Reduced expression of brain-enriched microRNAs in glioblastomas permits targeted regulation of a cell death gene.
Glioblastoma is a highly aggressive malignant tumor involving glial cells in the human brain. We used high-throughput sequencing to comprehensively profile the small RNAs expressed in glioblastoma and non-tumor brain tissues. MicroRNAs (miRNAs) made up the large majority of small RNAs, and we identified over 400 different cellular pre-miRNAs. No known viral miRNAs were detected in any of the samples analyzed. Cluster analysis revealed several miRNAs that were significantly down-regulated in glioblastomas, including miR-128, miR-124, miR-7, miR-139, miR-95, and miR-873. Post-transcriptional editing was observed for several miRNAs, including the miR-376 family, miR-411, miR-381, and miR-379. Using the deep sequencing information, we designed a lentiviral vector expressing a cell suicide gene, the herpes simplex virus thymidine kinase (HSV-TK) gene, under the regulation of a miRNA, miR-128, that was found to be enriched in non-tumor brain tissue yet down-regulated in glioblastomas, Glioblastoma cells transduced with this vector were selectively killed when cultured in the presence of ganciclovir. Using an in vitro model to recapitulate expression of brain-enriched miRNAs, we demonstrated that neuronally differentiated SH-SY5Y cells transduced with the miRNA-regulated HSV-TK vector are protected from killing by expression of endogenous miR-128. Together, these results provide an in-depth analysis of miRNA dysregulation in glioblastoma and demonstrate the potential utility of these data in the design of miRNA-regulated therapies for the treatment of brain cancers.


Simultaneous detection of Plasmodium vivax dhfr, dhps, mdr1 and crt-o resistance-associated mutations in the Colombian Amazonian region.
Malaria continues being a public health problem worldwide. Plasmodium vivax is the species causing the largest number of cases of malaria in Asia and South America. Due to the lack of a completely effective anti-malarial vaccine, controlling this disease has been based on transmission vector management, rapid diagnosis and suitable treatment. However, parasite resistance to anti-malarial drugs has become a major yet-to-be-overcome challenge. This study was thus aimed at determining pvmdr1, pvdhfr, pvdhps and pvcrt-o gene mutations and haplotypes from field samples obtained from an endemic area in the Colombian Amazonian region. Fifty samples of parasite DNA infected by a single P. vivax strain from symptomatic patients from the Amazonas department in Colombia were analysed by PCR and the pvdhfr, pvdhps, pvmdr1 and pvcrt-o genes were sequenced. Diversity estimators were calculated from the sequences and the haplotypes circulating in the Colombian Amazonian region were obtained. pvdhfr, pvdhps, pvmdr1 and pvcrt-o genes in the Colombian Amazonian region are characterized by low genetic diversity. Some resistance-associated mutations were found circulating in this population. New variants are also being reported. A selective sweep signal was located in pvdhfr and pvmdr1 genes, suggesting that these mutations (or some of them) could be providing an adaptive advantage.


Systems physiology of the baroreflex during orthostatic stress: from animals to humans.
The baroreflex is a key mechanism involved in the control of arterial pressure (AP) during orthostasis in humans. However, the baroreflex is a closed-loop feedback system, from baroreceptor pressure input to systemic AP, and therefore requires open-loop experiments to identify its system characteristics. The requirement limits our ability to identify baroreflex system characteristics in humans. Open-loop research in animals has revealed dynamic and static characteristics of the two baroreflex subsystems: the neural and peripheral arcs. The neural arc, from baroreceptor pressure input to sympathetic nerve activity (SNA), has high-pass dynamic characteristics, indicating that more rapid change in input AP causes greater response in SNA. In contrast, the peripheral arc, from SNA input to systemic AP, has low-pass characteristics. Orthostasis increases the gain of the neural arc, which compensates for the lower transfer gain of the peripheral arc and in turn maintains total baroreflex function. Here, I discuss the possibility that baroreflex subsystem characteristics identified in animals can be applicable to the human sympathetic response to orthostasis, with a focus on loading speed-dependence of orthostatic sympathetic activation.


Influence of near threshold visual distractors on perceptual detection and reaching movements.
Providing evidence against a dissociation between conscious vision for perception and unconscious vision for action, recent studies have suggested that perceptual and motor decisions are based on a unique signal but distinct decisional thresholds. The aim of the present study was to provide a direct test of this assumption in a perceptual-motor dual task involving arm movements. In 300 trials, 10 participants performed speeded pointing movements toward a highly visible target located at 10° from the fixation point and ± 45° from the body midline. The target was preceded by one or two close to threshold distractor(s) (80 ms stimulus onset asynchrony) presented ± 30° according to the target location. After each pointing movement, participants judged whether the distractor was present or not on either side of the target. Results showed a robust reaction time facilitation effect and a deviation toward the distractor when the distractor was both present and consciously perceived (Hit). A small reaction time facilitation was also observed when two distractors were physically present but undetected (double-miss)--this facilitation being highly correlated with the physical contrast of the distractors. These results are compatible with the theory proposing that perceptual and motor decisions are based on a common signal but emerge from a contrast dependent fixed threshold for motor responses and a variable context dependent criterion for perceptual responses. This paper thus extends to arm movement control previous findings related to oculomotor control.


Diet and lifestyle factors associated with miRNA expression in colorectal tissue.
MicroRNAs (miRNAs) are small non-protein-coding RNA molecules that regulate gene expression. Diet and lifestyle factors have been hypothesized to be involved in the regulation of miRNA expression. In this study it was hypothesized that diet and lifestyle factors are associated with miRNA expression. Data from 1,447 cases of colorectal cancer to evaluate 34 diet and lifestyle variables using miRNA expression in normal colorectal mucosa as well as for differential expression between paired carcinoma and normal tissue were used. miRNA data were obtained using an Agilent platform. Multiple comparisons were adjusted for using the false discovery rate q-value. There were 250 miRNAs differentially expressed between carcinoma and normal colonic tissue by level of carbohydrate intake and 198 miRNAs differentially expressed by the level of sucrose intake. Of these miRNAs, 166 miRNAs were differentially expressed for both carbohydrate intake and sucrose intake. Ninety-nine miRNAs were differentially expressed by the level of whole grain intake in normal colonic mucosa. Level of oxidative balance score was associated with 137 differentially expressed miRNAs between carcinoma and paired normal rectal mucosa. Additionally, 135 miRNAs were differentially expressed in colon tissue based on recent NSAID use. Other dietary factors, body mass index, waist and hip circumference, and long-term physical activity levels did not alter miRNA expression after adjustment for multiple comparisons. These results suggest that diet and lifestyle factors regulate miRNA level. They provide additional support for the influence of carbohydrate, sucrose, whole grains, NSAIDs, and oxidative balance score on colorectal cancer risk.


Molecular simulation of hydrophobin adsorption at an oil-water interface.
Hydrophobins are small, amphiphilic proteins expressed by strains of filamentous fungi. They fulfill a number of biological functions, often related to adsorption at hydrophobic interfaces, and have been investigated for a number of applications in materials science and biotechnology. In order to understand the biological function and applications of these proteins, a microscopic picture of the adsorption of these proteins at interfaces is needed. Using molecular dynamics simulations with a chemically detailed coarse-grained potential, the behavior of typical hydrophobins at the water-octane interface is studied. Calculation of the interfacial adsorption strengths indicates that the adsorption is essentially irreversible, with adsorption strengths of the order of 100 k(B)T (comparable to values determined for synthetic nanoparticles but significantly larger than small molecule surfactants and biomolecules). The protein structure at the interface is unchanged at the interface, which is consistent with the biological function of these proteins. Comparison of native proteins with pseudoproteins that consist of uniform particles shows that the surface structure of these proteins has a large effect on the interfacial adsorption strengths, as does the flexibility of the protein.


Molecule mechanism of stem cells in Arabidopsis thaliana.
Plants possess the ability to continually produce new tissues and organs throughout their life. Unlike animals, plants are exposed to extreme variations in environmental conditions over the course of their lives. The vitality of plants is so powerful that they can survive several hundreds of years or even more making it an amazing miracle that comes from plant stem cells. The stem cells continue to divide to renew themselves and provide cells for the formation of leaves, stems, and flowers. Stem cells are not only quiescent but also immortal, pluripotent and homeostatic. Stem cells are the magic cells that repair tissues and regenerate organs. During the past decade, scholars around the world have paid more and more attention toward plant stem cells. At present, the major challenge is in relating molecule action mechanism to root apical meristem, shoot apical meristem and vascular system. The coordination between stem cells maintenance and differentiation is critical for normal plant growth and development. Elements such as phytohormones, transcription factors and some other known or unknown genes cooperate to balance this process. In this review, Arabidopsis thaliana as a pioneer system, we highlight recent developments in molecule modulating, illustrating how plant stem cells generate new mechanistic insights into the regulation of plants growth and development.


The role of premorbid personality and cognitive factors in awareness of illness, memory, and behavioural functioning in Alzheimer's disease.
Research has suggested an association between personality factors and awareness in patients with dementia, yet valid measurement of premorbid personality is problematic. The present study aimed to better reveal the relationship between premorbid personality and awareness by using improved methodology. Moreover, the study aims to contrast the strength of the relationship of premorbid personality and awareness with that of cognitive factors. Awareness of illness, symptoms, mnemonic and behavioural impairments, and treatment compliance were measured in 27 patients with mild-to-moderate Alzheimer's disease (AD) diagnosed by standard criteria for probable AD. Participant premorbid personality was measured using average retrospective Neuroticism-Extroversion-Openness Inventory (NEO-FFI) scores from two informants. Correlations were performed to investigate the relationship between awareness and personality dimensions, as well as measures of cognitive style, neuropsychological function, mood, carer burden, and sociodemographic factors. There was little relationship between awareness and personality scores, but modest associations between awareness and mood, age, and age of onset of first symptoms. Awareness of memory was related to memory functioning. Increased carer burden was associated with lack of awareness of cognitive-behavioural deficits but there were only few and weak associations between awareness and measures of cognitive functioning. There was little support for an association between previous personality and awareness in dementia. However, increased carer burden was associated specifically with lack of awareness of cognitive-behavioural deficits not deficits in ADL, whereas lower awareness of ADL and not cognitive-behavioural deficits was associated with age. Awareness of memory appeared to be a metamemory capacity. Mood and age rather than personality and cognition are stronger predictors of awareness in early Alzheimer's disease.


