Natural killer cells, dendritic cells, and the alarmin high-mobility group box 1 protein: a dangerous trio in HIV-1 infection?
Natural killer (NK) cells promote antiviral immunity by producing proinflammatory cytokines and by lysing infected cells. In addition, NK cells can modulate dendritic cell functions. NK-dendritic cell crosstalk results in activation of both cell types, with dendritic cells promoting NK-cell activity and NK cells inducing further maturation of dendritic cells. Here we review the recent evidence suggesting that NK-dendritic cell crosstalk is disrupted during HIV-1 infection and we discuss the consequences on HIV persistence in dendritic cells. NK cell-mediated dendritic cell editing is compromised during HIV-1 infection, and NK cells from viremic individuals show a decreased ability to kill immature dendritic cells. The defect is associated with impaired NKp30 function. Moreover, the resistance of HIV-1-infected dendritic cells to NK-mediated lysis is associated with the upregulation of apoptosis inhibitors, thus protecting infected dendritic cells from TRAIL-dependent apoptosis. These inhibitors are upregulated by the high-mobility group box 1 protein (HMGB1), an alarmin produced at NK-dendritic cell synapse that is essential for NK-dependent dendritic cell maturation, but also promotes viral replication in infected dendritic cells. HIV-1-induced impairment of NK-dendritic cell crosstalk may significantly alter both innate and adaptive immunity. It may also contribute to HIV persistence in dendritic cells through an HMGB1-dependent mechanism.


Earlier time to hemostasis is associated with decreased mortality and rate of complications: Results from the Pragmatic Randomized Optimal Platelet and Plasma Ratio trial.
Clinicians intuitively recognize that faster time to hemostasis is important in bleeding trauma patients, but these times are rarely reported. Prospectively collected data from the Pragmatic Randomized Optimal Platelet and Plasma Ratios trial were analyzed. Hemostasis was predefined as no intraoperative bleeding requiring intervention in the surgical field or resolution of contrast blush on interventional radiology (IR). Patients who underwent an emergent (within 90 minutes) operating room (OR) or IR procedure were included. Mixed-effects Poisson regression with robust error variance (controlling for age, Injury Severity Score, treatment arm, injury mechanism, base excess on admission [missing values estimated by multiple imputation], and time to OR/IR as fixed effects and study site as a random effect) with modified Bonferroni corrections tested the hypothesis that decreased time to hemostasis was associated with decreased mortality and decreased incidence of acute kidney injury (AKI), acute respiratory distress syndrome (ARDS), multiple-organ failure (MOF), sepsis, and venous thromboembolism. Of 680 enrolled patients, 468 (69%) underwent an emergent procedure. Patients with decreased time to hemostasis were less severely injured, had less deranged base excess on admission, and lower incidence of blunt trauma (all p < 0.05). In 408 (87%) patients in whom hemostasis was achieved, every 15-minute decrease in time to hemostasis was associated with decreased 30-day mortality (RR, 0.97; 95% confidence interval [CI], 0.94-0.99), AKI (RR, 0.97; 95% CI, 0.96-0.98), ARDS (RR, 0.98; 95% CI, 0.97-0.99), MOF (RR, 0.94; 95% CI, 0.91-0.97), and sepsis (RR, 0.98; 95% CI, 0.96-0.99), but not venous thromboembolism (RR, 0.99; 95% CI, 0.96-1.03). Earlier time to hemostasis was independently associated with decreased incidence of 30-day mortality, AKI, ARDS, MOF, and sepsis in bleeding trauma patients. Time to hemostasis should be considered as an endpoint in trauma studies and as a potential quality indicator. Therapeutic/care management, level III.


Privacy in the age of psychological targeting.
Psychological targeting describes the practice of extracting people's psychological profiles from their digital footprints (e.g. their Facebook Likes, Tweets or credit card records) in order to influence their attitudes, emotions or behaviors through psychologically informed interventions at scale. We discuss how the increasingly blurred lines between public and private information, and the continuation of the outdated practices of notice and consent, challenge traditional conceptualizations of privacy in the context of psychological targeting. Drawing on the theory of contextual integrity, we argue that it is time to rethink privacy and move beyond the questions of who collects what data to how the data are being used. Finally, we suggest that regulations of psychological targeting should be accompanied by a mindset that fosters (1) privacy by design to make it easy for individuals to act in line with their privacy goals, as well as (2) disclosure by choice, to allow individuals to freely decide whether and when they might be willing to forsake their privacy for better service.


Acute effects of ambient temperature on hypotension hospital visits: A time-series analysis in seven metropolitan cities of Korea from 2011 to 2015.
Although blood pressure decreases in response to high ambient temperature, little is known about whether the ambient temperature can induce clinical hypotension events. Therefore, we conducted a time-series analysis to evaluate the association between hypotension hospital visits and ambient temperature in seven metropolitan cities of Korea. We used the National Health Insurance Database, which contains the complete hospital visit data of the entire Korean population. We collected hospital visit data of seven metropolitan cities and linked the number of daily hypotension hospital visits to city-level ambient temperature, relative humidity, and air pollution levels from 2011 to 2015. Time-series analysis using the Poisson generalized additive model was conducted for each metropolitan city and we meta-analyzed the time-series results using the random effect model. There were 132,097 hospital visits for hypotension during our study period. A 1 degree Celsius (°C) increase in ambient temperature was associated with 1.1% increase in hospital visits for hypotension on lag day 0. Effects of ambient temperature lasted for 7 days, showing greater effects in shorter lag days. Subgroup analysis by sex and income groups showed similar results, but effects of ambient temperature on hypotension hospital visits was higher in the younger age group compared to older age group (aged over 65 years old). The results were unchanged when we applied cumulative lags, different case definitions, degrees of freedom per year, and multi-pollutant model adjusting for air pollutants. Hospital visits for hypotension were positively associated with ambient temperature. Increased hypotension events in response to increased ambient temperature might explain the high cardiovascular mortality on hot days.


The international autoimmune hepatitis score in chronic hepatitis C.
Clinical and laboratory findings of autoimmunity are common in chronic hepatitis C. Autoimmune hepatitis (AIH), a disease of unknown cause, has been defined by use of the International Autoimmune Hepatitis Group Score (AIH score), which quantifies clinical and laboratory parameters. To further validate the specificity of the International AIH score and investigate the similarities between hepatitis C and AIH, we measured the International Autoimmune Hepatitis Group Score in patients with well-defined chronic hepatitis C. Thirty consecutive non-cirrhotic patients with chronic hepatitis C were evaluated. Scoring was performed using both components of the AIH score: a set of minimum required parameters including laboratory and historical data and a second set of additional parameters dominated by histological criteria. Autoantibodies were positive in 21 of 30 hepatitis C patients and associated (patient or first-degree relative) autoimmune diseases were present in eight of 30 patients. Histologically, chronic active hepatitis with periportal piecemeal necrosis was seen in 24 of 30 patients and lymphoid follicles in 16 of 30 patients. No patient scored as probable or definite AIH using the minimum required parameters of the AIH score. When histological parameters were included, four of 30 patients scored as probable AIH but none as definite AIH. Therefore, AIH was excluded by the minimal and additional criteria of the AIH score in 86% of patients with hepatitis C despite a high prevalence of autoantibodies in these patients. We conclude that the criteria set forth by the International AIH scoring system defines a distinct disease although it shares some features with chronic hepatitis C. Modification of the AIH scoring system to include other commonly accepted risk factors for hepatitis C and additional histological parameters would further improve its specificity.


Hypoxia induced tumor metabolic switch contributes to pancreatic cancer aggressiveness.
Pancreatic ductal adenocarcinoma remains one of the most lethal of all solid tumors with an overall five-year survival rate of only 3-5%. Its aggressive biology and resistance to conventional and targeted therapeutic agents lead to a typical clinical presentation of incurable disease once diagnosed. The disease is characterized by the presence of a dense stroma of fibroblasts and inflammatory cells, termed desmoplasia, which limits the oxygen diffusion in the organ, creating a strong hypoxic environment within the tumor. In this review, we argue that hypoxia is responsible for the highly aggressive and metastatic characteristics of this tumor and drives pancreatic cancer cells to oncogenic and metabolic changes facilitating their proliferation. However, the molecular changes leading to metabolic adaptations of pancreatic cancer cells remain unclear. Cachexia is a hallmark of this disease and illustrates that this cancer is a real metabolic disease. Hence, this tumor must harbor metabolic pathways which are probably tied in a complex inter-organ dialog during the development of this cancer. Such a hypothesis would better explain how under fuel source limitation, pancreatic cancer cells are maintained, show a growth advantage, and develop metastasis.


Ground Reaction Force Differences between Bionic Shoes and Neutral Running Shoes in Recreational Male Runners before and after a 5 km Run.
Running-related injuries are common among runners. Recent studies in footwear have shown that designs of shoes can potentially affect sports performance and risk of injury. Bionic shoes combine the functions of barefoot running and foot protection and incorporate traditional unstable structures based on bionic science. The purpose of this study was to investigate ground reaction force (GRF) differences for a 5 km run and how bionic shoes affect GRFs. Sixteen male recreational runners volunteered to participate in this study and finished two 5 km running sessions (a neutral shoe session and a bionic shoe session). Two-way repeated-measures ANOVAs were performed to determine the differences in GRFs. In the analysis of the footwear conditions of runners, bionic shoes showed significant decreases in vertical impulse, peak propulsive force, propulsive impulse, and contact time, while the braking impulse and vertical instantaneous loading rate (VILR) increased significantly compared to the neutral shoes. Main effects for a 5 km run were also observed at vertical GRFs and anterior-posterior GRFs. The increases of peak vertical impact force, vertical average loading rate (VALR), VILR, peak braking force and braking impulse were observed in post-5 km running trials and a reduction in peak propulsive force and propulsive impulse. The interaction effects existed in VILR and contact time. The results suggest that bionic shoes may benefit runners with decreasing injury risk during running. The findings of the present study may help to understand the effects of footwear design during prolonged running, thereby providing valuable information for reducing the risk of running injuries.


