Changes in air quality and tropospheric composition due to depletion of stratospheric ozone and interactions with changing climate: implications for human and environmental health.
UV radiation is an essential driver for the formation of photochemical smog, which includes ground-level ozone and particulate matter (PM). Recent analyses support earlier work showing that poor outdoor air quality is a major environmental hazard as well as quantifying health effects on regional and global scales more accurately. Greater exposure to these pollutants has been linked to increased risks of cardiovascular and respiratory diseases in humans and is associated globally with several million premature deaths per year. Ozone also has adverse effects on yields of crops, leading to loss of billions of US dollars each year. These detrimental effects also may alter biological diversity and affect the function of natural ecosystems. Future air quality will depend mostly on changes in emission of pollutants and their precursors, but changes in UV radiation and climate will contribute as well. Significant reductions in emissions, mainly from the energy and transportation sectors, have already led to improved air quality in many locations. Air quality will continue to improve in those cities/states that can afford controls, and worsen where the regulatory infrastructure is not available. Future changes in UV radiation and climate will alter the rates of formation of ground-level ozone and photochemically-generated particulate matter and must be considered in predictions of air quality. The decrease in UV radiation associated with recovery of stratospheric ozone will, according to recent global atmospheric model simulations, lead to increases in ground-level ozone at most locations. If correct, this will add significantly to future ground-level ozone trends. However, the spatial resolution of these global models is insufficient to inform policy at this time, especially for urban areas. UV radiation affects the atmospheric concentration of hydroxyl radicals, ˙OH, which are responsible for the self-cleaning of the atmosphere. Recent measurements confirm that, on a local scale, ˙OH radicals respond rapidly to changes in UV radiation. However, on large (global) scales, models differ in their predictions by nearly a factor of two, with consequent uncertainties for estimating the atmospheric lifetime and concentrations of key greenhouse gases and air pollutants. Projections of future climate need to consider these uncertainties. No new negative environmental effects of substitutes for ozone depleting substances or their breakdown-products have been identified. However, some substitutes for the ozone depleting substances will continue to contribute to global climate change if concentrations rise above current levels.


Determination of Plasma Concentration Reference Ranges for Risperidone and Paliperidone.
Schizophrenia is a common disease managed by a range of interventions, with the primary treatment being antipsychotic medications (APS). Inadequate response, lack of adherence, and/or adverse events often prevent optimal therapeutic effects or therapeutic efficiency. Monitoring APS plasma concentrations can be used together with a full clinical evaluation to help improve patient care or offer better treatment options for the patient. To enable interpretation of individual risperidone and paliperidone plasma concentrations, we developed "reference ranges," which consider the expected variability in plasma concentrations between subjects across the population, rather than representing a "therapeutic range" that relates to efficacy and/or safety outcomes. The reference ranges were derived from population pharmacokinetic models, which varied based upon administration route, dose, and time after dose. Good agreement between the proposed reference ranges and external data was obtained through graphical and numerical evaluations, indicating they could be reliably used in clinical practice.


Low subcutaneous adiposity associates with higher mortality in female patients with cirrhosis.
Two major body compartments, skeletal muscle and adipose tissue, exhibit independent functions. We aimed to explore the prognostic significance of skeletal muscle, visceral and subcutaneous adipose tissue, according to sex, in patients with cirrhosis assessed for liver transplantation (LT). CT images taken at the 3rd lumbar vertebra from 677 patients were quantified for three body composition indexes (cm2/m2), visceral adipose tissue index, subcutaneous adipose tissue index (SATI), and skeletal muscle index (SMI). Cox proportional and competing-risk analysis hazard models were conducted to assess associations between mortality and body composition. The majority of patients were male (67%) with a mean age of 57 ± 7 years, model for end-stage liver disease (MELD) score of 14 ± 8 and mean body mass index of 27 ± 6 kg/m2. Despite similar body mass index between the sexes, male patients had greater SMI (53 ± 12 vs. 45 ± 9 cm2/m2), whereas SATI (67 ± 52 vs. 48 ± 37 cm2/m2) was higher in females (p <0.001 for each). In sex stratified multivariate analyses after adjustment for MELD score and other confounding variables, SATI in females (hazard ratio [HR] 0.99; 95% CI 0.98-1.00; p = 0.01) and SMI in males (HR 0.98; 95% CI 0.96-1.00; p = 0.02) were significant predictors of mortality. Female patients with low SATI (<60 cm2/m2) had a higher risk of mortality (HR 2.06; 95% CI 1.08-3.91; p = 0.03). Using competitive risk analysis in female patients listed for LT, low SATI was also an independent predictor of mortality (subdistribution HR 2.80; 95% CI 1.28-6.12; p = 0.01) after adjusting for MELD, and other confounding factors. A lower SATI is associated with higher mortality in female patients with cirrhosis. Subcutaneous adipose tissue has a favorable metabolic profile - low SATI may reflect depletion of this major energy reservoir, leading to poor clinical outcomes. We looked at the importance of two of the main body compartments, skeletal muscle and adipose tissue (fat) on the prognosis of males and females with end-stage liver disease. Lower amounts of subcutaneous fat but not visceral fat (around internal organs), are associated with higher mortality in female patients with end-stage liver disease. However, low skeletal muscle predicts mortality in male patients with end-stage liver disease.


Early periprosthetic metastasis following total hip replacement in a patient with breast carcinoma: a case report and review of literature.
Early periprosthetic osteolysis following total hip replacement (THR) as a result of septic etiology has been well understood. Periprosthetic bone loss as a result of metastatic infiltration is an uncommon and infrequent cause of early, progressive loosening of joint replacement prosthesis. Proximal femur has been the most common site of involvement compared to acetabular prosthesis. The rarity of this clinical entity can lead to delay in definitive diagnosis and management, thus affecting the final outcome. Breast is the commonest site of carcinoma in female patients despite which not many cases of periprosthetic metastasis have been reported in the literature. We present the first case of extensive, isolated periacetabular bone destruction following a THR in a 59 years old female patient with a history of breast carcinoma. Patients with known primary malignancy should be screened thoroughly before operation and should be followed regularly after joint replacement surgery to detect any metastatic foci around the prosthesis. Periprosthetic metastasis; Total hip replacement; Breast carcinoma.


First Report of Tuber Rot Disease of Kala Zeera Caused by a Member of the Fusarium solani Species Complex in India.
Bunium persicum (Kala zeera, also black cumin) is an economically important culinary crop that is cultivated for its seed pods and its tuberlike roots. In India, high-altitude regions of Himachal Pradesh, including the Padder valley and the Gurez area of Jammu and Kashmir, are areas of kalazeera production (3). In 2008 to 2009, tuber rot disease of kala zeera was observed during the late spring season in the Padder valley. Symptomatic plants were distributed in localized areas in the field and the symptoms included drying of foliage and rotting of tubers. White mycelia were found on the tubers at the late stages of disease development. Incidence of infection in the surveyed area was 80 to 90%. Yield losses were 50 to 60%. To isolate the causal pathogen, we cultured tissues from symptomatic tubers. Small bits of the infected tissue were surface disinfested in 0.1% mercuric chloride, followed by rinsing three times in sterile distilled water. The surface disinfested tissues were plated on potato dextrose agar (PDA) and incubated at 27°C for 4 days. Pure cultures of the mycelium from the diseased tissues were transferred to a second set of PDA for species identification. The fungus produced three types of spores: small, one-celled, oval microconidia; large, slightly curved, septate macroconidia; and rounded, thick-walled chlamydospores. Microconidia were mostly non-septate and 8.91 to 15.73 × 2.3 to 3.5 μm, whereas macroconidia were three- to five-septate and were 35.55 to 54.74 × 3.91 to 6.5 μm. On the basis of morphological characteristics (1), the fungus was identified and deposited as a member of the Fusarium solani species complex in the Indian Type Culture Collection, New Delhi (ID No. 8422.11). To confirm pathogenicity, healthy tubers were submerged for 20 min in a conidial suspension of the isolated fungus (1 × 105 cfu/ml), which was prepared in potato dextrose broth, incubated for 10 days at 27°C, and centrifuged at 140 rpm. Noninoculated controls were submerged in distilled water. Inoculated and control tubers were then planted in separate pots filled with sterilized soil and kept in a shade house. Symptoms appeared on inoculated tubers 9 to 10 days after planting. Signs of the pathogen in the form of mycelia were present. The tubers rotted and died 12 to 15 days after inoculation. Control tubers did not display any symptoms. F. solani species complex was reisolated from inoculated tubers, fulfilling Koch's postulates. F. solani has been reported to cause corm rot on gladiolus and saffron (2). To our knowledge, this is the first report of the F. solani species complex as pathogenic to tubers of kalazeera in India. References: (1) C. Booth. The Genus Fusarium. 47, 1971. (2) L. Z. Chen et al. J. Shanghai Agric. College 12:240, 1994. (3) K. S. Panwar et al. Agriculture Situation in India. 48:151, 1993.


Asian consensus recommendations on optimizing the diagnosis and initiation of treatment of hepatitis B virus infection in resource-limited settings.
Asia has an intermediate-to-high prevalence of and high morbidity and mortality from hepatitis B virus (HBV) infection. Optimization of diagnosis and initiation of treatment is one of the crucial strategies for lowering disease burden in this region. Therefore, a panel of 24 experts from 10 Asian countries convened, and reviewed the literature, to develop consensus guidance on diagnosis and initiation of treatment of HBV infection in resource-limited Asian settings. The panel proposed 11 recommendations related to diagnosis, pre-treatment assessment, and indications of therapy of HBV infection, and management of HBV-infected patients with co-infections. In resource-limited Asian settings, testing for hepatitis B surface antigen may be considered as the primary test for diagnosis of HBV infection. Pre-treatment assessments should include tests for complete blood count, liver and renal function, hepatitis B e-antigen (HBeAg), anti-HBe, HBV DNA, co-infection markers and assessment of severity of liver disease. Noninvasive tests such as AST-to-platelet ratio index, fibrosis score 4 or transient elastography may be used as alternatives to liver biopsy for assessing disease severity. Considering the high burden of HBV infection in Asia, the panel adopted an aggressive approach, and recommended initiation of antiviral therapy in all HBV-infected, compensated or decompensated cirrhotic individuals with detectable HBV DNA levels, regardless of HBeAg status or alanine transaminase levels. The panel also developed a simple algorithm for guiding the initiation of treatment in noncirrhotic, HBV-infected individuals. The recommendations proposed herein, may help guide clinicians, to optimize the diagnosis and improvise the treatment rates for HBV infection in Asia.


