Microbiology and Nitrogen Cycle in the Benthic Sediments of a Glacial Oligotrophic Deep Andean Lake as Analog of Ancient Martian Lake-Beds.
Potential benthic habitats of early Mars lakes, probably oligotrophic, could range from hydrothermal to cold sediments. Dynamic processes in the water column (such as turbidity or UV penetration) as well as in the benthic bed (temperature gradients, turbation, or sedimentation rate) contribute to supply nutrients to a potential microbial ecosystem. High altitude, oligotrophic, and deep Andean lakes with active deglaciation processes and recent or past volcanic activity are natural models to assess the feasibility of life in other planetary lake/ocean environments and to develop technology for their exploration. We sampled the benthic sediments (down to 269 m depth) of the oligotrophic lake Laguna Negra (Central Andes, Chile) to investigate its ecosystem through geochemical, biomarker profiling, and molecular ecology studies. The chemistry of the benthic water was similar to the rest of the water column, except for variable amounts of ammonium (up to 2.8 ppm) and nitrate (up to 0.13 ppm). A life detector chip with a 300-antibody microarray revealed the presence of biomass in the form of exopolysaccharides and other microbial markers associated to several phylogenetic groups and potential microaerobic and anaerobic metabolisms such as nitrate reduction. DNA analyses showed that 27% of the Archaea sequences corresponded to a group of ammonia-oxidizing archaea (AOA) similar (97%) to Nitrosopumilus spp. and Nitrosoarchaeum spp. (Thaumarchaeota), and 4% of Bacteria sequences to nitrite-oxidizing bacteria from the Nitrospira genus, suggesting a coupling between ammonia and nitrite oxidation. Mesocosm experiments with the specific AOA inhibitor 2-Phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO) demonstrated an AOA-associated ammonia oxidation activity with the simultaneous accumulation of nitrate and sulfate. The results showed a rich benthic microbial community dominated by microaerobic and anaerobic metabolisms thriving under aphotic, low temperature (4°C), and relatively high pressure, that might be a suitable terrestrial analog of other planetary settings.


Lipid A from endotoxin: antigenic activities of purified fractions in liposomes.
Isolation of lipid A by acid hydrolysis of Shigella flexneri lipopolysaccharide resulted in a product that consisted of a heterogeneous mixture of bands when visualized by thin layer chromatography. Differential extraction with ethyl acetate and chloroform, or extraction with EDTA, followed by chloroform-methanol-water (Bligh-Dyer extraction), or a combination of both extraction schemes, resulted in partial purification of immunologically active lipid A. Eight fractions were purified further by preparative thin layer chromatography, and each of the fractions had phosphate, carbohydrate, and esterified fatty acids. Upon incorporation into liposomes, five of the eight purified fractions reacted with anti-lipid A serum, but the three fractions with the most number of esterified fatty acids failed to react with anti-lipid A serum. At least one fraction that originally was unreactive with anti-lipid A serum became reactive as a hapten inhibitor upon removal of esterified fatty acids by alkaline hydrolysis. Alkali-treated fractions from "unreactive" and "reactive" lipid A had similar activities as hapten inhibitors. Our data suggest that lipid A can exist in multiple forms that differ by the number and placement, and possibly by the type, of fatty acids linked to the carbohydrate of lipid A. Highly acylated forms of lipid A do not react with antiserum against the unpurified lipid A mixture, but removal of fatty acids does expose immunoreactive groups.


Pilates for Low Back Pain: Complete Republication of a Cochrane Review.
Systematic review. To determine the effects of the Pilates method for patients with nonspecific acute, subacute, or chronic low back pain. The Pilates method is one of the most common forms of intervention based on exercise used for treating patients with low back pain. However, its effectiveness is not well established. We conducted searches on CENTRAL, MEDLINE, EMBASE, CINAHL, PEDro, and SPORTDiscus up to March 2014. We included randomized controlled trials examining the effectiveness of Pilates in patients with acute, subacute, or chronic nonspecific low back pain. The outcomes evaluated were pain, disability, function, and global impression of recovery. Two independent reviewers screened for potentially eligible studies, assessed risk of bias, and extracted the data. We evaluated the overall quality of evidence using the GRADE approach and treatment effect sizes were described using mean differences and 95% confidence intervals. Searches retrieved 126 trials, of which 10 were included in the review (n = 510 participants). Seven studies were considered to have low risk of bias, and three were considered at high risk of bias. When compared to minimal intervention, Pilates reduces pain at short and intermediate term with low- to moderate-quality evidence and medium effect sizes. For disability, there is also a significant difference in favor to Pilates with low- to moderate-quality evidence and small effect size for short term and medium effect size for intermediate term compared with minimal intervention. It is unclear whether Pilates is better than other exercises for short-term pain, but there is low-quality evidence that Pilates reduces pain at intermediate term. For disability, there is moderate-quality evidence that there is no significant difference between Pilates and other exercises in either the short term or the intermediate term. There is low- to moderate-quality evidence that Pilates is more effective than minimal intervention with most of the effect sizes being considered medium. However, there is no conclusive evidence that Pilates is superior to other forms of exercises. 1.


Configuration of ripple domains and their topological defects formed under local mechanical stress on hexagonal monolayer graphene.
Ripples in graphene are extensively investigated because they ensure the mechanical stability of two-dimensional graphene and affect its electronic properties. They arise from spontaneous symmetry breaking and are usually manifested in the form of domains with long-range order. It is expected that topological defects accompany a material exhibiting long-range order, whose functionality depends on characteristics of domains and topological defects. However, there remains a lack of understanding regarding ripple domains and their topological defects formed on monolayer graphene. Here we explore configuration of ripple domains and their topological defects in exfoliated monolayer graphenes on SiO2/Si substrates using transverse shear microscope. We observe three-color domains with three different ripple directions, which meet at a core. Furthermore, the closed domain is surrounded by an even number of cores connected together by domain boundaries, similar to topological vortex and anti-vortex pairs. In addition, we have found that axisymmetric three-color domains can be induced around nanoparticles underneath the graphene. This fascinating configuration of ripple domains may result from the intrinsic hexagonal symmetry of two-dimensional graphene, which is supported by theoretical simulation using molecular dynamics. Our findings are expected to play a key role in understanding of ripple physics in graphene and other two-dimensional materials.


