Taxol and β-tubulins from endophytic fungi isolated from the Himalayan Yew, Taxus wallichiana Zucc.
Paclitaxel, better known as the anticancer drug Taxol®, has been isolated from several plant species and has been shown to be produced by fungi, actinomycetes, and even bacteria isolated from marine macroalgae. Given its cytostatic effect, studies conducted in the 1990's showed that paclitaxel was toxic to many pathogenic fungi and oomycetes. Further studies led to the idea that the differences in paclitaxel sensitivity exhibited by different fungi were due to differences in the β-tubulin protein sequence. With the recent isolation of endophytic fungi from the leaves and bark of the Himalayan Yew, Taxus wallichiana Zucc., and the availability of genomes from paclitaxel-producing fungi, we decided to further explore the idea that endophytic fungi isolated from Yews should be well-adapted to their environment by encoding β-tubulin proteins that are insensitive to paclitaxel. Our results found evidence of episodic positive/diversifying selection at 10 sites (default p-value threshold of 0.1) in the β-tubulin sequences, corresponding to codon positions 33, 55, 172, 218, 279, 335, 359, 362, 379, and 406. Four of these positions (i.e., 172, 279, 359, and 362) have been implicated in the binding of paclitaxel by β-tubulin or formed part of the binding pocket. As expected, all the fungal endophytes grew in different media regardless of the paclitaxel concentration tested. Furthermore, our results also showed that Taxomyces andreanae CBS 279.92, the first fungus shown to produce paclitaxel, is a Basidiomycete fungus as the two beta tubulins encoded by the fungus clustered together with other Basidiomycete fungi.


Nano-Biomechanical Study of Spatio-Temporal Cytoskeleton Rearrangements that Determine Subcellular Mechanical Properties and Endothelial Permeability.
The endothelial cell (EC) lining of the pulmonary vascular system forms a semipermeable barrier between blood and the interstitium and regulates various critical biochemical functions. Collectively, it represents a prototypical biomechanical system, where the complex hierarchical architecture, from the molecular scale to the cellular and tissue level, has an intimate and intricate relationship with its biological functions. We investigated the mechanical properties of human pulmonary artery endothelial cells (ECs) using atomic force microscopy (AFM). Concurrently, the wider distribution and finer details of the cytoskeletal nano-structure were examined using fluorescence microscopy (FM) and scanning transmission electron microscopy (STEM), respectively. These correlative measurements were conducted in response to the EC barrier-disrupting agent, thrombin, and barrier-enhancing agent, sphingosine 1-phosphate (S1P). Our new findings and analysis directly link the spatio-temporal complexities of cell re-modeling and cytoskeletal mechanical properties alteration. This work provides novel insights into the biomechanical function of the endothelial barrier and suggests similar opportunities for understanding the form-function relationship in other biomechanical subsystems.


[Baculovirus expression systems for recombinant protein production in insect and mammalian cells].
The baculovirus vector systems has been extensively used for the expression of foreign gene products in insect and mammalian cells. New advances increase the possibilities and applications of the baculovirus expression system, which has the capability to express multiple genes simultaneously within a single infected insect cells and to use recombinant virus with mammalian cell-active expression cassettes to permit expression of recombinant proteins in mammalian cells in vitro and in vivo. Future investigations of the baculovirus expression system designed for specific target cells, can open wide variety of applications. This review summarizes the recent achievements in applications the baculovirus vector systems and optimization recombinant protein expression in both insect and mammalian cell lines.


The 'ins and outs' of colonoscopy at Wits Donald Gordon Medical Centre, South Africa: A practice audit of the outpatient endoscopy unit.
In South Africa, there are no national guidelines for the conduct or quality assessment of colonoscopy, the gold standard for investigation and diagnosis of bowel pathology. To describe the clinical profile of patients and evaluate the practice of colonoscopy using procedural quality indicators at the Wits Donald Gordon Medical Centre (WDGMC) outpatient endoscopy unit (OEU). We conducted a prospective, clinical practice audit of colonoscopies performed on adults (≥18 years of age). A total of 1 643 patients were included in the study and variables that were collected enabled the assessment of adequacy of bowel preparation, length of withdrawal time and calculation of caecal intubation rate (CIR), polyp detection rate (PDR) and adenoma detection rate (ADR). We stratified PDR and ADR by sex, age, population group, withdrawal time and bowel preparation. CIR, PDR and ADR estimates were compared between patient groups by the χ2 test; Fisher's exact test was used for 2 × 2 tables. A p-value <0.05 was used. Benchmark recommendations by the American Society for Gastrointestinal Endoscopy (ASGE)/American College of Gastroenterology (ACG) Task Force on Colorectal Cancer (CRC) were used in this audit to assess individual endoscopist performance and that of the endoscopy unit as a whole. The mean age of patients was 55.7 (standard deviation (SD) 14.4; range 18 - 91) years, ~60% were female, and the majority (75.5%) were white. Of the outpatients, 77.6% had adequate bowel preparation (ASGE/ACG benchmark ≥85%). The CIR was 97.0% overall, and screening colonoscopy was 96.3% (ASGE/ACG benchmark ≥90% overall and ≥95% for screening colonoscopies). The median withdrawal time for negative-result screening colonoscopies was 5.7 minutes (interquartile range (IQR) 4.2 - 9.3; range 1.1 - 20.6) (ASGE/ACG benchmark ≥ 6minutes), and PDR and ADR were 27.6% and 15.6%, respectively (ASGE/ACG benchmark ADR ≥25%). We demonstrated a 23.7% increase in PDR and 14.1% increase in ADR between scopes that had mean withdrawal times of ≥6 minutes and <6 minutes, respectively. Although the number of black Africans in the study was relatively small, our results showed that they have similar ADRs and PDRs to the white population group, contradicting popular belief. The WDGMC OEU performed reasonably well against the international guidelines, despite some inadequacy in bowel preparation and lower than recommended median withdrawal times on negative-result colonoscopy. Annual auditing of clinical practice and availability of these data in the public domain will become standard of care, making this audit a baseline for longitudinal observation, assessing the impact of interventions, and contributing to the development of local guidelines.


Wnt-3a improves functional recovery after spinal cord injury by regulating the inflammatory and apoptotic response in rats via wnt/β-catenin signaling pathway.
The specific molecular mechanism of neuroprotective effects of wnt-3a on spinal cord injury (SCI) has not been elucidated. In our study, we evaluated the recovery of motor function after SCI by BBB, observed neuronal apoptosis by western blot and TUNEL, observed the changes of neuronal inflammation by western blot and immunofluorescence staining, and observed the changes of motoneurons and spinal cord area in the anterior horn of the spinal cord via Nissl and HE staining. We found that wnt-3a could significantly promote the recovery of motor function, reduce the loss of motor neurons in the anterior horn of the spinal cord, promote the recovery of injured spinal cord tissue, inhibit neuronal apoptosis and inflammatory response, and ultimately promote neuronal function after SCI. However, when XAV939 inhibits the wnt/β-catenin signaling pathway, the neuroprotective effects of wnt-3a are also significantly inhibited. The above results together indicated that wnt-3a exerts its neuroprotective effect on after SCI via activating the wnt/β-catenin signaling pathway.


Polygenic risk for circulating reproductive hormone levels and their influence on hippocampal volume and depression susceptibility.
Altered reproductive hormone levels have been associated with the pathophysiology of depressive disorders and this risk may be imparted by their modulatory effect upon hippocampal structure and function. Currently it is unclear whether altered levels of reproductive hormones are causally associated with hippocampal volume reductions and the risk of depressive disorders. Here, we utilize genome-wide association study (GWAS) summary statistics from a GWAS focusing on reproductive hormones, consisting of 2913 individuals. Using this data, we generated polygenic risk scores (PRS) for estradiol, progesterone, prolactin and testosterone in the European RADIANT cohort consisting of 176 postpartum depression (PPD) cases (100% female, mean age: 41.6 years old), 2772 major depressive disorder (MDD) cases (68.6% female, mean age: 46.9 years old) and 1588 control participants (62.5% female, mean age: 42.4 years old), for which there was also a neuroimaging subset of 111 individuals (60.4% female, mean age: 50.0 years old). Only the best-fit PRS for estradiol showed a significant negative association with hippocampal volume, as well as many of its individual subfields; including the molecular layer and granule cell layer of the dentate gyrus, subiculum, CA1, CA2/3 and CA4 regions. Interestingly, several of these subfields are implicated in adult hippocampal neurogenesis. When we tested the same estradiol PRS for association with case-control status for PPD or MDD there was no significant relationship observed. Here, we provide evidence that genetic risk for higher plasma estradiol is negatively associated with hippocampal volume, but this does not translate into an increased risk of MDD or PPD. This work suggests that the relationship between reproductive hormones, the hippocampus, and depression is complex, and that there may not be a clear-cut pathway for etiology or risk moderation.


Parkinson Disease.
Parkinson disease is a progressive neurodegenerative disorder with significant morbidity and mortality. Most patients consult with their primary care physician about Parkinson disease symptoms before seeking care from a specialist. The diagnosis of Parkinson disease is clinical, and key disease features are bradykinesia, rigidity, and tremor. The main diagnostic signs of Parkinson disease are motor symptoms; however, Parkinson disease is also associated with nonmotor symptoms, including autonomic dysfunction, depression, and hallucinations, which can make the initial diagnosis of Parkinson disease difficult. Disease progression is variable and clinical signs cannot be used to predict progression accurately. Therapies, including levodopa, have not demonstrated the ability to slow disease progression. Motor symptoms are managed with carbidopa/levodopa, monoamine oxidase-B inhibitors, and nonergot dopamine agonists. Prolonged use and higher doses of levodopa result in dyskinesias and motor symptom fluctuations over time. Deep brain stimulation surgery is performed for patients who do not achieve adequate control with levodopa therapy. Deep brain stimulation is most effective for significant motor fluctuations, dyskinesias, and tremors. Nonmotor symptom therapies target patient-specific conditions during the disease course. Interdisciplinary team care can alleviate multiple symptoms of Parkinson disease.