Mechanisms and roles of mitochondrial localisation and dynamics in neuronal function.
Neurons are highly polarised, complex and incredibly energy intensive cells, and their demand for ATP during neuronal transmission is primarily met by oxidative phosphorylation by mitochondria. Thus, maintaining the health and efficient function of mitochondria is vital for neuronal integrity, viability and synaptic activity. Mitochondria do not exist in isolation, but constantly undergo cycles of fusion and fission, and are actively transported around the neuron to sites of high energy demand. Intriguingly, axonal and dendritic mitochondria exhibit different morphologies. In axons mitochondria are small and sparse whereas in dendrites they are larger and more densely packed. The transport mechanisms and mitochondrial dynamics that underlie these differences, and their functional implications, have been the focus of concerted investigation. Moreover, it is now clear that deficiencies in mitochondrial dynamics can be a primary factor in many neurodegenerative diseases. Here, we review the role that mitochondrial dynamics play in neuronal function, how these processes support synaptic transmission and how mitochondrial dysfunction is implicated in neurodegenerative disease.


Estimating endurance shuttle walk test speed using the six-minute walk test in people with chronic obstructive pulmonary disease.
The objective of this study was to derive and validate an equation to estimate the speed for the endurance shuttle walk test (ESWT) using results from the six-minute walk test (6MWT) in patients with chronic obstructive pulmonary disease (COPD). Participants with diagnosed COPD (n = 84) performed two incremental shuttle walk tests (ISWTs) and two 6MWTs. ESWT speed was calculated from the ISWT results using the original published method. An equation was derived, which directly related six-minute walk distance (6MWD) to ESWT speed. The derived equation was validated in a different group of people with COPD (n = 52). There was a strong correlation between average 6MWD and the calculated ESWT speed (r = 0.88, p < 0.001). The ESWT speed (kilometre per hour) was estimated using the following equation: 0.4889 + (0.0083 × 6MWD). The mean difference (±limits of agreement) between ESWT speeds was calculated using the original published method and found to be 0.03 (±0.77) km/hour. When the ISWT is not the test of choice for clinicians, the 6MWT can be used to accurately estimate the speed for the ESWT.


Life Cycle Air Pollution, Greenhouse Gas, and Traffic Externality Benefits and Costs of Electrifying Uber and Lyft.
Transportation network companies (TNCs), such as Uber and Lyft, have pledged to fully electrify their ridesourcing vehicle fleets by 2030 in the United States. In this paper, we introduce AgentX, a novel agent-based model built in Julia for simulating ridesourcing services with high geospatial and temporal resolution. We then instantiate this model to estimate the life cycle air pollution, greenhouse gas, and traffic externality benefits and costs of serving rides based on Chicago TNC trip data from 2019 to 2022 with fully electric vehicles. We estimate that electrification reduces life cycle greenhouse gas emissions by 40-45% (9-10¢ per trip) but increases life cycle externalities from criteria air pollutants by 6-11% (1-2¢ per trip) on average across our simulations, which represent demand patterns on weekdays and weekends across seasons during prepandemic, pandemic, and post-vaccination periods. A novel finding of our work, enabled by our high resolution simulation, is that electrification may increase deadheading for TNCs due to additional travel to and from charging stations. This extra vehicle travel increases estimated congestion, crash risk, and noise externalities by 2-3% (2-3¢ per trip). Overall, electrification reduces net external costs to society by 3-11% (5-24¢ per trip), depending on the assumed social cost of carbon.


Observation of prethermalization in long-range interacting spin chains.
Although statistical mechanics describes thermal equilibrium states, these states may or may not emerge dynamically for a subsystem of an isolated quantum many-body system. For instance, quantum systems that are near-integrable usually fail to thermalize in an experimentally realistic time scale, and instead relax to quasi-stationary prethermal states that can be described by statistical mechanics, when approximately conserved quantities are included in a generalized Gibbs ensemble (GGE). We experimentally study the relaxation dynamics of a chain of up to 22 spins evolving under a long-range transverse-field Ising Hamiltonian following a sudden quench. For sufficiently long-range interactions, the system relaxes to a new type of prethermal state that retains a strong memory of the initial conditions. However, the prethermal state in this case cannot be described by a standard GGE; it rather arises from an emergent double-well potential felt by the spin excitations. This result shows that prethermalization occurs in a broader context than previously thought, and reveals new challenges for a generic understanding of the thermalization of quantum systems, particularly in the presence of long-range interactions.


No-reflow phenomenon in ST-segment elevation myocardial infarction: still the Achilles' heel of the interventionalist.
Improvements in systems, technology and pharmacotherapy have significantly changed the prognosis over recent decades in patients presenting with ST-segment elevation myocardial infarction. These clinical achievements have, however, begun to plateau and it is becoming increasingly necessary to consider novel strategies to further improve outcomes. Approximately a third of patients treated by primary percutaneous coronary intervention for ST-segment elevation myocardial infarction will suffer from coronary no-reflow (NR), a condition characterized by poor myocardial perfusion despite patent epicardial arteries. The presence of NR impacts significantly on clinical outcomes including left ventricular dysfunction, heart failure and death, yet conventional management algorithms neither assess the risk of NR nor treat NR. This review will provide a contemporary overview on the pathogenesis, diagnosis and treatment of NR.


The biomedical potential of genetically modified flax seeds overexpressing the glucosyltransferase gene.
Flax (Linum usitatissimum) is a potential source of many bioactive components that can be found in its oil and fibers, but also in the seedcake, which is rich in antioxidants. To increase the levels of medically beneficial compounds, a genetically modified flax type (named GT) with an elevated level of phenylopropanoids and their glycoside derivatives was generated. In this study, we investigated the influence of GT seedcake extract preparations on human fibroblast proliferation and migration, and looked at the effect on a human skin model. Moreover, we verified its activity against bacteria of clinical relevance. The GT flax used in this study is characterized by overexpression of the glucosyltransferase gene derived from Solanum sogarandinum. Five GT seedcake preparations were generated. Their composition was assessed using ultra pressure liquid chromatography and confirmed using the UPLC-QTOF method. For the in vitro evaluation, the influence of the GT seedcake preparations on normal human dermal fibroblast proliferation was assessed using the MTT test and the wound scratch assay. A human skin model was used to evaluate the potential for skin irritation. To assess the antimicrobial properties of GT preparations, the percentage of inhibition of bacterial growth was calculated. The GT seedcake extract had elevated levels of phenylopropanoid compounds in comparison to the control, non-transformed plants. Significant increases in the content of ferulic acid, p-coumaric acid and caffeic acid, and their glucoside derivatives, kaempferol, quercitin and secoisolariciresinol diglucoside (SDG) were observed in the seeds of the modified plants. The GT seedcake preparations were shown to promote the proliferation of normal human dermal fibroblasts and the migration of fibroblasts in the wound scratch assay. The superior effect of GT seedcake extract on fibroblast migration was observed after a 24-hour treatment. The skin irritation test indicated that GT seedcake preparations have no harmful effect on human skin. Moreover, GT seedcake preparations exhibited inhibitory properties toward two bacterial strains: Staphylococcus aureus and Escherichia coli. We suggest that preparations derived from the new GT flax are an effective source of phenylopropanoids and that their glycoside derivatives and might be promising natural products with both healing and bacteriostatic effects. This flax-derived product is a good candidate for application in the repair and regeneration of human skin and might also be an alternative to antibiotic therapy for infected wounds.


Autonomic Phenotypes in Chronic Fatigue Syndrome (CFS) Are Associated with Illness Severity: A Cluster Analysis.
In this study we set out to define the characteristics of autonomic subgroups of patients with Chronic Fatigue Syndrome (CFS). The study included 131 patients with CFS (Fukuda criteria). Participants completed the following screening symptom assessment tools: Chalder Fatigue Scale, Fatigue Impact Scale, Fatigue Severity Scale, Epworth Sleepiness Scales, the self-reported Composite Autonomic Symptom Scale. Autonomic parameters were measured at rest with a Task Force Monitor (CNS Systems) and arterial stiffness using an Arteriograph (TensioMed Kft.). Principal axis factor analysis yielded four factors: fatigue, subjective and objective autonomic dysfunction and arterial stiffness. Using cluster analyses, these factors were grouped in four autonomic profiles: 34% of patients had sympathetic symptoms with dysautonomia, 5% sympathetic alone, 21% parasympathetic and 40% had issues with sympathovagal balance. Those with a sympathetic-dysautonomia phenotype were associated with more severe disease, reported greater subjective autonomic symptoms with sympathetic over-modulation and had the lowest quality of life. The highest quality of life was observed in the balance subtype where subjects were the youngest, had lower levels of fatigue and the lowest values for arterial stiffness. Future studies will aim to design autonomic profile-specific treatment interventions to determine links between autonomic phenotypes CFS and a specific treatment.


New antituberculosis drugs, regimens, and adjunct therapies: needs, advances, and future prospects.
About 1·3 million people died of tuberculosis in 2012, despite availability of effective drug treatment. Barriers to improvements in outcomes include long treatment duration (resulting in poor patient adherence and loss of patients to follow-up), complex regimens that involve expensive and toxic drugs, toxic effects when given with antiretroviral therapy, and multidrug resistance. After 50 years of no antituberculosis drug development, a promising pipeline is emerging through the repurposing of old drugs, re-engineering of existing antibacterial compounds, and discovery of new compounds. A range of novel antituberculosis drugs are in preclinical development, several phase 2 and 3 trials are underway, and use of adjunct therapies is being explored for drug-sensitive and drug-resistant tuberculosis. Historical advances include approval of two new drugs, delamanid and bedaquiline. Combinations of new and existing drugs are being assessed to shorten the duration of therapy and to treat multidrug-resistant tuberculosis. There has also been progress in development of new antituberculosis drugs that are active against dormant or persister populations of Mycobacterium tuberculosis. In this Review, we discuss recent advances in antituberculosis drug discovery and development, clinical trial designs, laboratory methods, and adjunct host-directed therapies, and we provide an update of phase 3 trials of various fluoroquinolones (RIFAQUIN, NIRT, OFLOTUB, and REMoxTB). We also emphasise the need to engage the community in design, implementation, and uptake of research, to increase international cooperation between drug developers and health-care providers adopting new regimens.