Muffin-like lanthanide complexes with an N5O2-donor macrocyclic ligand showing field-induced single-molecule magnet behaviour.
Three mononuclear lanthanide complexes of a 2-pyridylmethyl pendant-armed 15-membered ligand {(3,12-bis(2-pyridylmethyl)-3,12,18-triaza-6,9-dioxabicyclo-[12.3.1]octadeca-1,14,16-triene); L} with general formula [Ln(L)(H2O)(NO3)](NO3)2 (Ln = Tb (1), Dy (2), and Er (3)) are reported. Based on X-ray diffraction analysis of 1 and 2, the central lanthanide atoms are nine-coordinated with the N5O4 donor set originating from the ligand L and one coordinated water molecule and one monodentate-bonded nitrato ligand. The coordination geometry of the [LnN5O4] cores can be described as a muffin-like shape. Magnetic measurements revealed that all three compounds show field-induced single-molecule magnet behaviour, with estimated energy barriers U ≈ 44-82 K. The experimental study was complemented by CASSCF calculations showing a trend of an increasing first excited energy gap (Tb → Dy → Er) within the muffin-like geometry with the lowest magnetization tunnelling probability for the DyIII complex 2.


Recommendations for the diagnosis and management of vaccine-induced immune thrombotic thrombocytopenia.
There have recently been safety concerns regarding an increased risk of vaccine-induced immune thrombotic thrombocytopenia (VITT) following administration of SARS-CoV-2 adenoviral vector vaccines. The Southern African Society of Thrombosis and Haemostasis reviewed the emerging literature on this idiosyncratic complication. A draft document was produced and revised by consensus agreement by a panel of professionals from various specialties. The recommendations were adjudicated by independent international experts to avoid local bias. We present concise, practical guidelines for the clinical management of VITT.


Van Buchem disease: clinical, biochemical, and densitometric features of patients and disease carriers.
Van Buchem disease (VBD) is a rare bone sclerosing dysplasia caused by the lack of a regulatory element of the SOST gene, which encodes for sclerostin, an osteocyte-derived negative regulator of bone formation. We studied the demographic, clinical, biochemical, and densitometric features of 15 patients with VBD (12 adults and 3 children) and 28 related carriers of the gene mutation. The most common clinical findings in patients were facial palsy (100%) and various degrees of hearing impairment (93%); raised intracranial pressure had been documented in 20%. The clinical course of the disease appeared to stabilize in adulthood, with the majority of patients reporting no progression of symptoms or development of complications with time. Carriers of the disease had none of the clinical features or complications of the disease. Sclerostin could be detected in the serum in all but 1 VBD patients (mean 8.0 pg/mL; 95% confidence interval [CI], 4.9-11.0 pg/mL), and were lower than those of carriers (mean 28.7 pg/mL; 95% CI, 24.5-32.9 pg/mL; p < 0.001) and healthy controls (mean 40.0 pg/mL; 95% CI, 34.5-41.0 pg/mL; p < 0.). Serum procollagen type 1 amino-terminal propeptide (P1NP) levels were also significantly higher in adult patients (mean 96.0; 95% CI, 54.6-137.4 ng/mL versus mean 47.8; 95% CI, 39.4-56.2 ng/mL, p = 0.003 in carriers and mean 37.8; 95% CI, 34.5-41.0 ng/mL, p = 0.028 in healthy controls) and declined with age. Bone mineral density (BMD) was markedly increased in all patients (mean Z-score 8.7 ± 2.1 and 9.5 ± 1.9 at the femoral neck and spine, respectively); BMD of carriers was significantly lower than that of patients but varied widely (mean Z-scores 0.9 ± 1.0 and 1.3 ± 1.5 at the femoral neck and spine, respectively). Serum sclerostin levels were inversely correlated with serum P1NP levels (r = -0.39, p = 0.018) and BMD values (femoral neck r = -0.69, p < 0.001; lumbar spine r = -0.78, p < 0.001). Our results show that there is a gene-dose effect of the VBD deletion on circulating sclerostin and provide further in vivo evidence of the role of sclerostin in bone formation in humans. The small amounts of sclerostin produced by patients with VBD may explain their milder phenotype compared to that of patients with sclerosteosis, in whom serum sclerostin is undetectable.


Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain.
Interleukin-12 (IL-12) is a heterodimeric molecule composed of p35 and p40 subunits. Analyses in vitro have defined IL-12 as an important factor for the differentiation of naive T cells into T-helper type 1 CD4+ lymphocytes secreting interferon-gamma (refs 1, 2). Similarly, numerous studies have concluded that IL-12 is essential for T-cell-dependent immune and inflammatory responses in vivo, primarily through the use of IL-12 p40 gene-targeted mice and neutralizing antibodies against p40. The cytokine IL-23, which comprises the p40 subunit of IL-12 but a different p19 subunit, is produced predominantly by macrophages and dendritic cells, and shows activity on memory T cells. Evidence from studies of IL-23 receptor expression and IL-23 overexpression in transgenic mice suggest, however, that IL-23 may also affect macrophage function directly. Here we show, by using gene-targeted mice lacking only IL-23 and cytokine replacement studies, that the perceived central role for IL-12 in autoimmune inflammation, specifically in the brain, has been misinterpreted and that IL-23, and not IL-12, is the critical factor in this response. In addition, we show that IL-23, unlike IL-12, acts more broadly as an end-stage effector cytokine through direct actions on macrophages.


Prevention of venous thromboembolism in critically ill medical patients: a Franco-Canadian cross-sectional study.
Medical intensive care unit (ICU) patients are at moderate risk of venous thromboembolism (VTE) and prophylaxis against VTE is recommended. To observe the range and frequency of VTE prophylaxis administered to medical ICU patients and to determine factors associated with different strategies in French and Canadian ICUs. Prospective cross-sectional observational study. 113/251 (45.0%) French and 29/30 (96.6%) Canadian ICUs agreed to participate. Of 1,222 critically ill medical patients, most were mechanically ventilated (62.5%). Overall, heparin VTE prophylaxis was administered to 63.9% patients, similarly between the 2 countries. Excluding patients with contraindications to heparin and those receiving therapeutic anticoagulation, 91.7% of medical ICU patients appropriately received either low dose unfractionated heparin (UFH) or low molecular weight heparin (LMWH) prophylaxis. Independent predictors of heparin prophylaxis were invasive mechanical ventilation (odds ratio [OR]; 95%CI, 2.4 (1.4-4.3) and obesity (OR 3.1; 1.1-8.8). LMWH was less likely to be prescribed for patients with renal failure (OR 0.1; 0.0009-0.9), or receiving antiembolic stockings (OR 0.4, 0.1-0.9), and much more likely to be prescribed in French ICUs (OR 9.2; 5.0-16.9); however, among patients receiving LMWH, high doses were more likely to be prescribed in Canadian ICUs (OR 8.7; 2.0-37.6). Patients who were pregnant or postpartum (OR 7.7, 1.3-44.3), had neurologic failure (OR 2.1, 1.3-3.4), or were Canadian (OR 3.0, 2.1-4.4) were most likely to receive mechanical VTE prophylaxis (with antiembolic stockings or pneumatic compression devices), whereas those who were already receiving heparin were less likely to receive mechanical prophylaxis (OR 0.5, 0.3-0.7). In this binational cross-sectional observational study of medical ICU patients, we found that 92% of eligible patients received either UFH or LWMH for VTE prophylaxis. Differences in prescribing between countries include significantly greater use of LMWH in France, but use of lower doses than in Canada, and greater use of mechanical VTE prophylaxis in Canada. More randomized trials of VTE prophylaxis in critically ill medical patients would better inform practice.


Characterization of auxotrophic mutants of Mycobacterium tuberculosis and their potential as vaccine candidates.
Auxotrophic mutants of Mycobacterium tuberculosis have been proposed as new vaccine candidates. We have analyzed the virulence and vaccine potential of M. tuberculosis strains containing defined mutations in genes involved in methionine (metB), proline (proC), or tryptophan (trpD) amino acid biosynthesis. The metB mutant was a prototrophic strain, whereas the proC and trpD mutants were auxotrophic for proline and tryptophan, respectively. Following infection of murine bone marrow-derived macrophages, H37Rv and the metB mutant strain survived intracellularly for over 10 days, whereas over 90% of proC and trpD mutants were killed during this time. In SCID mice, both H37Rv and the metB mutant were highly virulent, with mouse median survival times (MST) of 28.5 and 42 days, respectively. The proC mutant was significantly attenuated (MST, 130 days), whereas the trpD mutant was essentially avirulent in an immunocompromised host. Following infection of immunocompetent DBA mice with H37Rv, mice survived for a median of 83.5 days and the metB mutant now showed a clear reduction in virulence, with two of five infected mice surviving for 360 days. Both proC and trpD mutants were avirulent (MST of >360 days). In vaccination studies, prior infection with either the proC or trpD mutant gave protection equivalent (proC mutant) to or better (trpD mutant) than BCG against challenge with M. tuberculosis H37Rv. In summary, proC and trpD genes are essential for the virulence of M. tuberculosis, and mutants with disruptions in either of these genes show strong potential as vaccine candidates.


Hyperthermia-regulated immunogene therapy.
One of the key milestones that must be reached before gene therapy becomes feasible for clinical cancer treatment is to be able to regulate therapeutic gene expression. This is true for most current cancer gene therapy approaches, since the majority of therapeutic genes are toxic to both tumour and normal tissues. Among the wide array of strategies available for regulating gene expression, hyperthermia represents a unique approach. Hyperthermic regulation of gene therapy is feasible because of the widely conserved heat shock response, which allows therapeutic gene expression to be elevated to thousands of fold higher than background when temperature reaches 3-5 degrees C over physiological temperature (37 degrees C). In addition, because of the long history of experimental research on the use of hyperthermia as an approach for cancer therapy, it is now quite feasible to apply hyperthermia to a number of tumour sites and to achieve temperatures that are sufficient to induce a heat shock response. This review will attempt to discuss the current status of hyperthermia-regulated gene therapy, with special emphasis on hyperthermia-regulated immunogene therapy.