Efficacy of Area-Wide Inoculum Reduction and Vector Control on Temporal Progress of Huanglongbing in Young Sweet Orange Plantings.
Huanglongbing (HLB), caused by 'Candidatus Liberibacter' spp. and transmitted by the Asian citrus psyllid Diaphorina citri (ACP), is an important threat to citrus industries worldwide, causing significant yield loss. The current recommended strategies to manage HLB are to eliminate HLB-symptomatic trees to reduce sources of bacterial inoculum and to apply insecticides to reduce psyllid vector populations. The objective of this study was to assess the effectiveness and the importance of both strategies applied within young citrus plots (local management), in different frequencies and combinations, on HLB temporal progress. Two factorial field experiments, E1 and E2, were initiated in a new plantation of sweet orange in an HLB epidemic region of Sao Paulo, Brazil, in October 2005 and May 2006, respectively. Local inoculum reduction (tree removal) intervals for E1 were every 4, 8, and 16 weeks and, for E2, every 2, 4, 12, and 26 weeks. Local vector control strategies for E1 were no control, program A (PA), and program B (PB); and, for E2, no control and program C (PC), as follows. Psyllids were controlled with two 56-day-interval soil or drench applications of systemic insecticides concurrently with the rainy season each year and, during the rest of the year, with insecticide sprays every 28 days for PA and every 14 days for PB and PC. Regional HLB management was present for E1 and absent for E2. The beginning of the HLB epidemic was delayed for 10 months in E1 compared with appearance of the first diseased tree in E2 but wasn't affected by different local strategies in either experiment. After 60 (E1) and 53 (E2) months, the HLB incidence and progress rates were not affected by different frequencies of local inoculum reduction in either experiment, and were different only in plots with and without local vector control in E2. In E1, the disease incidence was reduced by 90% and the disease progress rate by 50% in plots both with and without vector control. These reductions were explained by smaller psyllid populations and lower frequency of bacterialiferous psyllids in E1 compared with E2. Annual productivity increased over time in E1, as expected for young plantings, but remained stable or decreased in E2. These results confirm that immigration of bacterialiferous ACP vectors plays a critical role in HLB epidemics and suggest that area-wide inoculum reduction and ACP management strongly affect HLB control.


Prospective predictive performance comparison between clinical gestalt and validated COVID-19 mortality scores.
Most COVID-19 mortality scores were developed at the beginning of the pandemic and clinicians now have more experience and evidence-based interventions. Therefore, we hypothesized that the predictive performance of COVID-19 mortality scores is now lower than originally reported. We aimed to prospectively evaluate the current predictive accuracy of six COVID-19 scores and compared it with the accuracy of clinical gestalt predictions. 200 patients with COVID-19 were enrolled in a tertiary hospital in Mexico City between September and December 2020. The area under the curve (AUC) of the LOW-HARM, qSOFA, MSL-COVID-19, NUTRI-CoV, and NEWS2 scores and the AUC of clinical gestalt predictions of death (as a percentage) were determined. In total, 166 patients (106 men and 60 women aged 56±9 years) with confirmed COVID-19 were included in the analysis. The AUC of all scores was significantly lower than originally reported: LOW-HARM 0.76 (95% CI 0.69 to 0.84) vs 0.96 (95% CI 0.94 to 0.98), qSOFA 0.61 (95% CI 0.53 to 0.69) vs 0.74 (95% CI 0.65 to 0.81), MSL-COVID-19 0.64 (95% CI 0.55 to 0.73) vs 0.72 (95% CI 0.69 to 0.75), NUTRI-CoV 0.60 (95% CI 0.51 to 0.69) vs 0.79 (95% CI 0.76 to 0.82), NEWS2 0.65 (95% CI 0.56 to 0.75) vs 0.84 (95% CI 0.79 to 0.90), and neutrophil to lymphocyte ratio 0.65 (95% CI 0.57 to 0.73) vs 0.74 (95% CI 0.62 to 0.85). Clinical gestalt predictions were non-inferior to mortality scores, with an AUC of 0.68 (95% CI 0.59 to 0.77). Adjusting scores with locally derived likelihood ratios did not improve their performance; however, some scores outperformed clinical gestalt predictions when clinicians' confidence of prediction was <80%. Despite its subjective nature, clinical gestalt has relevant advantages in predicting COVID-19 clinical outcomes. The need and performance of most COVID-19 mortality scores need to be evaluated regularly.


