoga interventions resulted in small to moderate improvements in both HRQOL and mental well-being in people aged 60+ years. Further, research is needed to determine the optimal dose of yoga to maximise health impact. (CRD42016052458).


Gateway-compatible yeast one-hybrid screens.
INTRODUCTIONProtein-DNA interactions (PDIs) between transcription factors (TFs) and their target genes form the backbone of transcription regulatory networks. Such PDIs can be identified with either a TF or a gene as a starting point. The Gateway-compatible yeast one-hybrid (Y1H) system provides a high-throughput, gene-centered method for the identification of interactions between a "DNA bait" (e.g., cis-regulatory DNA elements or gene promoters) and "protein preys" (e.g., TFs). The Y1H system is a genetic system to detect PDIs based on selection of reporter gene expression in yeast. DNA baits are fused by Gateway cloning to two reporter genes, HIS3 and lacZ, and the resulting DNA bait::reporter constructs are subsequently integrated into the genome of the host yeast strain. After integration, baits are examined for self-activation (i.e., their ability to drive reporter gene expression in the absence of an exogenous prey protein). Subsequently, each DNA bait is screened for interacting proteins by transforming a library of preys into the corresponding Y1H DNA bait yeast strain. Preys are hybrid proteins composed of a protein from the organism of interest and a heterologous transcription activation domain. When a prey protein binds to the DNA bait, the heterologous activation domain activates reporter gene expression. Thus, physical interactions between both repressors and activators and their DNA targets can be identified.


Remodeling sperm chromatin in Xenopus laevis egg extracts: the role of core histone phosphorylation and linker histone B4 in chromatin assembly.
We find that the remodeling of the condensed Xenopus laevis sperm nucleus into the paternal pronucleus in egg extracts is associated with phosphorylation of the core histones H2A, H2A.X and H4, and uptake of a linker histone B4 and a HMG 2 protein. Histone B4 is required for the assembly of chromatosome structures in the pronucleus. However neither B4 nor core histone phosphorylation are required for the assembly of spaced nucleosomal arrays. We suggest that the spacing of nucleosomal arrays is determined by interaction between adjacent histone octamers under physiological assembly conditions.


Human periodontal ligament fibroblast responses to compression in chronic periodontitis.
Test whether human periodontal ligament fibroblasts (PDLFs) retain homeostatic responses to a physiological compressive force during chronic periodontitis. Six cell lines were established from periodontally healthy individuals (H-PDLFs) and another six were cultured from patients diagnosed with chronic periodontitis (D-PDLFs). Compressive force at 150 psi was applied to H-and D-PDLFs for 3 h on 2 consecutive days. After compression, comparisons between H-and D-PDLFs were performed by gene expression analysis of IL-6, proteases and 84 inflammation-related targets using real-time PCR. Compression of H-PDLFs resulted in a significant increase only in MMP-1 mRNA. In contrast, the same compressive force on D-PDLFs produced significant increases in the expression of MMPs-1,-7,-9 and -16. Moreover, compression of H-PDLFs resulted in down-regulation of IL-6, while IL-6 was significantly up-regulated in compressed D-PDLFs. Compression of H-PDLFs slightly up-regulated 3 and significantly down-regulated 15 inflammation-related genes, while the same treatment strongly up-regulated 21 inflammation-related genes in D-PDLFs. These results suggest a fundamental difference in the inflammatory response of healthy versus diseased PDLFs under physiological compression. Maintenance of these characteristics in vitro suggests that these cells may be at least partly responsible for the persistence of inflammation and localized susceptibility in chronic periodontitis.


Cross sections for rotational decoherence of perturbed nitrogen measured via decay of laser-induced alignment.
We quantitatively determine cross sections for rotational decoherence from the decay of nonadiabatic laser-induced alignment in nitrogen and nitrogen-foreign gas mixtures in a temperature range between 80 K and room temperature. The cross section for rotational decoherence in pure nitrogen decreases from 102 A(2) at 80 K to 48 A(2) at 295 K, leading to long-lived coherences even at high temperatures. Comparison with the broadening of the transition lines of the Raman Q-branch reported in the literature shows that the decay of rotational coherence proceeds at the same rate as rotational depopulation. This is verified also for mixtures of nitrogen with hydrogen, helium, argon, and krypton. We discuss limits posed by a possible J-dependence of the cross sections and strategies for state resolved determination from the time-dependent alignment signal.