Accelerated mineralization of two organophosphate insecticides in the rhizosphere.
Numerous xenobiotic compounds, including the organophosphate insecticides O, O-diethyl-O-(2-isopropyl-6-methyl-4-pyrimidinyl) phosphorothioate (diazinon) and O, O-diethyl-O-p-nitrophenyl phosphorothioate (parathion), appear to be degraded in the soil environment by an initial cometabolic attack. Comparing the mineralization rates of radiolabeled diazinon and parathion in root-free and in rhizosphere soil, we tested our hypothesis that, because of the presence of root exudates, the rhizosphere is an especially favorable environment for such co-metabolic transformations. The insecticides were added individually at 5 mug/g to sealed flasks containing either soil permeated by the root system of a bush bean plant or identical soil without roots. Periodically, the flask atmospheres were flushed through traps and the evolved (14)CO(2) was quantitated. Bush bean plant roots without associated rhizosphere microorganisms failed to produce a significant amount of (14)CO(2). During 1 month of incubation, rhizosphere flasks mineralized 12.9 and 17.9% of the added diazinon and parathion radiocarbon, respectively, compared to 5.0 and 7.8% by the soil without roots. The mineralization of parathion but not of diazinon was stimulated in a similar manner when soil without roots was repeatedly irrigated with a root exudate produced in aseptic solution culture. Viable counts of microorganisms on soil extract agar were not significantly altered by root permeation or by root exudate treatment of the soil, leaving population selection and/or enhanced cometabolic activity as the most plausible interpretations for the observed stimulatory effects. Rhizosphere interactions may substantially shorten the predicted half-lives of some xenobiotic compounds in soil.


Chitosan coating silver nanoparticles as a promising feed additive in broilers chicken.
The present study aimed to evaluate the potential of chitosan coating silver nanoparticles to enhance the growth performance and immune status of broilers without inducing oxidative stress-related pathological lesions in any organs or leaving residues of silver in the edible parts. Five clusters of Cobb one-day-old chicks (n = 10/group in each replication) were given oral therapy, once a week for 36 days as follows: (1) distilled water, (2, 3) 0.5- and 5 ppm silver nanoparticles (AgNPs), respectively, (4, 5) 0.5- and 5 ppm chitosan/silver nanoconjugates (CS/Ag-NCs), respectively. The results demonstrated a marked elevation in the body weight gain with a decline in the food conversion ratio and marked improvement in feeding and drinking behavior of all nanoparticles treated groups, but higher in CS/Ag-NCs groups than AgNPs groups and control group. In contrast to the 0.5 ppm AgNPs receiving group, the group receiving 5 ppm AgNPs noticed remarkable histological changes in some organs, including the liver, kidneys, spleen, and heart. Moreover, the administration of CS/Ag-NCs at two dosage levels didn't influence any histological changes. The AgNPs groups' antibody titers against the ND and AI viruses were almost identical to those of the control group. Otherwise, CS/Ag-NCs groups recorded the highest antibody titers. Additionally, there was a significant increase in silver content in most edible organs of AgNPs groups at a dosage level of 5 ppm. Otherwise, the coating of AgNPs by CSNPs could decrease the aggregation of silver in the biological organs. Thus, we recommend utilizing 0.5 ppm CS/Ag-NCs in broiler farms to promote their growth performance and strengthen their immune defense.


Assessing site signal preservation in reference chronologies for dendro-provenancing.
Regional differences in tree growth can be used to approximate the geographical provenance of ring-width series ('dendro-provenancing'). This method relies on cross-dated ring-width series (reference chronologies) that are thought to represent the radial growth signal of trees in a given region. Reference chronologies are often established from ring-width series of living tree populations. Frequently, they are too short to allow for investigating the provenance of historical wood. Thus, references are extended by ring-width series from buildings and art-historical objects that exhibit best matching growth patterns with the living tree references. Yet, series from other provenances may erroneously be included. Thereby the local or regional growth signal of the references is progressively contaminated, but this has received little attention to date. I investigate this contamination risk using a simulation approach that allows for generating pseudo site chronologies that preserve the relevant statistical properties of the real site chronologies. While the exact provenance of historical wood is unknown, for simulated ring-width series the provenance is unambiguous. Hence, pseudo reference chronologies may be established while monitoring the signal mixture. Specifically, 15 site chronologies of Norway spruce (Picea abies (L.) H. Karst.) from northeastern Switzerland were used to generate 15 pseudo site growth signals that span 1000 years. The simulation demonstrates that quasi uncontaminated references can be established in ideal circumstances for the study area. However, the thresholds for the similarity in between-series correlation must be very high. Even then, contaminated pseudo references occurred in rare cases during the simulation. Yet, elevation-specific pseudo references were established with lower thresholds. Simulation currently offers the only approach for assessing the contamination risk of reference chronologies, and it allows for elucidating the conditions under which acceptable levels of contamination can be guaranteed. Therefore, the present approach paves the way towards a practical simulation tool for dendro-provenancing.


Macrophage immunomodulatory activity of polysaccharides isolated from Opuntia polyacantha.
Opuntia polyacantha (prickly pear cactus) has been used extensively for its nutritional properties; however, less is known regarding medicinal properties of Opuntia tissues. In the present study, we extracted polysaccharides from O. polyacantha and used size-exclusion chromatography to fractionate the crude polysaccharides into four polysaccharide fractions (designated as Opuntia polysaccharides C-I to C-IV). The average M(r) of fractions C-I through C-IV was estimated to be 733, 550, 310, and 168 kDa, respectively, and sugar composition analysis revealed that Opuntia polysaccharides consisted primarily of galactose, galacturonic acid, xylose, arabinose, and rhamnose. Analysis of the effects of Opuntia polysaccharides on human and murine macrophages demonstrated that all four fractions had potent immunomodulatory activity, inducing production of reactive oxygen species, nitric oxide, tumor necrosis factor alpha, and interleukin 6. Furthermore, modulation of macrophage function by Opuntia polysaccharides was mediated, at least in part, through activation of nuclear factor kappaB. Together, our results provide a molecular basis to explain a portion of the beneficial therapeutic properties of extracts from O. polyacantha and support the concept of using Opuntia polysaccharides as an immunotherapeutic adjuvant.


Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) colonization and infection in intravenous and inhalational opiate drug abusers.
Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen in hospital-acquired and community-acquired infections. We hypothesized that drug abuse is a risk factor for community-acquired MRSA (CA-MRSA) infection, and we employed a rapid multiplex PCR technique for MRSA identification. The study was conducted on MRSA isolates from 60 opiate addicts (intravenous and inhalational drug users) to detect the rate and location of MRSA colonization and infection among them in comparison to 60 non-addict patients and 15 healthy volunteer controls. The proportion of addicts with MRSA colonization (and/or infection) was significantly higher than non-addict patients with MRSA colonization. MRSA colonization was associated with infection in 58% of MRSA-colonized addicts. The MRSA nasal carriage in the addicts was significantly higher than MRSA carriage elsewhere, whether in the addicts group or in the non-addict patients group. Moreover, the nasopharyngeal carriage rate of MRSA among addicts was significantly higher than among the non-addict patients. Increasing the duration of addiction resulted in a significant increase in CA-MRSA colonization in opiate addicts. Both inhalational and intravenous drug use led to significant MRSA colonization in the addict population. In conclusion, this study demonstrated how drug abusers, a high-risk group for infections with MRSA, could be a source or a reservoir of CA-MRSA infection in the non-addict population.


Functional activation during reading in relation to the sulci of the angular gyrus region.
Neurological studies suggest that the angular gyrus region of the inferior parietal lobule may be critical for reading. However, unambiguous demonstration of angular gyrus involvement from lesion and functional neuroimaging studies is lacking, partly because of the absence of detailed morphological descriptions of this region. On the basis of our recent anatomical examination of this region and a tightly controlled functional magnetic resonance imaging paradigm, the present investigation demonstrated reading-related activity in the region of the angular gyrus that lies between the central and posterior branches of the caudal superior temporal sulcus, namely cytoarchitectonic area PG. Analysis of functional connectivity showed increased functional coupling during reading of area PG with the language areas of Broca and Wernicke, and a region previously identified as the visual word form area. Thus, the parietal reading area has been precisely localized, and its interactions with other cortical areas during reading have been demonstrated.


Foetal alcohol spectrum disorder: identifying the neurobehavioural phenotype and effective interventions.
Since the first description of the foetal damage of alcohol in 1967, numerous studies have outlined different aspects of neurodevelopmental dysfunction, adversely affecting the lives of children worldwide. Although the cause of the syndrome is sorted out, the pathogenesis of brain damage is far from being clear. In contrast to children exhibiting the full facial dysmorphology, who are relatively easy to diagnose, in those presenting only with alcohol-related neurodevelopmental damage diagnosis is much more challenging due to poor specificity of the brain dysfunction. Hence, identifying the neurodevelopmental phenotype of foetal alcohol spectrum disorder (FASD) is a major challenge. Recently, a behavioural phenotype of FASD has been described and validated using items from the Child Behaviour Checklist. This tool has high sensitivity and specificity in separating children with FASD from those with ADHD and from healthy controls. In parallel, a number of intervention studies show promise in improving the abilities of children and adolescents with the syndrome to cope with daily tasks and improve their quality of life. The neurobehavioural screening test can facilitate screening for FASD and is an official screening tool in the FASD toolkit of the Public Health Agency of Canada. Promising new interventions may attenuate the long-term outcome of these children.


Movement of a barley stripe mosaic virus chimera with a tobacco mosaic virus movement protein.
The tobacco mosaic virus (TMV) 30K movement protein (MP) gene was inserted into a full-length cDNA clone of barley stripe mosaic virus (BSMV) RNA beta replacing the triple gene block (TGB). The resulting recombinant ND-MPT genome, consisting of infectious wt transcripts of BSMV RNAs alpha and gamma, together with the hybrid RNA beta transcript, was inoculated onto test plants to study the functional compatibility between the BSMV TGB-adapted genetic system and the tobamovirus transport gene. ND-MPT infected the inoculated leaves of Nicotiana benthamiana and Chenopodium amaranticolor, which are common hosts for the parental viruses; the size, growth rate, and morphology of local lesions on C. amaranticolor were influenced by the foreign MP gene. However, the hybrid virus failed to infect barley, N. tabacum (var. Samsun), and N. clevelandii, the selective hosts. Thus, the TMV MP was able to functionally substitute for the BSMV TGB-coded MPs, i.e., the 30K MP functioned independently of any other BSMV sequences. However, the TMV MP gene promoted the cell-to-cell movement in a host-dependent manner.