Lens cell survival after exposure to stress in the closed capsular bag.
Despite recent improvements in IOL design posterior capsule opacification (PCO) remains a significant clinical problem after cataract surgery. The Perfect Capsule device (Milvella, Ltd., Epping, Australia) permits the introduction and subsequent removal of potentially toxic agents into the closed capsular bag. The present purpose was to compare the relative effectiveness of exposing cells within the human capsular bag with a range of stresses with clinical potential and to compare the response to the same agents when applied to rabbit capsular bags and to cultured human lens cells. Human capsular bags were prepared from donor eyes and sealed with the Perfect Capsule device. Distilled water, 3 M NaCl, 250 microg/mL, 25 mg/mL 5-fluorouracil and 100 microM thapsigargin (Tg) were introduced for 2 minutes. The bags were then perfused with Eagle's minimum essential medium (EMEM) and an IOL inserted before the bags were dissected and pinned to the base of plastic culture dishes. The bags were maintained in EMEM for 28 days and phase images were acquired throughout. Rabbit eyes were prepared and cultured in a similar manner, although tests were limited to 5-FU (25 mg/mL) and Tg (30-300 microM). FHL124 cells were cultured on plastic (EMEM supplemented with 5% FCS), and serum was removed for 24 hours before exposure to the same agents as human bags. Cell survival was assessed by quantifying Coomassie blue staining after 4 days. Initially, NaCl induced by far the most obvious signs of cell death, especially of anterior cells, followed by 5-FU>water>Tg. However, by 2 weeks, cell death became more apparent in the Tg-exposed bags, and, at the end of 4 weeks, there were no cells surviving. Cells on the posterior capsule were confluent in water-exposed bags (similar to unexposed controls), whereas after NaCl exposure, coverage was incomplete but greater than after 5-FU. In the rabbit bags, exposure to 25 mg/mL 5-FU totally eliminated cells, but 100 microM Tg was ineffective. At the end of a 4-day culture of FHL124 cells exposed for 2 minutes to NaCl, 5-FU or Tg, there were no cells surviving, whereas there was 50% cell survival compared with control cells after water treatment. Tissue-cultured human lens cells are much more sensitive to short-term hyperosmotic than hyposmotic stress, with a rapid onset of cell death of cultured cells exposed to 3 M NaCl. This finding was confirmed in human capsular bags although adherent fibers appear to offer additional protection to 5-FU which can be overcome by the very hydrophobic Tg. The application of the Perfect Capsule system in concert with thapsigargin provides a promising means of preventing PCO.


Thoracic and duodenopancreatic neuroendocrine tumors in multiple endocrine neoplasia type 1: natural history and function of menin in tumorigenesis.
Mutations of the multiple endocrine neoplasia type 1 (MEN1) gene lead to loss of function of its protein product menin. In keeping with its tumor suppressor function in endocrine tissues, the majority of the MEN1-related neuroendocrine tumors (NETs) show loss of heterozygosity (LOH) on chromosome 11q13. In sporadic NETs, MEN1 mutations and LOH are also reported, indicating common pathways in tumor development. Prevalence of thymic NETs (thNETs) and pulmonary carcinoids in MEN1 patients is 2-8%. Pulmonary carcinoids may be underreported and research on natural history is limited, but disease-related mortality is low. thNETs have a high mortality rate. Duodenopancreatic NETs (dpNETs) are multiple, almost universally found at pathology, and associated with precursor lesions. Gastrinomas are usually located in the duodenal submucosa while other dpNETs are predominantly pancreatic. dpNETs are an important determinant of MEN1-related survival, with an estimated 10-year survival of 75%. Survival differs between subtypes and apart from tumor size there are no known prognostic factors. Natural history of nonfunctioning pancreatic NETs needs to be redefined because of increased detection of small tumors. MEN1-related gastrinomas seem to behave similar to their sporadic counterparts, while insulinomas seem to be more aggressive. Investigations into the molecular functions of menin have led to new insights into MEN1-related tumorigenesis. Menin is involved in gene transcription, both as an activator and repressor. It is part of chromatin-modifying protein complexes, indicating involvement of epigenetic pathways in MEN1-related NET development. Future basic and translational research aimed at NETs in large unbiased cohorts will clarify the role of menin in NET tumorigenesis and might lead to new therapeutic options.


Enteral vs. intravenous ICU sedation management: study protocol for a randomized controlled trial.
A relevant innovation about sedation of long-term Intensive Care Unit (ICU) patients is the 'conscious target': patients should be awake even during the critical phases of illness. Enteral sedative administration is nowadays unusual, even though the gastrointestinal tract works soon after ICU admission. The enteral approach cannot produce deep sedation; however, it is as adequate as the intravenous one, if the target is to keep patients awake and adapted to the environment, and has fewer side effects and lower costs. A randomized, controlled, multicenter, single-blind trial comparing enteral and intravenous sedative treatments has been done in 12 Italian ICUs. The main objective was to achieve and maintain the desired sedation level: observed RASS = target RASS ± 1. Three hundred high-risk patients were planned to be randomly assigned to receive either intravenous propofol/midazolam or enteral melatonin/hydroxyzine/lorazepam. Group assignment occurred through online minimization process, in order to balance variables potentially influencing the outcomes (age, sex, SAPS II, type of admission, kidney failure, chronic obstructive pulmonary disease, sepsis) between groups. Once per shift, the staff recorded neurological monitoring using validated tools. Three flowcharts for pain, sedation, and delirium have been proposed; they have been designed to treat potentially correctable factors first, and, only once excluded, to administer neuroactive drugs. The study lasted from January 24 to December 31, 2012. A total of 348 patients have been randomized, through a centralized website, using a specific software expressly designed for this study. The created network of ICUs included a mix of both university and non-university hospitals, with different experience in managing enteral sedation. A dedicated free-access website was also created, in both Italian and English, for continuous education of ICU staff through CME courses. This 'educational research' project aims both to compare two sedative strategies and to highlight the need for a profound cultural change, improving outcomes by keeping critically-ill patients awake. Clinicaltrials.gov #NCT01360346.


Hydrogel Dressings for Chronic Wound Healing in Diabetes: Beyond Hydration.
Chronic wounds caused by diabetes are a significant medical challenge. Complications from non-healing can result in dire consequences for patients and cost the healthcare system billions of dollars annually. Non-healing in wounds for diabetic patient's results from a combination of factors which impair clearing of injured tissue, proliferation of healthy cell populations and increase risk of infection. Wound dressings continue to form the basis for the treatment of chronic wounds. Traditionally, these focused solely on hydration of the wound site and mitigating infection risk. Hydrogel systems are ready made to meet these basic requirements due to their intrinsic hydration properties and ability to deliver active ingredients. Flexibility in materials and methods of release allowed these systems to remain targets of research into the 21st century. Improved understanding of the wound environment and healing cascades has led to the development of more advanced systems which incorporate endogenous growth factors and living cells. Despite their promise, clinical efficacy of these systems has remained a challenge. Further, the regulatory pathways for approval add a layer of complexity to translate pre-clinical work into marketed products. In this review, we discuss systems currently in clinical use, pre-clinical directions and regulatory challenges for hydrogels in the treatment of diabetic chronic wounds.