A Survey on Knowledge Graphs: Representation, Acquisition, and Applications.
Human knowledge provides a formal understanding of the world. Knowledge graphs that represent structural relations between entities have become an increasingly popular research direction toward cognition and human-level intelligence. In this survey, we provide a comprehensive review of the knowledge graph covering overall research topics about: 1) knowledge graph representation learning; 2) knowledge acquisition and completion; 3) temporal knowledge graph; and 4) knowledge-aware applications and summarize recent breakthroughs and perspective directions to facilitate future research. We propose a full-view categorization and new taxonomies on these topics. Knowledge graph embedding is organized from four aspects of representation space, scoring function, encoding models, and auxiliary information. For knowledge acquisition, especially knowledge graph completion, embedding methods, path inference, and logical rule reasoning are reviewed. We further explore several emerging topics, including metarelational learning, commonsense reasoning, and temporal knowledge graphs. To facilitate future research on knowledge graphs, we also provide a curated collection of data sets and open-source libraries on different tasks. In the end, we have a thorough outlook on several promising research directions.


Chromosomal-level assembly of the blood clam, Scapharca (Anadara) broughtonii, using long sequence reads and Hi-C.
The blood clam, Scapharca (Anadara) broughtonii, is an economically and ecologically important marine bivalve of the family Arcidae. Efforts to study their population genetics, breeding, cultivation, and stock enrichment have been somewhat hindered by the lack of a reference genome. Herein, we report the complete genome sequence of S. broughtonii, a first reference genome of the family Arcidae. A total of 75.79 Gb clean data were generated with the Pacific Biosciences and Oxford Nanopore platforms, which represented approximately 86× coverage of the S. broughtonii genome. De novo assembly of these long reads resulted in an 884.5-Mb genome, with a contig N50 of 1.80 Mb and scaffold N50 of 45.00 Mb. Genome Hi-C scaffolding resulted in 19 chromosomes containing 99.35% of bases in the assembled genome. Genome annotation revealed that nearly half of the genome (46.1%) is composed of repeated sequences, while 24,045 protein-coding genes were predicted and 84.7% of them were annotated. We report here a chromosomal-level assembly of the S. broughtonii genome based on long-read sequencing and Hi-C scaffolding. The genomic data can serve as a reference for the family Arcidae and will provide a valuable resource for the scientific community and aquaculture sector.


Singing with reduced air sac volume causes uniform decrease in airflow and sound amplitude in the zebra finch.
Song of the zebra finch (Taeniopygia guttata) is a complex temporal sequence generated by a drastic change to the regular oscillations of the normal respiratory pattern. It is not known how respiratory functions, such as supply of air volume and gas exchange, are controlled during song. To understand the integration between respiration and song, we manipulated respiration during song by injecting inert dental medium into the air sacs. Increased respiratory rate after injections indicates that the reduction of air affected quiet respiration and that birds compensated for the reduced air volume. During song, air sac pressure, tracheal airflow and sound amplitude decreased substantially with each injection. This decrease was consistently present during each expiratory pulse of the song motif irrespective of the air volume used. Few changes to the temporal pattern of song were noted, such as the increased duration of a minibreath in one bird and the decrease in duration of a long syllable in another bird. Despite the drastic reduction in air sac pressure, airflow and sound amplitude, no increase in abdominal muscle activity was seen. This suggests that during song, birds do not compensate for the reduced physiological or acoustic parameters. Neither somatosensory nor auditory feedback mechanisms appear to effect a correction in expiratory effort to compensate for reduced air sac pressure and sound amplitude.


Parameter estimation of an asymmetric vocal-fold system from glottal area time series using chaos synchronization.
In this paper, we apply an iterative parameter adaption scheme based on chaos synchronization to estimate system parameters of the asymmetric vocal folds from glottal area time series. The original asymmetric vocal-fold system associated with recurrent laryngeal paralysis shows chaotic vibrations with positive Lyapunov exponents. Aperiodic glottal area time series from the original system will be applied as the feedback variable coupling the simulative and the original vocal-fold systems. The parameter adaption technique based on chaos synchronization is employed to manipulate the simulative system parameters. The chaotic vibrations, system parameters, and the bifurcation diagram of the original vocal-fold system can be exactly reproduced in the simulative system, and the two chaotic systems can be synchronized. Furthermore, the effects of noise, sampling rate, and equation difference due to nonlinear spring terms on vocal-fold parameter estimations are investigated. Despite large noise perturbations, large equation differences, and low sampling rate, the parameter adaption scheme can effectively estimate the original vocal-fold system parameters. This study provides a theoretical base to apply chaos synchronization to estimate the vocal-fold system parameters from the glottal area data and show its potential application in laryngeal physiology.


Dealing With Major Life Events and Transitions: A Systematic Literature Review on and Occupational Analysis of Spirituality.
This systematic literature review analyzed the construct of spirituality as perceived by people who have experienced or are experiencing a major life event or transition. The researchers investigated studies that used narrative analysis or a phenomenological methodology related to the topic. Thematic analysis resulted in three major themes: (1) avenues to and through spirituality, (2) the experience of spirituality, and (3) the meaning of spirituality. The results provide insights into the intersection of spirituality, meaning, and occupational engagement as understood by people experiencing a major life event or transition and suggest further research that addresses spirituality in occupational therapy and interdisciplinary intervention.


Synaptic connections of the auditory nerve in cats: relationship between endbulbs of held and spherical bushy cells.
This report focuses on a class of large synaptic endings, the endbulbs of Held. These endings are located in the anteroventral cochlear nucleus and arise from the axons of type I spiral ganglion neurons. Axons were stained with horseradish peroxidase (HRP) using intracellular injections of single fibers or extracellular injections into the auditory nerve. Individual endbulbs or pairs of endbulbs that converged onto the same spherical bushy cell were examined with the aid of a light microscope and subjected to morphometric analyses. Endbulbs of fibers having low spontaneous discharge rates (SR, less than or equal to 18 spikes/sec) have a more complex shape than those of high SR fibers (greater than 18 s/s), a feature represented by systematic differences in endbulb silhouette perimeter without differences in endbulb silhouette area. Consequently, the ratio, silhouette area divided by silhouette perimeter, yields a "form factor" separating endbulbs of high SR from those of low SR. High SR fibers had ratios greater than 0.52 (mean = 0.63 +/- 0.09), whereas low SR fibers had ratios less than 0.52 (mean = 0.45 +/- 0.06). Pairs of endbulbs with unknown physiological properties had similar form factor values, despite the wide range of values observed in the endbulb population. These data imply that endbulbs converging upon the cell body of a spherical bushy cell arise from fibers of the same SR group. Electron microscopic examination was conducted on the endbulb of one physiologically characterized and intracellularly stained auditory nerve fiber (CF = 1.4 kHz; SR = 55 s/s) and its unstained endbulb mate with the aid of serial ultrathin sections. In addition to the well-known axosomatic synapses, these endbulbs formed axodendritic synapses: 11.7% for the HRP-labeled endbulb and 13.3% for the unlabeled endbulb. The axodendritic synapses appear to occur on dendrites of nearby spherical bushy cells and may represent a mechanism whereby single endbulbs can disperse activity to multiple neurons in the cochlear nucleus. We propose that axosomatic synapses preserve fiber SR groupings, whereas axodendritic synapses may not.


Bedside testing for myasthenia gravis: the ice-test.
A 67-year-old man presented to the acute medical take with a history of droopy eyelids and difficulty swallowing. A diagnosis of myasthenia gravis was suspected from the clinical history. The patient's symptoms were progressive, raising concerns of aspiration pneumonia or respiratory compromise if untreated. Definitive diagnosis of myasthenia gravis relies upon a combination of serological and electrophysiological investigations, the results of which are not immediately available in an acute situation. Bedside tests are therefore of practical clinical importance to enable prompt and effective initiation of treatment, particularly where critical symptoms are present. The ice-test is a straightforward and non-invasive investigation with a high sensitivity for myasthenia gravis and is a useful alternative to edrophonium testing. It is especially helpful in patients with cardiac and respiratory co-morbidities for whom acetylcholinesterase inhibitors are contraindicated. We describe a case where this simple bedside test successfully expedited the correct treatment and improved patient care.


Lambert-Eaton myasthenic syndrome (LEMS): a rare autoimmune presynaptic disorder often associated with cancer.
Lambert-Eaton myasthenic syndrome (LEMS) is a rare autoimmune neuromuscular junction disorder that is related to the loss of functional P/Q-type voltage-gated calcium channels (VGCCs) on presynaptic nerve terminals. Up to 60% of cases occur as a paraneoplastic disorder (SCLC-LEMS), most commonly in association with small cell lung cancer. The remaining cases have an idiopathic non-tumor etiology but are associated with underlying autoimmune disease (NT-LEMS). Patients with LEMS invariably experience progressive proximal muscle weakness, often accompanied by general fatigue and autonomic symptoms. Some LEMS clinical symptoms overlap with those of other myasthenic syndromes, most commonly myasthenia gravis, which can contribute to misdiagnosis or delayed diagnosis. Prognosis is related to the presence of cancer or autoimmune disease and the severity/distribution of muscle weakness. Cause of death in patients with SCLC-LEMS is typically tumor progression, whereas NT-LEMS does not reduce life expectancy. LEMS diagnosis is supported by a threefold approach: clinical features, electromyography, and anti-VGCC antibody serology. LEMS is a clinically important early indicator of possible cancer; therefore, a LEMS diagnosis should immediately prompt rigorous oncological screening and surveillance. Symptomatic treatment of LEMS typically involves medications that improve neurotransmission (e.g., the potassium channel blocker amifampridine [3,4-diaminopyridine]), with addition of immunosuppressants/modulators (e.g., prednisone plus azathioprine) in individuals with persistent symptoms. Where a tumor is identified, oncological treatment should take priority. It should be remembered, however, that LEMS has a significant impact on a patient's quality of life and ability to perform daily activities, and therefore warrants timely diagnosis and appropriate treatment in and of itself.