Late cardiac effects after mantle radiotherapy in patients with Hodgkin's disease.
Twenty-five patients (21-45 years old) treated for Hodgkin's disease with mantle radiotherapy but no chemotherapy underwent cardiac testing with myocardial scintigraphy during exercise, Echo-Doppler cardiography and CT-examination, 10-20 years after treatment. Four of twenty-six (15%) young patients had serious cardiac complications after mantle therapy, and reduced systolic and/or diastolic function; and minor valvular disturbances were often found. One 36-year-old female died of myocardial infarction 4 years after therapy, one 39-year-old male had two non-lethal infarctions after 14 years, one 36-year-old male with no symptoms had severe reversible ischemia and three proximal coronary artery stenoses, and one 32-year-old female with constrictive pericarditis had pericardeictomy 14 years after therapy. In 23/24 patients the pericardial thickness was normal and no pericardial effusion was found. 23/24 patients had normal working capacity, but myocardial scintigraphy was normal in only 9 patients. 11/25 patients had reduced systolic function and in 12/24 patients the diastolic function was reduced. 11/25 patients had abnormal valvular or subvalvular structures. Valvular stenosis was not found but aortic, mitral and tricuspidal regurgitations were found in 1/25, 9/25 and 22/25, respectively. In all but two cases the regurgitations were mild. We conclude that mediastinal irradiation must be considered a risk factor for cardiac disease. It may be advisable to reduce other risk factors in these patients.


Evidence of a third ADPKD locus is not supported by re-analysis of designated PKD3 families.
Mutations to PKD1 and PKD2 are associated with autosomal dominant polycystic kidney disease (ADPKD). The absence of apparent PKD1/PKD2 linkage in five published European or North American families with ADPKD suggested a third locus, designated PKD3. Here we re-evaluated these families by updating clinical information, re-sampling where possible, and mutation screening for PKD1/PKD2. In the French-Canadian family, we identified PKD1: p.D3782_V3783insD, with misdiagnoses in two individuals and sample contamination explaining the lack of linkage. In the Portuguese family, PKD1: p.G3818A segregated with the disease in 10 individuals in three generations with likely misdiagnosis in one individual, sample contamination, and use of distant microsatellite markers explaining the linkage discrepancy. The mutation PKD2: c.213delC was found in the Bulgarian family, with linkage failure attributed to false positive diagnoses in two individuals. An affected son, but not the mother, in the Italian family had the nonsense mutation PKD1: p.R4228X, which appeared de novo in the son, with simple cysts probably explaining the mother's phenotype. No likely mutation was found in the Spanish family, but the phenotype was atypical with kidney atrophy in one case. Thus, re-analysis does not support the existence of a PKD3 in ADPKD. False positive diagnoses by ultrasound in all resolved families shows the value of mutation screening, but not linkage, to understand families with discrepant data.


Direct quantitative evaluation of disease symptoms on living plant leaves growing under natural light.
Leaf color is an important indicator when evaluating plant growth and responses to biotic/abiotic stress. Acquisition of images by digital cameras allows analysis and long-term storage of the acquired images. However, under field conditions, where light intensity can fluctuate and other factors (shade, reflection, and background, etc.) vary, stable and reproducible measurement and quantification of leaf color are hard to achieve. Digital scanners provide fixed conditions for obtaining image data, allowing stable and reliable comparison among samples, but require detached plant materials to capture images, and the destructive processes involved often induce deformation of plant materials (curled leaves and faded colors, etc.). In this study, by using a lightweight digital scanner connected to a mobile computer, we obtained digital image data from intact plant leaves grown in natural-light greenhouses without detaching the targets. We took images of soybean leaves infected by Xanthomonas campestris pv. glycines, and distinctively quantified two disease symptoms (brown lesions and yellow halos) using freely available image processing software. The image data were amenable to quantitative and statistical analyses, allowing precise and objective evaluation of disease resistance.


THe proton-per-electron stoicheiometry of 'site 1' of oxidative phosphorylation at high protonmotive force is close to 1.5.
The maximum redox potential difference between the NAD+/NADH couple and the succinate/fumarate couple generated during ATP-energized reduction of NAD+ by succinate in submitochondrial particles was measured, together with the electrochemical potential difference for protons (delta mu approximately H+). The presence of cyanide, the time-independence of the redox potential difference and the irrelevance of the initial redox state of the NAD+/NADH couple ensured that the experimental situation corresponded to a 'static-head condition' with delta mu approximately H+ as the input force and the redox potential difference as the output force, the flow of electrons having reached dynamic equilibrium. Consequently, the observed value of 1.6 for the ratio delta Ge/delta mu approximately H+ is interpreted as indicating that the leads to H+/e- stoicheiometry at 'site 1' is 1.5 and that therefore the mechanism of the proton pump at 'site 1' is not of the group-translocation type (no direct leads to e - leads to H+ coupling).


Potential Interactions between miRNAs and Hypoxia: A New Layer in Cancer Hypoxia.
Hypoxia pathway and aberrant miRNA expression profile play crucial roles in the development of various cancers. Recent studies have emphasized that there are many collaborations in this cases because both of them are involved in cancer cell processes, including differentiation, metastasis, and cell proliferation and signaling pathways. Further studies have elucidated that miRNAs affect the hypoxia route, and more interestingly, the hypoxia pathway also affects miRNAs expression profile. The literature review summarizes the fundamental roles of hypoxia-related miRNAs in different cancers. The mutual interactions between miRNAs and hypoxia are a new layer of complexity in cancer hypoxia, which might be helpful in controlling cancer progression. It is also possible that the hypoxia pathway is initiated by miRNAs. In contrast, the hypoxia pathway regulates the expression of a large group of miRNAs. In this review for the first time, we discussed the significant interactions between hypoxia and miRNAs in order to determine new perspectives for new therapeutic aims in the field of cancer.


HUMMR, a hypoxia- and HIF-1alpha-inducible protein, alters mitochondrial distribution and transport.
Mitochondrial transport is critical for maintenance of normal neuronal function. Here, we identify a novel mitochondria protein, hypoxia up-regulated mitochondrial movement regulator (HUMMR), which is expressed in neurons and is markedly induced by hypoxia-inducible factor 1 alpha (HIF-1alpha). Interestingly, HUMMR interacts with Miro-1 and Miro-2, mitochondrial proteins that are critical for mediating mitochondrial transport. Interestingly, knockdown of HUMMR or HIF-1 function in neurons exposed to hypoxia markedly reduces mitochondrial content in axons. Because mitochondrial transport and distribution are inextricably linked, the impact of reduced HUMMR function on the direction of mitochondrial transport was also explored. Loss of HUMMR function in hypoxia diminished the percentage of motile mitochondria moving in the anterograde direction and enhanced the percentage moving in the retrograde direction. Thus, HUMMR, a novel mitochondrial protein induced by HIF-1 and hypoxia, biases mitochondria transport in the anterograde direction. These findings have broad implications for maintenance of neuronal viability and function during physiological and pathological states.


[A questionnaire to capture the functional effects of daytime drowsiness on quality of life in case of obstructive sleep apnea syndrome. Functional Outcomes of Sleep Questionnaire (FOSQ)].
The Functional Outcome of Sleep Questionnaire (FOSQ) is a questionnaire for capturing the function-specific quality of life (impairment due to daytime drowsiness). It is subdivided into five subscales/sections, each comprising four degrees of severity. So far, no standard values for healthy persons without sleep disorders exist in the German-speaking countries. The aim of our study was therefore first of all to establish standard values for healthy subjects as well as to verify the three quality criteria required for testing procedures (objectivity, reliability, validity). Finally, a comparison between health subjects and OSAS patients was carried out. The study included 210 health subjects and 150 OSAS patients. Regarding standardisation of the FOSQ, the following standard values for healthy persons without sleep disorders were determined for the five subscales: activity 21.4 +/- 2.9; vigilance 20.5 +/- 4.3; intimacy 22.3 +/- 3.9; social QoL 22.6 +/- 3.3; general capability 22.3 +/- 2.7. Age and weight (BMI) influences could be excluded. Concerning verification of the three quality criteria, good values could be established for objectivity, reliability and validity. As a result, significant differences between health subjects and OSAS patients were found. In the five quality of life areas captured (activity 15.8 +/- 4.9; vigilance 14.6 +/- 6.4; intimacy 17.3 +/- 6.5; social QoL 18.9 +/- 5.5; general capability 17.9 +/- 5.2), the patients achieved values distinctly inferior to those of the health probands. In summary, for the German-speaking area, due to its test design, this questionnaire represents a simple procedure for capturing the function-related reduction in quality of life for all age groups both in the clinical field and in case of particular questions relating to expert opinions.


Protective effects of anthocyanins from blackberry in a rat model of acute lung inflammation.
Anthocyanins are a group of naturally occuring phenolic compounds related to the coloring of plants, flowers and fruits. These pigments are important as quality indicators, as chemotaxonomic markers and for their antioxidant activities. Here, we have investigated the therapeutic efficacy of anthocyanins contained in blackberry extract (cyanidin-3-O-glucoside represents about 80% of the total anthocyanin contents) in an experimental model of lung inflammation induced by carrageenan in rats. Injection of carrageenan into the pleural cavity elicited an acute inflammatory response characterized by fluid accumulation which contained a large number of neutrophils as well as an infiltration of polymorphonuclear leukocytes in lung tissues and subsequent lipid peroxidation, and increased production of nitrite/nitrate (NOx) and prostaglandin E2 (PGE2). All parameters of inflammation were attenuated in a dose-dependent manner by anthocyanins (10, 30 mg kg(-1) 30 min before carrageenan). Furthermore, carrageenan induced an upregulation of the adhesion molecule ICAM-1, nitrotyrosine and poly (ADP-ribose) synthetase (PARS) as determined by immunohistochemical analysis of lung tissues. The degree of staining was lowered by anthocyanins treatment. Thus, the anthocyanins contained in the blackberry extract exert multiple protective effects in carrageenan-induced pleurisy.