Counterpoint: Intraperitoneal chemotherapy: an investigational treatment in ovarian cancer.
Intraperitoneal (IP) chemotherapy in ovarian cancer has been studied since 1978. Numerous phase II trials have been performed, which have shown that higher levels can be obtained in the peritoneal cavity compared with systemic circulation after administration of cytoxic agents in a large volume via a semi-permanent catheter. Three randomized trials have been performed in patients with ovarian cancer comparing different IP regimens to standard therapy with intravenous agents. The last two trials from the Gynecologic Oncology Group (GOG) and the Southwest Oncology Group (SWOG) compared two different IP regimens versus standard therapy with intravenous cisplatin plus paclitaxel. Although an improvement in progression-free survival was reported for the IP regimens, they have been associated with unacceptable toxicity, and no IP regimen can be considered standard therapy. Maintenance therapy with IP cisplatin also failed to improve survival in patients who obtained complete remission after intravenous chemotherapy. The GOG is considering another phase III trial of IP therapy that will compare a carboplatin-based regimen versus standard therapy with intravenous paclitaxel plus carboplatin. Unless such a trial shows an improvement in clinical outcome, intravenous carboplatin plus paclitaxel remains the standard of care and IP chemotherapy should not be used outside of a clinical trial.


Utilization of electrocoagulation for the isolation of alkaloids from the aerial parts of Stemona aphylla and their mosquitocidal activities against Aedes aegypti.
The electrocoagulation (EC) technique is an alternative method of isolating natural products with the advantage of minimizing the amounts of organic solvents required for this process, which are often harmful to the environment. In this research, the EC and the conventional solvent extraction methods were used in the isolation of Stemona alkaloids from the aerial parts of Stemona aphylla. A comparison was made between the amounts of the isolated alkaloids and the solvents used. The isolated alkaloids were evaluated for their larvicidal, ovicidal and oviposition-deterrent activities against the dengue vector, Aedes aegypti. The morphology and histopatology of the alkaloid treated larvae were also investigated. Two Stemona alkaloids, (2'S)-hydroxystemofoline and stemofoline, were isolated from both the EC and the conventional method. The amounts of (2'S)-hydroxystemofoline from the EC method was about the same as that obtained from the conventional method. However, the amounts of stemofoline obtained from the EC method were about two times larger than those obtained from the conventional method. Importantly, the EC method required six times less total organic solvents. The larvicidal activity assays of (2'S)-hydroxystemofoline and stemofoline showed that these were highly effective against Aedes aegypti larvae with LC50 values of 3.91 μg/ml and 4.35 μg/ml, respectively. Whereas, the crude EC extract (LC50 = 11.86 μg/ml) showed greater larvicidal activity than the crude extract obtained from the conventional extraction method (LC50 = 53.40 μg/ml). The morphological observations of the (2'S)-hydroxystemofoline and the stemofoline treated larvae revealed that the anal gills were the sites of aberrations. A histopathological study showed that larvae treated with these alkaloids had cytopathological alterations to the epithelial cells of the midgut. At a concentration 40 μg/ml (2'S)-hydroxystemofoline showed 100% ovicidal activity on 24 h old eggs while stemofoline showed 97.2%. Furthermore, the oviposition-deterrent effects of (2'S)-hydroxystemofoline and stemofoline, at a concentration of 80 μg/ml were 99.5% and 97.2%, respectively.


Dimensions, discourses and differences: trainees conceptualising health care leadership and followership.
As doctors in all specialties are expected to undertake leadership within health care organisations, leadership development has become an inherent part of medical education. Whereas the leadership literature within medical education remains mostly focused on individual, hierarchical leadership, contemporary theory posits leadership as a group process, which should be distributed across all levels of health care organisation. This gap between theory and practice indicates that there is a need to understand what leadership and followership mean to medical trainees working in today's interprofessional health care workplace. Epistemologically grounded in social constructionism, this research involved 19 individual and 11 group interviews with 65 UK medical trainees across all stages of training and a range of specialties. Semi-structured interviewing techniques were employed to capture medical trainees' conceptualisations of leadership and followership. Interviews were audiotaped, transcribed verbatim and analysed using thematic framework analysis to identify leadership and followership dimensions which were subsequently mapped onto leadership discourses found in the literature. Although diversity existed in terms of medical trainees' understandings of leadership and followership, unsophisticated conceptualisations focusing on individual behaviours, hierarchy and personality were commonplace in trainees' understandings. This indicated the dominance of an individualist discourse. Patterns in understandings across all stages of training and specialties, and whether definitions were solicited or unsolicited, illustrated that context heavily influenced trainees' conceptualisations of leadership and followership. Our findings suggest that UK trainees typically hold traditional understandings of leadership and followership, which are clearly influenced by the organisational structures in which they work. Although education may change these understandings to some extent, changes in leadership practices to reflect contemporary theory are unlikely to be sustained if leadership experiences in the workplace continue to be based on individualist models.