Gravitropic response and circumnutation in pea (Pisum sativum) seedling roots.
Plant circumnutation is a helical movement of growing organs such as shoots and roots. Gravitropic response is hypothesized to act as an external oscillator in shoot circumnutation, although this is subject to debate. The relationship between circumnutational movement and gravitropic response in roots remains unknown. In this study, we analyzed circumnutation of agravitropic roots using the ageotropum pea (Pisum sativum) mutant, and compared it with that of wild-type (cv. Alaska) pea roots. We further examined the relationship of gravitropic response to circumnutation of Alaska seedling roots by removing the gravisensing tissue (the root cap) and by treating the roots with auxin transport inhibitors. Alaska roots displayed circumnutational movements with a period of approximately 150 min, whereas ageotropum roots did not exhibit distinct circumnutational movement. Removal of the root cap in Alaska roots reduced gravitropic response and circumnutational movements. Treatment of Alaska roots with auxin transport inhibitors, 2,3,5-triiodobenzoic acid (TIBA) and N-(1-naphthyl)phthalamic acid (NPA), dramatically reduced gravitropic response and circumnutational movements. These results suggest that a gravity-regulated auxin transport is involved in circumnutation of pea seedling roots.


External environment and internal state in relation to life-history behavioural profiles of adolescents in nine countries.
The external environment has traditionally been considered as the primary driver of animal life history (LH). Recent research suggests that animals' internal state is also involved, especially in forming LH behavioural phenotypes. The present study investigated how these two factors interact in formulating LH in humans. Based on a longitudinal sample of 1223 adolescents in nine countries, the results show that harsh and unpredictable environments and adverse internal states in childhood are each uniquely associated with fast LH behavioural profiles consisting of aggression, impulsivity, and risk-taking in adolescence. The external environment and internal state each strengthened the LH association of the other, but overall the external environment was more predictive of LH than was the internal state. These findings suggest that individuals rely on a multitude and consistency of sensory information in more decisively calibrating LH and behavioural strategies.


Using Systematic Anomalous Case Analysis to Examine Sexual and Reproductive Health Outcomes in the Philippines.
The Philippines is characterized by sustained economic growth and political stability, yet sexual and reproductive health indicators have stalled or even worsened in recent decades. We employed an innovative, mixed-methods approach-Systematic Anomalous Case Analysis-to gain insights into these worsening trends by examining sexual and reproductive decision-making among a cohort of young adults in Metro Cebu, Philippines. We first analyzed longitudinal data (1998-2009) to predict reproductive outcomes (i.e., age of first sex, number of living children) among participants in the Cebu Longitudinal Health and Nutrition Survey to identify cases (predicted and anomalous) with whom we subsequently conducted qualitative, in-depth interviews in 2013-14 (n = 48). Analysis of the qualitative data revealed unique social and contextual factors that shaped patterns of sexual and contraceptive decision-making across three, distinct reproductive life stages: (1) at first sex, (2) after the birth of first child, and (3) after the birth of several children. However, gendered roles and expectations exerted strong influences on sexual and reproductive outcomes across these life stages. Finally, we identified two constructs from our qualitative analysis-sexual fluidity and sexual agency-that deserve further examination and integration into theoretical and empirical models of sexual and reproductive decision-making.


Multiplication of microbes below 0.690 water activity: implications for terrestrial and extraterrestrial life.
Since a key requirement of known life forms is available water (water activity; aw ), recent searches for signatures of past life in terrestrial and extraterrestrial environments have targeted places known to have contained significant quantities of biologically available water. However, early life on Earth inhabited high-salt environments, suggesting an ability to withstand low water-activity. The lower limit of water activity that enables cell division appears to be ∼ 0.605 which, until now, was only known to be exhibited by a single eukaryote, the sugar-tolerant, fungal xerophile Xeromyces bisporus. The first forms of life on Earth were, though, prokaryotic. Recent evidence now indicates that some halophilic Archaea and Bacteria have water-activity limits more or less equal to those of X. bisporus. We discuss water activity in relation to the limits of Earth's present-day biosphere; the possibility of microbial multiplication by utilizing water from thin, aqueous films or non-liquid sources; whether prokaryotes were the first organisms able to multiply close to the 0.605-aw limit; and whether extraterrestrial aqueous milieux of ≥ 0.605 aw can resemble fertile microbial habitats found on Earth.