On fuzzy correlations.
A general framework for dealing with numerical measurements in an approximate, uncertain, or fuzzy environment is presented. A fuzzy measurement is defined. It possesses several unique properties, which arise from its physical nature and distinguish it from concepts such as the fuzzy number. These properties, which include the fuzzy correlation term and the fuzzy Equality relation, follow directly from physical considerations. The introduction of the fuzzy correlation term provides a mathematical tool for representing any correlation relations, which may exist between different fuzzy measurements. The main function of the fuzzy correlation term is to eliminate, or filter out, measurement values that are unlikely, given other fuzzy measurements. Thus, using the fuzzy correlation term, the range of possible measurement values is limited by physical realities. The information represented by the fuzzy correlation term is shown to be of great value in providing a wider picture of reality than it is possible to obtain by simply considering individual fuzzy measurements. Arithmetic operations on fuzzy measurements and functions of fuzzy measurements are also discussed, leading to the derivation of the fuzzy Riemann integral and its applications.


Inhibition of Clostridium difficile by faecal streptococci.
The inhibitory activity of seven strains of faecal streptococci against 34 strains of Clostridium difficile was examined in vitro after growth of the streptococci for 24 and 48 h. All strains of C. difficile were inhibited at 48 h but at 24 h the inhibition was variable. Streptococcus faecium, a group D streptococcus and an ungroupable streptococcus exhibited the most striking inhibitory activity. Lowering of pH of the medium occurred at the site of inhibition, but the pH change alone did not explain the inhibition of C. difficile. This antagonism may be related in vivo to the resistance to intestinal colonisation by C. difficile exhibited by the normal bowel flora, and in vitro to failures to isolate C. difficile from faecal specimens.


Fluctuations in brain temperature induced by lipopolysaccharides: central and peripheral contributions.
In this study, we examined changes in central (anterior-preoptic hypothalamus) and peripheral (temporal muscle and facial skin) temperatures in freely moving rats following intravenous administration of bacterial lipopolysaccharides (LPS) at low doses (1 and 10 μg/kg) at thermoneutral conditions (28°C). Recordings were made with high temporal resolution (5-s bin) and the effects of LPS were compared with those induced by a tail-pinch, a standard arousing somato-sensory stimulus. At each dose, LPS moderately elevated brain, muscle, and skin temperatures. In contrast to rapid, monophasic and relatively short hyperthermic responses induced by a tail-pinch, LPS-induced increases in brain and muscle temperatures occurred with ~40 min onset latencies, showed three not clearly defined phases, were slightly larger with the 10 μm/kg dose, and maintained for the entire 4-hour post-injection recording duration. Based on dynamics of brain-muscle and skin-muscle temperature differentials, it appears that the hyperthermic response induced by LPS at the lowest dose originates from enhanced peripheral heat production, with no evidence of brain metabolic activation and skin vasoconstriction. While peripheral heat production also appears to determine the first phase of brain and body temperature elevation with LPS at 10 μg/kg, a further prolonged increase in brain-muscle differentials (onset at ~100 min) suggests metabolic brain activation as a factor contributing to brain and body hyperthermia. At this dose, skin temperature increase was weaker than in temporal muscle, suggesting vasoconstriction as another contributor to brain/body hyperthermia. Therefore, although both LPS at low doses and salient sensory stimuli moderately increase brain and body temperatures, these hyperthermic responses have important qualitative differences, reflecting unique underlying mechanisms.


The urinary levels of prostanoid metabolites predict acute kidney injury in heterogeneous adult Japanese ICU patients: a prospective observational study.
Acute kidney injury (AKI) is frequently observed in critically ill patients in the intensive care unit (ICU) and is associated with increased mortality. Prostanoids regulate numerous biological functions, including hemodynamics and renal tubular transport. We herein investigated the ability of urinary prostanoid metabolites to predict the onset of AKI in critically ill adult patients. The current study was conducted as a prospective observational study. Urine of patients admitted to the ICU at Okayama University Hospital was collected and the urinary levels of prostaglandin E2 (PGE2), PGI2 metabolite (2,3-dinor-6-OXO-PGF1α), thromboxane A2 (TXA2) metabolite (11-dehydro-TXB2) were determined. Of the 93 patients, 24 developed AKI (AKIN criteria). Surgical intervention (93, 75 %) was the leading cause of ICU admission. Overall, the ratio of the level of serum Cr on Day 1 after ICU admission to that observed at baseline positively correlated with the urinary 2,3-dinor-6-OXO-PGF1α/Cr (r = 0.57, p < 0.0001) and 11-dehydro-TXB2/Cr (r = 0.47, p < 0.0001) ratios. In 16 cases of de novo AKI, the urinary 2,3-dinor-6-OXO-PGF1α/Cr and 11-dehydro-TXB2/Cr values were significantly elevated compared with that observed in the non-AKI group, whereas the urinary PGE2/Cr values were not. The urinary 2,3-dinor-6-OXO-PGF1α/Cr ratio exhibited the best diagnostic and predictive performance among the prostanoid metabolites according to the receiver operating characteristic (ROC) analysis [ROC-area under the curve (AUC): 0.75]. Taken together, these results demonstrate that the urinary 2,3-dinor-6-OXO-PGF1α/Cr and 11-dehydro-TXB2/Cr ratios are associated with the subsequent onset of AKI and poor outcomes in adult heterogeneous ICU patients.


Modulation of growth factors by growth hormone in children with chronic renal failure. The Southwest Pediatric Nephrology Study Group.
Anthropometric measurements and circulating growth factors were studied serially in 44 prepubertal children with growth failure and chronic renal failure (GFR = 10 to 40 ml/min/1.73 m2) who were randomized to receive either recombinant human growth hormone (rhGH; N = 30) or no treatment (N = 14). RhGH was given as Nutropin, 0.05 mg/kg/day, and the studies were carried out at baseline and after 3 and 12 months. At baseline, serum insulin-like growth factor binding protein (IGFBP)-1 and -2 levels were, while IGFBP-3 levels were not, higher than those of children with normal renal function. In addition, height SDS at baseline correlated inversely with serum IGFBP-2 levels (r = -0.461, P = 0.0016), but did not correlate significantly with any other factor. After 12 months of study, the 30 children receiving rhGH showed: (i) greater increase in height (9.1 +/- 2.8 vs. 5.5 +/- 1.9 cm, P < 0.0001); (ii) increases in serum levels of IGF-I, IGF-II, free IGF-I, IGFBP-3 and acid labile subunit (ALS); (iii) a greater decrease in serum IGFBP-1 levels; and (iv) no significant difference in serum IGFBP-2 levels, when compared to the 14 control patients. The change in height SDS after 12 months of rhGH (+0.8) in the 30 treated children correlated significantly and positively with serum ALS, IGFBP-3, total IGF, IGF-I, IGF-II and free IGF-I levels measured during treatment. These observations suggest that, in children with growth failure associated with chronic renal failure: (i) IGFBP-2, and not IGFBP-3, is likely to be a growth inhibitor; (ii) rhGH stimulates catch-up growth in part by increasing serum levels of IGF peptides; and (iii) linear growth is influenced by the balance between growth stimulating IGFs and growth inhibitory IGFBPs.


Integrase interactor 1 (Ini1/hSNF5) is a repressor of basal human immunodeficiency virus type 1 promoter activity.
Integrase interactor 1 (Ini1/hSNF5/BAF47/SMARCB1), the core subunit of the ATP-dependent chromatin-remodelling complex SWI/SNF, is a cellular interaction partner of the human immunodeficiency virus type 1 (HIV-1) integrase. Ini1/hSNF5 is recruited to HIV-1 pre-integration complexes before nuclear migration, suggesting a function in the integration process itself or a contribution to the preferential selection of transcriptionally active genes as integration sites of HIV-1. More recent evidence indicates, however, that, whilst Ini1/hSNF5 is dispensable for HIV-1 transduction per se, it may have an inhibitory effect on the early steps of HIV-1 replication but facilitates proviral transcription by enhancing Tat function. These partially contradictory observations prompted an investigation of the immediate and long-term effects of Ini1/hSNF5 depletion on the basal transcriptional potential of the virus promoter. Using small interfering RNAs, it was shown that Ini1/hSNF5-containing SWI/SNF complexes mediate transcriptional repression of the basal activity of the integrated HIV-1 long terminal repeat. Transient depletion of Ini1/hSNF5 during integration was accompanied by an early boost of HIV-1 replication. After the reappearance of Ini1/hSNF5, expression levels decreased and this was associated with increased levels of histone methylation at the virus promoter in the long term, indicative of epigenetic gene silencing. These results demonstrate the opposing effects of Ini1/hSNF5-containing SWI/SNF complexes on basal and Tat-dependent transcriptional activity of the HIV-1 promoter. It is proposed that Ini1/hSNF5 may be recruited to the HIV-1 pre-integration complex to initiate, immediately after integration, one of two mutually exclusive transcription programmes, namely post-integration latency or high-level, Tat-dependent gene expression.


Peppermint oil reduces gastric spasm during upper endoscopy: a randomized, double-blind, double-dummy controlled trial.
GI endoscopy without general anesthesia causes a hyperperistaltic state in the stomach, which frequently necessitates the use of antispasmodic agents, such as hyoscine-N-butylbromide, but these drugs have side effects. Peppermint oil is harmless and acts locally to inhibit GI smooth muscle contraction. A randomized double-blind, double-dummy, controlled trial was conducted in 100 patients to compare the antispasmodic effects of hyoscine-N-butylbromide administered intramuscularly and a placebo solution administered intraluminally by means of the endoscope, and also the effects of a placebo solution administered intramuscularly with those of a peppermint oil solution administered intraluminally. The percent change in diameter of the pyloric ring before and after the administrations was defined as the opening ratio, and the percent change in diameter between the maximally and minimally opened pyloric ring states was defined as the contraction ratio. Time until disappearance of the contraction ring(s) in the gastric antrum and side effects of the drugs were also determined. The opening ratio was significantly higher in the peppermint oil administration group than in the hyoscine-N-butylbromide injection group. The contraction ratio after peppermint oil administration was significantly lower than that after hyoscine-N-butylbromide injection. The time required for disappearance of the antral contraction ring(s) was shorter in the peppermint oil group (97.1 +/- 11.4) than in the hyoscine-N-butylbromide group (185.9 +/- 10.1 s; p < 0.0001). No significant side effects were associated with peppermint oil, whereas hyoscine-N-butylbromide injection produced side effects such as dry mouth, blurred vision, and urinary retention. Peppermint oil solution administered intraluminally can be used as an antispasmodic agent with superior efficacy and fewer side effects than hyoscine-N-butylbromide administered by intramuscular injection during upper endoscopy.