Genetically driven target tissue overexpression of CD40: a novel mechanism in autoimmune disease.
The CD40 gene, an important immune regulatory gene, is also expressed and functional on nonmyeloid-derived cells, many of which are targets for tissue-specific autoimmune diseases, including β cells in type 1 diabetes, intestinal epithelial cells in Crohn's disease, and thyroid follicular cells in Graves' disease (GD). Whether target tissue CD40 expression plays a role in autoimmune disease etiology has yet to be determined. In this study, we show that target tissue overexpression of CD40 plays a key role in the etiology of autoimmunity. Using a murine model of GD, we demonstrated that thyroidal CD40 overexpression augmented the production of thyroid-specific Abs, resulting in more severe experimental autoimmune GD (EAGD), whereas deletion of thyroidal CD40 suppressed disease. Using transcriptome and immune-pathway analyses, we showed that in both EAGD mouse thyroids and human primary thyrocytes, CD40 mediates this effect by activating downstream cytokines and chemokines, most notably IL-6. To translate these findings into therapy, we blocked IL-6 during EAGD induction in the setting of thyroidal CD40 overexpression and showed decreased levels of thyroid stimulating hormone receptor-stimulating Abs and frequency of disease. We conclude that target tissue overexpression of CD40 plays a key role in the etiology of organ-specific autoimmune disease.


[Whether or not ALS lesions spread contiguously?].
Using clinical information, it was investigated whether lesions in sporadic amyotrophic lateral sclerosis (sALS) spread contiguously from an onset site to the another regions in domino-like manner as hypothesized by prion-like propagation of pathogenic proteins. First, the data from medical records of 53 sALS patients with bulbar or lower limb onset showed that the symptom has noncontiguously spread from the bulbar region to the lower limbs or vice versa, skipping the upper limbs, in 18.9% of the patients. Second, in 18 patients with upper limb onset, correlation between the local progression speed of symptom severity in the onset limb and the interval from onset to involvement of the second region (lower limb) was investigated. The symptom severity was assessed by a score on "dressing and hygene", the subscale of the revised ALS functional rating scale. The two parameters should be positively correlated, if the lesion propagates contiguously from an initially affected motoneuron to the neighbouring ones within the same motoneuron pool (local progression) and then propagates to the another motoneuron pools (regional spread). However, the statistically significant correlation was not found, suggesting that there may be the different mechanisms between local progression and regional spread of ALS lesions.


Second harmonic generation in multi-domain χ2 media: from disorder to order.
We investigate systematically the evolution of second harmonic generation in strontium barium niobate with different degrees of disorder of its χ2 nonlinearity. These different degrees of disordered domain structures are achieved through electrically switching of the polarization at room temperature. The size and distribution of the domains change during the poling process and this in turn strongly affects the spatial distribution of the second harmonic signal. The degree of disorder can be determined by analyzing the angular distribution and wavelength dependence of the second harmonic emission patterns combined with measurements of the spontaneous polarization. We demonstrate evidence of the control of the second harmonic emission pattern by creating defined states of order, and successfully reproduce the resulting patterns theoretically.


Differences in health related quality of life in the randomised ARTSCAN study; accelerated vs. conventional radiotherapy for head and neck cancer. A five year follow up.
Health related quality of life (HRQoL) was assessed in the randomised, prospective ARTSCAN study comparing conventional radiotherapy (CF) with accelerated radiotherapy (AF) for head and neck cancer. 750 patients with squamous cell carcinoma (of any grade and stage) in the oral cavity, oro-, or hypopharynx or larynx (except T1-2, N0 glottic carcinoma) without distant metastases were randomised to either conventional fractionation (2 Gy/day, 5 days/week in 49 days, total dose 68 Gy) or accelerated fractionation (1.1+2.0 Gy/day, 5 days/week in 35 days, total dose 68 Gy). HRQoL was assessed with EORTC QLQ-C30, QLQ-H&N35 and HADS at baseline, at end of radiotherapy (eRT) and at 3 and 6 months and 1, 2 and 5 years after start of treatment. The AF group reported HRQoL was significantly lower at eRT and at 3 months for most symptoms, scales and functions. Few significant differences were noted between the groups at 6 months and 5 years. Scores related to functional oral intake never reached baseline. In comparison to CF, AF has a stronger adverse effect on HRQoL in the acute phase.


Awareness level of obstructive sleep apnea syndrome during routine unstructured interviews of a standardized patient by primary care physicians.
To assess the awareness level of primary care physicians of obstructive sleep apnea syndrome during patient-physician encounters. A prospective study using a standardized patient approach, conducted between December 2001 and March 2002. Ten sleep experts reviewed and approved the checklist questionnaire. Primary care clinics of Clalit Health Care Services, in the central region of Israel. Thirty physicians (100% compliance) randomly selected (matched by age, sex, board certification) from the 261 primary care givers in the region. A standardized patient incorporated into the physicians' daily practices. From the original checklist questionnaire, we identified 2 related question areas that at least 90% of sleep experts would pursue in light of the presenting scenario, "Do the patients snore, choke, or stop breathing in sleep?" and "Does the patient have sleepiness, unrefreshed sleep/fall asleep at undesirable times?" During the unstructured interview, only 10% of the physicians asked 3 or more questions. More than 85% of primary care physicians identified the need for polysomnography evaluation (27 physicians) or continuous positive airway pressure (26 physicians) treatment for obstructive sleep apnea syndrome. However, only 16% and 50% discussed possible complications of obstructive sleep apnea syndrome such as motor vehicle and work accidents and cardiovascular events, respectively. Primary care physicians cannot identify a common disorder associated with cardiovascular and neurobehavioral disease and could not identify the sleepiness as a source of dangerous driving. While understanding the algorithms for the diagnosis of sleep apnea, physicians cannot identify the patients for whom the diagnostics are needed. Education programs need to be developed to increase the level of suspicion of obstructive sleep apnea syndrome among practicing primary care physicians. Activities can be monitored and evaluated over time in the daily practice by standardized patients. Increased awareness level of obstructive sleep apnea syndrome among primary care physicians, through publications and educational programs, monitored by standardized patients.


Exposure to lipopolysaccharide and/or unconjugated bilirubin impair the integrity and function of brain microvascular endothelial cells.
Sepsis and jaundice are common conditions in newborns that can lead to brain damage. Though lipopolysaccharide (LPS) is known to alter the integrity of the blood-brain barrier (BBB), little is known on the effects of unconjugated bilirubin (UCB) and even less on the joint effects of UCB and LPS on brain microvascular endothelial cells (BMEC). Monolayers of primary rat BMEC were treated with 1 µg/ml LPS and/or 50 µM UCB, in the presence of 100 µM human serum albumin, for 4 or 24 h. Co-cultures of BMEC with astroglial cells, a more complex BBB model, were used in selected experiments. LPS led to apoptosis and UCB induced both apoptotic and necrotic-like cell death. LPS and UCB led to inhibition of P-glycoprotein and activation of matrix metalloproteinases-2 and -9 in mono-cultures. Transmission electron microscopy evidenced apoptotic bodies, as well as damaged mitochondria and rough endoplasmic reticulum in BMEC by either insult. Shorter cell contacts and increased caveolae-like invaginations were noticeable in LPS-treated cells and loss of intercellular junctions was observed upon treatment with UCB. Both compounds triggered impairment of endothelial permeability and transendothelial electrical resistance both in mono- and co-cultures. The functional changes were confirmed by alterations in immunostaining for junctional proteins β-catenin, ZO-1 and claudin-5. Enlargement of intercellular spaces, and redistribution of junctional proteins were found in BMEC after exposure to LPS and UCB. LPS and/or UCB exert direct toxic effects on BMEC, with distinct temporal profiles and mechanisms of action. Therefore, the impairment of brain endothelial integrity upon exposure to these neurotoxins may favor their access to the brain, thus increasing the risk of injury and requiring adequate clinical management of sepsis and jaundice in the neonatal period.


Evolutionary selection between alternative modes of gene regulation.
Microorganisms employ a wealth of gene regulatory mechanisms to adjust their growth programs to variations in the environment. It was pointed out long ago [Savageau M (1977) Proc Natl Acad Sci USA 74: 5647-5651] that the particular mode of gene regulation employed may be correlated with the "demand" on the regulated gene, i.e., how frequently the gene product is needed in its natural habitat. An evolutionary "use-it-or-lose-it" principle was proposed to govern the choice of gene regulatory strategies. Here, we examine quantitatively the forces selecting for and against two opposing modes of gene regulation, in the context of an evolutionary model that takes genetic drift, mutation, and time-dependent selection into account. We consider the effect of time-dependent selection, with periods of strong selection alternating with periods of neutral evolution. Using a variety of analytical methods, we find the effective population size and the typical time scale of environmental variations to be key parameters determining the fitness advantage of the different modes of regulation. Our results support Savageau's use-it-or-lose-it principle for small populations with long time scales of environmental variations and support a complementary "wear-and-tear" principle for the opposite situation.


Microbial communities of the Laurentian Great Lakes reflect connectivity and local biogeochemistry.
The Laurentian Great Lakes are a vast, interconnected freshwater system spanning strong physicochemical gradients, thus constituting a powerful natural laboratory for addressing fundamental questions about microbial ecology and evolution. We present a comparative analysis of pelagic microbial communities across all five Laurentian Great Lakes, focusing on Bacterial and Archaeal picoplankton characterized via 16S rRNA amplicon sequencing. We collected samples throughout the water column from the major basins of each lake in spring and summer over 2 years. Two oligotypes, classified as LD12 (Alphaproteobacteria) and acI-B1 (Actinobacteria), were among the most abundant in every sample. At the same time, microbial communities showed distinct patterns with depth during summer stratification. Deep hypolimnion samples were frequently dominated by a Chloroflexi oligotype that reached up to 19% relative abundance. Stratified surface communities differed between the colder, less productive upper lakes (Superior, Michigan, Huron) and warmer, more productive lower lakes (Erie, Ontario), in part due to an Actinobacteria oligotype (acI-C2) that averaged 7.7% of sequences in the lower lakes but <0.2% in the upper lakes. Together, our findings suggest that both hydrologic connectivity and local selective pressures shape microbial communities in the Great Lakes and establish a framework for future investigations.