Trait Emotional Intelligence and Its Correlates in Oman Medical Specialty Board Residents.
As part of the globalization of medical education, residency programs in Oman have adopted competency-based standards by the Accreditation Council for Graduate Medical Education International (ACGME-I). Correctly perceiving the emotions of others and managing one's own emotions are essential to high-quality patient care. We tested the reliability and construct validity of the Trait Emotional Intelligence Questionnaire-Short Form (TEIQue-SF), and assessed trait Emotional Intelligence (EI) in Oman Medical Specialty Board (OMSB) residents in multiple specialties. We explored for correlations with trainees' sociodemographic background data. We conducted a cross-sectional, observational study between February and August 2017. Participants were OMSB residents. We administered the TEIQue-SF and collected sociodemographic data from participants. Multiple linear regression analysis was conducted to identify independent predictors of trait EI. The present cohort scored high in the trait EI subscale of Well-being, followed by Sociability, Self-control, and Emotionality. Among sociodemographic factors, female gender and high income were significant predictors of TEIQue-SF's Well-being subscale and high income and living in a rented home were significant predictors of the Sociability subscale. This is the first study conducted among medical residents in Oman regarding trait EI and its correlates. Our findings of overall high EI and several socioeconomic predictors echo the literature on the assessment of EI in trainees. The findings add to the evidence of cross-cultural applicability of instruments to measure trait EI, and use assessments of EI in resident selection and education.


Attributional style of lonely and depressed people.
This article examines the attributional style of lonely and depressed people. Previous studies have suggested that both lonely and depressed individuals ascribe failure to characterological defects in themselves. However, the prototype of a lonely person and the prototype of a depressed person suggest that this characteristic attributional style should mainly hold for interpersonal failures. A questionnaire was formed, consisting of 20 hypothetical situations. Half of the situations were interpersonal, and half were not; half described successful outcomes, and half described failures. The subject selected an attributional alternative that best explained the outcome. The questionnaire was administered to 304 students, along with the Beck Depression Inventory and the UCLA Loneliness Scale. The results showed that lonely and depressed people ascribe interpersonal failures to unchangeable characterological defects in themselves (e.g., a lack of ability). Because the prototype of a lonely person is more singularly interpersonal than is the prototype of a depressed person, we hypothesized that loneliness would show higher correlations with the attributional style. This hypothesis was also confirmed. The findings were replicated using a modified version of the questionnaire.


[Infectious canine hepatitis in 4 dogs in Switzerland].
Infectious canine hepatitis in 4 Dogs in Switzerland. Four dogs presented with nonspecific symptoms of lethargy, vomiting, diarrhea, fever and weakness. Laboratory results were consistent with hepatopathy and disseminated intravascular coagulation. Three dogs died, one survived. In the three deceased dogs, a diagnosis of infectious canine hepatitis (ICH) was made based on histological findings and positive immunhistochemistry results for canine adenovirus-1 (CAV-1). In the surviving dog, an antemortem diagnosis of ICH was determined via positive polymerase chain reaction results from blood, occular, nasal and preputial discharge as well as from urine. Since the introduction of widespread vaccination, the incidence of CAV-1 infection in dogs is low. However, the disease has not been eradicated and should be considered when clinical signs consistent with ICH are present.


Improving the quality of medical care: the normativity of evidence-based performance standards.
Poor quality medical care is sometimes attributed to physicians' unwillingness to act on evidence about what works best. Evidence-based performance standards (EBPSs) are one response to this problem, and they are increasingly employed by health care regulators and payers. Evidence in this instance is judged according to the precepts of evidence-based medicine (EBM); it is probabilistic, and the randomized controlled trial (RCT) is the gold standard. This means that EBPSs suffer all the infirmities of EBM generally-well rehearsed problems with the external validity of research findings as well as the inferential leap from study results in the aggregate to individual patient care. These theoretical weaknesses promise to have a practical impact on the care of patients. To avoid this, EBPSs should be understood as guidelines indicative of average effectiveness rather than standards to be applied in every case.


Adipose tissue and the reproductive axis: biological aspects.
The discovery of leptin has clearly demonstrated a relationship between body fat and the neuroendocrine axis since leptin influences appetite and the reproductive axis. Since adipose tissue is a primary source of leptin, adipose tissue is no longer considered as simply a depot to store fat. Recent findings demonstrate that numerous other genes, i.e. neuropeptides, interleukins and other cytokines and biologically active substances such as leptin and insulin-like growth factors I and II, are also produced by adipose tissue, which could influence appetite and the reproductive axis. Targets of leptin in the hypothalamus include neuropeptide Y, proopiomelanocortin and kisspeptin. Transsynaptic connection of hypothalamic neurons to porcine adipose tissue may result in a direct influence of the hypothalamus on adipose tissue function. Nutritional signals such as leptin are detected by the central nervous system and translated by the neuroendocrine system into signals which ultimately regulates luteinizing hormone secretion. Furthermore, leptin directly affects gonadotropin-releasing hormone release from the hypothalamus, luteinizing hormone from the pituitary gland and ovarian follicular steroidogenesis. Although leptin is identified as a putative signal that links metabolic status and neuroendocrine control of reproduction, other adipocyte protein products may play key roles in regulating the reproductive axisin the pig.


Asiatic Acid Protects against Doxorubicin-Induced Cardiotoxicity in Mice.
The use of doxorubicin (DOX) can result in depression of cardiac function and refractory cardiomyopathy. Currently, there are no effective approaches to prevent DOX-related cardiac complications. Asiatic acid (AA) has been reported to provide cardioprotection against several cardiovascular diseases. However, whether AA could attenuate DOX-related cardiac injury remains unclear. DOX (15 mg/kg) was injected intraperitoneally into the mice to mimic acute cardiac injury, and the mice were given AA (10 mg/kg or 30 mg/kg) for 2 weeks for protection. The data in our study found that AA-treated mice exhibited attenuated cardiac injury and improved cardiac function in response to DOX injection. AA also suppressed myocardial oxidative damage and apoptosis without affecting cardiac inflammation in DOX-treated mice. AA also provided protection in DOX-challenged cardiomyocytes, improved cell viability, and suppressed intracellular reactive oxygen species (ROS) in vitro. Detection of signaling pathways showed that AA activated protein kinase B (AKT) signaling pathway in vivo and in vitro. Furthermore, we found that AA lost its protective effects in the heart with AKT inactivation. In conclusion, our results found that AA could attenuate DOX-induced myocardial oxidative stress and apoptosis via activation of the AKT signaling pathway.