The distribution of progesterone receptor immunoreactivity and mRNA in the preoptic area and hypothalamus of the ewe: upregulation of progesterone receptor mRNA in the mediobasal hypothalamus by oestrogen.
The distribution of progesterone receptors (PR) was mapped in the hypothalamus of the ewe using immunocytochemistry. These results were confirmed using in situ hybridization with a sheep-specific 35S-labelled riboprobe. In addition, the effect of oestrogen on the level of PR mRNA in the hypothalamus was examined in ovariectomized (OVX) ewes following treatment with an oestrogen implant or without treatment. PR immunoreactive (-ir) cells were readily detected in OVX animals. Labelled cells were observed in four main hypothalamic regions: the preoptic area (POA), including the organum vasculosum of the lamina terminalis, periventricular nucleus (PeVN), ventromedial nucleus (VMN) and the arcuate nucleus (ARC) (including the region ventral to the mamillary recess). In addition, lightly stained PR-ir cells were observed in the supraoptic nucleus and a few PR-ir cells were also found in the diagonal band of Broca. No PR-ir cells were found in the brainstem. PR mRNA-containing cells were found in the same hypothalamic regions as the PR-ir cells. Image analysis of emulsion-dipped slides following in situ hybridization indicated that oestrogen treatment increased (P<0.01) the mean number of silver grains/cell and the density of labelled cells in the VMN and ARC but had no effect on the level of PR mRNA expression in the POA or PeN. The distribution of PR-containing cells in the hypothalamus is similar to that described in other species and all cells were located in nuclei that contain large populations of oestrogen receptor-containing cells. These include regions implicated in the regulation of reproductive neuroendocrine function, and reproductive behaviour. Oestrogen and progesterone synergize to inhibit GnRH secretion and the present results suggest that these functions may involve cells of the VMN and ARC, with oestrogen acting to upregulate PR.


Tablet-Based Telerehabilitation Versus Conventional Face-to-Face Rehabilitation After Cochlear Implantation: Prospective Intervention Pilot Study.
Technologies allowing home-based rehabilitation may be a key means of saving financial resources while also facilitating people's access to treatment. After cochlear implantation, auditory training is necessary for the brain to adapt to new auditory signals transmitted by the cochlear implant (CI). To date, auditory training is conducted in a face-to-face setting at a specialized center. However, because of the COVID-19 pandemic's impact on health care, the need for new therapeutic settings has intensified. The aims of this study are to assess the feasibility of a novel teletherapeutic auditory rehabilitation platform in adult CI recipients and compare the clinical outcomes and economic benefits of this platform with those derived from conventional face-to-face rehabilitation settings in a clinic. In total, 20 experienced adult CI users with a mean age of 59.4 (SD 16.3) years participated in the study. They completed 3 weeks of standard (face-to-face) therapy, followed by 3 weeks of computer-based auditory training (CBAT) at home. Participants were assessed at three intervals: before face-to-face therapy, after face-to-face therapy, and after CBAT. The primary outcomes were speech understanding in quiet and noisy conditions. The secondary outcomes were the usability of the CBAT system, the participants' subjective rating of their own listening abilities, and the time required for completing face-to-face and CBAT sessions for CI users and therapists. Greater benefits were observed after CBAT than after standard therapy in nearly all speech outcome measures. Significant improvements were found in sentence comprehension in noise (P=.004), speech tracking (P=.004) and phoneme differentiation (vowels: P=.001; consonants: P=.02) after CBAT. Only speech tracking improved significantly after conventional therapy (P=.007). The program's usability was judged to be high: only 2 of 20 participants could not imagine using the program without support. The different features of the training platform were rated as high. Cost analysis showed a cost difference in favor of CBAT: therapists spent 120 minutes per week face-to-face and 30 minutes per week on computer-based sessions. For CI users, attending standard therapy required an average of approximately 78 (SD 58.6) minutes of travel time per appointment. The proposed teletherapeutic approach for hearing rehabilitation enables good clinical outcomes while saving time for CI users and clinicians. The promising speech understanding results might be due to the high satisfaction of users with the CBAT program. Teletherapy might offer a cost-effective solution to address the lack of human resources in health care as well as the global challenge of current or future pandemics.


Serous retinal detachments in thrombotic thrombocytopenic purpura.
Serous retinal detachments are a rare ocular complication of thrombotic thrombocytopenic purpura (TTP), with only six previous case reports in the literature. We have recently observed two patients with relapsing TTP who developed bilateral serous retinal detachments during acute exacerbations of their disease. In contrast to all but one of the previously described patients, both of our patients survived the episode during which retinal detachments occurred. Hypertension appears to contribute to the development of retinal detachments in TTP, and vigorous efforts at blood pressure control are indicated. Serous retinal detachments may be a more frequent component of chronic relapsing TTP than has been suspected.


In vitro and in vivo inhibitory effects of Carica papaya seed on α-amylase and α-glucosidase enzymes.
The present study was aimed to investigate the in vitro and in vivo inhibitory effects of Carica papaya seeds on α-amylase and α-glucosidase enzymes, as this is known to be an antidiabetic mechanism. Analysis of the extracts of the seeds for phytochemicals revealed the presence of a significant amount of saponins, alkaloids, flavonoids, phenols, terpenoids, and steroids. The seed extracts of Carica papaya exhibited good antioxidant capacity using 2, 2-diphenyl-1-picryl hydroxyl (DPPH), thiobarbituric reactive substance (TBARS) and ferric reducing antioxidant power (FRAP) method. The results of the inhibitory studies of the extracts revealed that the hexane extract followed by the ethyl acetate extract was the most potent inhibitor of α-amylase and α-glucosidase enzyme when compared to other extracts using their IC50 values. In the animal study, different doses (250, 500 and 1000 mg/kg/body weight) of the extracts of Carica papaya seed were administered orally for 120 min, to normal and streptozotocin-induced diabetic rats, and were compared with acarbose 100 mg/kg/body weight and control group for the effect on postprandial hyperglycemia. The extract of ethyl acetate (at doses of 250, 500 and 1000 mg/kg/body weight) significantly reduced postprandial glucose levels in these animals. The characterization of hexane and ethyl acetate extracts by GC-MS analysis revealed 20 bioactive compounds while the FTIR analysis confirmed the presence of this functional groups: -C=C, -C-Cl, -C-O, -O-H, -CH, -C=O, -C=C=C, -N=C=S, -O=C=O and -N-H in Carica papaya seed extracts. It was concluded that the inhibition of α-amylase and α-glucosidase enzymes and the prevention of oxidative stress in postprandial hyperglycemia could be some of the possible mechanisms by which they exert their anti-diabetic properties.


Exploring Factors Affecting Voluntary Adoption of Electronic Medical Records Among Physicians and Clinical Assistants of Small or Solo Private General Practice Clinics.
The health care reform initiative led by the Hong Kong government's Food and Health Bureau has started the implementation of an electronic sharing platform to provide an information infrastructure that enables public hospitals and private clinics to share their electronic medical records (EMRs) for improved access to patients' health care information. However, previous attempts to convince the private clinics to adopt EMRs to document health information have faced challenges, as the EMR adoption has been voluntary. The lack of electronic data shared by private clinics carries direct impacts to the efficacy of electronic record sharing between public and private healthcare providers. To increase the likelihood of buy-in, it is essential to proactively identify the users' and organizations' needs and capabilities before large-scale implementation. As part of the reform initiative, this study examined factors affecting the adoption of EMRs in small or solo private general practice clinics, by analyzing the experiences and opinions of the physicians and clinical assistants during the pilot implementation of the technology, with the purpose to learn from it before full-scale rollout. In-depth, semistructured interviews were conducted with 23 physicians and clinical assistants from seven small or solo private general practice clinics to evaluate their experiences, expectations, and opinions regarding the deployment of EMRs. Interview transcripts were content analyzed to identify key factors. Factors affecting the adoption of EMRs to record and manage health care information were identified as follows: system interface design; system functions; stability and reliability of hardware, software, and computing networks; financial and time costs; task and outcome performance, work practice, and clinical workflow; physical space in clinics; trust in technology; users' information technology literacy; training and technical support; and social and organizational influences. The factors are interrelated with the others. The adoption factors identified are multifaceted, ranging from technological characteristics, clinician-technology interactions, skills and knowledge, and the user-workflow-technology fit. Other findings, which have been relatively underrepresented in previous studies, contribute unique insights about the influence of work and social environment on the adoption of EMRs, including limited clinic space and the effects of physicians' decision to use the technology on clinical staffs' adoption decisions. Potential strategies to address the concerns, overcome adoption barriers, and define relevant policies are discussed.


Genetic Diversity within a Global Panel of Durum Wheat (Triticum durum) Landraces and Modern Germplasm Reveals the History of Alleles Exchange.
Durum wheat is the 10th most important crop in the world, and its use traces back to the origin of agriculture. Unfortunately, in the last century only part of the genetic diversity available for this species has been captured in modern varieties through breeding. Here, the population structure and genetic diversity shared among elites and landraces collected from 32 countries was investigated. A total of 370 entries were genotyped with Axiom 35K array to identify 8,173 segregating single nucleotide polymorphisms (SNPs). Of these, 500 were selected as highly informative with a PIC value above 0.32 and used to test population structure via DAPC, STRUCTURE, and neighbor joining tree. A total of 10 sub-populations could be identified, six constituted by modern germplasm and four by landraces of different geographical origin. Interestingly, genomic comparison among groups indicated that Middle East and Ethiopia had the lowest level of allelic diversity, while breeding programs and landraces collected outside these regions were the richest in rare alleles. Further, phylogenetic analysis among landraces indicated that Ethiopia might represent a second center of origin of durum wheat, rather than a second domestication site as previously believed. Together, the analyses carried here provide a global picture of the available genetic diversity for this crop and shall guide its targeted use by breeders.