IMI 2021 Yearly Digest.
The International Myopia Institute (IMI) Yearly Digest highlights new research considered to be of importance since the publication of the first series of IMI white papers. A literature search was conducted for articles on myopia between 2019 and mid-2020 to inform definitions and classifications, experimental models, genetics, interventions, clinical trials, and clinical management. Conference abstracts from key meetings in the same period were also considered. One thousand articles on myopia have been published between 2019 and mid-2020. Key advances include the use of the definition of premyopia in studies currently under way to test interventions in myopia, new definitions in the field of pathologic myopia, the role of new pharmacologic treatments in experimental models such as intraocular pressure-lowering latanoprost, a large meta-analysis of refractive error identifying 336 new genetic loci, new clinical interventions such as the defocus incorporated multisegment spectacles and combination therapy with low-dose atropine and orthokeratology (OK), normative standards in refractive error, the ethical dilemma of a placebo control group when myopia control treatments are established, reporting the physical metric of myopia reduction versus a percentage reduction, comparison of the risk of pediatric OK wear with risk of vision impairment in myopia, the justification of preventing myopic and axial length increase versus quality of life, and future vision loss. Large amounts of research in myopia have been published since the IMI 2019 white papers were released. The yearly digest serves to highlight the latest research and advances in myopia.


Feasibility and effectiveness of the implementation of a primary prevention programme for type 2 diabetes in routine primary care practice: a phase IV cluster randomised clinical trial.
The objective of this study is to perform an independent evaluation of the feasibility and effectiveness of an educational programme for the primary prevention of type 2 diabetes (DM2) in high risk populations in primary care settings, implanted within the Basque Health Service - Osakidetza. This is a prospective phase IV cluster clinical trial conducted under routine conditions in 14 primary health care centres of Osakidetza, randomly assigned to an intervention or control group. We will recruit a total sample of 1089 individuals, aged between 45 and 70 years old, without diabetes but at high risk of developing the condition (Finnish Diabetes Risk Score, FINDRISC ≥ 14) and follow them up for 2 years. Primary health care nursing teams of the intervention centres will implement DE-PLAN, a structured educational intervention program focused on changing healthy lifestyles (diet and physical activity); while the patients in the control centres will receive the usual care for the prevention and treatment of DM2 currently provided in Osakidetza. The effectiveness attributable to the programme will be assessed by comparing the changes observed in patients exposed to the intervention and those in the control group, with respect to the risk of developing DM2 and lifestyle habits. In terms of feasibility, we will assess indicators of population coverage and programme implementation. The aim of this study is to provide the scientific basis for disseminate the programme to the remaining primary health centres in Osakidetza, as a novel way of addressing prevention of DM2. The study design will enable us to gather information on the effectiveness of the intervention as well as the feasibility of implementing it in routine practice.


Job loss and reemployment after a cancer diagnosis in Koreans - a prospective cohort study.
Patients undergoing treatment for cancer have reported a variety of work-related problems. The aim of this study was to investigate the impact of a cancer diagnosis on employment status, and to identify relevant associated factors. This prospective cohort study was conducted at the National Cancer Center in Korea. Male patients newly diagnosed with stomach, liver, or colorectal cancers were recruited. Patients were interviewed and asked to complete an employment questionnaire every 3 months for 24 months. Clinical, sociodemographic and work-related factors were assessed. There were 305 male patients who had a primary diagnosis of cancer and underwent appropriate treatment. Of the 305 male patients who were employed at the time of diagnosis, 53% lost their job, and of these 23% later reemployed. In a multivariate Cox regression analysis, job loss was significantly associated with years of education, job characteristics and disease stage. Reemployment was significantly associated with disease stage and cancer site. Change of employment was common among cancer patients in Korea. With an understanding of the factors involved, it should be possible to reduce unnecessary work cessation, and increase the rate of employment of cancer survivors.