Danazol.
Danazol is a synthetic attenuated androgen that can interfere with normal interactions between the pituitary-hypothalamic axis and the gonads. These effects are mediated by complex mechanisms, including those in which danazol can compete with natural steroids in binding to androgen receptors or to sex hormone-binding globulin, possibly displacing natural steroids from this protein, and in binding to reactive sites of enzymes required for synthesis of natural steroids, thereby depressing synthesis. Because of danazol's impairment of the pituitary-hypothalamic interactions with gonads, it is an effective therapeutic agent for treatment of endometriosis and cystic disease of the breast. It is effective in the treatment of hereditary angioneurotic edema, but the mechanism of this therapeutic success is unclear. Danazol has been used, without universal success, in the treatment of other gynecologic and certain hematologic disorders.


A study of redundancy and neutrality in evolutionary optimization.
Some authors consider that evolutionary search may be positively influenced by the use of redundant representations, whereas others note that the addition of random redundancy to a representation could be useless in optimization. Given this lack of consensus, two new families of redundant binary representations are developed in this paper. The first family is based on linear transformations and is considered non-neutral. The second family of representations is designed to implement neutrality, and is based on the mathematical formulation of error control codes. A study aimed at assessing the influence of redundancy and neutrality on the performance of a simple evolutionary hillclimber is presented. The (1+1)-ES is modeled using Markov chains and is applied to NK fitness landscapes. The results indicate that the phenotypic neighborhood induced by a redundant representation dominates the behavior of the algorithm, affecting the search more strongly than neutrality, and the representations with better performance on NK fitness landscapes do not exhibit extreme values of any of the indicators of representation quality commonly adopted in the literature.


Immune surveillance of the lung by migrating tissue monocytes.
Monocytes are phagocytic effector cells in the blood and precursors of resident and inflammatory tissue macrophages. The aim of the current study was to analyse and compare their contribution to innate immune surveillance of the lung in the steady state with macrophage and dendritic cells (DC). ECFP and EGFP transgenic reporters based upon Csf1r and Cx3cr1 distinguish monocytes from resident mononuclear phagocytes. We used these transgenes to study the migratory properties of monocytes and macrophages by functional imaging on explanted lungs. Migratory monocytes were found to be either patrolling within large vessels of the lung or locating at the interface between lung capillaries and alveoli. This spatial organisation gives to monocytes the property to capture fluorescent particles derived from both vascular and airway routes. We conclude that monocytes participate in steady-state surveillance of the lung, in a way that is complementary to resident macrophages and DC, without differentiating into macrophages.


The inequitable distribution of tobacco outlet density: the role of income in two Black Mid-Atlantic geopolitical areas.
Studies have shown that communities with higher concentrations of low-income racial and ethnic minorities correlate with a greater presence of tobacco outlets. Community-level income has consistently been among the strongest predictors of tobacco outlet density. This study analyzes two Maryland geopolitical areas with similar racial concentrations yet differing income levels in an attempt to disentangle the race-income relationship with tobacco outlet density. In this cross-sectional examination of tobacco outlet and census tract-level sociodemographic data, Baltimore City, Maryland, and Prince George's County, Maryland, were geocoded to determine tobacco outlet density. Tobacco outlet density was defined as the mean number of tobacco outlets per 1000 persons per census tract. Comparisons of tobacco outlet density and sociodemographic variables were analysed via two-sample t-tests, and the direct effect of sociodemographic variables on tobacco outlet density for each area was analysed via spatial lag regressions. Prince George's County, the area with the higher income level ($77,190 vs $43,571), has a significantly lower tobacco outlet density than Baltimore City (P < 0.001). Prince George's County has a 67.5% Black population and an average of 3.94 tobacco outlets per 1000 persons per tract. By contrast, Baltimore City has a 65.3% Black population and an average of 7.95 tobacco outlets per 1000 persons per tract. Spatial lag regression model results indicate an inverse relationship between income and tobacco outlet density in Baltimore City and Prince George's County (β = -0.03, P < 0.01 &β = -0.01, P = 0.02, respectively), and a significant interaction term indicating a greater magnitude in the relationship between income and tobacco outlet density in Baltimore City (β = -0.05, P < 0.01). Results suggest that higher socio-economic status, even in primarily underrepresented racial and ethnic geopolitical areas, is linked to lower tobacco outlet density.


Radiation-associated lens opacities in catheterization personnel: results of a survey and direct assessments.
To estimate ocular radiation doses and prevalence of lens opacities in a group of interventional catheterization professionals and offer practical recommendations based on these findings to avoid future lens damage. Subjects included 58 physicians and 69 nurses and technicians attending an interventional cardiology congress and appropriate unexposed age-matched controls. Lens dose estimates were derived from combining experimental measurements in catheterization laboratories with questionnaire responses regarding workload, types of procedures, and use of eye protection. Lens opacities were observed by dilated slit lamp examination using indirect illumination and retroillumination. The frequency and severity of posterior lens changes were compared between the exposed and unexposed groups. The severity of posterior lens changes was correlated with cumulative eye dose. Posterior subcapsular lens changes characteristic of ionizing radiation exposure were found in 50% of interventional cardiologists and 41% of nurses and technicians compared with findings of similar lens changes in<10% of controls. Estimated cumulative eye doses ranged from 0.1-18.9 Sv. Most lens injuries result after several years of work without eye protection. A high prevalence of lens changes likely induced by radiation exposure in the study population suggests an urgent need for improved radiation safety and training, use of eye protection during catheterization procedures, and improved occupational dosimetry.


MapToGenome: a comparative genomic tool that aligns transcript maps to sequenced genomes.
Efforts to generate whole genome assemblies and dense genetic maps have provided a wealth of gene positional information for several vertebrate species. Comparing the relative location of orthologous genes among these genomes provides perspective on genome evolution and can aid in translating genetic information between distantly related organisms. However, large-scale comparisons between genetic maps and genome assemblies can prove challenging because genetic markers are commonly derived from transcribed sequences that are incompletely and variably annotated. We developed the program MapToGenome as a tool for comparing transcript maps and genome assemblies. MapToGenome processes sequence alignments between mapped transcripts and whole genome sequence while accounting for the presence of intronic sequences, and assigns orthology based on user-defined parameters. To illustrate the utility of this program, we used MapToGenome to process alignments between vertebrate genetic maps and genome assemblies 1) self/self alignments for maps and assemblies of the rat and zebrafish genome; 2) alignments between vertebrate transcript maps (rat, salamander, zebrafish, and medaka) and the chicken genome; and 3) alignments of the medaka and zebrafish maps to the pufferfish (Tetraodon nigroviridis) genome. Our results show that map-genome alignments can be improved by combining alignments across presumptive intron breaks and ignoring alignments for simple sequence length polymorphism (SSLP) marker sequences. Comparisons between vertebrate maps and genomes reveal broad patterns of conservation among vertebrate genomes and the differential effects of genome rearrangement over time and across lineages.


Resveratrol and liver disease: from bench to bedside and community.
Liver diseases incorporate several maladies, which can range from benign histological changes to serious life-threatening conditions. These may include inborn metabolic disease, primary and metastatic cancers, alcoholic cirrhosis, viral hepatitis and drug-induced hepatotoxicity. Liver disease remains a major cause of morbidity and mortality with significant economic and social costs. Several novel approaches are currently being studied which may provide a better therapeutic outcome. The use of naturally occurring phytochemicals, some of them obtained from dietary sources, in the amelioration of illness have recently gained considerable popularity. These agents, having anti-oxidant and anti-inflammatory properties, provide a safe and effective means of ameliorating chronic disease. Resveratrol, a grape polyphenol, has shown considerable promise as a therapeutic agent in the treatment of the aforementioned liver ailments. Several studies have highlighted the hepatoprotective properties of resveratrol. Resveratrol has been shown to prevent hepatic damage because of free radicals and inflammatory cytokines, induce anti-oxidant enzymes and elevate glutathione content. Resveratrol has also been shown to modulate varied signal transduction pathways implicated in liver diseases. This review critically examines the current preclinical in vitro and in vivo studies on the preventive and therapeutic effects of resveratrol in liver diseases. The review highlights the pharmacological mechanisms involved in mediating the aforementioned effects. Toxicity, pharmacokinetics and clinical bioavailability of resveratrol are also reviewed in this article. The challenges involved, future directions and novel approaches such as site-specific drug delivery in the use of resveratrol for the prevention and treatment of liver disease are also discussed.


PD-1 marks dysfunctional regulatory T cells in malignant gliomas.
Immunotherapies targeting the immune checkpoint receptor programmed cell death protein 1 (PD-1) have shown remarkable efficacy in treating cancer. CD4+CD25hiFoxP3+ Tregs are critical regulators of immune responses in autoimmunity and malignancies, but the functional status of human Tregs expressing PD-1 remains unclear. We examined functional and molecular features of PD-1hi Tregs in healthy subjects and patients with glioblastoma multiforme (GBM), combining functional assays, RNA sequencing, and cytometry by time of flight (CyTOF). In both patients with GBM and healthy subjects, circulating PD-1hi Tregs displayed reduced suppression of CD4+ effector T cells, production of IFN-γ, and molecular signatures of exhaustion. Transcriptional profiling of tumor-resident Tregs revealed that several genes coexpressed with PD-1 and associated with IFN-γ production and exhaustion as well as enrichment in exhaustion signatures compared with circulating PD-1hi Tregs. CyTOF analysis of circulating and tumor-infiltrating Tregs from patients with GBM treated with PD-1-blocking antibodies revealed that treatment shifts the profile of circulating Tregs toward a more exhausted phenotype reminiscent of that of tumor-infiltrating Tregs, further increasing IFN-γ production. Thus, high PD-1 expression on human Tregs identifies dysfunctional, exhausted Tregs secreting IFN-γ that exist in healthy individuals and are enriched in tumor infiltrates, possibly losing function as they attempt to modulate the antitumoral immune responses.