Reaching kinematics to measure motor changes after mental practice in stroke.
To examine mental practice (MP) efficacy using a new kinematics reaching model. This was a prepost, case series conducted at an outpatient rehabilitation hospital of 5 patients who experienced stroke >1 year before study entry (3 males; mean age = 52.6 +/- 15.4 years [range, 38-76 years]; mean time since stroke = 51.2 months [range, 13-126 months]) exhibiting upper limb hemiparesis on their dominant sides. Participants received 30-minute therapy sessions emphasizing activities of daily living (ADLs) using their affected arms, which occurred 2 days/week for 6 weeks. After therapy, participants received 30-minute MP sessions requiring MP of the ADLs. The main outcome measure was 3-D Motion Analysis (kinematics), in which patients performed 2 functional reaching tasks consisting of reaching and grasping a plastic cylinder positioned at either elbow height (reach out) or shoulder height (reach up). Dependent variables included horizontal reaching distance, hand velocity, elbow range of motion, and shoulder range of motion. Prior to intervention, the mean horizontal reaching distance was 8.3 +/- 1.7 cm and 10.9 +/- 2.2 cm for the reach-up and reach-out tasks, respectively. Upon completion of the intervention, ability to reach up significantly improved to 9.9 +/- 1.6 cm (p <.001). Horizontal reach distance also improved during the reach-out task (11.7 +/- 2.2 cm, p = .366). No statistically significant change was observed in linear hand velocity. Patients also exhibited greater shoulder flexion and elbow extension during both the posttest reach-up and posttest reach-out tasks. Kinematics appears to offer a precise, objective way of quantifying MP-induced motor changes during ADL performance. MP appears to improve several aspects of affected arm reaching.


First Report of Tomato Gray Leaf Spot Disease Caused by Stemphylium solani in Malaysia.
In June 2011, tomatoes (Solanum lycopersicum) in major growing areas of the Cameron Highlands and the Johor state in Malaysia were affected by a leaf spot disease. Disease incidence exceeded 80% in some severely infected regions. Symptoms on 50 observed plants initially appeared on leaves as small, brownish black specks, which later became grayish brown, angular lesions surrounded by a yellow border. As the lesions matured, the affected leaves dried up and became brittle and later developed cracks in the center of the lesions. A survey was performed in these growing areas and 27 isolates of the pathogen were isolated from the tomato leaves on potato carrot agar (PCA). The isolates were purified by the single spore technique and were transferred onto PCA and V8 agar media for conidiophore and conidia production under alternating light (8 hours per day) and darkness (16 hours per day) (4). Colonies on PCA and V8 agar exhibited grey mycelium and numerous conidia were formed at the terminal end of conidiophores. The conidiophores were up to 240 μm long. Conidia were oblong with 2 to 11 transverse and 1 to 6 longitudinal septa and were 24 to 69.6 μm long × 9.6 to 14.4 μm wide. The pathogen was identified as Stemphylium solani on the basis of morphological criteria (2). In addition, DNA was extracted and the internal transcribed spacer region (ITS) was amplified by universal primers ITS5 and ITS4 (1). The PCR product was purified by the commercial PCR purification kit and the purified PCR product sequenced. The resulting sequences were 100% identical to published S. solani sequences (GenBank Accestion Nos. AF203451 and HQ840713). The amplified ITS region was deposited with NCBI GenBank under Accession No. JQ657726. A representative isolate of the pathogen was inoculated on detached 45-day-old tomato leaves of Malaysian cultivar 152177-A for pathogenicity testing. One wounded and two nonwounded leaflets per leaf were used in this experiment. The leaves were wounded by applying pressure to leaf blades with the serrated edge of a forceps. A 20-μl drop of conidial suspension containing 105 conidia/ml was used to inoculate these leaves (3). The inoculated leaves were placed on moist filter paper in petri dishes and incubated for 48 h at 25°C. Control leaves were inoculated with sterilized distilled water. After 7 days, typical symptoms for S. solani similar to those observed in the farmers' fields developed on both wounded and nonwounded inoculated leaves, but not on noninoculated controls, and S. solani was consistently reisolated. To our knowledge, this is the first report of S. solani causing gray leaf spot of tomato in Malaysia. References: (1) M. P. S. Camara et al. Mycologia 94:660, 2002. (2) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiversity Series 6:775, 2007.


Integrating large-scale neuroimaging research datasets: Harmonisation of white matter hyperintensity measurements across Whitehall and UK Biobank datasets.
Large scale neuroimaging datasets present the possibility of providing normative distributions for a wide variety of neuroimaging markers, which would vastly improve the clinical utility of these measures. However, a major challenge is our current poor ability to integrate measures across different large-scale datasets, due to inconsistencies in imaging and non-imaging measures across the different protocols and populations. Here we explore the harmonisation of white matter hyperintensity (WMH) measures across two major studies of healthy elderly populations, the Whitehall II imaging sub-study and the UK Biobank. We identify pre-processing strategies that maximise the consistency across datasets and utilise multivariate regression to characterise study sample differences contributing to differences in WMH variations across studies. We also present a parser to harmonise WMH-relevant non-imaging variables across the two datasets. We show that we can provide highly calibrated WMH measures from these datasets with: (1) the inclusion of a number of specific standardised processing steps; and (2) appropriate modelling of sample differences through the alignment of demographic, cognitive and physiological variables. These results open up a wide range of applications for the study of WMHs and other neuroimaging markers across extensive databases of clinical data.


Esophageal atresia and severe respiratory failure--cuffed pediatric tracheal tubes as an additional therapeutic option?
This article describes the use of a cuffed pediatric endotracheal tube occluding selectively the air leakage resulting from tracheoesophageal fistula in a preterm neonate with esophageal atresia and severe respiratory failure due to respiratory distress syndrome. The gastric distension resolved completely within 4 hours. Surgical correction was performed on the third day of life after respiratory stabilization.


Safety and efficacy of oral levosimendan in people with amyotrophic lateral sclerosis (the REFALS study): a randomised, double-blind, placebo-controlled phase 3 trial.
There is an urgent unmet need for new therapies in amyotrophic lateral sclerosis. In a clinical study with healthy volunteers, levosimendan, a calcium sensitiser, was shown to improve neuromechanical efficiency and contractile function of the human diaphragm. We aimed to evaluate the safety and efficacy of oral levosimendan in people with amyotrophic lateral sclerosis, with a focus on respiratory function. The REFALS study is a randomised, double-blind, placebo-controlled phase 3 trial at 99 amyotrophic lateral sclerosis specialist centres in 14 countries worldwide. People with amyotrophic lateral sclerosis were eligible for participation if they were at least 18 years of age and had a sitting slow vital capacity (SVC) of 60-90% predicted. Participants were randomly assigned (2:1) by interactive web-response system to receive either levosimendan or placebo. The capsules for oral administration were identical in appearance to maintain blinding of participants and investigators. The primary endpoint was the change from baseline in supine SVC at 12 weeks, assessed as the percentage of predicted normal sitting SVC. The key secondary endpoint was the combined assessment of function and survival (CAFS) up to 48 weeks. Analyses were done in the intention-to-treat population, comprising all participants who were randomly assigned. This trial is registered at ClinicalTrials.gov (NCT03505021) and has been completed. An extension study (REFALS-ES; NCT03948178) has also been completed, but will be reported separately. Between June 21, 2018, and June 28, 2019, 871 people were screened for the study, of whom 496 were randomly assigned either levosimendan (n=329) or placebo (n=167). Participants were followed up between June 27, 2018 and June 26, 2020, for a median duration of 50·1 (IQR 37·5-51·1) weeks. The median duration of treatment was 47·9 (IQR 26·4-48·1) weeks. Change from baseline in supine SVC at 12 weeks was -6·73% with levosimendan and -6·99% with placebo, with no significant difference between the treatments (estimated treatment difference 0·26%, 95% CI -2·03 to 2·55, p=0·83). Similarly, at week 48, CAFS did not differ between treatment groups (least squares mean change from baseline 10·69, 95% CI -15·74 to 37·12; nominal p value=0·43). The most frequent adverse events were increased heart rate (106 [33%] of 326 receiving levosimendan vs 12 [7%] of 166 receiving placebo), fall (85 [26%] vs 48 [29%]), headache (93 [29%] vs 36 [22%]), and dyspnoea (59 [18%] vs 32 [19%]). 33 (10%) participants allocated levosimendan and 20 (12%) assigned placebo died during the trial, mainly due to respiratory failure or progression of amyotrophic lateral sclerosis. Levosimendan was not superior to placebo in maintaining respiratory function in a broad population with amyotrophic lateral sclerosis. Although levosimendan was generally well tolerated, increased heart rate and headache occurred more frequently with levosimendan than with placebo. The possibility of a clinically relevant subgroup of responsive individuals requires further evaluation. Orion Corporation.


Clinical and cellular characterisation of Hermansky-Pudlak syndrome type 6.
In the last decade, Hermansky-Pudlak syndrome (HPS) has arisen as an instructive disorder for cell biologists to study the biogenesis of lysosome related organelles (LROs). Of the eight human HPS subtypes, only subtypes 1 through 5 are well described. To characterise extensively the HPS-6 subtype, caused by defects in HPS6, a subunit of the biogenesis of lysosome related organelles complex-2 (BLOC-2). Mutation analysis for the HPS6 gene was performed on DNA from our group of unclassified HPS patients. The clinical phenotype of patients with HPS6 mutations was then carefully ascertained, and their cultured dermal melanocytes were employed for cellular immunofluorescence studies. Molecular studies showed a variety of mutations in the single exon HPS6 gene, including frame shift, missense, and nonsense mutations as well as a approximately 20 kb deletion spanning the entire HPS6 genomic region. Cellular studies revealed that the melanogenic proteins tyrosinase and tyrosinase related protein 1 failed to be efficiently delivered to the melanosomes of HPS-6 patients, explaining their hypopigmentation. Clinical studies indicated that HPS-6 patients exhibit oculocutaneous albinism and a bleeding diathesis. Importantly, granulomatous colitis and pulmonary fibrosis, debilitating features present in HPS subtypes 1 and 4, were not detected in our HPS-6 patients. The HPS-6 subtype resembles other BLOC-2 defective subtypes (that is, HPS-3 and HPS-5) in its molecular, cellular and clinical findings. These findings are not only important for providing a prognosis to newly diagnosed HPS-6 patients, but also for further elucidation of HPS function in the biogenesis of LROs.