One year mortality of patients treated with an emergency department based early goal directed therapy protocol for severe sepsis and septic shock: a before and after study.
Early structured resuscitation of severe sepsis has been suggested to improve short term mortality; however, no previous study has examined the long-term effect of this therapy. We sought to determine one year outcomes associated with implementation of early goal directed therapy (EGDT) in the emergency department (ED) care of sepsis. We performed a longitudinal analysis of a prospective before and after study conducted at a large urban ED. Adult patients were enrolled if they had suspected infection, 2 or more systemic inflammatory response criteria, and either systolic blood pressure (SBP) <90 mmHg after a fluid bolus or lactate >4 mM. Exclusion criteria were: age <18 years, no aggressive care desired, or need for immediate surgery. Clinical and outcomes data were prospectively collected on consecutive eligible patients for 1 year before and 2 years after implementing EGDT. Patients in the pre-implementation phase received non-protocolized care at attending physician discretion. The primary outcome was mortality at one year. 285 subjects, 79 in the pre- and 206 in the post-implementation phases, were enrolled. Compared to pre-implementation, post-implementation subjects had a significantly lower ED SBP (72 vs. 85 mm Hg, P < 0.001) and higher sequential organ failure assessment score (7 vs. 5, P = 0.0004). The primary outcome of 1 year mortality was observed in 39/79 (49%) pre-implementation subjects and 77/206 (37%) post-implementation subjects (difference 12%; P = 0.04). Implementation of EGDT for the treatment of ED patients with severe sepsis and septic shock was associated with significantly lower mortality at one year.


Maintenance of cytomegalovirus-specific CD4pos T-cell response in rheumatoid arthritis patients receiving anti-tumor necrosis factor treatments.
Anti-tumor necrosis factor (TNF)-α biotherapies have considerably changed the treatment of rheumatoid arthritis (RA). However, serious infections are a major concern in patients with rheumatic diseases treated with anti-TNF-α. Little is known about viral, especially latent, infections in anti-TNF-α treatments. Infections by cytomegalovirus (CMV), a β-herpes virus, are frequent and induce a strong CD4pos T-cell immunity, which participates in the control of infection. We thus have chosen to analyze the CD4pos T-cell response to CMV antigens as a model of antiviral response in RA patients treated with anti-TNF-α. CD28 expression was evaluated. We have measured the CD4pos response to CMV antigens in RA patients, before and after initiation of treatment with an anti-TNF-α agent. The intracellular production of interferon (IFN)-γ in total and CD28neg CD4pos T cells in response to CMV antigens (Ags) was evaluated with flow cytometry. The proliferation of total CD4pos T cells in the presence of CMV antigens was measured with 3H-thymidine incorporation. Anti-TNF-α treatments impaired neither the anti-CD4pos anti-CMV IFN-γ response nor the proliferative response in patients. The percentage of CD28neg CD4pos cells remained constant. Our data suggest that the CD4pos T-cell response against CMV is not altered by anti-TNF-α treatments and that infection remains controlled in treated RA patients latently infected with CMV. Our observation brings new insight into the current knowledge of the risks of infection in patients treated with anti-TNF-α biotherapies.


Hepatic involvement in hereditary hemorrhagic telangiectasia: helical computed tomography features in 24 consecutive patients.
Among the various organs that may be affected by hereditary hemorrhagic telangiectasia (HHT), the liver can show various degrees of vascular and parenchymal involvement. The purpose of this prospective study comprising a large series of patients was to reassess the computed tomography (CT) features of hepatic involvement in HHT using helical CT. Twenty-four consecutive patients with HHT had prospective helical CT of the liver, including noncontrast, arterial-dominant, and portal-dominant phases. The CT images were analyzed by 2 readers in consensus to determine the presence of vascular and parenchymal abnormalities. The diameter of the proper hepatic artery in these 24 patients was compared with that in 24 healthy subjects (Student t test). Helical CT was normal in 5 patients (21%) and abnormal in 19 patients (79%). Vascular abnormalities were found in 16 patients (67%), consisting of marked dilatation of the hepatic artery (n = 16), intrahepatic telangiectases (n = 12), arteriovenous shunting (n = 5), and arterioportal shunting (n=3). The diameter of the proper hepatic artery was greater in the patients with HHT than in control subjects (6.12 +/- 2.52 mm vs. 3.29 +/- 0.65 mm, respectively; P < 0.05). Helical CT showed nodular hyperplasia in 1 patient with vascular and parenchymal abnormalities, cavernous hemangiomas in 2 patients (1 in a patient with an enlarged hepatic artery, intrahepatic telangiectases, and arteriovenous shunting and 1 in a patient with an isolated enlarged hepatic artery), and biliary cysts in 3 patients (2 biliary cysts were present in 2 patients with an enlarged hepatic artery and intrahepatic telangiectases, and 1 biliary cyst was present without any manifestations in the third patient). Liver involvement in HHT is associated with a constellation of findings on helical CT, including significant dilatation of the proper hepatic artery, telangiectases, arteriovenous shunting, and focal liver lesions. Familiarity with these findings will result in more accurate diagnosis and allows better therapeutic options if necessary.