Study on Current Levels of Physical Activity and Sedentary Behavior among Middle School Students in Beijing, China.
This study aimed to determine current levels of physical activity and sedentary behavior in middle school students on the basis of grade, sex, student attitudes toward physical education, and residence location. In 2013, a cross-sectional study of 1793 students aged 12 to 15 years was conducted across eight middle schools in Beijing, China. Four schools were selected from an urban district and another four schools were from a suburban district. Physical activity and sedentary behavior data were collected using the commonly used school-based Chinese version of the China Health and Nutrition Survey. The mean age of sampled students was 13.3 ± 1.0 years; 51.5% were boys. Approximately 76.6% of students reported having three 45-minute physical education classes every week. A total of 35.6% students spent ≥ 1 h/day performing moderate to vigorous physical activity (MVPA) during school, and 34.9% spent ≥ 1 h/day in MVPA outside school time. Approximately half (49.7%) of the students engaged in reading, writing, or drawing for ≥ 2 h/day, and 42.9% reported screen time for ≥ 2 h/day. Although boys spent more time engaged in physical activity than girls did, they also spent more time exhibiting sedentary behavior. Each 10-unit increase in attitudes toward physical education was associated with an increased odds of 1.15 (95%CI: 1.09-1.20) for spending more than 1 h/day on MVPA. Students in suburban schools reported engaging in physical activity less when compared with those in urban schools. The majority of our students did not meet the current physical activity recommendations, and about half of the students spent excessive time engaging in sedentary behaviors. Findings from this study highlight a positive association between student attitudes toward physical education and physical activity. Studies are needed to further explore the role of student attitudes toward physical education in promoting physical activity among Chinese students.


Transforming machine translation: a deep learning system reaches news translation quality comparable to human professionals.
The quality of human translation was long thought to be unattainable for computer translation systems. In this study, we present a deep-learning system, CUBBITT, which challenges this view. In a context-aware blind evaluation by human judges, CUBBITT significantly outperformed professional-agency English-to-Czech news translation in preserving text meaning (translation adequacy). While human translation is still rated as more fluent, CUBBITT is shown to be substantially more fluent than previous state-of-the-art systems. Moreover, most participants of a Translation Turing test struggle to distinguish CUBBITT translations from human translations. This work approaches the quality of human translation and even surpasses it in adequacy in certain circumstances.This suggests that deep learning may have the potential to replace humans in applications where conservation of meaning is the primary aim.


Malaria severity and human nitric oxide synthase type 2 (NOS2) promoter haplotypes.
Nitric oxide (NO) mediates host resistance to severe malaria and other infectious diseases. NO production and mononuclear cell expression of the NO producing enzyme-inducible nitric oxide synthase (NOS2) have been associated with protection from severe falciparum malaria. The purpose of this study was to identify single nucleotide polymorphisms (SNPs) and haplotypes in the NOS2 promoter, to identify associations of these haplotypes with malaria severity and to test the effects of these polymorphisms on promoter activity. We identified 34 SNPs in the proximal 7.3 kb region of the NOS2 promoter and inferred NOS2 promoter haplotypes based on genotyping 24 of these SNPs in a population of Tanzanian children with and without cerebral malaria. We identified 71 haplotypes; 24 of these haplotypes comprised 82% of the alleles. We determined whether NOS2 promoter haplotypes were associated with malaria severity in two groups of subjects from Dar es Salaam (N = 185 and N = 250) and in an inception cohort of children from Muheza-Tanga, Tanzania (N = 883). We did not find consistent associations of NOS2 promoter haplotypes with malaria severity or malarial anemia, although interpretation of these results was potentially limited by the sample size of each group. Furthermore, cytokine-induced NOS2 promoter activity determined using luciferase reporter constructs containing the proximal 7.3 kb region of the NOS2 promoter and the G-954C or C-1173T SNPs did not differ from NOS2 promoter constructs that lacked these polymorphisms. Taken together, these studies suggest that the relationship between NOS2 promoter polymorphisms and malaria severity is more complex than previously described.


Skeletal muscle tissue in movement and health: positives and negatives.
The history of muscle physiology is a wonderful lesson in 'the scientific method'; our functional hypotheses have been limited by our ability to decipher (observe) muscle structure. The simplistic understanding of how muscles work made a large leap with the remarkable insights of A. V. Hill, who related muscle force and power to shortening velocity and energy use. However, Hill's perspective was largely limited to isometric and isotonic contractions founded on isolated muscle properties that do not always reflect how muscles function in vivo. Robert Josephson incorporated lengthening contractions into a work loop analysis that shifted the focus to dynamic muscle function, varying force, length and work done both by and on muscle during a single muscle work cycle. It became apparent that muscle is both a force generator and a spring. Titin, the missing filament in the sliding filament model, is a muscle spring, which functions very differently in cardiac versus skeletal muscle; its possible role in these two muscle types is discussed relative to their contrasting function. The good news for those of us who choose to work on skeletal muscle is that muscle has been reluctant to reveal all of its secrets.


Curcuma longa extract supplementation reduces oxidative stress and attenuates aortic fatty streak development in rabbits.
This study evaluates the effect of a Curcuma longa extract on the development of experimental atherosclerosis (fatty streak) in rabbits and its interaction with other plasmatic antioxidants. Two experimental groups of male New Zealand White rabbits, a control group and a curcuma-extract (CU) group, were fed an atherogenic diet. Additionally, the CU group received an oral curcuma hydroalcoholic extract. Six animals from each experimental group were killed after 10, 20, and 30 days. Compared with the CU group, the control group showed significantly higher plasma lipid peroxide at all experimental times (10, 20, and 30 days) and significantly lower alpha-tocopherol and coenzyme Q levels at 20 and 30 days. Histological results for the fatty streak lesions revealed damage in the thoracic and abdominal aorta that was significantly lower in the CU group than in the control group at 30 days. Supplementation with Curcuma longa reduces oxidative stress and attenuates the development of fatty streaks in rabbits fed a high cholesterol diet.


Action naming with impaired semantics: Neuropsychological evidencecontrasting naming and reading for objects and verbs.
We contrast naming from pictures, and reading words, for objects and verbs (actions relating to the objects) in a patient with a large, posterior left-hemisphere lesion. We present evidence for spared picture naming for verbs relative to objects, whilst the opposite pattern of sparing occurred in reading. Objects were also spared relative to verbs in tasks requiring that written words be matched to either pictures or auditory words, in the presence of semantically related or unrelated distractors. We conclude that verb semantics were more impaired than semantic knowledge for objects, and that the better semantic knowledge for object names supported word reading. With pictures, however, action verb retrieval was maintained through a nonsemantic route from vision to action, or though preserved right-hemisphere "action semantics."


A comprehensive review of health benefits of qigong and tai chi.
Research examining psychological and physiological benefits of Qigong and Tai Chi is growing rapidly. The many practices described as Qigong or Tai Chi have similar theoretical roots, proposed mechanisms of action, and expected benefits. Research trials and reviews, however, treat them as separate targets of examination. This review examines the evidence for achieving outcomes from randomized controlled trials (RCTs) of both. The key words Tai Chi, Taiji, Tai Chi Chuan, and Qigong were entered into electronic search engines for the Cumulative Index for Allied Health and Nursing (CINAHL), psychological literature (PsycINFO), PubMed, Cochrane database, and Google Scholar. STUDY INCLUSION CRITERIA: RCTs reporting on the results of Qigong or Tai Chi interventions and published in peer-reviewed journals from 1993 to 2007. Country, type and duration of activity, number/type of subjects, control conditions, and reported outcomes were recorded for each study. Outcomes related to Qigong and Tai Chi practice were identified and evaluated. Seventy-seven articles met the inclusion criteria. The nine outcome category groupings that emerged were bone density (n = 4), cardiopulmonary effects (n = 19), physical function (n = 16), falls and related risk factors (n = 23), quality of life (n = 17), self-efficacy (n = 8), patient-reported outcomes (n = 13), psychological symptoms (n = 27), and immune function (n = 6). Research has demonstrated consistent, significant results for a number of health benefits in RCTs, evidencing progress toward recognizing the similarity and equivalence of Qigong and Tai Chi.


Functional Implications of the CLOCK 3111T/C Single-Nucleotide Polymorphism.
Circadian rhythm disruptions are prominently associated with bipolar disorder (BD). Circadian rhythms are regulated by the molecular clock, a family of proteins that function together in a transcriptional-translational feedback loop. The CLOCK protein is a key transcription factor of this feedback loop, and previous studies have found that manipulations of the Clock gene are sufficient to produce manic-like behavior in mice (1). The CLOCK 3111T/C single-nucleotide polymorphism (SNP; rs1801260) is a genetic variation of the human CLOCK gene that is significantly associated with increased frequency of manic episodes in BD patients (2). The 3111T/C SNP is located in the 3'-untranslated region of the CLOCK gene. In this study, we sought to examine the functional implications of the human CLOCK 3111T/C SNP by transfecting a mammalian cell line (mouse embryonic fibroblasts isolated from Clock(-/-) knockout mice) with pcDNA plasmids containing the human CLOCK gene with either the T or C SNP at position 3111. We then measured circadian gene expression over a 24-h time period. We found that the CLOCK3111C SNP resulted in higher mRNA levels than the CLOCK 3111T SNP. Furthermore, we found that Per2, a transcriptional target of CLOCK, was also more highly expressed with CLOCK 3111C expression, indicating that the 3'-UTR SNP affects the expression, function, and stability of CLOCK mRNA.