Quantification of optic disc edema during exposure to high altitude shows no correlation to acute mountain sickness.
The study aimed to quantify changes of the optic nerve head (ONH) during exposure to high altitude and to assess a correlation with acute mountain sickness (AMS). This work is related to the Tuebingen High Altitude Ophthalmology (THAO) study. A confocal scanning laser ophthalmoscope (cSLO, Heidelberg Retina Tomograph, HRT3®) was used to quantify changes at the ONH in 18 healthy participants before, during and after rapid ascent to high altitude (4559 m). Slitlamp biomicroscopy was used for clinical optic disc evaluation; AMS was assessed with Lake Louise (LL) and AMS-cerebral (AMS-c) scores; oxygen saturation (SpO₂) and heart rate (HR) were monitored. These parameters were used to correlate with changes at the ONH. After the first night spent at high altitude, incidence of AMS was 55% and presence of clinical optic disc edema (ODE) 79%. Key stereometric parameters of the HRT3® used to describe ODE (mean retinal nerve fiber layer [RNFL] thickness, RNFL cross sectional area, optic disc rim volume and maximum contour elevation) changed significantly at high altitude compared to baseline (p<0.05) and were consistent with clinically described ODE. All changes were reversible in all participants after descent. There was no significant correlation between parameters of ODE and AMS, SpO₂ or HR. Exposure to high altitude leads to reversible ODE in the majority of healthy subjects. However, these changes did not correlate with AMS or basic physiologic parameters such as SpO₂ and HR. For the first time, a quantitative approach has been used to assess these changes during acute, non-acclimatized high altitude exposure. In conclusion, ODE presents a reaction of the body to high altitude exposure unrelated to AMS.


Trends and transformation of nutrients and pesticides in a coastal plain aquifer system, United States.
Four local-scale sites in areas with similar corn (Zea mays L.) and soybean [Glycine max (L.) Merr.] agriculture were studied to determine the effects of different hydrogeologic settings of the Northern Atlantic Coastal Plain (NACP) on the transport of nutrients and pesticides in groundwater. Settings ranged from predominantly well-drained soils overlying thick, sandy surficial aquifers to predominantly poorly drained soils with complex aquifer stratigraphy and high organic matter content. Apparent age of groundwater, dissolved gases, N isotopes, major ions, selected pesticides and degradates, and geochemical environments in groundwater were studied. Agricultural chemicals were the source of most dissolved ions in groundwater. Specific conductance was strongly correlated with reconstructed nitrate (the sum of N in nitrate and N gas) (R(2) = 0.81, p < 0.0001), and is indicative of the relative degree of agricultural effects on groundwater. Trends in nitrate were primarily related to changes in manure and fertilizer use at the well-drained sites where aquifer conditions were consistently oxic. Nitrate was present in young groundwater but completely removed over time through denitrification at the poorly drained sites where there were variations in chemical input and in geochemical environment. Median concentrations of atrazine (6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine), metolachlor (2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl)acetamide), and some of their common degradates were higher at well-drained sites than at poorly drained sites, with concentrations of degradates generally higher than those of the parent compounds at all sites. An increase in the percentage of deethylatrazine to total atrazine over time at one well-drained site may be related to changes in manure application.


First report of barley root-knot nematode, Meloidogyne naasi from turfgrass in Idaho, with multigene molecular characterization.
Barley root-knot nematode, Meloidogyne naasi Franklin, 1965, is one of the most important pest nematodes infecting monocots (Franklin, 1965). Two-inch core soil samples collected from a golf course in Ada County, Idaho were submitted for identification in November of 2019. A high number of Meloidogyne sp. juveniles were recovered from both soil samples using sieving and decantation followed by the sugar centrifugal flotation method. They were examined by light microscopy, morphometric measurements, and multiple molecular markers, including the ribosomal 28S D2-D3 and intergenic spacer 2 (IGS-2) regions, mitochondrial markers cytochrome oxidase I (COI) and the interval from COII to 16S, and the protein-coding gene Hsp90. Morphometrics as well as BlastN comparisons with other root-knot nematode sequences from GenBank were consistent with identification as M. naasi. Phylogenetic trees inferred from 28S, IGS-2, COI, or Hsp90 alignments each separated the Idaho population into a strongly supported clade with other populations of M. naasi, while the COII-16S interval could not resolve M. naasi from M. minor. This report represents the first morphological and molecular characterization of Meloidogyne naasi from turfgrass in Idaho.


Western spruce budworm outbreaks did not increase fire risk over the last three centuries: a dendrochronological analysis of inter-disturbance synergism.
Insect outbreaks are often assumed to increase the severity or probability of fire occurrence through increased fuel availability, while fires may in turn alter susceptibility of forests to subsequent insect outbreaks through changes in the spatial distribution of suitable host trees. However, little is actually known about the potential synergisms between these natural disturbances. Assessing inter-disturbance synergism is challenging due to the short length of historical records and the confounding influences of land use and climate changes on natural disturbance dynamics. We used dendrochronological methods to reconstruct defoliator outbreaks and fire occurrence at ten sites along a longitudinal transect running from central Oregon to western Montana. We assessed synergism between disturbance types, analyzed long-term changes in disturbance dynamics, and compared these disturbance histories with dendroclimatological moisture availability records to quantify the influence of moisture availability on disturbances. After approximately 1890, fires were largely absent and defoliator outbreaks became longer-lasting, more frequent, and more synchronous at our sites. Fires were more likely to occur during warm-dry years, while outbreaks were most likely to begin near the end of warm-dry periods. Our results show no discernible impact of defoliation events on subsequent fire risk. Any effect from the addition of fuels during defoliation events appears to be too small to detect given the overriding influence of climatic variability. We therefore propose that if there is any relationship between the two disturbances, it is a subtle synergistic relationship wherein climate determines the probability of occurrence of each disturbance type, and each disturbance type damps the severity, but does not alter the probability of occurrence, of the other disturbance type over long time scales. Although both disturbance types may increase in frequency or extent in response to future warming, our records show no precedent that western spruce budworm outbreaks will increase future fire risk.


A prospective, multi-institutional study of flexible ureteroscopy for proximal ureteral stones smaller than 2 cm.
Flexible ureteroscopy is rapidly becoming a first line therapy for many patients with renal and ureteral stones. However, current understanding of treatment outcomes in patients with isolated proximal ureteral stones is limited. Therefore, we performed a prospective, multi-institutional study of ureteroscopic management of proximal ureteral stones smaller than 2 cm to better define clinical outcomes associated with this approach. Adult patients with proximal ureteral calculi smaller than 2 cm were prospectively identified. Patients with concomitant ipsilateral renal calculi or prior ureteral stenting were excluded from study. Flexible ureteroscopy, holmium laser lithotripsy and ureteral stent placement was performed. Ureteral access sheath use, laser settings and other details of perioperative and postoperative management were based on individual surgeon preference. Stone clearance was determined by the results of renal ultrasound and plain x-ray of the kidneys, ureters and bladder 4 to 6 weeks postoperatively. Of 71 patients 44 (62%) were male and 27 (38%) were female. Mean age was 48.2 years. ASA(®) score was 1 in 12 cases (16%), 2 in 41 (58%), 3 in 16 (23%) and 4 in 2 (3%). Mean body mass index was 31.8 kg/m(2), mean stone size was 7.4 mm (range 5 to 15) and mean operative time was 60.3 minutes (range 15 to 148). Intraoperative complications occurred in 2 patients (2.8%), including mild ureteral trauma. Postoperative complications developed in 6 patients (8.7%), including urinary tract infection in 3, urinary retention in 2 and flash pulmonary edema in 1. The stone-free rate was 95% and for stones smaller than 1 cm it was 100%. Flexible ureteroscopy is associated with excellent clinical outcomes and acceptable morbidity when applied to patients with proximal ureteral stones smaller than 2 cm.


A long-duration dihydroorotate dehydrogenase inhibitor (DSM265) for prevention and treatment of malaria.
Malaria is one of the most significant causes of childhood mortality, but disease control efforts are threatened by resistance of the Plasmodium parasite to current therapies. Continued progress in combating malaria requires development of new, easy to administer drug combinations with broad-ranging activity against all manifestations of the disease. DSM265, a triazolopyrimidine-based inhibitor of the pyrimidine biosynthetic enzyme dihydroorotate dehydrogenase (DHODH), is the first DHODH inhibitor to reach clinical development for treatment of malaria. We describe studies profiling the biological activity, pharmacological and pharmacokinetic properties, and safety of DSM265, which supported its advancement to human trials. DSM265 is highly selective toward DHODH of the malaria parasite Plasmodium, efficacious against both blood and liver stages of P. falciparum, and active against drug-resistant parasite isolates. Favorable pharmacokinetic properties of DSM265 are predicted to provide therapeutic concentrations for more than 8 days after a single oral dose in the range of 200 to 400 mg. DSM265 was well tolerated in repeat-dose and cardiovascular safety studies in mice and dogs, was not mutagenic, and was inactive against panels of human enzymes/receptors. The excellent safety profile, blood- and liver-stage activity, and predicted long half-life in humans position DSM265 as a new potential drug combination partner for either single-dose treatment or once-weekly chemoprevention. DSM265 has advantages over current treatment options that are dosed daily or are inactive against the parasite liver stage.


Fetal responses to acute maternal cocaine injection in sheep.
Maternal cocaine abuse has been associated with neonatal neurological and neurobehavioral problems of unknown pathogenesis. We administered a single intravenous dose of cocaine (2 mg/kg) to 12 unanesthetized pregnant sheep; their fetuses had been catheterized in utero 2 days before the study. We measured fetal cerebral blood flow (CBF), cerebral metabolic rate of O2 (CMRO2), mean arterial blood pressure (MAP), and blood gases before and 2, 5, 15, and 30 min after maternal cocaine injection. Fetal CBF increased by 37 +/- 33% (mean +/- SD) at 5 min and returned to baseline by 15 min. Regional brain blood flow changes paralleled CBF changes with the greatest increases occurring in cerebellum (54 +/- 43%) and brain stem (54 +/- 52%). Cerebral vascular resistance was decreased for cerebellum (22%) and brain stem (19%) but was unchanged for cerebral hemispheres and caudate. Increased CBF at 5 min was associated with a 20 +/- 9% increase in fetal MAP and a 38 +/- 13% decrease in fetal arterial O2 content. Fetal CMRO2 was unchanged. There was a decrease in fetal intestinal blood flow at 2 min, an increase in myocardial, adrenal, and renal blood flow at 5 min, and no change in placental blood flow. Maternal cocaine injection causes fetal hypoxemia, hypertension, and increased CBF. Possible mechanisms for cerebral vasodilation (in some areas) include hypoxemia, impaired autoregulatory response to increased blood pressure, and/or direct or indirect vascular effects of cocaine or its metabolites.


Terminal coma affects messenger RNA detection in post mortem human temporal cortex.
In situ hybridization histochemistry has been used to study the amount of M1 muscarinic receptor mRNA in temporal cortex from subjects with Alzheimer's disease and other neurodegenerative disorders, where the duration of terminal coma was known. Total polyadenylated mRNA and glutamate decarboxylase activity were also measured. Both muscarinic receptor mRNA and enzyme activity showed a significant decline with increasing duration of terminal coma, but were not related to diagnosis. Polyadenylated mRNA signal did not show an association with coma. These data indicate the need to consider the nature of the terminal illness in post mortem studies of mRNA as well as for neurochemical research.