Dynamics in multi-domain protein recognition of RNA.
Protein-RNA interactions play essential roles in gene regulation and RNA metabolism. While high-resolution structures have revealed principles of RNA recognition by individual RNA binding domains (RBDs), the presence of multiple RBDs in many eukaryotic proteins suggests additional modes of RNA recognition by combination and cooperation of these interactions. Recent structures, together with biochemical and biophysical studies have revealed novel principles of RNA recognition by multi-domain proteins. These examples highlight an important role for dynamics in RNA recognition, with mechanisms including fly-casting and conformational selection, and advocate the use of solution techniques for their analysis.


Spatiotemporal conversion of auditory information for cochleotopic mapping.
Auditory communication signals such as monkey calls are complex FM vocal sounds and in general induce action potentials in different timing in the primary auditory cortex. Delay line scheme is one of the effective ways for detecting such neuronal timing. However, the scheme is not straightforwardly applicable if the time intervals of signals are beyond the latency time of delay lines. In fact, monkey calls are often expressed in longer time intervals (hundreds of milliseconds to seconds) and are beyond the latency times observed in the brain (less than several hundreds of milliseconds). Here, we propose a cochleotopic map similar to that in vision known as a retinotopic map. We show that information about monkey calls could be mapped on a cochleotopic cortical network as spatiotemporal firing patterns of neurons, which can then be decomposed into simple (linearly sweeping) FM components and integrated into unified percepts by higher cortical networks. We suggest that the spatiotemporal conversion of auditory information may be essential for developing the cochleotopic map, which could serve as the foundation for later processing, or monkey call identification by higher cortical areas.


Nitrogen Surplus Benchmarks for Controlling N Pollution in the Main Cropping Systems of China.
Nitrogen (N) surplus is a useful indicator for improving agricultural N management and controlling N pollution. Few studies have developed benchmark values for cropping systems in China, a country with the largest N fertilizer use in the world. We established N surplus benchmarks for 13 main cropping systems, at optimal N management, using results from >4500 on-farm field experiments and a soil surface balance approach. These cropping systems accounted for about 50% of total N fertilizer consumption in Chinese agriculture in 2009. The results showed that N surplus benchmarks for single cropping systems ranged from 40 to 100 kg N ha-1 yr-1 (average 73 kg N ha-1 yr-1), and for double cropping systems from 110 to 190 kg N ha-1 yr-1 (average 160 kg N ha-1 yr-1), roughly twice that of single cropping systems. These N surplus benchmarks may be further refined, following further decreases in N deposition rates and reactive N losses as a result of strict implementation of "4R-nutrient stewardship" and improvements in fertilization techniques and agronomic managements. Our N surplus benchmarks could serve as realistic targets to improve the N management of current conventional practices, and thereby could lay the foundations for a more sustainable N management in China.


Anticaries effects of polyphenolic compounds from Japanese green tea.
The dental caries inhibiting effect of the extract from Japanese green tea, one of the most popular drinks in Japan, was studied both in vitro and in vivo. The crude tea polyphenolic compounds (designated Sunphenon) from the leaf of Camellia sinensis were found to effectively inhibit the attachment of Streptococcus mutans strain JC-2 (serotype c) to saliva-coated hydroxyapatide discs. Sunphenon was also inhibitory to water-insoluble glucan formation from sucrose by crude glucosyltransferase of S. mutans JC-2 (c). Among the tea catechins tested, (-)-epigallocatechin gallate and (-)-epicatechin gallate showed the most potent inhibition of the glucosyltransferase activity. Finally, significantly lower caries scores were observed in specific pathogen free rats infected with S. mutans JC-2 (c) and fed a cariogenic diet and/or drinking water containing 0.05% Sunphenon as compared with control rats not receiving polyphenolic compounds.


Climate change and unequal phenological changes across four trophic levels: constraints or adaptations?
1. Climate change has been shown to affect the phenology of many organisms, but interestingly these shifts are often unequal across trophic levels, causing a mismatch between the phenology of organisms and their food. 2. We consider two alternative hypotheses: consumers are constrained to adjust sufficiently to the lower trophic level, or prey species react more strongly than their predators to reduce predation. We discuss both hypotheses with our analyses of changes in phenology across four trophic levels: tree budburst, peak biomass of herbivorous caterpillars, breeding phenology of four insectivorous bird species and an avian predator. 3. In our long-term study, we show that between 1988 and 2005, budburst advanced (not significantly) with 0.17 d yr(-1), while between 1985 and 2005 both caterpillars (0.75 d year(-1)) and the hatching date of the passerine species (range for four species: 0.36-0.50 d year(-1)) have advanced, whereas raptor hatching dates showed no trend. 4. The caterpillar peak date was closely correlated with budburst date, as were the passerine hatching dates with the peak caterpillar biomass date. In all these cases, however, the slopes were significantly less than unity, showing that the response of the consumers is weaker than that of their food. This was also true for the avian predator, for which hatching dates were not correlated with the peak availability of fledgling passerines. As a result, the match between food demand and availability deteriorated over time for both the passerines and the avian predators. 5. These results could equally well be explained by consumers' insufficient responses as a consequence of constraints in adapting to climate change, or by them trying to escape predation from a higher trophic level, or both. Selection on phenology could thus be both from matches of phenology with higher and lower levels, and quantifying these can shed new light on why some organisms do adjust their phenology to climate change, while others do not.