TREATMENT OF PROGRESSION OF DIFFUSE ASTROCYTOMA BY HERBAL MEDICINE: CASE REPORT.
The paper presents the results of the use of phytotherapy in a 33-year-old woman who, after finishing the oncological treatment of diffuse astrocytoma, had tumour progression. Phytotherapy was introduced after the tumour had progressed. It consisted of 4 types of herbal medicine which the subject was taking in form of tea once a day at regular intervals. The patient started phytotherapy along with temozolomide, which was the only oncological treatment she was under after the tumour had progressed. Following the finished chemotherapy, the patient continued the treatment with herbal medicine only. She regularly took phytotherapy without interruption and to the fullest extent for 30 months, and the results of treatment were monitored by periodic scanning using nuclear magnetic resonance technique. The control scanning that was conducted after the end of combined treatment with temozolomide and phytotherapy showed tumour regression. The patient continued with phytotherapy after finishing chemotherapy and, during the following 24 months, it was the sole treatment option. In that period, the regression of the tumour continued, until a control examination 30 months after the introduction of phytotherapy showed no clinical and radiological signs of tumour. The results presented in this research paper clearly indicate the potential of phytotherapy in the treatment of some types of brain tumours. A complete regression of tumour following the treatment with nothing but herbal medicine offers support for such claim. Future research should demonstrate the effectiveness of phytotherapy, as a supplementary form of brain tumour treatment, and the results of this research should be compared with the existing information on the effectiveness of the protocols currently used in the treatment of these types of tumour.


Multispike interactions in a stochastic model of spike-timing-dependent plasticity.
Recently we presented a stochastic, ensemble-based model of spike-timing-dependent plasticity. In this model, single synapses do not exhibit plasticity depending on the exact timing of pre- and postsynaptic spikes, but spike-timing-dependent plasticity emerges only at the temporal or synaptic ensemble level. We showed that such a model reproduces a variety of experimental results in a natural way, without the introduction of various, ad hoc nonlinearities characteristic of some alternative models. Our previous study was restricted to an examination, analytically, of two-spike interactions, while higher-order, multispike interactions were only briefly examined numerically. Here we derive exact, analytical results for the general n-spike interaction functions in our model. Our results form the basis for a detailed examination, performed elsewhere, of the significant differences between these functions and the implications these differences have for the presence, or otherwise, of stable, competitive dynamics in our model.


General outbreaks of infectious intestinal disease (IID) in hospitals, England and Wales, 1992-2000.
Between 1992 and 2000, 26.6% (1,396/5,257) of all general outbreaks of infectious intestinal disease (IID) reported to the Public Health Laboratory Service (PHLS) Communicable Disease Surveillance Centre (CDSC) occurred in hospitals. Over 29,000 patients and staff were affected and the mortality risk was higher than for outbreaks in other settings [relative risk 2.00 (95% CI: 1.52-2.63) P<0.001]. Person-to-person spread was the predominant mode of transmission. The mortality risk was highest in foodborne disease outbreaks [relative risk 3.22 (95% CI: 1.41-7.36); P=0.003]. Most outbreaks occurred between November and April. The pathogens most frequently reported were Norwalk-like virus (NLV) (54%) and Clostridium difficile (12.6%). These findings emphasize the public health importance of outbreaks of IID in hospitals, especially during the winter when pressures on hospitals are at their height.


A theoretical model of the evolution of maternal effects under parent-offspring conflict.
The evolution of maternal effects on offspring phenotype should depend on the extent of parent-offspring conflict and costs and constraints associated with maternal and offspring strategies. Here, we develop a model of maternal effects on offspring dispersal phenotype under parent-offspring conflict to evaluate such dependence. In the absence of evolutionary constraints and costs, offspring evolve dispersal rates from different patch types that reflect their own, rather than the maternal, optima. This result also holds true when offspring are unable to assess their own environment because the maternal phenotype provides an additional source of information. Consequently, maternal effects on offspring diapause, dispersal, and other traits that do not necessarily represent costly resource investment are more likely to maximize offspring than maternal fitness. However, when trait expression was costly, the evolutionarily stable dispersal rates tended to deviate from those under both maternal and offspring control. We use our results to (re)interpret some recent work on maternal effects and their adaptive value and provide suggestions for future work.


Mathematical models of dorsal closure.
Dorsal closure is a model cell sheet movement that occurs midway through Drosophila embryogenesis. A dorsal hole, filled with amnioserosa, closes through the dorsalward elongation of lateral epidermal cell sheets. Closure requires contributions from 5 distinct tissues and well over 140 genes (see Mortensen et al., 2018, reviewed in Kiehart et al., 2017 and Hayes and Solon, 2017). In spite of this biological complexity, the movements (kinematics) of closure are geometrically simple at tissue, and in certain cases, at cellular scales. This simplicity has made closure the target of a number of mathematical models that seek to explain and quantify the processes that underlie closure's kinematics. The first (purely kinematic) modeling approach recapitulated well the time-evolving geometry of closure even though the underlying physical principles were not known. Almost all subsequent models delve into the forces of closure (i.e. the dynamics of closure). Models assign elastic, contractile and viscous forces which impact tissue and/or cell mechanics. They write rate equations which relate the forces to one another and to other variables, including those which represent geometric, kinematic, and or signaling characteristics. The time evolution of the variables is obtained by computing the solution of the model's system of equations, with optimized model parameters. The basis of the equations range from the phenomenological to biophysical first principles. We review various models and present their contribution to our understanding of the molecular mechanisms and biophysics of closure. Models of closure will contribute to our understanding of similar movements that characterize vertebrate morphogenesis.


Urinary beta-2 microglobulin in renal dysfunction associated with hypothyroidism.
Hypothyroidism is associated with a number of functional renal disorders primarily affecting water and salt handling. In this study, we aim to investigate functional renal disorders in overt and subclinical hypothyroidism. We utilised urinary beta-2 microglobulin (beta2M) as a sensitive marker of tubular dysfunction. Urinary beta2M excretion and tests for renal functions were repeated before and after T4 replacement therapy. Forty-four patients (28 with overt disease, 16 with subclinical disease) and 31 healthy control subjects were involved in the study. There was a significant increase in urinary beta2M in both overt and subclinical hypothyroidism (p < 0.05 in both). TSH levels were correlated with beta2M excretion. Effects of hypothyroidism on renal functions were readily reversible by 3 weeks of thyroid hormone replacement therapy. Urinary beta2M was a sensitive marker of renal tubular dysfunction associated with overt and subclinical hypothyroidism.


Home-Based Exercise Prehabilitation During Preoperative Treatment for Pancreatic Cancer Is Associated With Improvement in Physical Function and Quality of Life.
Purpose: To investigate relationships among physical activity, changes in physical function, and health-related quality of life (HRQOL) among patients with pancreatic adenocarcinoma enrolled in a home-based exercise prehabilitation program. Methods: Patients with resectable pancreatic adenocarcinoma receiving preoperative chemotherapy and/or chemoradiation were enrolled on this prospective, single-arm trial and were advised to perform ≥60 minutes each of moderate-intensity aerobic exercise and strengthening exercise weekly. Activity was measured via self-report and accelerometers, including moderate-to-vigorous physical activity (MVPA), light physical activity (LPA), and sedentary activity (SA). Physical function measures at baseline and restaging follow-up included 6-minute walk test (6MWT), 5 times sit-to-stand (5×STS), handgrip strength (HGS), 3-m walk for gait speed (GS), and the PROMIS Physical Function Short Form. HRQOL was measured via the FACT-Hep questionnaire. Results: Fifty participants with mean age 66 years (standard deviation = 8 years) were enrolled. The 6MWT, 5×STS, and GS significantly improved from baseline to restaging follow-up (P=.001, P=.049, and P=.009, respectively). Increases in self-reported aerobic exercise, weekly MVPA, and LPA were associated with improvement in 6MWT (β=.19, P=.048; β=.18, P=.03; and β=.08, P=.03, respectively) and self-reported physical functioning (β=.02, P=.03; β=.03, P=.005; and β=.01, P=.02, respectively). Increased weekly LPA was associated with increased HRQOL (β=.03, P=.02). Increased SA was associated with decreased HRQOL (β=-.02,P=.01). Conclusions: Patients with potentially resectable pancreatic cancer exhibit meaningful improvement in physical function with prehabilitation; physical activity was associated with improved physical function and HRQOL. These data highlight the importance of physical activity during treatment for pancreatic cancer.


Observation and control in a model of a cell population affected by radiation.
The effect of radiation on a cell population is described by a two-dimensional nonlinear system of differential equations. If the radiation rate is not too high, the system is known to have an asymptotically stable equilibrium. First, for the monitoring of this effect, the concept of observability is applied. For the case when the total number of cells is observed, without distinction between healthy and affected cells, a so-called observer system is constructed, which, at least near the equilibrium state, makes it possible to recover the dynamics of both the healthy and the affected cells, from the observation of the total number of cells without distinction. Results of simulations with illustrative data are also presented. If we want to control the system into a required new equilibrium state, and maintain this new equilibrium by a constant control, a technique of theory of optimal control can be applied to construct a feedback control system.


Viral infection and serum antibodies to heat shock protein 70 in the acute phase of Ménière's disease.
Ménière's disease (MD) is an idiopathic inner-ear disorder characterized by fluctuating hearing loss, episodic vertigo, and tinnitus. Though MD's etiology is unknown, growing evidence suggests that autoimmunity may be involved in its development. The aim of this prospective study was to investigate the presence of anti-heat shock protein 70 (anti-HSP70) antibodies during the acute phase of MD and to relate its presence to the antibody pattern. We examined the sera of 13 patients by Western blot immunoassays for reactivity to bovine inner-ear antigen (anti-HSP70) antibodies. The presence of viral antibodies and autoantibodies (herpes simplex, types 1, 2; herpes zoster; cytomegalovirus; Epstein-Barr; IgM; IgG; cardiolipin; thyroglobulin and thyroperoxidase; and antinuclear, antimitochondrial, and anti-smooth-cell antibodies) were also tested. We found reactivity to HSP70 in only 1 of the 13 MD patients (7.7%), and it occurred during herpes zoster reactivation. We found no relationship between the presence of antibodies to HSP70 and immunological or viral testing.


Identification of cytochemical reaction products by scanning x-ray emission microanalysis.
The primary, secondary, and final reaction products of the cytochemical staining method for alkaline phosphatase have been demonstrated by x-ray emission microanalysis. The advantages and limitations of the technique in measuring reaction products are discussed. Scanning x-ray emission analysis provides a rapid way of detecting and measuring the amounts of individual elements in cytochemical reaction products. It has a resolution of 1 micro and a sensitivity of 0.01 per cent (w/v). It will provide information on the elementary composition of the products in situ and will permit study of the efficiency of conversion of one product to another. It will also yield information on the diffusion of reaction products in the tissue.