Anti-inflammatory functions of Houttuynia cordata Thunb. and its compounds: A perspective on its potential role in rheumatoid arthritis.
The aim of this review was to take a look at the anti-inflammatory functions of Houttuynia cordata Thunb. (HCT) that have been illustrated in the literature and to explore new fields in which HCT could be used in the future. The use of HCT has been described in broad inflammatory domains, where it has exhibited a variety of activities, including antiviral, antibacterial, antiparasitic and immunostimulant activity, with high efficiency, mild features and definite therapeutic effects. The numerous anti-inflammatory functions of HCT have demonstrated that HCT has wide application prospects. New uses of HCT and the full extent of its utilization await further investigation. The basic pathological change of rheumatoid arthritis (RA) is synovial proliferation which leads to joint destruction in the long-term. There are types of drugs that have been used clinically for patients with RA, however, due to their side-effects or high prices their broad usage is limited. A safe and low-cost drug is urgently required to be developed for the clinical usage of patients with RA. Thus, HCT has the potential to be a good candidate in the treatment of rheumatoid arthritis.


Effect of melatonin on tinnitus.
Evaluate melatonin as a treatment for subjective tinnitus. Randomized, prospective, double-blind, placebo-controlled crossover trial. Patients were given 3.0 mg melatonin, which was taken nightly for 30 days followed or preceded by a placebo nightly for 30 days, with a 7-day washout period between medications. Outpatient, private, neurotology practice. Thirty patients with subjective tinnitus. Tinnitus matching, Tinnitus Handicap Inventory (THI), patient questionnaire and interview. The average pretreatment THI score was 33.91 as compared with 26.43 after the placebo and 26.09 after melatonin. The difference in the THI scores between melatonin and placebo treatment were not statistically significant. The average pretreatment THI score for patients who reported overall improvement with melatonin was statistically higher (P = 0.02) than the average pretreatment THI score for patients who reported no improvement with melatonin. Among subjects reporting difficulty sleeping attributable to their tinnitus, 46.7% reported an overall improvement after melatonin compared with 20.0% for placebo (P = 0.04). There was also a statistically significant difference in improvement with melatonin for those patients with bilateral tinnitus compared with those with unilateral tinnitus (P = 0.02). Melatonin has been shown to be useful in the treatment of subjective tinnitus. Patients with high THI scores and/or difficulty sleeping are most likely to benefit from treatment with melatonin. In light of its minimal side effects, melatonin should be a part of the physician's armamentarium in the treatment of tinnitus.


rs4711751 and rs1999930 are not associated with neovascular age-related macular degeneration or polypoidal choroidal vasculopathy in the Chinese population.
rs1999930 and rs4711751 have recently been identified as novel variants associated with advanced age-related macular degeneration (AMD) in populations of European ancestry. We aimed to investigate whether these two single nucleotide polymorphisms (SNPs) were associated with neovascular AMD (nAMD) or with polypoidal choroidal vasculopathy (PCV), a variant of AMD in Asians, using a Chinese case-control study. A total of 900 subjects, including 300 controls, 300 cases with nAMD and 300 cases with PCV, were included in the present study. Genomic DNA was extracted from venous blood leukocytes. The allelic variants of rs1999930 and rs4711751 were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The differences in allele distribution between cases and controls were tested by a χ(2) test, with additional adjustments for age and gender using logistic regression. The statistical power was also calculated. Values of p < 0.05 were considered statistically significant. No statistically significant association was observed between the two polymorphisms of nAMD or PCV phenotype (p > 0.05 for all comparisons). The difference remained insignificant after correction for age and gender (p > 0.05 for all comparisons). The statistical powers to detect the association between these two SNPs and nAMD or PCV range from 0.05 to 0.36, assuming conventional levels of statistical significance. In the present study, we could not replicate the reported association of these two SNPs and either nAMD or PCV in a Chinese population, suggesting that they are unlikely to be a major AMD and PCV susceptibility gene locus in the Chinese population. Considering the low power value, a large sample size is required to draw more reliable conclusions.


The occurrence of dermatosparaxis in a commercial Drakensberger cattle herd in South Africa.
Dermatosparaxis is a heritable collagen dysplasia causing skin extensibility and fragility. In Belgian Blue cattle this mutation has been described as a 3 base pair (bp) change followed by a 17bp deletion in the gene coding for procollagen 1 N-Proteinase (pNPI). An outbreak in a commercial Drakensberger herd in South Africa followed the introduction in late 2000 of a 3-year-old bull that developed skin lesions in 2001 and was culled in 2002. Some of his offspring were similarly affected, 1 of which was kept as a breeding bull after his sire's death. Two affected calves were referred to the Onderstepoort Veterinary Academic Hospital in October 2005. Detailed examination revealed only skin abnormalities limited to the lateral extremities of the thorax, abdomen and pelvis, viz. either acute lacerations of varying sizes, slow healing defects or thin scars in chronic cases. During a subsequent farm visit, 13 animals with similar wounds were seen in the herd of 146 animals. Electron microscopic examination of skin biopsies revealed haphazard arrangement and loose packing of dermal collagen fibrils within collagen fibres. The fibrils showed size variation and slightly irregular outlines on cross-section, consistent with mild dermatosparaxis. DNA samples of affected calves were analysed using primers designed to amplify the region of the pNPI gene that contained the mutation described in Belgian Blue cattle, but this mutation could not be demonstrated in any of the animals tested. It is concluded that a form of dermatosparaxis with a different gene mutation from that described in Belgian Blue cattle exists in Drakensberger cattle in South Africa. This possibly also explains the milder and more delayed clinical signs and the milder dermal collagen ultrastructural abnormalities.


The M50I polymorphic substitution in association with the R263K mutation in HIV-1 subtype B integrase increases drug resistance but does not restore viral replicative fitness.
First-generation integrase strand-transfer inhibitors (INSTIs), such as raltegravir (RAL) and elvitegravir (EVG), have been clinically proven to be effective antiretrovirals for the treatment of HIV-positive patients. However, their relatively low genetic barrier for resistance makes them susceptible to the emergence of drug resistance mutations. In contrast, dolutegravir (DTG) is a newer INSTI that appears to have a high genetic barrier to resistance in vivo. However, the emergence of the resistance mutation R263K followed by the polymorphic substitution M50I has been observed in cell culture. The M50I polymorphism is also observed in 10-25% of INSTI-naïve patients and has been reported in combination with R263K in a patient failing treatment with RAL. Using biochemical cell-free strand-transfer assays and resistance assays in TZM-bl cells, we demonstrate that the M50I polymorphism in combination with R263K increases resistance to DTG in tissue culture and in biochemical assays but does not restore the viral fitness cost associated with the R263K mutation. Since the combination of the R263K mutation and the M50I polymorphism results in a virus with decreased viral fitness and limited cross-resistance, the R263K resistance pathway may represent an evolutionary dead-end. Although this hypothesis has not yet been proven, it may be more advantageous to treat HIV-positive individuals with DTG in first-line than in second or third-line therapy.


Social Drinking on Social Media: Content Analysis of the Social Aspects of Alcohol-Related Posts on Facebook and Instagram.
Alcohol is often consumed in social contexts. An emerging social context in which alcohol is becoming increasingly apparent is social media. More and more young people display alcohol-related posts on social networking sites such as Facebook and Instagram. Considering the importance of the social aspects of alcohol consumption and social media use, this study investigated the social content of alcohol posts (ie, the evaluative social context and presence of people) and social processes (ie, the posting of and reactions to posts) involved with alcohol posts on social networking sites. Participants (N=192; mean age 20.64, SD 4.68 years, 132 women and 54 men) gave researchers access to their Facebook and/or Instagram profiles, and an extensive content analysis of these profiles was conducted. Coders were trained and then coded all screenshotted timelines in terms of evaluative social context, presence of people, and reactions to post. Alcohol posts of youth frequently depict alcohol in a positive social context (425/438, 97.0%) and display people holding drinks (277/412, 67.2%). In addition, alcohol posts were more often placed on participants' timelines by others (tagging; 238/439, 54.2%) than posted by participants themselves (201/439, 45.8%). Furthermore, it was revealed that such social posts received more likes (mean 35.50, SD 26.39) and comments than nonsocial posts (no people visible; mean 10.34, SD 13.19, P<.001). In terms of content and processes, alcohol posts on social media are social in nature and a part of young people's everyday social lives. Interventions aiming to decrease alcohol posts should therefore focus on the broad social context of individuals in which posting about alcohol takes place. Potential intervention strategies could involve making young people aware that when they post about social gatherings in which alcohol is visible and tag others, it may have unintended negative consequences and should be avoided.


The immune response of the mouse to lymphocytic choriomeningitis virus. V. High numbers of cytolytic T lymphocytes are generated in the spleen during acute infection.
In the spleens of C57BL/6J (B6) and CBA/J (CBA) mice undergoing acute infection with lymphocytic choriomeningitis (LCM) virus, lymphocytes with the ability to develop in vitro into LCM virus-specific cytolytic clones were enumerated by use of the limiting dilution method. At intervals after virus inoculation, defined numbers of cells were cultivated with virus-infected syngeneic stimulator cells and T cell growth factor in multiple wells of microculture plates. After 7 days, individual cell cultures were tested for their ability to cause release of 51Cr from infected and uninfected syngeneic target cells. In cultures seeded with spleen cells from uninfected mice or from mice infected 3 days previously, no cytolytic activity was observed. On day 5, cells developing into LCM virus-specific cytolytic effector cells were detected. They rose in numbers, and on days 8 to 9 after infection, values of approximately 1/10 and 1/200 in B6 and CBA mice, respectively, were calculated. A low proportion of microcultures proved cytolytic also for noninfected syngeneic target cells, but the counts thus released were consistently much lower than the counts set free from infected targets, and no regular dose-response relationships existed between seeded cells and positive cultures. Determination of cell surface antigens of responder cells by negative and positive selection procedures disclosed that they were predominantly T lymphocytes and expressed Lyt-2 but not L3T4 surface markers. Lysis by the great majority of LCM virus-specific clones was restricted by products of the major histocompatibility gene complex (MHC), but a few lysed, in addition, allogeneic infected or uninfected targets; however, a consistent pattern of alloreactivity was not observed. Furthermore, cells of a proportion of the cultures also lysed uninfected YAC cells. Probably this natural killer-like activity was acquired by T lymphocytes during prolonged cultivation. We conclude that most spleen cells that during acute infection with LCM virus attained the ability to develop in vitro into LCM virus-specific cytolytic clones were derived from MHC-restricted Lyt-2+, L3T4- antigen-specific cytolytic T lymphocytes and their activated precursors.