WE-E-BRB-10: DosCheck - an Electronic Chart Checking Tool for Dosimetrists.
In addition to treatment planning, dosimetrists have to prepare documentation and manually enter data in treatment management system (TMS) which did not transfer or setup automatically. The required documents and data are dependent on the disease site, treatment machine and clinical workflow. Errors and inconsistencies can cause redundant work, treatment delays and potentially treatment errors. To address these issues, an electronic checking software tool, DosCheck was clinically implemented to check the existence of necessary documentations and the integrity of manually-entered data. The purpose of this software is to reduce the frequency of human errors and to improve efficiency. DosCheck reads data and documents from 1) TMS, 2) Pinnacle TPS, and 3) DICOM plan files stored in a DICOM-RT PACS. It processes documents in Word and PDF format, treatment plan data in Pinnacle native format and DICOM format, and Mosaiq data in database records. The software cross-checks data accuracy and consistency by following rules that are pre-defined according to the clinical requirements and treatment sties. It interacts with dosimetrists and presents instantaneous results via graphical user interface. DosCheck has been implemented in C#. It performs a full check for a patient with 20 seconds. It has been clinically commissioned and is used daily by all dosimetrists at our institution. Retrospective analysis shows that DosCheck identifies 30% to 40% of previously reported dosimetrist human errors. Additional ∼30% errors are checked by other tools that could be integrated DosCheck in the near future. As an electronic data checking tool, DosCheck can obtain and process data and documents from multiple clinical computer systems in the radiation oncology department, and perform checks according to clinical rules. It is able to improve the accuracy and efficiency of clinical data and document process, and therefore to reduce any potential inconsistencies and errors.


Histone deacetylase inhibitors SAHA and sodium butyrate block G1-to-S cell cycle progression in neurosphere formation by adult subventricular cells.
Histone deacetylases (HDACs) are enzymes that modulate gene expression and cellular processes by deacetylating histones and non-histone proteins. While small molecule inhibitors of HDAC activity (HDACi) are used clinically in the treatment of cancer, pre-clinical treatment models suggest they also exert neuroprotective effects and stimulate neurogenesis in neuropathological conditions. However, the direct effects of HDACi on cell cycle progression and proliferation, two properties required for continued neurogenesis, have not been fully characterized in adult neural stem cells (NSCs). In this study, we examined the effects of two broad class I and class II HDACi on adult mouse NSCs, the hydroxamate-based HDACi suberoylanilide hydroxamic acid (vorinostat, SAHA) and the short chain fatty acid HDACi sodium butyrate. We show that both HDACi suppress the formation of neurospheres by adult mouse NSCs grown in proliferation culture conditions in vitro. DNA synthesis is significantly inhibited in adult mouse NSCs exposed to either SAHA or sodium butyrate and inhibition is associated with an arrest in the G1 phase of the cell cycle. HDACi exposure also resulted in transcriptional changes in adult mouse NSCs. Cdk inhibitor genes p21 and p27 transcript levels are increased and associated with elevated H3K9 acetylation levels at proximal promoter regions of p21 and p27. mRNA levels for notch effector Hes genes and Spry-box stem cell transcription factors are downregulated, whereas pro-neural transcription factors Neurog1 and Neurod1 are upregulated. Lastly, we show HDAC inhibition under proliferation culture conditions leads to long-term changes in cell fate in adult mouse NSCs induced to differentiate in vitro. SAHA and sodium butyrate directly regulate cdk inhibitor transcription to control cell cycle progression in adult mouse NSCs. HDAC inhibition results in G1 arrest in adult mouse NSCs and transcriptional changes associated with activation of neuronal lineage commitment programs and a reduction of stem/progenitor state. Changes in differentiated cell state in adult mouse NSCs treated with HDACi under proliferation culture conditions suggests an intrinsic relationship between multipotency, cell cycle progression and HDAC activity in these cells.