Lack of autologous mixed lymphocyte reaction in patients with chronic lymphocytic leukemia: evidence for autoreactive T-cell dysfunction not correlated with phenotype, karyotype, or clinical status.
In the present study, there was a complete lack of autologous MLR between responding T cells or T subsets and unirradiated or irradiated leukemic B cells or monocytes in all 20 patients with CLL, regardless of disease status, stage, phenotype, or karyotype of the disease. The stimulating capacity of unirradiated CLL B cells and CLL monocytes or irradiated CLL B cells was significantly depressed as compared to that of respective normal B cells and monocytes in allogeneic MLR. The responding capacity of CLL T cells was also variably lower than that of normal T cells against unirradiated or irradiated normal allogeneic B cells and monocytes. The depressed allogeneic MLR between CLL B cells or CLL monocytes and normal T cells described in the present study could be explained on the basis of a defect in the stimulating antigens of leukemic B cells or monocytes. The decreased allogeneic MLR of CLL T cells might simply be explained by a defect in the responsiveness of T lymphocytes from patients with CLL. However, these speculations do not adequately explain the complete lack of autologous MLR in these patients. When irradiated CLL B cells or irradiated CLL T cells were cocultured with normal T cells and irradiated normal B cells, it was found that there was no suppressor cell activity of CLL B cells or CLL T cells on normal autologous MLR. Our data suggest that the absence or dysfunction of autoreactive T cells within the Tnon-gamma subset account for the lack of autologous MLR in patients with CLL. The possible significance of the autologous MLR, its relationship to in vivo immunoregulatory mechanisms, and the possible role of breakdown of autoimmunoregulation in the oncogenic process of certain lymphoproliferative and autoimmune diseases in man are discussed.


Helical computed tomographic angiography of canine portosystemic shunts.
Helical computed tomographic (CT) angiography was performed in 16 dogs with known or suspected portosystemic shunts. Fifteen portosystemic shunts were detected including five single intrahepatic shunts, five single extrahepatic shunts, and five multiple extrahepatic shunts. One dog had a normal CT examination. All diagnoses were confirmed by one or several alternate methods including ultrasound, surgery, necropsy, angiography, and liver biopsy. CT detected the origin of 13 of 15 portosystemic shunts and insertion of 13 of 15 shunts. Limitations included inability to resolve two vessels originating very close to each other, and identification of vessels that traveled parallel to the axial image plane. CT angiography is a promising, minimally invasive method of diagnosing a variety of portosystemic shunts in dogs.


Caveolin-1-dependent infectious entry of human papillomavirus type 31 in human keratinocytes proceeds to the endosomal pathway for pH-dependent uncoating.
High-risk human papillomaviruses (HPVs) are small nonenveloped DNA viruses with a strict tropism for squamous epithelium. The viruses are causative agents of cervical cancer and some head and neck cancers, but their differentiation-dependent life cycles have made them difficult to study in simple cell culture. Thus, many aspects of early HPV infection remain mysterious. We recently showed the high-risk HPV type 31 (HPV31) enters its natural host cell type via caveola-dependent endocytosis, a distinct mechanism from that of the closely related HPV16 (Smith et al., J. Virol. 81:9922-9931, 2007). Here, we determined the downstream trafficking events after caveolar entry of HPV31 into human keratinocytes. After initial plasma membrane binding, HPV31 associates with caveolin-1 and transiently localizes to the caveosome before trafficking to the early endosome and proceeding through the endosomal pathway. Caveosome-to-endosome transport was found to be Rab5 GTPase dependent. Although HPV31 capsids were observed in the lysosome, Rab7 GTPase was dispensable for HPV31 infection, suggesting that viral genomes escape from the endosomal pathway prior to Rab7-mediated capsid transport. Consistent with this, the acidic pH encountered by HPV31 within the early endosomal pathway induces a conformational change in the capsid resulting in increased DNase susceptibility of the viral genome, which likely aids in uncoating and/or endosomal escape. The entry and trafficking route of HPV31 into human keratinocytes represents a unique viral pathway by which the virions use caveolar entry to eventually access a low-pH site that appears to facilitate endosomal escape of genomes.


Saethre-Chotzen syndrome associated with balanced translocations involving 7p21: three further families.
We describe three families segregating different reciprocal chromosome translocations, t(7;18)(p21.2;q23), t(2;7)(q21.1;p21.2), and t(5;7)(p15.3;p21.2). A total of seven apparently balanced carriers have been identified and all manifest features of the Saethre-Chotzen syndrome, although only two have overt craniosynostosis. In one family the carriers are immediately recognisable by their unusual ears, and clefts of the hard or soft palate are present in all three families. These observations extend previous linkage and cytogenetic evidence that a locus for Saethre-Chotzen syndrome resides in band 7p21.2.


Diagnostic Criteria for Somatosensory Tinnitus: A Delphi Process and Face-to-Face Meeting to Establish Consensus.
Since somatic or somatosensory tinnitus (ST) was first described as a subtype of subjective tinnitus, where altered somatosensory afference from the cervical spine or temporomandibular area causes or changes a patient's tinnitus perception, several studies in humans and animals have provided a neurophysiological explanation for this type of tinnitus. Due to a lack of unambiguous clinical tests, many authors and clinicians use their own criteria for diagnosing ST. This resulted in large differences in prevalence figures in different studies and limits the comparison of clinical trials on ST treatment. This study aimed to reach an international consensus on diagnostic criteria for ST among experts, scientists and clinicians using a Delphi survey and face-to-face consensus meeting strategy. Following recommended procedures to gain expert consensus, a two-round Delphi survey was delivered online, followed by an in-person consensus meeting. Experts agreed upon a set of criteria that strongly suggest ST. These criteria comprise items on somatosensory modulation, specific tinnitus characteristics, and symptoms that can accompany the tinnitus. None of these criteria have to be present in every single patient with ST, but in case they are present, they strongly suggest the presence of ST. Because of the international nature of the survey, we expect these criteria to gain wide acceptance in the research field and to serve as a guideline for clinicians across all disciplines. Criteria developed in this consensus paper should now allow further investigation of the extent of somatosensory influence in individual tinnitus patients and tinnitus populations.


Simultaneous velocity and temperature measurements in gaseous flowfields using the vibrationally excited nitric oxide monitoring technique: a comprehensive study.
The performance of the vibrationally excited nitric oxide monitoring (VENOM) technique for simultaneous velocity and temperature measurements in gaseous flowfields is presented. Two different schemes were investigated, employing different methods to "write" a transient NO grid in the flow using the 355 nm photolysis of NO(2), which was subsequently probed by planar laser induced fluorescence imaging to extract velocity maps. We find that only one scheme provides full-frame temperature maps. The most accurate velocity measurement was attained by writing an NO pattern in the flow using a microlens array and then comparing the line displacement with respect to a reference image. The demonstrated uncertainty of this approach was 1.0%, corresponding to 7 m/s in a 705 m/s uniform flow. We found that the uncertainty associated with the instantaneous temperature measurements using the NO two-line thermometry technique was largely determined by the shot-to-shot power fluctuations of the probe lasers and, for the flows employed, were determined to range from 6% to 7% of the mean freestream temperature. Finally, simultaneous and local velocity/temperature measurements were performed in the wake of a cylinder in a uniform Mach 4.6 flowfield. The mean and fluctuation velocity and temperature maps were computed from 5000 single-shot measurements. The wake temperature and velocity fluctuations, with respect to the freestream values, were 15% to 30% and 5% to 20%, respectively. The spatial distributions agree with the results of computational fluid dynamics (CFD) simulations. Our results suggest that the VENOM technique holds promise for interrogating high-speed unsteady flowfields.


ERK1/2 regulation of CD44 modulates oral cancer aggressiveness.
Carcinogen-induced oral cavity squamous cell carcinoma (OSCC) incurs significant morbidity and mortality and constitutes a global health challenge. To gain further insight into this disease, we generated cell line models from 7,12-dimethylbenz(a)anthracene-induced murine primary OSCC capable of tumor formation upon transplantation into immunocompetent wild-type mice. Whereas several cell lines grew rapidly and were capable of metastasis, some grew slowly and did not metastasize. Aggressively growing cell lines displayed ERK1/2 activation, which stimulated expression of CD44, a marker associated with epithelial to mesenchymal transition and putative cancer stem cells. MEK (MAP/ERK kinase) inhibition upstream of ERK1/2 decreased CD44 expression and promoter activity and reduced cell migration and invasion. Conversely, MEK1 activation enhanced CD44 expression and promoter activity, whereas CD44 attenuation reduced in vitro migration and in vivo tumor formation. Extending these findings to freshly resected human OSCC, we confirmed a strict relationship between ERK1/2 phosphorylation and CD44 expression. In summary, our findings identify CD44 as a critical target of ERK1/2 in promoting tumor aggressiveness and offer a preclinical proof-of-concept to target this pathway as a strategy to treat head and neck cancer.


Suppression of EGFR/STAT3 activity by lupeol contributes to the induction of the apoptosis of human non‑small cell lung cancer cells.
The aim of this study was to investigate the underlying mechanisms responsible for the anticancer effects of lupeol on human non‑small cell lung cancer (NSCLC). MTT assay and Trypan blue exclusion assay were used to evaluate the cell viability. DAPI staining and flow cytometric analysis were used to detect apoptosis. Molecular docking and western blot analysis were performed to determine the target of lupeol. We found that lupeol suppressed the proliferation and colony formation of NSCLC cells in a dose‑dependent manner. In addition, lupeol increased chromatin condensation, poly(ADP‑ribose) polymerase (PARP) cleavage, sub‑G1 cell populations, and the proportion of Annexin V‑positive cells, indicating that lupeol triggered the apoptosis of NSCLC cells. Notably, lupeol inhibited the phosphorylation of epithelial growth factor receptor (EGFR). A docking experiment revealed that lupeol directly bound to the tyrosine kinase domain of EGFR. We observed that the signal transducer and activator of transcription 3 (STAT3), a downstream molecule of EGFR, was also dephosphorylated by lupeol. Lupeol suppressed the nuclear translocation and transcriptional activity of STAT3 and downregulated the expression of STAT3 target genes. The constitutive activation of STAT3 by STAT3 Y705D overexpression suppressed lupeol‑induced apoptosis, demonstrating that the inhibition of STAT3 activity contributed to the induction of apoptosis. The anticancer effects of lupeol were consistently observed in EGFR tyrosine kinase inhibitor (TKI)‑resistant H1975 cells (EGFR L858R/T790M). Taken together, the findings of this study suggest that lupeol may be used, not only for EGFR TKI‑naïve NSCLC, but also for advanced NSCLC with acquired resistance to EGFR TKIs.