Cine magnetic resonance imaging after surgical repair in patients with transposition of the great arteries.
Cine magnetic resonance imaging (MRI) was used for postoperative evaluation of eight patients who underwent intra-atrial baffle procedure for surgical repair of D-transposition of the great arteries (D-TGA). Their ages ranged from 9 months to 8 years. Younger patients were sedated with chloral hydrate (80 to 100 mg/kg) orally. MRI was performed with use of a General Electric Signa system operating at a field strength of 1.5 tesla. A body or head coil was used depending on the size of the patient. Images were obtained by use of a technique of gradient-recalled acquisition in steady state (GRASS) that utilizes a low flip angle and shorter repetition and echo times. Five patients had widely patent venae cavae and three had superior vena caval obstruction at the junction of the right atrium with a dilated azygos vein. There was no evidence of pulmonary venous obstruction in any of the patients. Right ventricular function was assessed in four patients and their ejection fractions ranged from 58% to 81%. Tricuspid and mitral regurgitation were observed in three and two patients, respectively. Both right and left ventricular outflow tracts were well visualized and showed no evidence of obstruction. Cine MRI is an entirely noninvasive, nonionizing, and safe procedure in young patients and appears to be a valuable alternative method for evaluating patients after surgical repair of D-TGA. With advancing technologies and an accumulation of experience with cine MRI, it appears that this new technique will play an important role in patient care for children with congenital heart disease.


Connecting through summer camp: Youth with visual impairments find a sense of community.
The social meaning of a one-week residential summer sports camp to young people with visual impairments is described. The experiences of 13 youths (7 females and 6 males) with visual impairments (3 B1, 1 B2, and 9 B3) between 9 and 15 years of age were gathered using the phenomenological methods of focus groups, conversational interviews, and field notes. The thematic analysis revealed three themes: connected, reaching out, and resisting and acquiescing. Experiences of group membership and shared emotional connection to others with visual impairments surfaced in a supportive sport context although resistance to others' assumptions of ability was evident. The theory of psychological sense of community (McMillan & Chivas, 1986) provided the conceptual framework for interpreting the findings.


Musculoskeletal mnemonics: differentiating features.
Mnemonics are often used in musculoskeletal radiology to help radiologists remember long differential diagnoses. However, unless the specific appearance of each entity on a differential is also recalled, mnemonics become useless. This article presents 8 mnemonics with their corresponding differential diagnoses and distinguishing features. Bubbly lucent lesions of bone are recalled with the FEGNOMASHIC mnemonic, but when only lucent lesions of the diaphysis are included, a more appropriate mnemonic is FEMALE. The lucent lesions of bone differentials often can be narrowed based on specific characteristics of the lesion but radiographic findings elsewhere and clinical information often help. Osseous metastases may present as lucent or sclerotic lesions; when sclerotic, the differential is best remembered with the mnemonic 5 "BEES" Like Pollen. The mnemonic for Wormian bones is PORKCHOPS. The Wormian bones in most of these entities are indistinguishable, so one must rely on radiographic findings outside the skull for diagnosis. By contrast, differentiating causes of acro-osteolysis is often possible with findings seen only on the hand radiographs; the mnemonic for acro-osteolysis is RADSHIP. In skeletally immature patients with frayed metaphyses, the mnemonic is CHARMS. Although the appearance of the fraying is seldom diagnostic, findings in the adjacent portions of the long bones may be characteristic. FETISH is the mnemonic used to remember the entities for the differential diagnosis of vertebra plana. Age of the patient, clinical history, and findings in the adjacent spine often help to provide the specific diagnosis. Nearly all the entities on the differential diagnosis for distal clavicle erosion (mnemonic: SHIRT Pocket) are included in other differentials in this article.


Randomized Control Trial Investigating the Effects of Kinesiology Tape on Shoulder Proprioception.
Athletes participating in upper-extremity-dominant sports such as softball and volleyball are at increased risk for glenohumeral-joint pain and injury. For these athletes, an integral part of many injury-prevention and -rehabilitation programs includes improving joint proprioception. One way to measure joint proprioception is through the reproduction of joint angles, or joint-reposition sense (JRS). Kinesiology tape is purported to enhance neuromuscular feedback; therefore, it may influence JRS. However, conflicting findings and the lack of research in the upper extremity warrant further investigation. To determine the effects of kinesiology tape on shoulder-joint proprioception by actively reproducing joint angles, or measurement of JRS. Randomized controlled trial. College laboratory. 9 men and 7 women 24 ± 3 y old. SpiderTech kinesiology tape precut Shoulder Spider was applied to the shoulder of participants block randomized to the experimental group, following product-specific instructions, to measure its influence on JRS compared with a control group. JRS-error scores in shoulder flexion, extension, internal rotation, and external rotation (ER). There was a significant interaction between groups pre- to postintervention resulting in decreased JRS errors in flexion (P = .04) and ER (P = .03) in the experimental compared with the control group. The 95% confidence intervals suggest a clinically relevant difference in the variability of JRS errors between postintervention movements for the experimental group in flexion and ER, such that the control group demonstrated much more variability in JRS errors than the experimental group. After the application of kinesiology tape the JRS errors were smaller in flexion and ER. This may be of clinical significance in improving proprioception and thus improving joint stability. Additional research should determine the effectiveness of kinesiology tape in reducing joint injury.


The Value of Workshops on Psychological Flexibility for Early Childhood Special Education Staff.
High stress and burnout are common for early childhood special educators, contributing to high rates of attrition, diminished educational effectiveness, and high turnover. Acceptance and Commitment Therapy (ACT) is a promising approach for the prevention and treatment of a wide variety of problems. Using a randomized wait-list control design, this pilot study evaluated whether ACT workshops delivered to preschool teachers who serve children with developmental disabilities would improve stress-related problems of teachers (i.e., stress, depression, and burnout) and increase collegial support. At pretest, measures of experiential avoidance (EA) and mindful awareness (MA) showed significant relationships to reports of depression, stress, and burnout. The intervention reduced staff members' EA, increased teachers' MA and valued living (VL), and improved teachers' sense of efficacy. This suggests that ACT workshops can help influence factors affecting depression, stress, and burnout in an early childhood special education setting.


In vivo uptake kinetics of aminoglycosides in the kidney cortex of rats.
The renal cortical uptake kinetics of four aminoglycosides were studied in vivo. Gentamicin, netilmicin, tobramycin or amikacin were administered to rats by continuous infusion over 6 hr achieving constant serum levels ranging from 0.2 to 100 micrograms/ml. Renal cortical concentrations at the end of the infusion were plotted against the steady-state serum levels. Steady-state elevations of serum gentamicin and netilmicin were associated with nonlinear increases in cortical levels, suggesting saturable uptake. Analysis of the data using Michaelis-Menten kinetics indicates that the apparent Km for gentamicin and netilmicin were 15.01 and 23.84 micrograms/ml and Vmax 149.83 and 178.36 micrograms/g of cortex per hr, respectively. The "initial" rate of uptake (at serum levels below 15 micrograms/ml) was highest for gentamicin. The cortical uptake of tobramycin was linearly related to elevations in serum levels [cortex concentration (conc) = 9.24 + 1.40 serum conc]. The initial rate of tobramycin uptake was considerably lower than that for gentamicin and netilmicin. For amikacin, the initial rate of uptake followed Michaelis Menten kinetics and the second phase of the titration curve was linear. The equation for total amikacin uptake was: cortex conc = 12.98 + 1.71 serum conc. Aminoglycosides exhibit differing kinetics for renal cortical uptake in the rat during constant infusions. These results indicate that more than one mechanism probably mediates the uptake of each aminoglycoside. Depending on which mechanism predominates, the kinetic pattern may be saturable, linear or mixed.


EEGLAB: an open source toolbox for analysis of single-trial EEG dynamics including independent component analysis.
We have developed a toolbox and graphic user interface, EEGLAB, running under the crossplatform MATLAB environment (The Mathworks, Inc.) for processing collections of single-trial and/or averaged EEG data of any number of channels. Available functions include EEG data, channel and event information importing, data visualization (scrolling, scalp map and dipole model plotting, plus multi-trial ERP-image plots), preprocessing (including artifact rejection, filtering, epoch selection, and averaging), independent component analysis (ICA) and time/frequency decompositions including channel and component cross-coherence supported by bootstrap statistical methods based on data resampling. EEGLAB functions are organized into three layers. Top-layer functions allow users to interact with the data through the graphic interface without needing to use MATLAB syntax. Menu options allow users to tune the behavior of EEGLAB to available memory. Middle-layer functions allow users to customize data processing using command history and interactive 'pop' functions. Experienced MATLAB users can use EEGLAB data structures and stand-alone signal processing functions to write custom and/or batch analysis scripts. Extensive function help and tutorial information are included. A 'plug-in' facility allows easy incorporation of new EEG modules into the main menu. EEGLAB is freely available (http://www.sccn.ucsd.edu/eeglab/) under the GNU public license for noncommercial use and open source development, together with sample data, user tutorial and extensive documentation.


The Role of Mucosal Associated Invariant T Cells in Antimicrobial Immunity.
Mucosal associated invariant T (MAIT) cells are an innate-like T cell subset prevalent in humans and distributed throughout the blood and mucosal sites. Human MAIT cells are defined by the expression of the semi-invariant TCRα chain TRAV1-2/TRAJ12/20/33 and are restricted by the non-polymorphic major histocompatibility complex (MHC) class I-like molecule, MHC-related protein 1, MR1. MAIT cells are activated by small organic molecules, derived from the riboflavin biosynthesis pathway of bacteria and fungi, presented by MR1. Traditionally, MAIT cells were thought to recognize a limited number of antigens due to usage of an invariant TCRα chain and restriction by a non-polymorphic MHC molecule. However, recent studies demonstrate that the TCR repertoire of MAIT cells is more heterogeneous, suggesting there is a more diverse array of MR1 antigens that MAIT cells can recognize. In response to infected cells, MAIT cells produce the pro-inflammatory cytokines, IFN-γ and TNF, and are cytolytic. Studies performed in MR1-deficient mice suggest that MAIT cells can provide anti-bacterial control within the first few days post-infection, as well as contribute to enhanced adaptive immunity in murine models of respiratory infections. In humans, the role of MAIT cells is unclear; however, evidence points to interplay between MAIT cells and microbial infections, including Mycobacterium tuberculosis. Given that MAIT cells are pro-inflammatory, serve in early control of bacterial infections, and appear enriched at tissue sites where microbes interface and gain access to the body, we postulate that they play an important role in antimicrobial immune responses. In this review, we discuss the most recent studies on the function and phenotype of MAIT cells, including their TCR diversity and antigenic repertoire, with a focus on the contribution of human MAIT cells in the immune response to microbial infection.