The role of rostral prefrontal cortex in prospective memory: a voxel-based lesion study.
Patients with lesions in rostral prefrontal cortex (PFC) often experience problems in everyday-life situations requiring multitasking. A key cognitive component that is critical in multitasking situations is prospective memory, defined as the ability to carry out an intended action after a delay period filled with unrelated activity. The few functional imaging studies investigating prospective memory have shown consistent activation in both medial and lateral rostral PFC but also in more posterior prefrontal regions and non-frontal regions. The aim of this study was to determine regions that are necessary for prospective memory performance, using the human lesion approach. We designed an experimental paradigm allowing us to assess time-based (remembering to do something at a particular time) and event-based (remembering to do something in a particular situation) prospective memory, using two types of material, words and pictures. Time estimation tasks and tasks controlling for basic attention, inhibition and multiple instructions processing were also administered. We examined brain-behaviour relationships with a voxelwise lesion method in 45 patients with focal brain lesions and 107 control subjects using this paradigm. The results showed that lesions in the right polar prefrontal region (in Brodmann area 10) were specifically associated with a deficit in time-based prospective memory tasks for both words and pictures. This deficit could not be explained by impairments in basic attention, detection, inhibition or multiple instruction processing, and there was also no deficit in event-based prospective memory conditions. In addition to their prospective memory difficulties, these polar prefrontal patients were significantly impaired in time estimation ability compared to other patients. The same region was found to be involved using both words and pictures, suggesting that right rostral PFC plays a material nonspecific role in prospective memory. This is the first lesion study showing that rostral PFC is crucial for time-based prospective memory. The findings suggest that time-based and event-based prospective memory might be supported at least in part by distinct brain regions. Two particularly plausible explanations for the deficit rest upon a possible role for polar prefrontal structures in supporting in time estimation, and/or in retrieving an intention to act. More broadly, the results are consistent with the view that the deficit of rostral patients in multitasking situations might at least in part be explained by a deficit in prospective memory.


Farnesoid X nuclear receptor ligand obeticholic acid for non-cirrhotic, non-alcoholic steatohepatitis (FLINT): a multicentre, randomised, placebo-controlled trial.
The bile acid derivative 6-ethylchenodeoxycholic acid (obeticholic acid) is a potent activator of the farnesoid X nuclear receptor that reduces liver fat and fibrosis in animal models of fatty liver disease. We assessed the efficacy of obeticholic acid in adult patients with non-alcoholic steatohepatitis. We did a multicentre, double-blind, placebo-controlled, parallel group, randomised clinical trial at medical centres in the USA in patients with non-cirrhotic, non-alcoholic steatohepatitis to assess treatment with obeticholic acid given orally (25 mg daily) or placebo for 72 weeks. Patients were randomly assigned 1:1 using a computer-generated, centrally administered procedure, stratified by clinical centre and diabetes status. The primary outcome measure was improvement in centrally scored liver histology defined as a decrease in non-alcoholic fatty liver disease activity score by at least 2 points without worsening of fibrosis from baseline to the end of treatment. A planned interim analysis of change in alanine aminotransferase at 24 weeks undertaken before end-of-treatment (72 weeks) biopsies supported the decision to continue the trial (relative change in alanine aminotransferase -24%, 95% CI -45 to -3). A planned interim analysis of the primary outcome showed improved efficacy of obeticholic acid (p=0·0024) and supported a decision not to do end-of-treatment biopsies and end treatment early in 64 patients, but to continue the trial to obtain the 24-week post-treatment measures. Analyses were done by intention-to-treat. This trial was registered with ClinicalTrials.gov, number NCT01265498. Between March 16, 2011, and Dec 3, 2012, 141 patients were randomly assigned to receive obeticholic acid and 142 to placebo. 50 (45%) of 110 patients in the obeticholic acid group who were meant to have biopsies at baseline and 72 weeks had improved liver histology compared with 23 (21%) of 109 such patients in the placebo group (relative risk 1·9, 95% CI 1·3 to 2·8; p=0·0002). 33 (23%) of 141 patients in the obeticholic acid developed pruritus compared with nine (6%) of 142 in the placebo group. Obeticholic acid improved the histological features of non-alcoholic steatohepatitis, but its long-term benefits and safety need further clarification. National Institute of Diabetes and Digestive and Kidney Diseases, Intercept Pharmaceuticals.