On predicting time to completion for the first stage of spontaneous labor at term in multiparous women.
Labor that progresses faster than anticipated may lead to unplanned out-of-hospital births. With the aim to improve planning of transportation to birthing institutions, this study investigated predictors of time to completion for the first stage of labor conditional on cervical opening (conditional time) in multiparous women at term. We performed a retrospective analysis of partograms for women in Robson's group 3 who delivered at one hospital from 2003 to 2013. A generalized additive mixed model was fitted, accounting for possible non-linear relationships between the predictor variables and outcome, e.g. the time from each cervical measurement to full dilation, using multiple measurements for each woman. The following predictors were included: cervical dilation (cm), parity (1, 2, or ≥3 previous vaginal births), oxytocin infusion (no/yes), epidural (no/yes), maternal age (years), maternal height (cm), body mass index (BMI, kg/m2), birthweight (kg), spontaneous rupture of membranes (no/yes). A modified regression model with gestational age (days) instead of birthweight was used to predict conditional time to full cervical dilation for combinations of the most relevant predictors. A total of 1753 partograms were included in the analysis. The strongest predictors were birthweight, epidural and oxytocin use, and spontaneous rupture of membranes, along with cervical measurements. For birthweight, there was an almost 40% increase in time to full cervical dilation for each 1-kg increment. Conditional time was on average 23% longer in cases with epidural use and 53% longer in cases requiring oxytocin augmentation. Spontaneous rupture of the membranes shortened conditional time by 31%. Maternal age was not associated with the outcome, while increasing BMI and parity modestly reduced conditional time. Higher parity, lower fetal weight (gestational age), and spontaneous rupture of the membranes are associated with more rapid labor.


Human corneal endothelial cells employ phosphorylation of p27(Kip1) at both Ser10 and Thr187 sites for FGF-2-mediated cell proliferation via PI 3-kinase.
FGF-2 stimulates cell proliferation of rabbit corneal endothelial cells (rCECs) by degrading the cyclin-dependent kinase inhibitor p27(Kip1) (p27) through its phosphorylation mechanism. The authors investigated whether the cell proliferation of human CECs (hCECs) is also induced by FGF-2 stimulation through the p27 phosphorylation pathway. Expression and activation of protein were analyzed by immunoblotting. Cell proliferation was measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Transfection of hCECs with small interference RNA (siRNA) was performed using a transfection reagent. FGF-2 stimulated cell proliferation in hCECs; the FGF-2 action was completely blocked by pathway-specific inhibitors for PI 3-kinase (LY294002) and MEK1/2 (U0126), respectively. Using immunoblotting, the authors showed that FGF-2 induced phosphorylation of p27 at both serine 10 (Ser10) and threonine 187 (Thr187) sites. These effects were also completely blocked by LY294002 or U0126. The authors then determined cross-talk between PI 3-kinase and extracellular signal-regulated kinase (ERK)1/2; blocking of ERK1/2 activation by LY294002 indicated that in hCECs ERK1/2 works as a downstream effector to PI 3-kinase for cell proliferation induced by FGF-2, whereas the ERK1/2 pathway in rCECs is parallel to the PI 3-kinase pathway. However, the downstream mechanism involved in cell cycle progression in hCECs is identical to that of rCECs: phosphorylation of p27 at Ser10 was mediated by kinase-interacting stathmin (KIS), confirmed with siRNA to KIS, and phosphorylation of p27 at Thr187 was mediated by cell division cycle 25A (Cdc25A), confirmed using Cdc25A inhibitor. CONCLUSIONS; FGF-2 stimulates proliferation of hCECs through PI 3-kinase and its downstream target ERK1/2 pathways. This linear signal transduction significantly downregulates p27 through its phosphorylation at both Ser10 and Thr187 sites mediated by KIS and Cdc25A, respectively.


Protecting and improving breastfeeding practices during a major emergency: lessons learnt from the baby tents in Haiti.
The 2010 earthquake in Haiti displaced about 1.5 million people, many of them into camps for internally displaced persons. It was expected that disruption of breastfeeding practices would lead to increased infant morbidity, malnutrition and mortality. Haiti's health ministry and the United Nations Children's Fund, in collaboration with local and international nongovernmental organizations, established baby tents in the areas affected by the earthquake. The tents provided a safe place for mothers to breastfeed and for non-breastfed infants to receive ready-to-use infant formula. Such a large and coordinated baby tent response in an emergency context had never been mounted before anywhere in the world. Baby tents were set up in five cities but mainly in Port-au-Prince, where the majority of Haiti's 1555 camps for displaced persons had been established. Between February 2010 and June 2012, 193 baby tents were set up; 180 499 mother-infant pairs and 52 503 pregnant women were registered in the baby tent programme. Of infants younger than 6 months, 70% were reported to be exclusively breastfed and 10% of the "mixed feeders" moved to exclusive breastfeeding while enrolled. In 2010, 13.5% of registered infants could not be breastfed. These infants received ready-to-use infant formula. Thanks to rapid programme scale-up, breastfeeding practices remained undisrupted. However, better evaluation methods and comprehensive guidance on the implementation and monitoring of baby tents are needed for future emergencies, along with a clear strategy for transitioning baby tent activities into facility and community programmes.


Multi-generational carcinogenesis from diethylstilbestrol investigated by blastocyst transfers in mice.
Previous studies have shown that a carcinogenic effect can be transmitted from female mice exposed prenatally to diethylstilbestrol (DES) to their female offspring. Furthermore, male mice exposed pre-natally to DES can transmit a carcinogenic effect to their offspring through their germ cells. To study how multi-generational carcinogenesis is transmitted through females exposed pre-natally to DES, the technique of blastocyst transfer was utilized. Blastocysts from strain CD-1 mice exposed pre-natally to vehicle were transferred to mice exposed pre-natally to DES. Among 143 offspring from these transfers, there were 10 ovarian adenomas and 10 uterine adenocarcinomas. Among 92 offspring from blastocyst transfers between mice exposed pre-natally to vehicle only, there was 1 ovarian adenoma and 1 uterine adenocarcinoma. Thus the pre-natal exposure of the host to DES produced a maternal environment which increased the incidence of ovarian and uterine tumors. The reverse type of transfer was also performed, in which blastocysts from female mice exposed pre-natally to DES were transferred into mice exposed to vehicle only pre-natally. Among 99 offspring derived from DES-exposed germ cells, 6 developed ovarian adenomas and 16 developed uterine adenocarcinomas. Thus DES also has a multi-generational effect transmitted through the blastocyst, which is consistent with fetal germ cell mutation from DES.