Nigella sativa oil for prevention of chronic cyclosporine nephrotoxicity: an experimental model.
Nephrotoxicity is the main secondary effect of cyclosporine A (CsA) treatment. The antioxidant action of Nigella sativa oil (NSO) may explain the protective effect of these agents against various hepatotoxic and nephrotoxic models in vivo and in vitro. This study was designed to investigate the possible protective effects of NSO, in prevention of chronic CsA-induced nephrotoxicity in rats. Animals were randomly divided into four experimental groups: the control group received sunflower oil, the other groups were treated with CsA (25 mg/kg/day b.w. orally) or NSO (2 ml/kg orally) or CsA + NSO, respectively. Urine and serum creatinine levels, tissue superoxide dismutase, glutathione peroxidase and catalase enzyme activities, and nitric oxide and malondialdehyde levels were measured, and histological examination was performed. In our study, CsA caused a significant deterioration in the renal function, morphology and gave rise to severe oxidative stress in the kidney. NSO significantly improved the functional and histological parameters and attenuated the oxidative stress induced by CsA. In conclusion, our study demonstrated for the first time that NSO protects kidney tissue against oxygen free radicals, preventing renal dysfunction and morphological abnormalities associated with chronic CsA administration.


The colonic epithelium plays an active role in promoting colitis by shaping the tissue cytokine profile.
Inflammatory bowel disease (IBD) is a chronic condition driven by loss of homeostasis between the mucosal immune system, the commensal gut microbiota, and the intestinal epithelium. Our goal is to understand how these components of the intestinal ecosystem cooperate to control homeostasis. By combining quantitative measures of epithelial hyperplasia and immune infiltration with multivariate analysis of inter- and intracellular signaling, we identified epithelial mammalian target of rapamycin (mTOR) signaling as a potential driver of inflammation in a mouse model of colitis. A kinetic analysis of mTOR inhibition revealed that the pathway regulates epithelial differentiation, which in turn controls the cytokine milieu of the colon. Consistent with our in vivo analysis, we found that cytokine expression of organoids grown ex vivo, in the absence of bacteria and immune cells, was dependent on differentiation state. Our study suggests that proper differentiation of epithelial cells is an important feature of colonic homeostasis because of its effect on the secretion of inflammatory cytokines.


The effect of blood contamination on the compressive strength and surface microstructure of mineral trioxide aggregate.
To investigate the effects of whole, fresh human blood contamination on compressive strength and surface microstructure of grey and tooth-coloured mineral trioxide aggregate (MTA). The materials investigated were grey ProRoot MTA Original (Dentsply Tulsa Dental, Johnson City, TN, USA) and tooth-coloured ProRoot MTA (Dentsply Tulsa Dental). Three groups of 10 custom-made cylindrical moulds (internal dimensions 6 +/- 0.1 mm length and 4 +/- 0.1 mm diameter) were filled with tooth-coloured MTA. In the control group, MTA was mixed with water and exposed to water. In the second group, MTA was mixed with water and exposed to whole, fresh human blood. In the third group, MTA was mixed with and exposed to whole, fresh human blood. These three groups were then duplicated using grey MTA, creating a total of 60 samples. A predetermined amount of MTA and appropriate liquid were triturated in a plastic mixing capsule then subjected to ultrasonic energy after placement in the moulds. After 4 days of incubation, specimens were subjected to compressive strength testing. The surface microstructure of one extra specimen in each group was examined using scanning electron microscopy. Data were subjected to a two-way anova. Regardless of MTA type, the mean compressive strength values of both experimental groups, which were in contact with blood, were significantly less than that of the control groups (P < 0.0001). In experimental groups in which MTA was mixed with water and exposed to blood, there was a significant difference (P < 0.0001) in compressive strength between tooth-coloured MTA (30.37 +/- 10.16 MPa) and grey MTA (13.92 +/- 3.80 MPa). When blood becomes incorporated into MTA, its compressive strength is reduced. In clinical situations in which blood becomes mixed with MTA, its physical properties are likely to be compromised.


Brief Report: Service Use and Associated Expenditures Among Adolescents with Autism Spectrum Disorder Transitioning to Adulthood.
This study compared Medicaid service utilization and expenditures among adolescents with autism spectrum disorder (ASD) to adolescents with intellectual disability (ID) as they aged into adulthood. Medicaid Analytic eXtract (MAX) data was used to identify a national cohort. Winsorization was utilized to control for expenditure outliers. A greater proportion of adolescents with ASD utilized most services. Decreases in the use of key services, including psychiatric outpatient services, were observed for both groups. Changes in medical services, such as increases in inpatient and long term care services, among the ASD cohort suggest medical needs of adolescents with ASD change as they age. Information remains lacking on changing ASD symptom presentation during the transition to adolescence.


Behavioural comorbidity in Tanzanian children with epilepsy: a community-based case-control study.
The aim of this study was to define the prevalence of and risk factors for behavioural disorders in children with epilepsy from a rural district of Tanzania by conducting a community-based case-control study. One hundred and twelve children aged 6 to 14 years (55 males, 57 females; median age 12 y) with active epilepsy (at least two unprovoked seizures in the last 5 y) were identified in a cross-sectional survey and included in this study. Children who were younger than 6 years were excluded in order to eliminate febrile seizures. Behaviour was assessed using the Rutter scale; children who scored 13 or more were considered to have disordered behaviour. A comparison group was made up of age- and sex-matched children without epilepsy (n = 113; 57 males, 56 females; median age 12 y). Behavioural disorders were diagnosed in 68 of 103 (66%) children with epilepsy and in 19 of 99 (19%) controls. Disordered behaviour was significantly more common in children with epilepsy than in the comparison group (univariate odds ratio 8.2; 95% confidence interval [CI] 4.3-15.6; p < 0.001) and frequent seizures and poor scholastic attainment were associated in children with epilepsy. Behavioural disorders were not associated with antiepileptic drug usage. Attention problems were present in 48 of 91 (53%) children with epilepsy and 16 of 97 (17%) controls (univariate odds ratio 5.7; 95% CI 2.9-11.1; p < 0.001). In children with epilepsy, attention problems were significantly more common in males and were associated with frequent seizures. Children with epilepsy in a rural area of sub-Saharan Africa have a high prevalence of behavioural disorders and attention problems, both of which are associated with frequent seizures. Providing behaviour assessment and appropriate intervention programmes for children with epilepsy may reduce the burden of behaviour disorders in this setting.


Tacrolimus ointment for the treatment of atopic dermatitis in adult patients: part I, efficacy.
A total of 632 adults with atopic dermatitis applied tacrolimus ointment (0.03% or 0.1%) or vehicle twice daily for up to 12 weeks in two randomized, double-blind studies. This report focuses on the efficacy of tacrolimus ointment in these studies. The mean percent body surface area (%BSA) affected at baseline was 45%, and 56% of patients had severe atopic dermatitis. Evaluations included a physician's global evaluation of clinical response, %BSA affected, individual signs of atopic dermatitis, the Eczema Area and Severity Index (EASI) score, and the patient's assessment of pruritus. A 90% or greater improvement from baseline in disease status was observed for 6.6%, 27.5%, and 36.8% of patients in the vehicle, 0.03% tacrolimus ointment, and 0.1% tacrolimus ointment groups, respectively (P<.001), and 50% or better improvement was observed for 19.8%, 61.6%, and 72.7% of patients, respectively. Tacrolimus ointment-treated patients showed significantly greater improvement than vehicle-treated patients for all efficacy parameters evaluated, including the %BSA affected, the total score and individual scores for signs of atopic dermatitis, the patient's assessment of pruritus, and EASI score. The 0.1% concentration was more effective than the 0.03% concentration, particularly in patients with severe disease and/or extensive BSA involvement at baseline and in African Americans. Tacrolimus ointment is an effective therapy for the treatment of adult patients with atopic dermatitis on all skin regions including the head and neck.


Exploring person-centredness in emergency departments: A literature review.
Person-centred approaches to care delivery have been increasingly promoted in international policy and strategy, but despite this there is evidence of failings within healthcare systems that negatively impact on the care experience for patients and staff. This paper explores the international literature on person-centredness within emergency departments (EDs). The Person-centred Practice Framework was used as the underpinning theoretical framework. This theory contends that staff must possess certain attributes to manage the care environment appropriately to deliver effective care processes in order to achieve effective person-centred outcomes for patients and staff. An initial search of the literature identified no relevant papers that discussed person-centredness as a concept within EDs. A further search using terms drawn from a definition of person-centredness revealed literature that reflected components of person-centredness. Themes that emerged included medical-technical intervention, a culture of worthiness, managing the patient journey, nurse/doctor relationships, patients' and relatives' experience of care, and ED as a stressful environment. The themes can be mapped onto the Person-centred Practice Framework, suggesting that components of person-centred practice have emerged from studies in a fragmented fashion, without consideration of person-centredness as a whole within an ED context.


Identification of a crenarchaeal orthologue of Elf1: implications for chromatin and transcription in Archaea.
The transcription machineries of Archaea and eukaryotes are similar in many aspects, but little is understood about archaeal chromatin and its role in transcription. Here, we describe the identification in hyperthermophilic Crenarchaeota and a Korarchaeon of an orthologue of the eukaryotic transcription elongation factor Elf1, which has been shown to function in chromatin structure maintenance of actively transcribed templates. Our discovery has implications for the relationship of chromatin and transcription in Archaea and the evolution of these processes in eukaryotes.