Miniaturized isothermal nucleic acid amplification, a review.
Micro-Total Analysis Systems (µTAS) for use in on-site rapid detection of DNA or RNA are increasingly being developed. Here, amplification of the target sequence is key to increasing sensitivity, enabling single-cell and few-copy nucleic acid detection. The several advantages to miniaturizing amplification reactions and coupling them with sample preparation and detection on the same chip are well known and include fewer manual steps, preventing contamination, and significantly reducing the volume of expensive reagents. To-date, the majority of miniaturized systems for nucleic acid analysis have used the polymerase chain reaction (PCR) for amplification and those systems are covered in previous reviews. This review provides a thorough overview of miniaturized analysis systems using alternatives to PCR, specifically isothermal amplification reactions. With no need for thermal cycling, isothermal microsystems can be designed to be simple and low-energy consuming and therefore may outperform PCR in portable, battery-operated detection systems in the future. The main isothermal methods as miniaturized systems reviewed here include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), and strand displacement amplification (SDA). Also, important design criteria for the miniaturized devices are discussed. Finally, the potential of miniaturization of some new isothermal methods such as the exponential amplification reaction (EXPAR), isothermal and chimeric primer-initiated amplification of nucleic acids (ICANs), signal-mediated amplification of RNA technology (SMART) and others is presented.


Art of disaster preparedness in European union: a survey on the health systems.
Naturally occurring and man-made disasters have been increasing in the world, including Europe, over the past several decades. Health systems are a key part of any community disaster management system. The success of preparedness and prevention depends on the success of activities such as disaster planning, organization and training. The aim of this study is to evaluate health system preparedness for disasters in the 27 European Union member countries. A cross-sectional analysis study was completed between June-September 2012. The checklist used for this survey was a modified from the World Health Organization toolkit for assessing health-system capacity for crisis management. Three specialists from each of the 27 European Union countries were included in the survey. Responses to each survey question were scored and the range of preparedness level was defined as 0-100%, categorized in three levels as follows: Acceptable; Transitional; or Insufficient. Response rate was 79.1%. The average level of disaster management preparedness in the health systems of 27 European Union member states was 68% (Acceptable). The highest level of preparedness was seen in the United Kingdom, Luxemburg, and Lithuania. Considering the elements of disaster management system, the highest level of preparedness score was at health information elements (86%), and the lowest level was for hospitals, and educational elements (54%). This survey study suggests that preparedness level of European Union countries in 2012 is at an acceptable level but could be improved. Elements such as hospitals and education and training suffer from insufficient levels of preparedness. The European Union health systems need a collective strategic plan, as well as enough resources, to establish a comprehensive and standardized disaster management strategy plan. A competency based training curriculum for managers and first responders is basic to accomplishing this goal. Disaster medicine; Disaster preparedness; Disaster epidemiology; Health systems; European Union.