Nonbeneficial treatment Canada: definitions, causes, and potential solutions from the perspective of healthcare practitioners*.
Many healthcare workers are concerned about the provision of nonbeneficial treatment in the acute care setting. We sought to explore the perceptions of acute care practitioners to determine whether they perceived nonbeneficial treatment to be a problem, to generate an acceptable definition of nonbeneficial treatment, to learn about their perceptions of the impact and causes of nonbeneficial treatment, and the ways that they feel could reduce or resolve nonbeneficial treatment. National, bilingual, cross-sectional survey of a convenience sample of nursing and medical staff who provide direct patient care in acute medical wards or ICUs in Canada. We received 688 responses (response rate 61%) from 11 sites. Seventy-four percent of respondents were nurses. Eighty-two percent of respondents believe that our current means of resolving nonbeneficial treatment are inadequate. The most acceptable definitions of nonbeneficial treatment were "advanced curative/life-prolonging treatments that would almost certainly result in a quality of life that the patient has previously stated that he/she would not want" (88% agreement) and "advanced curative/life-prolonging treatments that are not consistent with the goals of care (as indicated by the patient)" (83% agreement). Respondents most commonly believed that nonbeneficial treatment was caused by substitute decision makers who do not understand the limitations of treatment, or who cannot accept a poor prognosis (90% agreement for each cause), and 52% believed that nonbeneficial treatment was "often" or "always" continued until the patient died or was discharged from hospital. Respondents believed that nonbeneficial treatment was a common problem with a negative impact on all stakeholders (> 80%) and perceived that improved advance care planning and communication training would be the most effective (92% and 88%, respectively) and morally acceptable (95% and 92%, respectively) means to resolve the problem of nonbeneficial treatment. Canadian nurses and physicians perceive that our current means of resolving nonbeneficial treatment are inadequate, and that we need to adopt new techniques of resolving nonbeneficial treatment. The most promising strategies to reduce nonbeneficial treatment are felt to be improved advance care planning and communication training for healthcare professionals.


Collaborative design of a health care experience survey for persons with disability.
When assessing results of health care delivery system reforms targeting persons with disability, quality metrics must reflect the experiences and perspectives of this population. For persons with disability and researchers to develop collaboratively a survey that addresses critical quality questions about a new Massachusetts health care program for persons with disability dually-eligible for Medicare and Medicaid. Persons with significant physical disability or serious mental health diagnoses participated fully in all research activities, including co-directing the study, co-moderating focus groups, performing qualitative analyses, specifying survey topics, cognitive interviewing, and refining survey language. Several sources informed survey development, including key informant interviews, focus groups, and cognitive testing. We interviewed 18 key informants from key stakeholder groups, including disability advocates, health care providers, and governmental agencies. We conducted 12 total English- and Spanish-language focus groups involving 87 participants (38 with physical disability, 49 with mental health diagnoses). Although some details differed, focus group findings were similar across the two disability groups. Analyses by collaborators with disability identified 29 questions for persons with physical disability and 38 for persons with mental health diagnoses. After cognitive testing, the final survey includes questions on topics ranging from independent living principles to health care delivery system concerns. The Persons with Disabilities Quality Survey (PDQ-S) captures specific quality concerns of Massachusetts residents with physical or mental health disability about an integrated health plan. PDQ-S requires further testing elsewhere to determine its value for quality assessment more generally and to other populations with disability.


Protocol for a prospective, observational, longitudinal study in paediatric patients with moderate-to-severe atopic dermatitis (PEDISTAD): study objectives, design and methodology.
Atopic dermatitis (AD) is a chronic inflammatory skin disease often associated with atopic comorbidities and has significant impact on children and their families. There is a lack of robust and longitudinal long-term data on disease characteristics and typical clinical practice with currently available treatments in children with moderate-to-severe AD. Hence, an observational study is needed to evaluate AD characteristics and progression in paediatric patients with moderate-to-severe AD. Pediatric Study in Atopic Dermatitis (PEDISTAD) is a prospective, observational, longitudinal study in paediatric patients with moderate-to-severe AD who are currently receiving systemic or topical treatment and whose disease is not adequately controlled by topical prescription therapies or for whom those therapies are not medically advisable. 1300 children at 100-150 sites in approximately 20 countries worldwide will be enrolled and followed for 5 years. AD therapy is at the discretion of the investigator. Data collected will include: AD disease characteristics and comorbidities; current therapy for AD and initiation of new treatments/changes in current treatment; patient-reported/caregiver-reported outcomes; days missed from school/work for the patient/caregiver; healthcare professional visits; safety and biomarkers. This study is conducted in accordance with the principles established by the 18th World Medical Assembly and all subsequent amendments and the guidelines for Good Epidemiology Practice. Each individual country assures that ethics approval has been received and local regulatory requirements are met. Ethics approval has been obtained in all countries currently participating in PEDISTAD. Study data will be disseminated in manuscripts submitted to peer-reviewed medical journals as well as in abstracts submitted to congresses and in the resulting posters and presentations. NCT03687359; pre-results.


Absolute quantitation of individual SARS-CoV-2 RNA molecules provides a new paradigm for infection dynamics and variant differences.
Despite an unprecedented global research effort on SARS-CoV-2, early replication events remain poorly understood. Given the clinical importance of emergent viral variants with increased transmission, there is an urgent need to understand the early stages of viral replication and transcription. We used single-molecule fluorescence in situ hybridisation (smFISH) to quantify positive sense RNA genomes with 95% detection efficiency, while simultaneously visualising negative sense genomes, subgenomic RNAs, and viral proteins. Our absolute quantification of viral RNAs and replication factories revealed that SARS-CoV-2 genomic RNA is long-lived after entry, suggesting that it avoids degradation by cellular nucleases. Moreover, we observed that SARS-CoV-2 replication is highly variable between cells, with only a small cell population displaying high burden of viral RNA. Unexpectedly, the B.1.1.7 variant, first identified in the UK, exhibits significantly slower replication kinetics than the Victoria strain, suggesting a novel mechanism contributing to its higher transmissibility with important clinical implications.


Travelling-wave nuclear magnetic resonance.
Nuclear magnetic resonance (NMR) is one of the most versatile experimental methods in chemistry, physics and biology, providing insight into the structure and dynamics of matter at the molecular scale. Its imaging variant-magnetic resonance imaging (MRI)-is widely used to examine the anatomy, physiology and metabolism of the human body. NMR signal detection is traditionally based on Faraday induction in one or multiple radio-frequency resonators that are brought into close proximity with the sample. Alternative principles involving structured-material flux guides, superconducting quantum interference devices, atomic magnetometers, Hall probes or magnetoresistive elements have been explored. However, a common feature of all NMR implementations until now is that they rely on close coupling between the detector and the object under investigation. Here we show that NMR can also be excited and detected by long-range interaction, relying on travelling radio-frequency waves sent and received by an antenna. One benefit of this approach is more uniform coverage of samples that are larger than the wavelength of the NMR signal-an important current issue in MRI of humans at very high magnetic fields. By allowing a significant distance between the probe and the sample, travelling-wave interaction also introduces new possibilities in the design of NMR experiments and systems.


Sources of carbon for hepatic glycogen synthesis in the conscious dog.
To identify the source(s) of carbon for the indirect pathway of hepatic glycogen synthesis, we studied nine 42-h fasted conscious dogs given a continuous intraduodenal infusion of glucose, labeled with [1-13C]glucose and [3-3H]glucose, at 8 mg.kg-1.min-1 for 240 min. Glycogen formation by the direct pathway was measured by 13C-NMR. Net hepatic balances of glucose, gluconeogenic amino acids, lactate, and glycerol were determined using the arteriovenous difference technique. During the steady-state period (the final hour of the infusion), 81% of the glucose infused was absorbed as glucose. Net gut output of lactate and alanine accounted for 5% and 3% of the glucose infused, respectively. The cumulative net hepatic uptakes were: glucose, 15.5 +/- 3.8 g; gluconeogenic amino acids, 32.2 +/- 2.2 mmol (2.9 +/- 0.2 g of glucose equivalents); and glycerol, 6.1 +/- 0.9 mmol (0.6 +/- 0.1 g of glucose equivalents). The liver produced a net of 29.2 +/- 9.6 mmol of lactate (2.6 +/- 0.8 g of glucose equivalents). Net hepatic glycogen synthesis totaled 9.3 +/- 2.5 g (1.8 +/- 0.4 g/100 g liver), with the direct pathway being responsible for 57 +/- 10%. Thus, net hepatic glucose uptake was sufficient to account for all glycogen formed by both the direct and indirect pathways. Total net hepatic uptake of gluconeogenic precursors (gluconeogenic amino acids, glycerol, and lactate) was able to account for only 20% of net glycogen synthesis by the indirect pathway. In a net sense, our data are consistent with an intrahepatic origin for most of the three-carbon precursors used for indirect glycogen synthesis.


The effect of visual cues on auditory stream segregation in musicians and non-musicians.
The ability to separate two interleaved melodies is an important factor in music appreciation. This ability is greatly reduced in people with hearing impairment, contributing to difficulties in music appreciation. The aim of this study was to assess whether visual cues, musical training or musical context could have an effect on this ability, and potentially improve music appreciation for the hearing impaired. Musicians (N = 18) and non-musicians (N = 19) were asked to rate the difficulty of segregating a four-note repeating melody from interleaved random distracter notes. Visual cues were provided on half the blocks, and two musical contexts were tested, with the overlap between melody and distracter notes either gradually increasing or decreasing. Visual cues, musical training, and musical context all affected the difficulty of extracting the melody from a background of interleaved random distracter notes. Visual cues were effective in reducing the difficulty of segregating the melody from distracter notes, even in individuals with no musical training. These results are consistent with theories that indicate an important role for central (top-down) processes in auditory streaming mechanisms, and suggest that visual cues may help the hearing-impaired enjoy music.