Development of a comparative genetic linkage map for Armigeres subalbatus using Aedes aegypti RFLP markers.
One of the causative agents of lympahtic filariasis is the nematode parasite Brugia malayi that requires a competent mosquito vector for its development and transmission. Armigeres subalbatus mosquitoes rapidly destroy invading B. malayi microfilariae via a defense response known as melanotic encapsulation. We have constructed a genetic linkage map for this mosquito species using RFLP markers from Aedes aegypti. This heterologous approach was possible because of the conserved nature of the coding sequences used as markers and provided an experimental framework to evaluate the hypothesis that linkage and gene order are conserved between these mosquito species. Of the 56 Ae. aegypti markers tested, 77% hybridize to genomic DNA digests of Ar. subalbatus under stringent conditions, with 53% of these demonstrating strain-specific polymorphisms. Twenty-six Ae. aegypti markers have been mapped using an F2- segregating Ar. subalbatus population derived from a cross of strains originating in Japan and Malaysia. Linear order of these marker loci is highly conserved between the two species. Only 1 of these markers, LF92, was not linked in the manner predicted by the Ae. aegypti map. In addition, the autosomal sex-determination locus that occurs in linkage group 1 in Ae. aegypti resides in group 3 in Ar. subalbatus. The Ar. subalbatus map provides a basic genetic context that can be utilized in further genetic studies to clarify the genetic basis of parasite resistance in this mosquito and is a necessary precursor to the identification of genome regions that carry genes that determine the encapsulation phenotype. [The composite map and sequence database information for Ae. aegypti markers can be retrieved directly from the Ae. aegypti Genome Database through the World Wide Web: http://klab.agsci.colostate.edu.]


Observations of ground-level ozone and NO2 in northernmost Sweden, including the Scandian Mountain Range.
Ozone was measured using passive diffusion samplers at alpine Latnjajaure (980 m above sea level [asl]) in the northern Scandian Mountain Range during spring and summer 2006-2008, and year-round at three further sites in northernmost Sweden 2004-2008. These observations were compared with ozone concentrations from three permanent monitoring stations using ultraviolet absorption instruments. Ozone concentrations at Latnjajaure were higher than at the closest monitoring site, illustrating the importance of high elevation for ozone. At the northern sites the ozone spring peak was more pronounced, higher, and earlier (April maximum) compared to a site in south Sweden (May maximum). During summer, ozone concentrations were higher in south Sweden. Presently, the growing season largely starts after the ozone spring peak in north Sweden but is likely to start earlier in the future climate. This could lead to an increased risk for ozone effects on vegetation if the current yearly ozone cycle persists.


Deficiency of α-actinin-3 is associated with increased susceptibility to contraction-induced damage and skeletal muscle remodeling.
Sarcomeric α-actinins (α-actinin-2 and -3) are a major component of the Z-disk in skeletal muscle, where they crosslink actin and other structural proteins to maintain an ordered myofibrillar array. Homozygosity for the common null polymorphism (R577X) in ACTN3 results in the absence of fast fiber-specific α-actinin-3 in ∼20% of the general population. α-Actinin-3 deficiency is associated with decreased force generation and is detrimental to sprint and power performance in elite athletes, suggesting that α-actinin-3 is necessary for optimal forceful repetitive muscle contractions. Since Z-disks are the structures most vulnerable to eccentric damage, we sought to examine the effects of α-actinin-3 deficiency on sarcomeric integrity. Actn3 knockout mouse muscle showed significantly increased force deficits following eccentric contraction at 30% stretch, suggesting that α-actinin-3 deficiency results in an increased susceptibility to muscle damage at the extremes of muscle performance. Microarray analyses demonstrated an increase in muscle remodeling genes, which we confirmed at the protein level. The loss of α-actinin-3 and up-regulation of α-actinin-2 resulted in no significant changes to the total pool of sarcomeric α-actinins, suggesting that alterations in fast fiber Z-disk properties may be related to differences in functional protein interactions between α-actinin-2 and α-actinin-3. In support of this, we demonstrated that the Z-disk proteins, ZASP, titin and vinculin preferentially bind to α-actinin-2. Thus, the loss of α-actinin-3 changes the overall protein composition of fast fiber Z-disks and alters their elastic properties, providing a mechanistic explanation for the loss of force generation and increased susceptibility to eccentric damage in α-actinin-3-deficient individuals.


A double-monoclonal immunoenzymometric assay for alpha 1-fetoprotein modified for increased analytical precision.
We modified a one-step, two-site, double monoclonal immunoenzymometric assay (Abbott Laboratories) for serum alpha1-fetoprotein (AFP) to increase its sensitivity and improve test precision at the low end. We increased sample size, incubation interval, and reaction time and temperature, and decreased the final reaction volume. Interassay CVs for the modified method ranged from 6.2 to 8.0% for mean concentrations of AFP in serum of 5.2 to 34.2 micrograms/L--substantially better than those for the unmodified monoclonal method--and agreed well with those of the comparison method (modified Abbott polyclonal immunoenzymometric assay). AFP values by the modified monoclonal procedure (y) correlated well with results by the polyclonal method (x): y = 0.983x + 1.84 micrograms/L (r = 0.927, n = 59). The detection limit of the modified monoclonal test was 0.2 microgram/L, as compared with 1.0 and 1.4 micrograms/L, respectively, for the modified polyclonal and the unmodified monoclonal procedures. We recommend using the modified monoclonal method for monitoring cancer patients with low tumor burden.


Genome-Wide Fitness and Genetic Interactions Determined by Tn-seq, a High-Throughput Massively Parallel Sequencing Method for Microorganisms.
The lagging annotation of bacterial genomes and the inherent genetic complexity of many phenotypes is hindering the discovery of new drug targets and the development of new antimicrobial agents and vaccines. This unit presents Tn-seq, a method that has made it possible to quantitatively determine fitness for most genes in a microorganism and to screen for quantitative genetic interactions on a genome-wide scale and in a high-throughput fashion. Tn-seq can thus direct studies on the annotation of genes and untangle complex phenotypes. The method is based on the construction of a saturated transposon insertion library. After library selection, changes in the frequency of each insertion mutant are determined by sequencing flanking regions en masse. These changes are used to calculate each mutant's fitness. The method was originally developed for the Gram-positive bacterium Streptococcus pneumoniae, a causative agent of pneumonia and meningitis, but has now been applied to several different microbial species.


[Determination of azathiopurine concentration in human serum by second derivative spectra method].
A sensitive and rapid method for the determination of azathiopurine concentration in human serum was established. By using ultraviolet second derivative spectra, the method eliminated background interference of biological sample and coexisting drugs. The serum sample was extracted with ethyl acetate, the extract was evaporated to dryness with a gentle N2 stream, and 0.25 ml of methanol was added to dissolve the residue. Quantitative calculation was made with analytical wavelength of 274 nm and 293 nm, and peak- to- valley method. The minimum detectable concentration of AZP in serum was 25 ng.ml-1 and average recovery was 99.3% +/- 5.9%.


Economically motivated relationships and transactional sex among unmarried African American and white women: results from a U.S. national telephone survey.
We explored links among economically motivated relationships, transactional sex, and risk behavior for human immunodeficiency virus (HIV) and sexually transmitted infections (STIs) among unmarried African American and white women. We drew on data from 1,371 unmarried African American and white women aged 20 to 45 years that we collected via a random-digit-dial telephone survey in the U.S. Of all respondents, 33.3% (95% confidence interval 28.8, 38.0) reported staying in a relationship longer than they wanted to because of economic considerations. African American women were more likely than white women to report starting a relationship due to economic considerations (21.6% vs. 10.5%) and having transactional sex with someone who was not a regular partner (13.1% vs. 2.9%). These behaviors were all associated with lack of education, experience of economic hardship, need to care for dependents, and increased levels of HIV/STI risk. All three behaviors were associated with having more sexual partners. Staying in a sexual relationship because of economic considerations was also associated with anal sex, reduced condom use, and concurrent sexual partnerships. Transactional sex with non-regular partners was associated with concurrent sexual partnerships, binge drinking, drug use, perceived concurrency by main partner, and having high-risk sexual partners. HIV/STI risk-reduction policies and programs in the U.S. need to explicitly address overall economic disempowerment among women, as well as racial disparities in poverty. These economic disparities likely contribute both to increasing rates of HIV among women in the U.S. and to the extraordinary racial disparities in HIV/STI risk among American women.


Towards a definition of "difference" in osteoarthritis.
To assess existing information regarding detectable differences in osteoarthritis (OA), a systematic literature search was conducted up to December 1999. Thirty-three articles were considered methodologically relevant to the definition and categorization of detectable differences in OA. It was determined that the musculoskeletal literature contains a wealth of information that relates to observed changes, much of which is derived from the clinical trials literature, but there have been relatively few methodological studies that have systematically evaluated the nature, categorization, and relevance of the change. Furthermore, most of those that have been published take the perspective of an individual or groups of experts other than that of the patient. This summary of the current literature reveals that the diverse sources of information go part way towards developing an understanding of detectable differences and their importance in the area of OA research and clinical practice. Stakeholders' interests as well as factors that modulate perceptions of importance need to be taken under consideration. In particular, the patient's perspective of the importance of change at an individual level requires further evaluation. This area of clinical research is relatively underdeveloped, but there is considerable opportunity for progress.


Recognition of spoken words by native and non-native listeners: talker-, listener-, and item-related factors.
In order to gain insight into the interplay between the talker-, listener-, and item-related factors that influence speech perception, a large multi-talker database of digitally recorded spoken words was developed, and was then submitted to intelligibility tests with multiple listeners. Ten talkers produced two lists of words at three speaking rates. One list contained lexically "easy" words (words with few phonetically similar sounding "neighbors" with which they could be confused), and the other list contained lexically "hard" words (words with many phonetically similar sounding "neighbors"). An analysis of the intelligibility data obtained with native speakers of English (experiment 1) showed a strong effect of lexical similarity. Easy words had higher intelligibility scores than hard words. A strong effect of speaking rate was also found whereby slow and medium rate words had higher intelligibility scores than fast rate words. Finally, a relationship was also observed between the various stimulus factors whereby the perceptual difficulties imposed by one factor, such as a hard word spoken at a fast rate, could be overcome by the advantage gained through the listener's experience and familiarity with the speech of a particular talker. In experiment 2, the investigation was extended to another listener population, namely, non-native listeners. Results showed that the ability to take advantage of surface phonetic information, such as a consistent talker across items, is a perceptual skill that transfers easily from first to second language perception. However, non-native listeners had particular difficulty with lexically hard words even when familiarity with the items was controlled, suggesting that non-native word recognition may be compromised when fine phonetic discrimination at the segmental level is required. Taken together, the results of this study provide insight into the signal-dependent and signal-independent factors that influence spoken language processing in native and non-native listeners.