Therapeutic Targets for Treatment of Heart Failure: Focus on GRKs and β-Arrestins Affecting βAR Signaling.
Heart failure (HF) is a heart disease that is classified into two main types: HF with reduced ejection fraction (HFrEF) and HF with preserved ejection fraction (HFpEF). Both types of HF lead to significant risk of mortality and morbidity. Pharmacological treatment with β-adrenergic receptor (βAR) antagonists (also called β-blockers) has been shown to reduce the overall hospitalization and mortality rates and improve the clinical outcomes in HF patients with HFrEF but not HFpEF. Although, the survival rate of patients suffering from HF continues to drop, the management of HF still faces several limitations and discrepancies highlighting the need to develop new treatment strategies. Overstimulation of the sympathetic nervous system is an adaptive neurohormonal response to acute myocardial injury and heart damage, whereas prolonged exposure to catecholamines causes defects in βAR regulation, including a reduction in the amount of βARs and an increase in βAR desensitization due to the upregulation of G protein-coupled receptor kinases (GRKs) in the heart, contributing in turn to the progression of HF. Several studies show that myocardial GRK2 activity and expression are raised in the failing heart. Furthermore, β-arrestins play a pivotal role in βAR desensitization and, interestingly, can mediate their own signal transduction without any G protein-dependent pathway involved. In this review, we provide new insight into the role of GRKs and β-arrestins on how they affect βAR signaling regarding the molecular and cellular pathophysiology of HF. Additionally, we discuss the therapeutic potential of targeting GRKs and β-arrestins for the treatment of HF.


Phosphoproteomic network analysis in the sea urchin Strongylocentrotus purpuratus reveals new candidates in egg activation.
Fertilization triggers a dynamic symphony of molecular transformations induced by a rapid rise in intracellular calcium. Most prominent are surface alterations, metabolic activation, cytoskeletal reorganization, and cell-cycle reentry. While the activation process appears to be broadly evolutionarily conserved, and protein phosphorylation is known to play a key role, the signaling networks mediating the response to fertilization are not well described. To address this gap, we performed a time course phosphoproteomic analysis of egg activation in the sea urchin Strongylocentrotus purpuratus, a system that offers biochemical tractability coupled with exquisite synchronicity. By coupling large-scale phosphopeptide enrichment with unbiased quantitative MS, we identified striking changes in global phosphoprotein patterns at 2- and 5-min postfertilization as compared to unfertilized eggs. Overall, we mapped 8796 distinct phosphosite modifications on 2833 phosphoproteins, of which 15% were differentially regulated in early egg activation. Activated kinases were identified by phosphosite mapping, while enrichment analyses revealed conserved signaling cascades not previously associated with egg activation. This work represents the most comprehensive study of signaling associated with egg activation to date, suggesting novel mechanisms that can be experimentally tested and providing a valuable resource for the broader research community. All MS data have been deposited in the ProteomeXchange with identifier PXD002239 (http://proteomecentral.proteomexchange.org/dataset/PXD002239).


The DNA and RNA sugar-phosphate backbone emerges as the key player. An overview of quantum-chemical, structural biology and simulation studies.
Knowledge of geometrical and physico-chemical properties of the sugar-phosphate backbone substantially contributes to the comprehension of the structural dynamics, function and evolution of nucleic acids. We provide a side by side overview of structural biology/bioinformatics, quantum chemical and molecular mechanical/simulation studies of the nucleic acids backbone. We highlight main features, advantages and limitations of these techniques, with a special emphasis given to their synergy. The present status of the research is then illustrated by selected examples which include classification of DNA and RNA backbone families, benchmark structure-energy quantum chemical calculations, parameterization of the dihedral space of simulation force fields, incorporation of arsenate into DNA, sugar-phosphate backbone self-cleavage in small RNA enzymes, and intricate geometries of the backbone in recurrent RNA building blocks. Although not apparent from the current literature showing limited overlaps between the QM, simulation and bioinformatics studies of the nucleic acids backbone, there in fact should be a major cooperative interaction between these three approaches in studies of the sugar-phosphate backbone.


Photochromic histone deacetylase inhibitors based on dithienylethenes and fulgimides.
Histone deacetylases (HDACs) play a crucial role in numerous biological processes and therefore are targeted in anticancer research and in the field of epigenetics. Dithienylethenes (DTEs) and fulgimides were functionalized with hydroxamic acids, which is a known moiety binding to zinc dependent HDACs, to gain photoswitchable HDAC inhibitors. The new DTE based inhibitors showed moderate photochromic properties in polar solvents and the inhibitory activity changes up to a factor of four. The photochromic performance of the prepared fulgimide inhibitors was very good, even in aqueous buffer. They achieved a maximum three-fold difference in inhibitory activity. Docking experiments using the crystal structures of the tested enzymes gave a rationale for the observed moderate differences in the activities of the inhibitors.