Pathogenic Vibrio Species Are Associated with Distinct Environmental Niches and Planktonic Taxa in Southern California (USA) Aquatic Microbiomes.
Interactions between vibrio bacteria and the planktonic community impact marine ecology and human health. Many coastal Vibrio spp. can infect humans, representing a growing threat linked to increasing seawater temperatures. Interactions with eukaryotic organisms may provide attachment substrate and critical nutrients that facilitate the persistence, diversification, and spread of pathogenic Vibrio spp. However, vibrio interactions with planktonic organisms in an environmental context are poorly understood. We quantified the pathogenic Vibrio species V. cholerae, V. parahaemolyticus, and V. vulnificus monthly for 1 year at five sites and observed high abundances, particularly during summer months, with species-specific temperature and salinity distributions. Using metabarcoding, we established a detailed profile of both prokaryotic and eukaryotic coastal microbial communities. We found that pathogenic Vibrio species were frequently associated with distinct eukaryotic amplicon sequence variants (ASVs), including diatoms and copepods. Shared environmental conditions, such as high temperatures and low salinities, were associated with both high concentrations of pathogenic vibrios and potential environmental reservoirs, which may influence vibrio infection risks linked to climate change and should be incorporated into predictive ecological models and experimental laboratory systems. IMPORTANCE Many species of coastal vibrio bacteria can infect humans, representing a growing health threat linked to increasing seawater temperatures. However, their interactions with surrounding microbes in the environment, especially eukaryotic organisms that may provide nutrients and attachment substrate, are poorly understood. We quantified three pathogenic Vibrio species monthly for a duration of 1 year, finding that all three species were abundant and exhibited species-specific temperature and salinity distributions. Using metabarcoding, we investigated associations between these pathogenic species and prokaryotic and eukaryotic microbes, revealing genus and amplicon sequence variant (ASV)-specific relationships with potential functional implications. For example, pathogenic species were frequently associated with chitin-producing eukaryotes, such as diatoms in the genus Thalassiosira and copepods. These associations between high concentrations of pathogenic vibrios and potential environmental reservoirs should be considered when predicting infection risk and developing ecologically relevant model systems.


Pet dogs as promoters of wellbeing.
The rise in life expectancy requires strategies to enable healthy ageing and the promotion of a high quality of life in old age. Poor mental health including depression and social isolation can blight older people's lives. Despite the positive benefits of physical activity for both mental and physical health, only a minority of those over 65 years are attaining the recommended levels of physical activity. The evidence relating to the benefits of pet dogs as promoters of wellbeing is set out in this article, although meeting their care needs may place an additional strain on an older person and/or their carer who has limited resources and physical capabilities.


Two cases of failed back surgery syndrome after correction and stabilization surgery for scoliosis with Harrington instrumentation.
We report two cases of adjacent segment degeneration in patients with idiopathic scoliosis who underwent surgical treatment with Harrington instrumentation in young age. Both patients developed a symptomatic degeneration of the disk immediately under the last stabilized level and were treated with decompression and stabilization. Clinical and radiological results are satisfactory at the follow-up.


The neural basis of economic decision-making in the Ultimatum Game.
The nascent field of neuroeconomics seeks to ground economic decision making in the biological substrate of the brain. We used functional magnetic resonance imaging of Ultimatum Game players to investigate neural substrates of cognitive and emotional processes involved in economic decision-making. In this game, two players split a sum of money;one player proposes a division and the other can accept or reject this. We scanned players as they responded to fair and unfair proposals. Unfair offers elicited activity in brain areas related to both emotion (anterior insula) and cognition (dorsolateral prefrontal cortex). Further, significantly heightened activity in anterior insula for rejected unfair offers suggests an important role for emotions in decision-making.


Transgenic overexpression of GTP cyclohydrolase 1 in cardiomyocytes ameliorates post-infarction cardiac remodeling.
GTP cyclohydrolase 1 (GCH1) and its product tetrahydrobiopterin play crucial roles in cardiovascular health and disease, yet the exact regulation and role of GCH1 in adverse cardiac remodeling after myocardial infarction are still enigmatic. Here we report that cardiac GCH1 is degraded in remodeled hearts after myocardial infarction, concomitant with increases in the thickness of interventricular septum, interstitial fibrosis, and phosphorylated p38 mitogen-activated protein kinase and decreases in left ventricular anterior wall thickness, cardiac contractility, tetrahydrobiopterin, the dimers of nitric oxide synthase, sarcoplasmic reticulum Ca2+ release, and the expression of sarcoplasmic reticulum Ca2+ handling proteins. Intriguingly, transgenic overexpression of GCH1 in cardiomyocytes reduces the thickness of interventricular septum and interstitial fibrosis and increases anterior wall thickness and cardiac contractility after infarction. Moreover, we show that GCH1 overexpression decreases phosphorylated p38 mitogen-activated protein kinase and elevates tetrahydrobiopterin levels, the dimerization and phosphorylation of neuronal nitric oxide synthase, sarcoplasmic reticulum Ca2+ release, and sarcoplasmic reticulum Ca2+ handling proteins in post-infarction remodeled hearts. Our results indicate that the pivotal role of GCH1 overexpression in post-infarction cardiac remodeling is attributable to preservation of neuronal nitric oxide synthase and sarcoplasmic reticulum Ca2+ handling proteins, and identify a new therapeutic target for cardiac remodeling after infarction.


Butyrate-induced apoptotic cascade in colonic carcinoma cells: modulation of the beta-catenin-Tcf pathway and concordance with effects of sulindac and trichostatin A but not curcumin.
Short-chain fatty acids play a critical role in colonic homeostasis because they stimulate pathways of growth arrest, differentiation, and apoptosis. These effects have been well characterized in colonic cell lines in vitro. We investigated the role of beta-catenin-Tcf signaling in these responses to butyrate and other well-characterized inducers of apoptosis of colonic epithelial cells. Unlike wild-type APC, which down-regulates Tcf activity, butyrate, as well as sulindac and trichostatin A, all inducers of G0-G1 cell cycle arrest and apoptosis in the SW620 colonic carcinoma cell line, up-regulate Tcf activity. In contrast, structural analogues of butyrate that do not induce cell cycle arrest or apoptosis and curcumin, which stimulates G2-M arrest without inducing apoptosis, do not alter Tcf activity. Similar to the cell cycle arrest and apoptotic cascade induced by butyrate, the up-regulation of Tcf activity is dependent upon the presence of a mitochondrial membrane potential, unlike the APC-induced down-regulation, which is insensitive to collapse of the mitochondrial membrane potential. Moreover, the butyrate-induced increase in Tcf activity, which is reflected in an increase in beta-catenin-Tcf complex formation, is independent of the down-regulation caused by expression of wild-type APC. Thus, butyrate and wild-type APC have different and independent effects on beta-catenin-Tcf signaling. These data are consistent with other reports that suggest that the absence of wild-type APC, associated with the up-regulation of this signaling pathway, is linked to the probability of a colonic epithelial cell entering an apoptotic cascade.


'At the grass roots level it's about sitting down and talking': exploring quality improvement through case studies with high-improving Aboriginal and Torres Strait Islander primary healthcare services.
Improving the quality of primary care is an important strategy to improve health outcomes. However, responses to continuous quality improvement (CQI) initiatives are variable, likely due in part to a mismatch between interventions and context. This project aimed to understand the successful implementation of CQI initiatives in Aboriginal and Torres Strait Islander health services in Australia through exploring the strategies used by 'high-improving' Indigenous primary healthcare (PHC) services. This strengths-based participatory observational study used a multiple case study method with six Indigenous PHC services in northern Australia that had improved their performance in CQI audits. Interviews with healthcare providers, service users and managers (n=134), documentary review and non-participant observation were used to explore implementation of CQI and the enablers of quality improvement in these contexts. Services approached the implementation of CQI differently according to their contexts. Common themes previously reported included CQI systems, teamwork, collaboration, a stable workforce and community engagement. Novel themes included embeddedness in the local historical and cultural contexts, two-way learning about CQI and the community 'driving' health improvement. These novel themes were implicit in the descriptions of stakeholders about why the services were improving. Embeddedness in the local historical and cultural context resulted in 'two-way' learning between communities and health system personnel. Practical interventions to strengthen responses to CQI in Indigenous PHC services require recruitment and support of an appropriate and well prepared workforce, training in leadership and joint decision-making, regional CQI collaboratives and workable mechanisms for genuine community engagement. A 'toolkit' of strategies for service support might address each of these components, although strategies need to be implemented through a two-way learning process and adapted to the historical and cultural community context. Such approaches have the potential to assist health service personnel strengthen the PHC provided to Indigenous communities.


First Report of Xanthomonas citri pv. citri Pathotype A Causing Asiatic Citrus Canker on Grapefruit and Mexican Lime in Senegal.
In February 2010, grapefruit (Citrus paradisi) and Mexican lime (C. aurantifolia) leaves with erumpent callus-like lesions were collected in Senegal in the Sebikotane area between Dakar and Thies. Similar symptoms have been observed by local farmers since 2008, and lesions were morphologically similar to those of citrus canker caused by Xanthomonas citri pv. citri (Asiatic canker) and X. citri pv. aurantifolii (South American canker). Lesions were primarily reported from grapefruit (cv. Shambar), which is the most frequent citrus species produced in this area, and Mexican lime, which is also commonly grown. Both species are very susceptible to X. citri pv. citri pathotype A, and Mexican lime is susceptible to X. citri pv. citri pathotype A* and X. citri pv. aurantifolii (4). Fifteen Xanthomonas-like strains were isolated from disease samples using KC semiselective medium (3). PCR with primer pair 4/7 (2) revealed that all the Senegalese strains and the X. citri pv. citri strain CFBP 2525 from New Zealand, used as a positive control, generated the expected DNA fragment, whereas no fragment was observed for negative controls (distilled water instead of the template). Insertion sequence ligation-mediated (IS-LM)-PCR analysis (1) of X. citri pv. citri strains from Senegal and reference strains of X. citri pv. citri pathotypes A and A* (1), with MspI and four primer pairs (unlabelled MspI primer and four 5'-labelled insertion sequence-specific primers targeting three IS elements), indicated that the strains from Senegal were related to X. citri pv. citri but not to pv. aurantifolii. They were closely related to X. citri pv. citri pathotype A strains, with a broad host range, present in the Indian subcontinent and Mali (C. Vernière, unpublished data). Multilocus sequence analysis of four partial housekeeping gene sequences (atpD, dnaK, efp, and gyrB) confirmed that four Senegalese strains were not related to X. citri pv. aurantifolii and showed a full sequence identity to X. citri pv. citri sequence type ST3 (2), fully consistent with IS-LM-PCR. Using a detached leaf assay (4), Duncan grapefruit, Pineapple sweet orange, and Mexican lime leaves inoculated with all strains from Senegal developed typical erumpent, callus-like tissue at wound sites 2 weeks after the inoculations. Xanthomonas-like colonies were reisolated and PCR amplification with the primer pair 4/7 produced the same 468-nt DNA fragment. This represents the fourth outbreak of citrus canker reported from Africa within the last 5 years, the other documented reports were from Ethiopia (2007) and Mali and Somalia (2008). High disease prevalence was observed in Senegal with incidence exceeding 90% in the orchards where lime and grapefruit were infected for 3 years, indicating the suitability of environmental conditions in this region for the development of Asiatic citrus canker. The origin of the inoculum associated with the reported canker outbreak in Senegal is currently unknown and the precise distribution of the pathogen needs to be thoroughly assessed. To our knowledge, this is the first documented report of the presence of Asiatic citrus canker in Senegal and this occurrence increases the threat to citriculture in West Africa. References: (1) L. Bui Thi Ngoc et al. FEMS Microbiol. Lett. 288:33, 2008. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (4) C. Vernière et al. Eur. J. Plant Pathol. 104:477, 1998.