Left ventricular function depends on previous beat ejection but not previous beat pressure load.
Previous beat contraction history, in which the performance of the left ventricle on any one beat is influenced by the mechanical events of the previous beat, may be important in the beat-to-beat regulation of left ventricular performance in the intact cardiovascular system. Prior studies of this phenomenon have established that mechanical events of the previous beat influence the function of the current beat, but it is not known whether the important mechanical influence is exerted by previous beat ejection or previous beat pressure. In addition, the magnitude of the effect of previous beat contraction history on left ventricular performance is unknown. To make these determinations, we performed experiments in six isolated rabbit left ventricle preparations buffer perfused at 30 degrees C. Left ventricular pressure and volume were controlled precisely with a servo-controlled linear motor system. After steady-state ejecting conditions were established by clamping left ventricular ejection pressure at 60% of peak isovolumic pressure, single test beats, which were pressure clamped at 40%, 60%, 80%, and 100% of peak isovolumic pressure, were introduced and followed by an isovolumic reference beat. As the level of pressure clamp decreased from 100% to 40%, developed pressure on the isovolumic beat following the single test beats increased from 139 +/- 15 (mean +/- SD) to 151 +/- 13 mm Hg. Similarly, peak positive left ventricular dP/dt increased from 1,718 +/- 209 to 1,864 +/- 181 mm Hg.sec-1 (both p less than 0.01). Multiple regression analysis showed that this increase in left ventricular function was related to previous beat ejection but not to previous beat pressure load or relaxation.(ABSTRACT TRUNCATED AT 250 WORDS)


Holocene shifts in the assembly of plant and animal communities implicate human impacts.
Understanding how ecological communities are organized and how they change through time is critical to predicting the effects of climate change. Recent work documenting the co-occurrence structure of modern communities found that most significant species pairs co-occur less frequently than would be expected by chance. However, little is known about how co-occurrence structure changes through time. Here we evaluate changes in plant and animal community organization over geological time by quantifying the co-occurrence structure of 359,896 unique taxon pairs in 80 assemblages spanning the past 300 million years. Co-occurrences of most taxon pairs were statistically random, but a significant fraction were spatially aggregated or segregated. Aggregated pairs dominated from the Carboniferous period (307 million years ago) to the early Holocene epoch (11,700 years before present), when there was a pronounced shift to more segregated pairs, a trend that continues in modern assemblages. The shift began during the Holocene and coincided with increasing human population size and the spread of agriculture in North America. Before the shift, an average of 64% of significant pairs were aggregated; after the shift, the average dropped to 37%. The organization of modern and late Holocene plant and animal assemblages differs fundamentally from that of assemblages over the past 300 million years that predate the large-scale impacts of humans. Our results suggest that the rules governing the assembly of communities have recently been changed by human activity.


Neutrophil Recruitment and Articular Hyperalgesia in Antigen-Induced Arthritis are Modulated by the Cholinergic Anti-Inflammatory Pathway.
The cholinergic anti-inflammatory pathway (CAP) is a complex neuroimmune mechanism triggered by the central nervous system to regulate peripheral inflammatory responses. Understanding the role of CAP in the pathogenesis of rheumatoid arthritis (RA) could help develop new therapeutic strategies for this disease. Therefore, we investigated the participation of this neuroimmune pathway on the progression of experimental arthritis. Using antigen-induced arthritis (AIA) model, we investigated in mice the effects of vagotomy or the pharmacological treatments with hexamethonium (peripheral nicotinic receptor antagonist), methylatropine (peripheral muscarinic receptor antagonist) or neostigmine (peripheral acetylcholinesterase inhibitor) on AIA progression. Unilateral cervical vagotomy was performed 1 week before the immunization protocol with methylated bovine serum albumin (mBSA), while drug administration was conducted during the period of immunization. On day 21, 6 hr after the challenge with mBSA injection in the femur-tibial joint, the local neutrophil migration and articular mechanical hyperalgesia were assessed. Herein, we observed that vagotomy or blockade of peripheral nicotinic (but not muscarinic) receptors exacerbated the clinical parameters of this disease. Moreover, peripheral acetylcholinesterase inhibition by neostigmine treatment promoted a reduction of neutrophil recruitment in the knee joint and articular hyperalgesia. Our results demonstrated that peripheral activation of CAP modulates experimental arthritis, providing a pre-clinical evidence of a potential therapeutic strategy for RA.


Nuclear transfer of synchronized african wild cat somatic cells into enucleated domestic cat oocytes.
The African wild cat is one of the smallest wild cats and its future is threatened by hybridization with domestic cats. Nuclear transfer, a valuable tool for retaining genetic variability, offers the possibility of species continuation rather than extinction. The aim of this study was to investigate the ability of somatic cell nuclei of the African wild cat (AWC) to dedifferentiate within domestic cat (DSH) cytoplasts and to support early development after nuclear transplantation. In experiment 1, distributions of AWC and DSH fibroblasts in each cell-cycle phase were assessed by flow cytometry using cells cultured to confluency and disaggregated with pronase, trypsin, or mechanical separation. Trypsin (89.0%) and pronase (93.0%) yielded higher proportions of AWC nuclei in the G0/G1 phase than mechanical separation (82.0%). In contrast, mechanical separation yielded higher percentages of DSH nuclei in the G0/G1 phase (86.6%) than pronase (79.7%) or trypsin (74.2%) treatments. In both species, pronase induced less DNA damage than trypsin. In experiment 2, the effects of serum starvation, culture to confluency, and exposure to roscovitine on the distribution of AWC and DSH fibroblasts in various phases of the cell cycle were determined. Flow cytometry analyses revealed that the dynamics of the cell cycle varied as culture conditions were modified. Specifically, a higher percentage of AWC and DSH nuclei were in the G0/G1 phase after cells were serum starved (83% vs. 96%) than were present in cycling cells (50% vs. 64%), after contact inhibition (61% vs. 88%), or after roscovitine (56% vs. 84%) treatment, respectively. In experiment 3, we evaluated the effects of cell synchronization and oocyte maturation (in vivo vs. in vitro) on the reconstruction and development of AWC-DSH- and DSH-DSH-cloned embryos. The method of cell synchronization did not affect the fusion and cleavage rate because only a slightly higher percentage of fused couplets cleaved when donor nuclei were synchronized by serum starvation (83.0%) than after roscovitine (80.0%) or contact-inhibition (80.0%). The fusion efficiency of in vivo and in vitro matured oocytes used as recipient cytoplasts of AWC donor nuclei (86.6% vs. 85.2%) was similar to the rates obtained with DSH donor nuclei, 83.7% vs. 73.0%, respectively. The only significant effect of source of donor nucleus (AWC vs. DSH) was on the rate of blastocyst formation in vitro. A higher percentage of the embryos derived from AWC nuclei developed to the blastocyst stage than did embryos produced from DSH nuclei, 24.2% vs. 3.3%, respectively (P < 0.05). In experiment 4, the effect of calcium in the fusion medium on induction of oocyte activation and development of AWC-DSH-cloned embryos was determined. The presence of calcium in the fusion medium induced a high incidence of cleavage of DSH oocytes (54.3%), while oocyte cleavage frequency was much lower in the absence of calcium (16.6%). The presence or absence of calcium in the fusion medium did not affect the fusion, cleavage, and blastocyst development of AWC-DSH-cloned embryos. In experiment 5, AWC-DSH-cloned embryos were transferred to the uteri of 11 synchronized domestic cat recipients on Day 6 or 7 after oocyte aspiration. Recipients were assessed by ultrasonography on Day 21 postovulation, but no pregnancies were observed. In the present study, after NT, AWC donor nuclei were able to dedifferentiate in DSH cytoplasts and support high rates of blastocyst development in vitro. Incomplete reprogramming of the differentiated nucleus may be a major constraint to the in vivo developmental potential of the embryos.


Colonization of paediatric lower respiratory tract with genital Mycoplasma species.
Recently, much attention has been given to the possible role played by pathogens that colonize neonatal or paediatric airway and their potential involvement in chronic respiratory disease. The goal of the current study was to evaluate the prevalence of Mycoplasma organisms in the BAL fluid of paediatric patients suffering from a variety of chronic respiratory diseases to determine if there was any clear disease association with bacterial presence. We examined 319 paediatric BAL samples for the presence of M.genitalium, M.hominis, U.urealyticum, U.parvum and M.pneumoniae DNA with species-specific PCR. Mycoplasma DNA was found in 32.6% (104/319) of patient samples; 10% (32/319) for M.pneumoniae, 8.8% for U.parvum, 2.8% for U.urealyticum; 4.7% for M.hominis and 9.1% for M.genitalium. There were no significant clinical and laboratory differences except serum IgE in asthmatics according to Mycoplasma colonization or not. Elevated levels of IgE were found more often in Mycoplasma DNA-negative patients than patients with bacterial DNA, 85/109 versus 24/109 respectively (P<0.0001). There was no difference in the frequency of Mycoplasmas between the asthmatics and the non-asthmatics; 30.6% (69/225) versus 37.2% (35/94) for Mycoplasma 16S DNA, and 8% versus 14.9% for M.pneumoniae, respectively. Our data indicate that in addition to M.pneumoniae, urogenital Mycoplasma species may colonize the airway of patients with chronic respiratory diseases. There was, however, no association between chronic asthma diagnosis and Mycoplasma colonization in this study.


A Qualitative Study Exploring Perceptions of Patients With Gout.
Gout is a chronic disease that is on a rising trend and greatly affects one's physical and psychosocial well-being. The aim of this study was to explore patients' perceptions of living with gout. A descriptive qualitative study was conducted and 15 adults with gout were interviewed face-to-face between December 2014 and January 2015. Thematic analysis was used to analyze the transcribed data. The experiences of patients with gout were found to revolve around four themes: emotional experiences with gout, disruptions in daily lives, interactions with doctor, and coping with gout using internal and external resources. The in-depth understanding of the patients' experiences indicates a need to provide holistic patient education and to involve family members to create nurse-led support groups and to raise public awareness regarding gout.