Thymus-dependent in vivo suppression of IgE synthesis in a murine IgE-secreting hybridoma.
In previous studies we demonstrated that BALB/c mice bearing ascitic tumors of the IgE-secreting hybridoma B53 (epsilon, kappa, anti-dinitrophenyl) developed large numbers of Lyt-1-2+ Fc epsilon R(+) T lymphocytes (T cells with membrane Fc receptors) in response to the elevated serum IgE concentration. The development of Fc epsilon R(+) T lymphocytes was followed by a progressive decrease in the levels of serum IgE in spite of continued proliferation of the hybridoma cells. This sequence of events suggested that the IgE-secreting hybridoma triggered a suppressive immunoregulatory circuit of the host that inhibited IgE expression by the hybridoma cells. The present studies were undertaken to investigate the basis for the subsequent decline in serum IgE levels in mice with B53 tumors and to identify host factors that might be involved in this process. We observed that ascitic B53 cells recovered at increasing time points from BALB/c mice exhibited a selective decline in steady state levels and rates of synthesis of epsilon-heavy chain protein and mRNA. The expression of kappa-light chain protein and mRNA appeared relatively unchanged. The decrease in epsilon-heavy chain gene expression did not occur when B53 tumors were passaged in nu+/nu+ mice or in BALB/c mice depleted of Lyt-2+ cells (suppressor/cytotoxic cell lineage), but did occur in nu+/nu+ mice reconstituted with neonatal BALB/c thymus and in BALB/c mice depleted of L3T4+ cells (helper/inducer cell lineage). That Fc epsilon R(+) T lymphocytes were directly involved in the inhibition of IgE expression was supported by the earlier and more pronounced inhibition of B53 IgE in mice infused with Fc epsilon R(+) T lymphocytes. We conclude from these findings that: 1) the decline in serum IgE levels that occurs toward the end of each generation of in vivo passage of the B53 hybridoma is due to decreased production of IgE by the hybridoma cells, 2) the decreased production of IgE is due to a selective loss of epsilon mRNA expression, 3) the decrease production of IgE by B53 cells is dependent on the presence of Lyt-2+ cells, and 4) Fc epsilon R(+) T lymphocytes participate in the mechanism by which IgE production is suppressed.


Stable Isotope Probing Techniques and Methodological Considerations Using 15N.
Nitrogen fixation and assimilation processes are vital to the functioning of any ecosystem. Nevertheless, studying these processes using 15N-based stable isotope probing was so far limited because of technical challenges related to the relative rarity of nitrogen in nucleic acids and proteins compared to carbon, and because of its absence in lipids. However, the recent adoption of high-throughput sequencing and statistical modelling methods to SIP studies increased the sensitivity of the method and enabled overcoming some of the challenges. This chapter describes in detail how to perform DNA- and RNA-SIP using 15N.


Self-incompatibility systems: barriers to self-fertilization in flowering plants.
Flowering plants (angiosperms) are the most prevalent and evolutionarily advanced group of plants. Success of these plants is owed to several unique evolutionary adaptations that aid in reproduction: the flower, the closed carpel, double fertilization, and the ultimate products of fertilization, seeds enclosed in the fruit. Angiosperms exhibit a vast array of reproductive strategies, including both asexual and sexual, the latter of which includes both self-fertilization and cross-fertilization. Asexual reproduction and self-fertilization are important reproductive strategies in a variety of situations, such as when mates are scarce or when the environment remains relatively stable. However, reproductive strategies promoting cross-fertilization are critical to angiosperm success, since they contribute to the creation of genetically diverse populations, which increase the probability that at least one individual in a population will survive given changing environmental conditions. The evolution of several physical and genetic barriers to self-fertilization or fertilization among closely related individuals is thus widespread in angiosperms. A major genetic barrier to self-fertilization is self-incompatibility (SI), which allows female reproductive cells to discriminate between "self" and "non-self" pollen, and specifically reject self pollen. Evidence for the importance of SI in angiosperm evolution lies in the highly diverse set of mechanisms used by various angiosperm families for recognition of self pollen tube development and preventing self-fertilization.


Long-term health and wellbeing of people affected by the 2002 Bali bombing.
To examine the physical and mental health status of individuals directly affected by the 2002 Bali bombing, 8 years after the incident. Cross-sectional study of people directly exposed to and/or bereaved by the 2002 Bali bombing who had participated in a New South Wales Health therapeutic support program. Telephone interviews were conducted during July - November 2010. The sample was weighted to reflect the population of interest, registered participants in the program (n = 115). Self-rated physical health, personal resilience (Connor-Davidson Resilience Scale), past-03 psychological distress and daily functioning (Kessler Psychological Distress Scale), and traumatic stress-related symptoms (Primary Care PTSD Screen). Of 81 individuals contacted, 55 responded (68%). Mean age of respondents was 50 years (range, 20-73 years), 32 were female, and seven were physically injured in the bombing. Most (45/55) reported good physical health, but 12 were experiencing high or very high levels of psychological distress. Being injured in the attack was associated with current functional impairment (P = 0.04) and very high levels of distress (P = 0.005). Lower distress was associated with perceived family support (P> = 0.03) and being in a marital or de facto relationship (P = 0.02). Complicated grief factors were consistently associated with high psychological distress, traumatic stress-related symptoms and lower personal resilience. Eight years after the bombing, directly affected individuals had good physical health but relatively high rates of psychological distress. Marital or de facto relationships and perceived family support appear to be protective factors against long-term distress. Bereavement factors were the strongest correlates of trauma symptoms and distress. Outreach and screening programs incorporating complicated grief items may be useful in the longer-term support of such individuals.


Media coverage and public reaction to a celebrity cancer diagnosis.
Celebrity diagnoses can have important effects on public behaviour. UK television celebrity Jade Goody died from cervical cancer in 2009. We investigated the impact of her illness on media coverage of cervical cancer prevention, health information seeking behaviour and cervical screening coverage. National UK newspaper articles containing the words 'Jade Goody' and 'cancer' were examined for public health messages. Google Insights for Search was used to quantify Internet searches as a measure of public health information seeking. Cervical screening coverage data were examined for temporal associations with this story. Of 1203 articles, 116 (9.6%) included a clear public health message. The majority highlighted screening (8.2%). Fewer articles provided advice about vaccination (3.0%), number of sexual partners (1.4%), smoking (0.6%) and condom use (0.4%). Key events were associated with increased Internet searches for 'cervical cancer' and 'smear test', although only weakly with searches for 'HPV'. Cervical screening coverage increased during this period. Increased public interest in disease prevention can follow a celebrity diagnosis. Although media coverage sometimes included public health information, articles typically focused on secondary instead of primary prevention. There is further potential to maximize the public health benefit of future celebrity diagnoses.


Short report: Diagnostic testing for hemorrhagic fevers in Pakistan: 2007-2013.
Crimean-Congo hemorrhagic fever virus (CCHFV) and dengue virus (DENV) are endemic to Pakistan. Patients presenting with symptoms of fever, bleeding, and rash cannot be distinguished without appropriate testing. We report data on 354 samples tested for CCHFV at The Aga Khan University Hospital in Pakistan between 2007 and 2013. All samples were tested for the presence of CCHFV RNA. Some samples were also tested for DENV RNA, NS-1 antigen, and/or reactive immunoglobulin M antibodies. Of 354 clinical specimens screened for CCHFV, 52 specimens were positive, with 24 cases in 2013 alone. Most cases were from Sindh and Baluchistan, which border other CCHFV-endemic regions: Iran and Afghanistan. Among CCHFV-negative samples, 168 samples were tested for DENV, and 36% of these samples were found to be DENV-positive. Rapid differentiation of CCHFV and DENV can prevent nosocomial transmission and result in time and cost savings for patients and healthcare workers.


Iteratively Developing an mHealth HIV Prevention Program for Sexual Minority Adolescent Men.
Five activities were implemented between November 2012 and June 2014 to develop an mHealth HIV prevention program for adolescent gay, bisexual, and queer men (AGBM): (1) focus groups to gather acceptability of the program components; (2) ongoing development of content; (3) Content Advisory Teams to confirm the tone, flow, and understandability of program content; (4) an internal team test to alpha test software functionality; and (5) a beta test to test the protocol and intervention messages. Findings suggest that AGBM preferred positive and friendly content that at the same time, did not try to sound like a peer. They deemed the number of daily text messages (i.e., 8-15 per day) to be acceptable. The Text Buddy component was well received but youth needed concrete direction about appropriate discussion topics. AGBM determined the self-safety assessment also was acceptable. Its feasible implementation in the beta test suggests that AGBM can actively self-determine their potential danger when participating in sexual health programs. Partnering with the target population in intervention development is critical to ensure that a salient final product and feasible protocol are created.


Specific and Sensitive Detection of Enterobacter mori Using Reliable RT-PCR.
Enterobacter mori, the causal agent of bacterial wilt in mulberry, is becoming a serious disease in mulberry orchards in China. Because no effective control strategy has been devised for this disease, the reliable screening of mulberry material for latent infection became necessary. Hence, a fast polymerase chain reaction (PCR) assay for the detection of E. mori was developed in this study. The primers were designed within regions of the RNA polymerase β-subunit (rpoB) gene. The method is fast and simple and showed 100% sensitivity (no false negatives) and 100% specificity (no false positives), which was tested with 4 representative E. mori strains, 9 Enterobacter type strains, 2 strains of the other major mulberry bacterial pathogens (Ralstonia solanacearum and Pseudomonas syringae pv. mori) in China, 7 strains of other plant-associated pathogens, and 50 unidentified epiphytic bacterial isolates from mulberry plants. The real-time PCR assays reliably detected the DNA at at least 10 fg/μl and the bacterial cells at 102 CFU/ml from mulberry shoots and roots suspension. The strong positive reaction in testing of all symptomatic plants (with 100% positive) and parts of asymptomatic latent infected plant samples (with 36.4% positive) provided proof that this method is reliable and sensitive and suitable for screening plant material with latent infections of E. mori.