Are randomized controlled trials in urology being conducted with justification?
Considering the substantial increase in research funding in the field of urology, minimizing research waste should be a top priority. Systematic reviews (SRs) compile available evidence regarding a clinical question into a single critical resource. If properly utilized, SRs can help minimize redundant studies, focus attention to unsubstantiated treatments, and reduce research waste. To appraise the use of SRs as justification for conducting randomized controlled trials (RCTs) published in high impact urology journals, and to report the ways SRs were incorporated into RCT manuscripts published in the top four urology journals by h5 index. On December 13, 2019, a PubMed search was conducted for RCTs published in the top four urology journals according to the Google Scholar h5-index: European Urology, BJU International, The Journal of Urology, and Urology. For an article to be eligible for inclusion in this study, it must have been a full length RCT, published between November 30, 2014, and November 30, 2019 in one of the identified journals, reported only human subjects, and been accessible in English. The following data points were extracted independently by select investigators from each included RCT: manuscript title, year of publication, journal title, type of intervention (drug, medical device, procedure, other), funding source (government, hospital/university, industry, mixed) type of trial (parallel groups, crossover, cluster), and total number of participants reported in each RCT. The included RCTs were searched for reference to an SR, which was then recorded as "yes - verbatim," "yes - inferred," or "not used as justification" and the location in the manuscript where the SR was cited was recorded. Of the 566 articles retrieved, 276 were included. Overall, 150 (54.3%) RCTs cited an SR as either verbatim (108; 39.1%) or inferred (42; 15.2%) trial justification, while 126 (45.7%) did not use an SR for RCT justification. Of those 126, 107 (84.9%) RCTs did not cite an SR to any extent. A significant association was noted between verbatim justification and type of intervention (x2=20.23, p=0.017), with 18 of 31 (58.1%) "other" interventions (i.e. psychosocial intervention, exercise programs, and online therapy) having an SR cited as verbatim justification. Only 39 of 118 (33.1%) pharmaceutical trials referenced an SR as verbatim justification. Of 403 systematic review citations, 205 (50.8%) appeared in the Discussion section, while 15 (3.7%) were in the Methods section. We found that RCTs published in four high impact urology journals inconsistently referenced an SR as justification and 39.1% of our entire sample did not reference an SR at all. These findings indicate that a divide exists between the instruction and implementation of evidence based medicine in the field of urology concerning RCTs published in the top four journals. Educating clinicians and researchers on the use of SR as justification for RCTs in urology may reduce research waste and increase the quality of RCTs in the field.


Nonidet P-40 extraction of lymphocyte plasma membrane. Characterization of the insoluble residue.
Purified preparations of lymphocyte plasma membrane were extracted exhaustively with Nonidet P-40 in Dulbecco's phosphate-buffered saline medium. The insoluble fraction, as defined by sedimentation at 10(6) g-min, contained about 10% of the membrane protein as well as cholesterol and phospholipid. The lipid/protein ratio, cholesterol/phospholipid ratio and sphingomyelin content were increased in the residue. Density-gradient centrifugation suggested that the lipid and protein form a common entity. As judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the Nonidet P-40-insoluble fractions of the plasma membranes of human B lymphoblastoid cells and pig mesenteric lymph-node lymphocytes possessed similar qualitative polypeptide compositions but differed quantitatively. Both residues comprised major polypeptides of Mr 28 000, 33 000, 45 000 and 68 000, together with a prominent band of Mr 120 000 in the human and of Mr 200 000 in the pig. The polypeptides of Mr 28 000, 33 000, 68 000 and 120 000 were probably located exclusively in the Nonidet P-40-insoluble residue, which also possessed a 4-fold increase in 5'-nucleotidase specific activity. The results indicate that a reproducible fraction of lymphocyte plasma membrane is insoluble in non-ionic detergents and that this fraction possesses a unique polypeptide composition. By analogy with similar studies with erythrocyte ghosts, it appears likely that the polypeptides are located on the plasma membrane's cytoplasmic face.


Water Pipeline Leakage Detection Based on Machine Learning and Wireless Sensor Networks.
The detection of water pipeline leakage is important to ensure that water supply networks can operate safely and conserve water resources. To address the lack of intelligent and the low efficiency of conventional leakage detection methods, this paper designs a leakage detection method based on machine learning and wireless sensor networks (WSNs). The system employs wireless sensors installed on pipelines to collect data and utilizes the 4G network to perform remote data transmission. A leakage triggered networking method is proposed to reduce the wireless sensor network's energy consumption and prolong the system life cycle effectively. To enhance the precision and intelligence of leakage detection, we propose a leakage identification method that employs the intrinsic mode function, approximate entropy, and principal component analysis to construct a signal feature set and that uses a support vector machine (SVM) as a classifier to perform leakage detection. Simulation analysis and experimental results indicate that the proposed leakage identification method can effectively identify the water pipeline leakage and has lower energy consumption than the networking methods used in conventional wireless sensor networks.


On the phase reduction and response dynamics of neural oscillator populations.
We undertake a probabilistic analysis of the response of repetitively firing neural populations to simple pulselike stimuli. Recalling and extending results from the literature, we compute phase response curves (PRCs) valid near bifurcations to periodic firing for Hindmarsh-Rose, Hodgkin-Huxley, FitzHugh-Nagumo, and Morris-Lecar models, encompassing the four generic (codimension one) bifurcations. Phase density equations are then used to analyze the role of the bifurcation, and the resulting PRC, in responses to stimuli. In particular, we explore the interplay among stimulus duration, baseline firing frequency, and population-level response patterns. We interpret the results in terms of the signal processing measure of gain and discuss further applications and experimentally testable predictions.


Development of Antiviral Innate Immunity During In Vitro Differentiation of Mouse Embryonic Stem Cells.
The innate immunity of embryonic stem cells (ESCs) has recently emerged as an important issue in ESC biology and in ESC-based regenerative medicine. We have recently reported that mouse ESCs (mESCs) do not have a functional type I interferon (IFN)-based antiviral innate immunity. They are deficient in expressing IFN in response to viral infection and have limited ability to respond to IFN. Using fibroblasts (FBs) as a cell model, the current study investigated the development of antiviral mechanisms during in vitro differentiation of mESCs. We demonstrate that mESC-differentiated FBs (mESC-FBs) share extensive similarities with naturally differentiated FBs in morphology, marker expression, and growth pattern, but their development of antiviral mechanisms lags behind. Nonetheless, the antiviral mechanisms are inducible during mESC differentiation as demonstrated by the transition of nuclear factor kappa B (NFκB), a key transcription factor for IFN expression, from its inactive state in mESCs to its active state in mESC-FBs and by increased responses of mESC-FBs to viral stimuli and IFN during their continued in vitro propagation. Together with our previously published study, the current data provide important insights into molecular basis for the deficiency of IFN expression in mESCs and the development of antiviral innate immunity during mESC differentiation.


Effects of acute alcohol consumption on executive cognitive functioning in naturalistic settings.
Laboratory studies have demonstrated that acute alcohol intoxication can disrupt performance on neuropsychological tests of executive cognitive functioning such as the Wisconsin Card Sorting Test (WCST). However, the generalizability of such findings to typical self-regulated alcohol intake in social settings can be questioned. In the present study, 86 young adults were recruited at Australian bars to perform a computer version of the WCST. Participants displayed blood alcohol concentrations (BACs) across a range from 0 to 0.15%. Although self-report measures of typical alcohol consumption, impulsivity, and frontal lobe related everyday functioning were all intercorrelated in line with other recent findings, multiple regression indicated that these measures did not predict perseverative errors (PE) nor non-perseverative errors (NPE) on the WCST, whereas BAC uniquely predicted PE but not NPE. The results were consistent with a dose-dependent selective disruption of prefrontal cortical functioning by alcohol. There were no differences in performance between participants tested on the ascending limb of the BAC curve and those tested on the descending limb. Alcohol-associated perseveration may reflect the inhibitory effect of alcohol-induced dopamine release in the prefrontal cortex.


Characterization of the abomasal transcriptome for mechanisms of resistance to gastrointestinal nematodes in cattle.
The response of the abomasal transcriptome to gastrointestinal parasites was evaluated in parasite-susceptible and parasite-resistant Angus cattle using RNA-seq at a depth of 23.7 million sequences per sample. These cattle displayed distinctly separate resistance phenotypes as assessed by fecal egg counts. Approximately 65.3% of the 23,632 bovine genes were expressed in the fundic abomasum. Of these, 13,758 genes were expressed in all samples tested and likely represent core components of the bovine abomasal transcriptome. The gene (BT14427) with the most abundant transcript, accounting for 10.4% of sequences in the transcriptome, is located on chromosome 29 and has unknown functions. Additionally, PIGR (1.6%), Complement C3 (0.7%), and Immunoglobulin J chain (0.5%) were among the most abundant transcripts in the transcriptome. Among the 203 genes impacted, 64 were significantly over-expressed in resistant animals at a stringent cutoff (FDR < 5%). Among the 94 224 splice junctions identified, 133 were uniquely present: 90 were observed only in resistant animals, and 43 were present only in susceptible animals. Gene Ontology (GO) enrichment of the genes under study uncovered an association with lipid metabolism, which was confirmed by an independent pathway analysis. Several pathways, such as FXR/RXR activation, LXR/RXR activation, LPS/IL-1 mediated inhibition of RXR function, and arachidonic acid metabolism, were impacted in resistant animals, which are potentially involved in the development of parasite resistance in cattle. Our results provide insights into the development of host immunity to gastrointestinal nematode infection and will facilitate understanding of mechanism underlying host resistance.