Pain is not associated with cognitive decline in older adults: A four-year longitudinal study.
The finding of a potential association between pain and cognitive decline is limited to a few cross-sectional studies with relatively samples. We therefore aimed to investigate whether the presence and severity of pain at baseline could predict a decline in cognitive function over four years of follow-up in the English Longitudinal Study of Ageing. At baseline, participants with no dementia who were "often troubled by pain" were considered to have pain. Pain severity was categorized as mild, moderate, or severe. Cognitive function was explored through verbal fluency (assessed by asking how many different animals the participants could name in 60 s), memory (sum of immediate and delayed verbal memory) and processing speed (number of target letters correctly identified on the letter cancellation task). Multivariable linear regression was used (exposure: pain; outcomes: cognitive change between follow-up and baseline, based on standardized residuals). Altogether, 6515 community-dwelling people with a mean age of 65 years (women = 57.3%) were included. Over a 4-year follow-up, after adjusting for 26 potential confounders, no association between pain (yes vs. no) and verbal fluency (beta = 0.02; 95%CI: -0.15 to 0.18), memory (0.05; 95%CI: -0.28 to 0.38), or processing speed (0.55; 95%CI: -18.4 to 2.93) at follow-up was found. Only severe pain was associated with greater decline in memory (-0.36; 95%CI: -0.68 to -0.04). In conclusion, in older people, pain was not associated with worsening in cognition, except for severe pain, which was marginally associated with worsening in memory tests. Further longitudinal studies are needed to confirm or refute our findings.


Need for Enhanced Environmental Representation in the Implementation of One Health.
Issues of global environmental change, global health, emerging disease, and sustainability present some of the most complex challenges of the twenty-first century. Individual disciplines cannot address these issues in isolation. Proactive, innovative, and trans-disciplinary solutions are required. Recognizing the inherent connectedness of humans, animals, plants, and their shared environment, One Health encourages the collaboration of many disciplines-including human and veterinary medicine, public health, social science, public policy, environmental science, and others-to address global and local health challenges. Despite great progress in this shift toward transdisciplinarity, the environmental component of the One Health paradigm remains underrepresented in One Health discourse. Human and animal health issues are commonly discussed under the umbrella of the One Health paradigm, while upstream environmental drivers and solutions are less prominent. We assessed the current integration of environmental issues in One Health publications and leadership. There is room for enhanced integration of environmental knowledge in the implementation of One Health approaches. We discuss the potential benefits from the collaboration between One Health and ecohealth, and explore strategies for increased environmental involvement.


Hepatic muscarinic acetylcholine receptors are not critically involved in maintaining glucose homeostasis in mice.
An increase in the rate of hepatic glucose production is the major determinant of fasting hyperglycemia in type 2 diabetes. A better understanding of the signaling pathways and molecules that regulate hepatic glucose metabolism is therefore of great clinical importance. Recent studies suggest that an increase in vagal outflow to the liver leads to decreased hepatic glucose production and reduced blood glucose levels. Since acetylcholine (ACh) is the major neurotransmitter of the vagus nerve and exerts its parasympathetic actions via activation of muscarinic ACh receptors (mAChRs), we examined the potential metabolic relevance of hepatocyte mAChRs. We initially demonstrated that the M(3) mAChR is the only mAChR subtype expressed by mouse liver/hepatocytes. To assess the physiological role of this receptor subtype in regulating hepatic glucose fluxes and glucose homeostasis in vivo, we used gene targeting and transgenic techniques to generate mutant mice lacking or overexpressing M(3) receptors in hepatocytes only. Strikingly, detailed in vivo phenotyping studies failed to reveal any significant metabolic differences between the M(3) receptor mutant mice and their control littermates, independent of whether the mice were fed regular or a high-fat diet. Moreover, the expression levels of genes for various key transcription factors, signaling molecules, and enzymes regulating hepatic glucose fluxes were not significantly altered in the M(3) receptor mutant mice. This rather surprising finding suggests that the pronounced metabolic effects mediated by activation of hepatic vagal nerves are mediated by noncholinergic signaling pathways.


Broad-spectrum aptamer inhibitors of HIV reverse transcriptase closely mimic natural substrates.
A detailed understanding of how aptamers recognize biological binding partners is of considerable importance in the development of oligonucleotide therapeutics. For antiviral nucleic acid aptamers, current models predict a correlation between broad-spectrum inhibition of viral proteins and suppression of emerging viral resistance, but there is little understanding of how aptamer structures contribute to recognition specificity. We previously established that two independent single-stranded DNA aptamers, R1T and RT1t49(-5), are potent inhibitors of reverse transcriptases (RTs) from diverse branches of the primate lentiviral family, including HIV-1, HIV-2 and SIV(cpz). In contrast, class 1 RNA pseudoknots, such as aptamer T1.1, are specific for RTs from only a few viral clades. Here, we map the binding interfaces of complexes formed between RT and aptamers R1T, RT1t49(-5) and T1.1, using mass spectrometry-based protein footprinting of RT and hydroxyl radical footprinting of the aptamers. These complementary methods reveal that the broad-spectrum aptamers make contacts throughout the primer-template binding cleft of RT. The double-stranded stems of these aptamers closely mimic natural substrates near the RNase H domain, while their binding within the polymerase domain significantly differs from RT substrates. These results inform our perspective on how sustained, broad-spectrum inhibition of RT can be achieved by aptamers.


Catheter induced aortocoronary dissection during percutaneous coronary intervention: successful aortoostial stenting.
Iatrogenic acute dissection of the ascending aorta is a serious but rare complication of cardiac catheterization and percutaneous coronary intervention (PCI). Although treatment of ascending aortic dissection may require surgical repair, stenting at the origin of the dissection area may be sufficient in aortocoronary dissection (AD) following PCI. Described is the case of a 64-year-old female patient who was treated with PCI for a critical lesion of the right coronary artery (RCA). Immediate stenting at the source of the dissection was sufficient to repair the RCA lesions and successfully seal the site of retrograde propagation of the dissection. The ascending aortic dissection was monitored with computed tomography and the dissection was spontaneously repaired within 72 hours.


Extracellular vesicles as a platform for membrane-associated therapeutic protein delivery.
Membrane proteins are of great research interest, particularly because they are rich in targets for therapeutic application. The suitability of various membrane proteins as targets for therapeutic formulations, such as drugs or antibodies, has been studied in preclinical and clinical studies. For therapeutic application, however, a protein must be expressed and purified in as close to its native conformation as possible. This has proven difficult for membrane proteins, as their native conformation requires the association with an appropriate cellular membrane. One solution to this problem is to use extracellular vesicles as a display platform. Exosomes and microvesicles are membranous extracellular vesicles that are released from most cells. Their membranes may provide a favourable microenvironment for membrane proteins to take on their proper conformation, activity, and membrane distribution; moreover, membrane proteins can cluster into microdomains on the surface of extracellular vesicles following their biogenesis. In this review, we survey the state-of-the-art of extracellular vesicle (exosome and small-sized microvesicle)-based therapeutics, evaluate the current biological understanding of these formulations, and forecast the technical advances that will be needed to continue driving the development of membrane protein therapeutics.


Intervention efficacy and malignant transformation to oral cancer among patients with leukoplakia (Review).
Elimination of leukoplakia is one of the strategies for prevention of oral cancer. However, the efficacy of reducing malignant transformation for the treated leukoplakia has rarely been reported despite a number of studies addressing its malignant transformation to oral cancer after intervention. The obstacle to do so is partially due to different lengths of follow-up and partially due to lack of information pertaining to the disease natural history that can be taken as a standard group for comparison. This work aimed to quantify the progression from leukoplakia to carcinoma with and without intervention after systematic literature review. The overall comparison of the efficacy of intervention across studies was therefore made. The literature between 1934 and 2001 was first reviewed, including two studies addressing the disease natural history model and 15 studies pertaining to intervention model in reducing malignant transformation. The simple constant-incidence exponential model and non-constant incidence models (Weibull and Quadratic models) were therefore applied to estimate annual malignant transformation after intervention in various studies. Annual transition rates pertaining to the disease natural history were also estimated by Markov models. Intervention efficacy index using 10-year cumulative incidence from both models was also developed to assess intervention efficacy across studies. For the disease natural history, the estimates of annual transition rate of leukoplakia, annual transition rate from leukoplakia to oral cancer in the PCDP, and annual transition rate from the PCDP to clinical phase were 0.00121 (0.00019-0.00150), 0.0605 (0.0436-0.0755), and 1.8797 (0.13242-2.4352), respectively. Similar findings were observed in another retrospective study. Annual malignant transformation rates after intervention range from 0.0003 to 0.062 assuming constant incidence. Studies assuming non-constant incidence show different patterns of increasing or decreasing risk with time. The estimates regarding the efficacy of intervention for each study with different follow-up periods ranged from 42.9 to 99.3%. The heterogeneity of intervention efficacy due to a wide range of annual rates of malignant transformation was demonstrated in this review. This implies that prevention program for reducing malignant transformation may vary with different areas. Factors need to be considered including different diagnosis criteria, different histological type, distributions of risk factor, different patient resources, and different compliance rates or insufficient medical intervention.


A method of inducing global cerebral ischemia.
The four-vessel occlusion (4-VO) method of global forebrain cerebral ischemia mimics the human clinical condition of cardiac arrest. It results in selective neuronal damage and is a useful experimental system to dissect underlying mechanisms behind ischemic phenomena such as the differential susceptibility of CA1 compared to the CA3 region of the hippocampus. It also provides a "proof-of-principle" system for testing out potential agents for neuroprotection.


Fever in returned travelers: results from the GeoSentinel Surveillance Network.
Fever is a marker of potentially serious illness in returned travelers. Information about causes of fever, organized by geographic area and traveler characteristics, can facilitate timely, appropriate treatment and preventive measures. Using a large, multicenter database, we assessed how frequently fever is cited as a chief reason for seeking medical care among ill returned travelers. We defined the causes of fever by place of exposure and traveler characteristics. Of 24,920 returned travelers seen at a GeoSentinel clinic from March 1997 through March 2006, 6957 (28%) cited fever as a chief reason for seeking care. Of patients with fever, 26% were hospitalized (compared with 3% who did not have fever); 35% had a febrile systemic illness, 15% had a febrile diarrheal disease, and 14% had fever and a respiratory illness. Malaria was the most common specific etiologic diagnosis, found in 21% of ill returned travelers with fever. Causes of fever varied by region visited and by time of presentation after travel. Ill travelers who returned from sub-Saharan Africa, south-central Asia, and Latin America whose reason for travel was visiting friends and relatives were more likely to experience fever than any other group. More than 17% of travelers with fever had a vaccine-preventable infection or falciparum malaria, which is preventable with chemoprophylaxis. Malaria accounted for 33% of the 12 deaths among febrile travelers. Fever is common in ill returned travelers and often results in hospitalization. The time of presentation after travel provides important clues toward establishing a diagnosis. Preventing and promptly treating malaria, providing appropriate vaccines, and identifying ways to reach travelers whose purpose for travel is visiting friends and relatives in advance of travel can reduce the burden of travel-related illness.