Bromocriptine and dopamine mediate independent and synergistic apoptotic pathways in pituitary cells.
Dopamine (DA) agonists are the primary treatment choice for prolactinoma, effectively suppressing prolactin expression and reducing tumour size. However, the intracellular pathway(s) through which either DA or its agonists impact on proliferation or lead to tumour shrinkage are incompletely understood. To identify the mediators in the apoptotic cascades after DA or DA agonist challenges we used a well-characterized model system, the rodent somatolactotroph cell line GH3. In these cells, we show that apoptosis induced by the DA agonist bromocriptine (BC), but not DA, is initiated through activation of the JNK pathway. However, both DA and BC activate the terminal effector caspase, caspase-3. Kinetic studies and chemical inhibitor co-incubation experiments support a role for JNK activation preceding caspase-9 activation in BC challenged cells, however, engagement of these mediators was not apparent in DA challenge cells. These studies suggest that apoptosis induced by BC or DA is mediated through distinct and independent pathways that converge with activation of the terminal caspase, caspase-3. These observations were further reinforced by our findings that DA and BC, in co-incubation experiments, synergistically induce apoptosis. These findings raise the possibility that drugs acting through the same pathway as DA may be clinically beneficial when combined with BC.


Frontier orbitals of photosubstitutionally active ruthenium complexes: an experimental study of the spectator ligands' electronic properties influence on photoreactivity.
The synthesis and characterization of [Ru(tpy)(R2bpy)(L)](X)n complexes (tpy = 2,2':6',2''-terpyridine, R2bpy = 4,4'-dimethyl-2,2'-bipyridine (dmbpy), or 4,4'-bis(trifluoromethyl)-2,2'-bipyridine (tfmbpy), X = Cl- or PF6-, and n = 1 or 2) are described. The dmbpy and tfmbpy bidentate ligands allow for investigating the effects of electron-donating and electron-withdrawing ligands, respectively, on the frontier orbital energetics as well as the photoreactivity of these ruthenium polypyridyl complexes for five prototypical monodentate ligands L = Cl-, H2O, CH3CN, 2-(methylthio)ethanol (Hmte), or pyridine. According to spectroscopic and electrochemical studies, the dmbpy analogues displayed a singlet metal-to-ligand charge transfer (1MLCT) transition at higher energy than the tfmbpy analogues. The shift of the 1MLCT to higher energy results from the lowest unoccupied molecular orbital (LUMO) for the dmbpy analogues being tpy-based, whereas for the tfmbpy analogues orbital inversion occurs resulting in a tfmbpy-based LUMO. The energy level of the highest occupied molecular orbital (HOMO) was considerably affected by the nature of the monodentate ligand. Visible light irradiation of the complexes demonstrated that the tfmbpy analogue increased the rate and quantum yields of photosubstitution reactions, compared to the dmbpy analogue, suggesting that the electron-withdrawing substituents allowed better thermal accessibility of the triplet metal-centered (3MC) state from the photochemically generated triplet metal-to-ligand charge transfer (3MLCT) excited state. A correlation between the photolability of the monodentate ligands and the electrochemical reversibility of the metal-based oxidation is also reported.


Small scale membrane mechanics.
Large scale changes to lipid bilayer shapes are well represented by the Helfrich model. However, there are membrane processes that take place at smaller length scales that this model cannot address. In this work, we present a one-dimensional continuum model that captures the mechanics of the lipid bilayer membrane at the length scale of the lipids themselves. The model is developed using the Cosserat theory of surfaces with lipid orientation, or 'tilt', as the fundamental degree of freedom. The Helfrich model can be recovered as a special case when the curvatures are small and the lipid tilt is everywhere zero. We use the tilt model to study local membrane deformations in response to a protein inclusion. Parameter estimates and boundary conditions are obtained from a coarse-grained molecular model using dissipative particle dynamics (DPD) to capture the same phenomenon. The continuum model is able to reproduce the membrane bending, stretch and lipid tilt as seen in the DPD model. The lipid tilt angle relaxes to the bulk tilt angle within 5-6 nm from the protein inclusion. Importantly, for large tilt gradients induced by the proteins, the tilt energy contribution is larger than the bending energy contribution. Thus, the continuum model of tilt accurately captures behaviors at length scales shorter than the membrane thickness.


Variation in human performance in the hypoxic mountain environment.
Ascent to altitude is associated with a fall in barometric pressure, and with it a decline in the partial pressure of atmospheric (and thus alveolar) oxygen. As a result, a variety of adaptive physiological processes are engaged to mitigate the fall in tissue convective oxygen delivery which might otherwise occur. The magnitude and nature of such changes is also modified with time, a process known as acclimatization. However, other phenomena are at work; the ability to perform physical work at altitude falls in a manner which is not wholly related to changes in arterial oxygen content. Indeed, alterations in local skeletal muscle blood flow and metabolism may play an axial role. Thus, for those who are not native to high altitude, the ability to compete at altitude is likely to be impaired. The magnitude of such impairment in performance, however, differs greatly between individuals, and it seems that genetic variation underpins much of this difference. The identification of the relevant genetic elements is in its infancy in humans, but ongoing work is likely to help us gain an increasing understanding of how humans adapt to altitude and to develop mitigating interventions.


Minimally invasive repair of pectus excavatum: a novel morphology-tailored, patient-specific approach.
Minimally invasive repair of pectus excavatum, introduced by Nuss in 1998, has undergone a serious learning curve because of a lack of understanding on morphologies and repair techniques. To summarize the current status of minimally invasive repair of pectus excavatum, we reviewed and appraised our 10-year experience with a novel approach, a morphology-tailored technique, including diverse bar shaping, bar fixation, and techniques for adults. We analyzed the data of 1170 consecutive patients with pectus excavatum who underwent minimally invasive repair between August 1999 and September 2008. All pectus repairs were performed by the primary author (H.J.P.) with our modified technique. The mean age was 10.3 years (range, 16 months to 51 years). There were 331 adult patients (>15 years) (28.3%). A total of 576 patients (49.2%) had bar removal after a mean of 2.5 years (range, 10 days to 7 years). The asymmetry index change (1.10-1.02, P < .001) demonstrated post-repair symmetry. Complication rates decreased through the 3 time periods (1999-2002 [n = 335]; 2003-2005 [n = 441]; 2006-2008 [n = 394]) as follows: pneumothorax rate (7.5% vs 4.3% vs 0.8%; P < .001) and bar displacement rate (3.8% vs 2.3% vs 0.5%; P = .002). Reoperation rate also decreased (4.8% vs 2.5% vs 0.8%; P = .002). Satisfaction outcomes were excellent in 92.7%, good in 5.9%, and fair in 1.4% of patients. After bar removal, 3 patients (0.6%) had minor recurrences. Minimally invasive repair of pectus excavatum based on a novel morphology-tailored, patient-specific approach is effective for quality repair of the full spectrum of pectus excavatum, including asymmetry and adult patients. Continuous technical refinements have significantly decreased the complication rates and postoperative morbidity.


Effect of polymer concentration on camouflaging of pancreatic islets with mPEG-succinimidyl carbonate.
Modifying the surface of living cells with specially selected polymers is a new method of protecting them from the immune system. Covalent attachment of polyethylene glycol (PEG) derivatives on the capsule of pancreatic islets in aqueous media under moderate conditions is an example. This study aimed to find out whether higher polymer concentration could protect PEGylated islets from immune systems more efficiently without affecting their viability and morphology. Methoxy polyethylene glycol succinimidyl carbonate (mPEG-SC) with molecular weight of 5 kDa at different concentrations was grafted onto islets capsules. It was found that at polymer concentrations higher than 22 mg/mL, islets lost their viability with changes in their surface structure as compared to free islets. At mPEG-SC concentrations less than 22 mg/mL, there was no change in morphology and viability of PEGylated islets, but PEGylation of islets at polymer concentration 7 mg/mL was not sufficient for their camouflaging. Polymer concentration of 22 mg/mL was more effective in weakening the immunogenicity of islets. It was concluded that increasing the 5kDa mPEG-SC concentration up to 22 mg/ml protected islets from the immune cells more efficiently without affecting their viability and functionality.


[Furin and its biological role].
The survey deals with structure, properties and biological role of furin, the calcium-dependent serine endoprotease, which is expressed in all tissues and cell lines examined. It is the best-characterized representative of the mammalian subtilisin-like family of proprotein convertases, which is capable to cleave precursors of a wide variety of proteins, including hormones, growth factors, serum proteins, proteases of the blood-clotting system, matrix metalloproteinases, receptors, viral envelope glycoproteins, and bacterial exotoxins, and so on. The enzyme plays the essential role in embryogenesis, homeostasis; it is also involved in tumor metastasis, activation and virulence of many bacterial and viral pathogens and in neurodegenerative processes associated with Alzheimer's disease. Furin is a very specific enzyme: it recognizes the cleavage-site sequence Arg-Xaa-Lys/Arg-Arg and catalyzes the hydrolysis of the precursors, containing a pair of basic amino acids Arg-Arg or Lys-Arg. The enzyme is a multidomain protein which is initially synthesized as 100 kDa glycosylated profurin consisting of 794 amino acid residues (for human furin) and including a N-terminal signal peptide, propeptide, catalytic domain, possessing approximately 30% homology to subtilisin, a well-conserved P-domain, Cys-rich domain, transmembrane domain and cytoplasmic domain. The active site cleft of furin differs considerably from that of subtilisin with respect to the depth, shape and molecule charge. In furin it is a canyon-like groove with clusters of negatively charged residues along it. Furin inhibitors are rather promising therapeutic agents for treating furin-mediated diseases and bacterial infections. Most furin inhibitors belong to proteins or peptides. The protein-based inhibitors include both naturally occurring and bioengineered variants of protease inhibitors. The peptide-based inhibitors are represented by polyarginines, peptidyl chloromethyl ketones, aminomethyl ketones or ketomethylene pseudopeptides, isostere-containing cyclic peptides, short peptides derived from the prosegments of furin or al-PDX-related peptides. The nonpeptide inhibitors are natural products of the andrographolide family, certain metal complexes of pyridine derivatives and small molecules derived from 2.5-dideoxystreptamine. The furin inhibitors may be used not only as valuable tools for studying furin action, but also as therapeutic agents for furin-dependent diseases.