A novel mutation in the putative DNA helicase XH2 is responsible for male-to-female sex reversal associated with an atypical form of the ATR-X syndrome.
We describe a pedigree presenting X-linked severe mental retardation associated with multiple congenital abnormalities and 46,XY gonadal dysgenesis, leading in one family member to female gender assignment. Female carriers are unaffected. The dysmorphic features are similar to those described in the alpha-thalassemia and mental retardation (ATR-X) syndrome, although there is no clinical evidence of alpha-thalassemia in this family. In addition, the family had other clinical features not previously observed in the ATR-X syndrome, including partial optic-nerve atrophy and partial ocular albinism. Mutations in a putative DNA helicase, termed XH2, have been reported to give rise to the ATR-X syndrome. We screened the XH2 gene for mutations in affected members of the family and identified a 4-bp deletion at an intron/exon boundary that removes an invariant 3' splice-acceptor site. The mutation cosegregates with the syndrome. The genomic deletion causes missplicing of the pre-mRNA, which results in the loss of 8 bp of coding sequence, thereby generating a frameshift and a downstream premature stop codon. Our finding increases the range of clinical features associated with mutations in the XH2 gene.


Mechanisms of aging-induced impairment of endothelium-dependent relaxation: role of tetrahydrobiopterin.
Oxidative stress has been implicated as an important mechanism of vascular endothelial dysfunction induced by aging. Previous studies suggested that tetrahydrobiopterin (BH4), an essential cofactor of endothelial NO synthase, could be a molecular target for oxidation. We tested the hypothesis that oxidative stress, in particular oxidation of BH4, may contribute to attenuation of endothelium-dependent relaxation in aged mice. Vasomotor function of isolated carotid arteries was studied using a video dimension analyzer. Vascular levels of BH4 and its oxidation products were measured via HPLC. In aged mice (age, 95 +/- 2 wk), endothelium-dependent relaxation to ACh (10(-5) to 10(-9) M) as well as endothelium-independent relaxation to the NO donor diethylammonium (Z)-1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA-NONOate, 10(-5) to 10(-9) M) were significantly reduced compared with relaxation detected in young mice (age, 23 +/- 0.5 wk). Incubation of aged mouse carotid arteries with the cell-permeable SOD mimetic Mn(III)tetra(4-benzoic acid)porphyrin chloride normalized relaxation to ACh and DEA-NONOate. Furthermore, production of superoxide anion in aorta and serum levels of amyloid P component, which is the murine analog of C-reactive protein, was increased in old mice. In aorta, neither the concentration of BH4 nor the ratio of reduced BH4 to the oxidation products were different between young and aged mice. Our results demonstrate that in mice, aging impairs relaxation mediated by NO most likely by increased formation of superoxide anion. Oxidation of BH4 does not appear to be an important mechanism underlying vasomotor dysfunction in aged mouse arteries.


Markov random field model for segmenting large populations of lipid vesicles from micrographs.
Giant unilamellar lipid vesicles, artificial replacements for cell membranes, are a promising tool for in vitro assessment of interactions between products of nanotechnologies and biological membranes. However, the effect of nanoparticles can not be derived from observations on a single specimen, vesicle populations should be observed instead. We propose an adaptation of the Markov random field image segmentation model which allows detection and segmentation of numerous vesicles in micrographs. The reliability of this model with different lighting, blur, and noise characteristics of micrographs is examined and discussed. Moreover, the automatic segmentation is tested on micrographs with thousands of vesicles and the result is compared to that of manual segmentation. The segmentation step presented is part of a methodology we are developing for bio-nano interaction assessment studies on lipid vesicles.


Can we teach an old drug new tricks?
Although resistance to chloroquine (CQ) has relegated it from modern chemotherapeutic strategies to treat Plasmodium falciparum malaria, new evidence suggests that higher doses of the drug may exert a different killing mechanism and offers this drug a new lease of life. Whereas the established antimalarial mechanisms of CQ are usually associated with nanomolar levels of the drug, micromolar levels of CQ trigger a distinct cell death pathway involving the permeabilization of the digestive vacuole of the parasite and a release of hydrolytic enzymes. In this paper, we propose that this pathway is a promising antimalarial strategy and suggest that revising the CQ treatment regimen may elevate blood drug levels to trigger this pathway without increasing the incidence of adverse reactions.


Fluorescent whole-cell hybridization with 16S rRNA-targeted oligonucleotide probes to identify Brucella spp. by flow cytometry.
A whole-cell hybridization assay with fluorescent oligonucleotide probes derived from the 16S rRNA sequence of Brucella abortus in combination with flow cytometry has been developed. With the three fluorescent probes selected, a positive signal was observed with all the representative strains of the species and biovars of Brucella and with a total of nine different Brucella clinical isolates. Using the B9 probe in the hybridization assay, it was possible to discriminate between Brucella suis biovars 2, 3, 4, and 5 and almost all the other Brucella spp. On the basis of differences in fluorescence intensities, no discrimination was established between Brucella spp. and other phylogenetically related microorganisms. No positive fluorescence signals were detected with any of the bacteria showing serological cross-reactions with Brucella spp. and with a total of 17 clinical isolates not belonging to the genus Brucella. These results suggest that the 16S rRNA whole-cell hybridization technique could be a valuable diagnostic tool for the detection and identification of Brucella spp.


Diagnosis and management of red eye in primary care.
Red eye is the cardinal sign of ocular inflammation. The condition is usually benign and can be managed by primary care physicians. Conjunctivitis is the most common cause of red eye. Other common causes include blepharitis, corneal abrasion, foreign body, subconjunctival hemorrhage, keratitis, iritis, glaucoma, chemical burn, and scleritis. Signs and symptoms of red eye include eye discharge, redness, pain, photophobia, itching, and visual changes. Generally, viral and bacterial conjunctivitis are self-limiting conditions, and serious complications are rare. Because there is no specific diagnostic test to differentiate viral from bacterial conjunctivitis, most cases are treated using broad-spectrum antibiotics. Allergies or irritants also may cause conjunctivitis. The cause of red eye can be diagnosed through a detailed patient history and careful eye examination, and treatment is based on the underlying etiology. Recognizing the need for emergent referral to an ophthalmologist is key in the primary care management of red eye. Referral is necessary when severe pain is not relieved with topical anesthetics; topical steroids are needed; or the patient has vision loss, copious purulent discharge, corneal involvement, traumatic eye injury, recent ocular surgery, distorted pupil, herpes infection, or recurrent infections.


Graphene oxide and carbon dots as broad-spectrum antimicrobial agents - a minireview.
Due to the increasing global population, growing contamination of water and air, and wide spread of infectious diseases, antibiotics are extensively used as a major antibacterial drug. However, many microbes have developed resistance to antibiotics through mutation over time. As an alternative to antibiotics, antimicrobial nanomaterials have attracted great attention due to their advantageous properties and unique mechanisms of action toward microbes. They inhibit bacterial growth and destroy cells through complex mechanisms, making it difficult for bacteria to develop drug resistance, though some health concerns related to biocompatibility remain for practical applications. Among various antibacterial nanomaterials, carbon-based materials, especially graphene oxide (GO) and carbon dots (C-Dots), are promising candidates due to the ease of production and functionalization, high dispersibility in aqueous media, and promising biocompatibility. The antibacterial properties of these nanomaterials can be easily adjusted by surface modification. They are promising materials for future applications against multidrug-resistant bacteria based on their strong capacity in disruption of microbial membranes. Though many studies have reported excellent antibacterial activity of carbon nanomaterials, their impact on the environment and living organisms is of concern due to the accumulatory and cytotoxic effects. In this review, we discuss antimicrobial applications of the functional carbon nanomaterials (GO and C-Dots), their antibacterial mechanisms, factors affecting antibacterial activity, and concerns regarding cytotoxicity.


Does the pandemic medical early warning score system correlate with disposition decisions made at patient contact by emergency care practitioners?
To assess the performance of the pandemic medical early warning score (PMEWS) in a cohort of adult patients seen in the community by emergency care practitioners (ECP) and its correlation with ECP decision-making to either 'treat and leave' or transfer for hospital assessment. Cases attended by ECP in South Yorkshire in 2007 in which the final ECP working diagnosis was a respiratory condition were retrospectively identified from the Yorkshire Ambulance Service database. The patient report forms were reviewed for the PMEWS variables and scores calculated using the PMEWS system. The outcome measure was management in the community versus transport to hospital. Receiver operating characteristics (ROC) curves were calculated to assess the discrimination of PMEWS. A cohort of 300 patients was assessed. 217 (72%) were aged 65 years or over, and 272 (91%) had either comorbid disease or impaired functional status. 98 (33%) were deemed to need hospital assessment or admission. The ROC curves suggested that there is good correlation between the PMEWS score and the decision to discharge. PMEWS correlates well with decisions to admit to hospital or leave at home made by extended role practitioners in the patient group studied; however, further prospective work is required to further validate early warning scoring systems in prehospital care.


Experimental ocular tuberculosis in guinea pigs.
To develop an animal model of intraocular tuberculosis (TB) with features of pulmonary TB and extrapulmonary dissemination to the eye. Hartley strain guinea pigs were infected via an aerosol route with virulent Mycobacterium tuberculosis. One group of guinea pigs was infected with a relatively low bacterial inoculum and received no treatment. A second group of guinea pigs received high-dose infection and were treated with the first-line anti-TB drugs isoniazid, rifampin, and pyrazinamide. Development of ocular TB lesions was documented by histological analysis, acid-fast staining, and real-time polymerase chain reaction for M tuberculosis DNA. Untreated guinea pigs developed pulmonary and extrapulmonary TB. Ocular TB, primarily involving the uvea, developed in 5 of 12 eyes (42%). Uveal granulomatous lesions showed the presence of acid-fast organisms and M tuberculosis DNA. In treated animals, none of the 8 eyes examined revealed the presence of acid-fast organisms; however, there was mild nongranulomatous uveitis in 4 eyes. Mycobacterium tuberculosis delivered via aerosol to guinea pigs results in extrapulmonary dissemination to the eye. Of significance, intraocular changes in this model include granulomatous inflammation and the presence of acid-fast organisms, as seen in human cases of ocular TB. Clinical Relevance The guinea pig model may provide greater insight into the pathogenesis of intraocular TB and assist in the development of novel modalities to treat this global infectious disease.