Biomechanical demands on posterior fusion instrumentation during lordosis restoration procedures.
OBJECTIVE The goal of this study was to investigate the forces placed on posterior fusion instrumentation by 3 commonly used intraoperative techniques to restore lumbar lordosis: 1) cantilever bending; 2) in situ bending; and 3) compression and/or distraction of screws along posterior fusion rods. METHODS Five cadaveric torsos were instrumented with pedicle screws at the L1-5 levels. Specimens underwent each of the 3 lordosis restoration procedures. The pedicle screw pullout force was monitored in real time via strain gauges that were mounted unilaterally at each level. The degree of correction was noted through fluoroscopic imaging. The peak loads experienced on the screws during surgery, total demand on instrumentation, and resting loads after corrective maneuvers were measured. RESULTS A mean overall lordotic correction of 10.9 ± 4.7° was achieved. No statistically significant difference in lordotic correction was observed between restoration procedures. In situ bending imparted the largest loads intraoperatively with an average of 1060 ± 599.9 N, followed by compression/distraction (971 ± 534.1 N) and cantilever bending (705 ± 413.0 N). In situ bending produced the largest total demand and postoperative loads at L-1 (1879 ± 1064.1 and 487 ± 118.8 N, respectively), which were statistically higher than cantilever bending and compression/distraction (786 ± 272.1 and 138 ± 99.2 N, respectively). CONCLUSIONS In situ bending resulted in the highest mechanical demand on posterior lumbar instrumentation, as well as the largest postoperative loads at L-1. These results suggest that the forces generated with in situ bending indicate a greater chance of intraoperative instrumentation failure and postoperative proximal pedicle screw pullout when compared with cantilever bending and/or compression/distraction options. The results are aimed at optimizing correction and fusion strategies in lordosis restoration cases.


Even aberration measurement of lithographic projection optics based on intensity difference of adjacent peaks in alternating phase-shifting mask image.
We propose an in situ technique for measuring an even aberration of lithographic projection optics. By using the Hopkins theory of partially coherent imaging and the thick-mask model, the linear relationship between the intensity difference of adjacent peaks in an alternating phase-shifting mask image and an even aberration is established by equations and verified by numerical results. The sensitivity of measuring the even aberration of lithographic projection optics based on this linear relationship is analyzed, and the measurement mark is designed accordingly. Measurement performance of the present technique is evaluated using the lithographic simulator PROLITH, which shows that the present technique is capable of measuring the even aberration of lithographic projection optics with ultrahigh measurement accuracy.


Breeding structure of the sand fly Lutzomyia longipalpis (Lutz & Neiva) in Brazil.
Eleven populations of Lutzomyia longipalpis (Lutz & Neiva), the sand fly vector of Leishmania chagasi, from different areas of Brazil were analyzed for genetic variation at 16 enzyme loci. In this region, the prevalence of visceral leishmaniasis (VL) caused by L. chagasi is spotty and reproductive isolation among populations of Lu. longipalpis has been reported. It is thought that morphologically similar cryptic species with varying vectorial capacity may be responsible for the discontinuous distribution of VL. The aim was to study the genetic structure of populations within this region and to identify demes that may represent sibling species. Genotypic frequencies within populations were in close compliance to Hardy-Weinberg expectations, suggesting there are no sympatric species among these 11 populations. Levels of genetic distance between pairs of populations were very low (< 0.03), consistent with local populations within a single sand fly species. When genotypic frequency data for all populations were pooled, 9 of the 13 polymorphic loci deviated from Hardy-Weinberg expectations, indicating some degree of genetic substructuring. Estimates of effective migration rates (N(e)m) among all populations were low, 2.73, suggesting that gene flow is restricted among populations, which is probably the reason for the observed genetic substructuring.


[Exposure to aflatoxin B1 in experimental animals and its public health significance].
The presence of AFB1 in human beings was detected in Mexico in 1996 both as a mutation of the gene p53 in hepatocellular carcinomas in Monterrey, Mexico, and as the adduct AFB1-lysine in serum from patients in Matamoros, Mexico in 2003. Aflatoxin B1 has been classified as a carcinogenic agent to humans by the International Agency for Research on Cancer. The compound is a natural contaminant produced by Aspergillus flavus and/or A. parasiticus when these fungi grow on different food products. At the molecular level, this review covers the carcinogenic, mutagenic and toxic properties of these mycotoxins and their risk to humans. It also gives insight into the causal relationship between aflatoxins and hepatocellular carcinoma. Information is provided about AFB1-formamidopyrimidine, which is a determinant of the carcinogenic and mutagenic capabilities. The results suggest that the Mexican population ingests food containing low amounts of AFB1. Analyses is presented of AFB1 toxicity, which is a consequence of the carcinogenic activity in liver cells.


The Colletotrichum lagenariu Ste12-like gene CST1 is essential for appressorium penetration.
Colletotrichum lagenarium is the causal agent of anthracnose of cucumber. This fungus produces a darkly melanized infection structure, appressoria, to penetrate the host leaves. The C. lagenarium CMK1 gene, a homologue of the Saccharomyces cerevisiae FUS3/KSS1 mitogen-activated protein (MAP) kinase genes, was shown to regulate conidial germination, appressorium formation, and invasive growth. In S. cerevisiae, Ste12p is known to be a transcriptional factor downstream of Fus3p/Kss1p MAP kinases. To evaluate the CMK1 MAP kinase pathway, we isolated the Ste12 homologue CST1 gene from C. lagenarium and characterized. The cst1delta strains were nonpathogenic on intact host leaves, but could form lesions when inoculated on wounded leaves. Conidia of the cst1delta strains could germinate and form melanized appressoria on both host leaf surface and artificial cellulose membrane, but could not produce infectious hyphae from appressoria, suggesting that CST1 is essential for appressorium penetration in C. lagenarium. In addition, matured appressoria of the cst1delta strains contained an extremely low level of lipid droplets compared with that of the wild-type strain. Lipid droplets were abundant in conidia of the cst1delta strains, but rapidly disappeared during appressorium formation. This misscheduled lipid degradation might be related to the failure of appressorium penetration in the cst1delta strain.


Topoisomerase II regulates yeast genes with singular chromatin architectures.
Eukaryotic topoisomerase II (topo II) is the essential decatenase of newly replicated chromosomes and the main relaxase of nucleosomal DNA. Apart from these general tasks, topo II participates in more specialized functions. In mammals, topo IIα interacts with specific RNA polymerases and chromatin-remodeling complexes, whereas topo IIβ regulates developmental genes in conjunction with chromatin remodeling and heterochromatin transitions. Here we show that in budding yeast, topo II regulates the expression of specific gene subsets. To uncover this, we carried out a genomic transcription run-on shortly after the thermal inactivation of topo II. We identified a modest number of genes not involved in the general stress response but strictly dependent on topo II. These genes present distinctive functional and structural traits in comparison with the genome average. Yeast topo II is a positive regulator of genes with well-defined promoter architecture that associates to chromatin remodeling complexes; it is a negative regulator of genes extremely hypo-acetylated with complex promoters and undefined nucleosome positioning, many of which are involved in polyamine transport. These findings indicate that yeast topo II operates on singular chromatin architectures to activate or repress DNA transcription and that this activity produces functional responses to ensure chromatin stability.


Simple phase-shifting lateral shearing interferometer.
A phase-shifting electronic speckle pattern shearing interferometer with a very simple shearing device is proposed. Two partially reflective glass plates are used to introduce the shear in this new interferometer. The reflection coefficients of the coatings on the two plates are 0.3 and 0.7. The distance between the two glass plates controls the size of the shear. The proposed new interferometric system is simple, flexible, and low cost.


The dynamical interplay between a megadalton peptide nanocage and solutes probed by microsecond atomistic MD; implications for design.
Understanding the assembly and dynamics of protein-based supramolecular capsids and cages is of fundamental importance and could lead to applications in synthetic biology and biotechnology. Here we present long and large atomistic molecular dynamics simulations of de novo designed self-assembling protein nanocages (SAGEs) in aqueous media. Microsecond simulations, comprised of ≈42 million atoms for three pre-formed SAGEs of different charges, in the presence of solutes and solvent have been completed. Here, the dynamics, stability and porosity of the peptide networks are explored along with their interactions with ions, small molecules and macromolecular solutes. All assemblies are stable over the μs timescale, and the solutes show a mixture of transport behaviour across or adherence to the fabric of the SAGE particles. Solute proteins largely retained native-like conformation on contact with SAGE. Certain residues of the SAGE peptides are identified as "repeat offenders" for contacting many different solutes, which suggest modifications to reduce non-specific binding. These studies highlight how molecular dynamics can aid the design process of SAGE and similar assemblies for potential applications as diverse as platforms for drug and vaccine delivery and nanoreactors to encapsulate enzyme pathways.


Controversies in the Science of Sedentary Behaviour and Health: Insights, Perspectives and Future directions from the 2018 Queensland Sedentary Behaviour Think Tank.
The development in research concerning sedentary behaviour has been rapid over the past two decades. This has led to the development of evidence and views that have become more advanced, diverse and, possibly, contentious. These include the effects of standing, the breaking up of prolonged sitting and the role of moderate-to-vigorous physical activity (MVPA) in the association between sedentary behaviour and health outcomes. The present aim is to report the views of experts (n = 21) brought together (one-day face-to-face meeting in 2018) to consider these issues and provide conclusions and recommendations for future work. Each topic was reviewed and presented by one expert followed by full group discussion, which was recorded, transcribed and analysed. The experts concluded that (a). standing may bring benefits that accrue from postural shifts. Prolonged (mainly static) standing and prolonged sitting are both bad for health; (b). 'the best posture is the next posture'. Regularly breaking up of sitting with postural shifts and movement is vital; (c). health effects of prolonged sitting are evident even after controlling for MVPA, but high levels of MVPA can attenuate the deleterious effects of prolonged sitting depending on the health outcome of interest. Expert discussion addressed measurement, messaging and future directions.