Chemoprophylaxis is effective in the prevention of leprosy in endemic countries: a systematic review and meta-analysis. MILEP2 Study Group. Mucosal Immunology of Leprosy.
To quantify the efficacy of chemoprophylaxis against leprosy. Literature searching of Medline and Embase databases, hand-searching of references and correspondence with investigators. published papers relating to the prevention of leprosy and the use of chemotherapy in leprosy were identified for critical appraisal. Trials were selected and grouped into three categories according to study design and control groups. the relative risks (RR) with 95% confidence intervals were calculated from the original data using a random effects model. To assess the cost-effectiveness of chemoprophylaxis, a further analysis of the rates of disease in the trial and control groups was done based on the numbers needed to be treated (NNT) to prevent one new case of leprosy. A total of 14 trials were identified from 127 published papers on chemoprophylaxis of leprosy. The trials were categorized into randomized controlled trials, non-randomized controlled trials, and uncontrolled trials. The overall results of the meta-analysis shows that chemoprophylaxis gives around 60% protection against leprosy. The NNT are low in trials of household contacts. The evidence shows that chemoprophylaxis against leprosy is an effective way to reduce the incidence of leprosy, particularly in household contacts. The role of chemoprophylaxis needs to be re-examined using newer drugs given the continuing case detection rates globally.


Cardiopulmonary and acid-base effects of desflurane and sevoflurane in spontaneously breathing cats.
The cardiopulmonary effects of desflurane and sevoflurane anesthesia were compared in cats breathing spontaneously. Heart (HR) and respiratory (RR) rates; systolic (SAP), diastolic (DAP) and mean arterial (MAP) pressures; partial pressure of end tidal carbon dioxide (PETCO2), arterial blood pH (pH), arterial partial pressure of oxygen (PaO2) and carbon dioxide (PaCO2); base deficit (BD), arterial oxygen saturation (SaO2) and bicarbonate ion concentration (HCO3) were measured. Anesthesia was induced with propofol (8+/-2.3mg/kg IV) and maintained with desflurane (GD) or sevoflurane (GS), both at 1.3 MAC. Data were analyzed by analysis of variance (ANOVA), followed by the Tukey test (P<0.05). Both anesthetics showed similar effects. HR and RR decreased when compared to the basal values, but remained constant during inhalant anesthesia and PETCO2 increased with time. Both anesthetics caused acidemia and hypercapnia, but BD stayed within normal limits. Therefore, despite reducing HR and SAP (GD) when compared to the basal values, desflurane and sevoflurane provide good stability of the cardiovascular parameters during a short period of inhalant anesthesia (T20-T60). However, both volatile anesthetics cause acute respiratory acidosis in cats breathing spontaneously.


Musculoskeletal examination--an ignored aspect. Why are we still failing the patients?
Over the past two decades, rheumatologists from around the world have not only championed the musculoskeletal system examination but also modified the undergraduate teaching curriculum. This has led to the development and adoption of the gait, arms, legs and spine (GALS) screening along with regional examination techniques. The purpose of this study is to review current practice, determining the frequency of patient exposure to appropriate examination and confidence of junior doctors when dealing with MSK conditions. Two district-general hospitals (non-teaching) and one teaching hospital in North-East London were chosen. At each site, 50 patient notes were reviewed from the acute admission wards for medicine and surgery and the medical assessment unit. Factors considered included whether GALS screenings had taken place, documentation of MSK examinations and assessment of confidence of junior doctors in assessing MSK conditions. GALS screenings were performed for 4% of patients on the medical assessment unit, 7% of acute medical and 0% of acute surgical patients on admission. Examination of the MSK system yielded better results with 16%, 22% and 10% on each of the respective wards. Interviews with junior doctors found 10% routinely screening for MSK conditions, despite 87% feeling confident in taking MSK histories. This prospective audit of clinical practice highlights that patients failed to have a minimal assessment of the MSK system through GALS screenings. When examining the MSK system, results were somewhat better, although still fewer than expected. It is curious that the majority of junior doctors in training felt confident in dealing with MSK disease but few did it in practice. This begs the question of whether current teaching curricula and strategies are adequate. At a time where there is ever-increasing national momentum to address issues on obesity and cardiovascular health, our patients are still deprived of a standard MSK examination by the medical faculty.


Study on the optimization of the deposition rate of planetary GaN-MOCVD films based on CFD simulation and the corresponding surface model.
Metal-organic chemical vapour deposition (MOCVD) is a key technique for fabricating GaN thin film structures for light-emitting and semiconductor laser diodes. Film uniformity is an important index to measure equipment performance and chip processes. This paper introduces a method to improve the quality of thin films by optimizing the rotation speed of different substrates of a model consisting of a planetary with seven 6-inch wafers for the planetary GaN-MOCVD. A numerical solution to the transient state at low pressure is obtained using computational fluid dynamics. To evaluate the role of the different zone speeds on the growth uniformity, single factor analysis is introduced. The results show that the growth rate and uniformity are strongly related to the rotational speed. Next, a response surface model was constructed by using the variables and the corresponding simulation results. The optimized combination of the matching of different speeds is also proposed as a useful reference for applications in industry, obtained by a response surface model and genetic algorithm with a balance between the growth rate and the growth uniformity. This method can save time, and the optimization can obtain the most uniform and highest thin film quality.


Pediatric Intensive Care Unit related Sleep and Circadian Dysregulation: a focused review.
The pediatric intensive care unit (PICU) is bright, loud, and disruptive to children. Strategies to improve the sleep of adults in the ICU have improved delirium and mortality rates. Children need more sleep than adults for active growth, healing, and development when well; this is likely true when they are critically ill. This review was performed to describe what we know in this area to date with the intent to identify future directions for research in this field. Since the 1990s, 16 articles on 14 observational trials have been published investigating the sleep on a total of 312 critically ill children and the melatonin levels of an additional 144. Sleep measurements occurred in 9 studies through bedside observation (n = 2), actigraphy (n = 2), electroencephalogram (n = 1) and polysomnography (n = 4), of which polysomnography is the most reliable. Children in the PICU sleep more during the day, have fragmented sleep and disturbed sleep architecture. Melatonin levels may be elevated and peak later in critically ill children. Early data suggest there are at-risk subgroups for sleep and circadian disruption in the PICU including those with sepsis, burns, traumatic brain injury and after cardiothoracic surgery. The available literature describing the sleep of critically ill children is limited to small single-center observational studies with varying measurements of sleep and inconsistent findings. Future studies should use validated measurements and standardized definitions to begin to harmonize this area of medicine to build toward pragmatic interventional trials that may shift the paradigm of care in the pediatric intensive care unit.


Snake trajectories in ultraclean graphene p-n junctions.
Snake states are trajectories of charge carriers curving back and forth along an interface. There are two types of snake states, formed by either inverting the magnetic field direction or the charge carrier type at an interface. The former has been demonstrated in GaAs-AlGaAs heterostructures, whereas the latter has become conceivable only with the advance of ballistic graphene where a gap-less p-n interface governed by Klein tunnelling can be formed. Such snake states were hidden in previous experiments due to limited sample quality. Here we report on magneto-conductance oscillations due to snake states in a ballistic suspended graphene p-n junction, which occur already at a very small magnetic field of 20 mT. The visibility of 30% is enabled by Klein collimation. Our finding is firmly supported by quantum transport simulations. We demonstrate the high tunability of the device and operate it in different magnetic field regimes.


Linkage of familial dilated cardiomyopathy with conduction defect and muscular dystrophy to chromosome 6q23.
Inherited cardiomyopathies may arise from mutations in genes that are normally expressed in both heart and skeletal muscle and therefore may be accompanied by skeletal muscle weakness. Phenotypically, patients with familial dilated cardiomyopathy (FDC) show enlargement of all four chambers of the heart and develop symptoms of congestive heart failure. Inherited cardiomyopathies may also be accompanied by cardiac conduction-system defects that affect the atrioventricular node, resulting in bradycardia. Several different chromosomal regions have been linked with the development of autosomal dominant FDC, but the gene defects in these disorders remain unknown. We now characterize an autosomal dominant disorder involving dilated cardiomyopathy, cardiac conduction-system disease, and adult-onset limb-girdle muscular dystrophy (FDC, conduction disease, and myopathy [FDC-CDM]). Genetic linkage was used to exclude regions of the genome known to be linked to dilated cardiomyopathy and muscular dystrophy phenotypes and to confirm genetic heterogeneity of these disorders. A genomewide scan identified a region on the long arm of chromosome 6 that is significantly associated with the presence of myopathy (D6S262; maximum LOD score [Z(max)] 4.99 at maximum recombination fraction [theta(max)] .00), identifying FDC-CDM as a genetically distinct disease. Haplotype analysis refined the interval containing the genetic defect, to a 3-cM interval between D6S1705 and D6S1656. This haplotype analysis excludes a number of striated muscle-expressed genes present in this region, including laminin alpha2, laminin alpha4, triadin, and phospholamban.


High Aspect Ratio Nanoscale Pores through BCP-Based Metal Oxide Masks and Advanced Dry Etching.
The reliable and regular modification of the surface properties of substrates plays a crucial role in material research and the development of functional surfaces. A key aspect of this is the development of the surface pores and topographies. These can confer specific advantages such as high surface area as well as specific functions such as hydrophobic properties. Here, we introduce a combination of nanoscale self-assembled block-copolymer-based metal oxide masks with optimized deep reactive ion etching (DRIE) of silicon to permit the fabrication of porous topographies with aspect ratios of up to 50. Following the evaluation of our procedure and involved parameters using various techniques, such as AFM or SEM, the suitability of our features for applications relying on high light absorption as well as efficient thermal management is explored and discussed in further detail.


The southern megalopolis: using the past to predict the future of urban sprawl in the Southeast U.S.
The future health of ecosystems is arguably as dependent on urban sprawl as it is on human-caused climatic warming. Urban sprawl strongly impacts the urban ecosystems it creates and the natural and agro-ecosystems that it displaces and fragments. Here, we project urban sprawl changes for the next 50 years for the fast-growing Southeast U.S. Previous studies have focused on modeling population density, but the urban extent is arguably as important as population density per se in terms of its ecological and conservation impacts. We develop simulations using the SLEUTH urban growth model that complement population-driven models but focus on spatial pattern and extent. To better capture the reach of low-density suburban development, we extend the capabilities of SLEUTH by incorporating street-network information. Our simulations point to a future in which the extent of urbanization in the Southeast is projected to increase by 101% to 192%. Our results highlight areas where ecosystem fragmentation is likely, and serve as a benchmark to explore the challenging tradeoffs between ecosystem health, economic growth and cultural desires.