Oxidative potential of smoke from burning wood and mixed biomass fuels.
More than half the world's population still rely on burning biomass fuels to heat and light their homes and cook food. Household air pollution, a common component of which is inhalable particulate matter (PM), emitted from biomass burning is associated with increased vulnerability to respiratory infection and an enhanced risk of developing chronic obstructive pulmonary disease. In the light of an emerging hypothesis linking chronic PM exposure during childhood and increased vulnerability to respiratory diseases in adulthood, in a chain of events involving oxidative stress, reduced immunity and subsequent infection, the aim of this study was to characterise the oxidative potential (OP) of PM collected during the burning of wood and mixed biomass, whilst cooking food in the Kathmandu Valley, Nepal. Our assessments were based on the capacity of the particles to deplete the physiologically relevant antioxidants from a validated, synthetic respiratory tract lining fluid (RTLF). Incubation of mixed biomass and wood smoke particles suspensions with the synthetic RTLF for 4 h resulted in a mean loss of ascorbate of 64.76 ± 16.83% and 83.37 ± 14.12% at 50 μg/ml, respectively. Reduced glutathione was depleted by 49.29 ± 15.22% in mixed biomass and 65.33 ± 13.01% in wood smoke particles under the same conditions. Co-incubation with the transition metal chelator diethylenetriaminepentaacetate did not inhibit the rate of ascorbate oxidation, indicating a negligible contribution by redox-active metals in these samples. The capacity of biomass smoke particles to elicit oxidative stress certainly has the potential to contribute towards negative health impacts associated with traditional domestic fuels in the developing world.


Experimental radioimmunotherapy of HeLa tumours in nude mice with 131I-labeled monoclonal antibodies.
The radioimmunotherapeutic potential of 131I-labeled monoclonal antibodies was investigated in 36 nude mice (BALB/c nu/nu) inoculated s.c. with the HeLa Hep 2 human adenocarcinoma cell line. The membrane bound tumour associated antigen placental alkaline phosphatase and several intracellular cytokeratins served as targets for the antibodies. The specific radioactivity in each organ was determined after i.p. injection of the 131I-labeled antibodies (0.2-0.3 mg, approximately 15 MBq/animal), and high localization to the tumours was seen. Significant growth inhibition was observed after injection of the radiolabeled monoclonal antibody H7 against the placental alkaline phosphatase, which reduced the tumour growth to only 12% during a 3 week period compared to a growth of more than 100% for the controls. Animal weight losses were seen. Synthesis of endogenous antibodies to the target antigens was found to be significant. Morphometric evaluation of the relations between stroma, tumour cells and necrotic areas in the tumours after radioimmunotherapy demonstrated a significant increase of the mean relative connective tissue volume and a significant decreased mean of relative volume of tumour cells in the group treated with iodinated antiplacental alkaline phosphatase antibody. This therapeutic principle is encouraging and may offer new possibilities for future treatment of some malignant diseases.


Isolation and characterization of Escherichia coli mutants that lack the heat shock sigma factor sigma 32.
The product of the Escherichia coli rpoH (htpR) gene, sigma 32, is required for heat-inducible transcription of the heat shock genes. Previous studies on the role of sigma 32 in growth at low temperature and in gene expression involved the use of nonsense and missense rpoH mutations and have led to ambiguous or conflicting results. To clarify the role of sigma 32 in cell physiology, we have constructed loss-of-function insertion and deletion mutations in rpoH. Strains lacking sigma 32 are extremely temperature sensitive and grow only at temperatures less than or equal to 20 degrees C. There is no transcription from the heat shock promoters preceding the htpG gene or the groESL and dnaKJ operons; however, several heat shock proteins are produced in the mutants. GroEL protein is present in the rpoH null mutants, but its synthesis is not inducible by a shift to high temperature. The low-level synthesis of GroEL results from transcription initiation at a minor sigma 70-controlled promoter for the groE operon. DnaK protein synthesis cannot be detected at low temperature, but can be detected after a shift to 42 degrees C. The mechanism of this heat-inducible synthesis is not known. We conclude that sigma 32 is required for cell growth at temperatures above 20 degrees C and is required for transcription from the heat shock promoters. Several heat shock proteins are synthesized in the absence of sigma 32, indicating that there are additional mechanisms controlling the synthesis of some heat shock proteins.


CD23 expression in mantle cell lymphoma: clinicopathologic features of 18 cases.
The distinction between small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) and mantle cell lymphoma (MCL) has important clinical implications. Typically, SLL/CLL is CD23+, whereas MCL is CD23-. However, CD23 is expressed in a subset of MCLs, and the clinicopathologic features of patients with these neoplasms are not well described. We report 18 CD23+ MCLs, detected by flow cytometry in all cases (dim intensity, 16; bright intensity, 2), 5 (28%), also positive by immunohistochemical analysis. There were 13 men and 5 women (median age, 56 years), 5 of whom died (median survival, 46 months). Seventeen (94%) had bone marrow involvement. Lymphadenopathy (14 cases [78%]), splenomegaly (11 cases [61%]), and leukemic involvement (10 cases [56%]) were common. Five cases (28%) had blastoid morphologic features. The frequency of CD23 expression by MCL is method-dependent, being typically dim and most commonly detected by flow cytometry. In this small study group, bone marrow and leukemic involvement were relatively common.


Effect of amino acids and oxygen on chemotaxis in Escherichia coli.
Adler, Julius (University of Wisconsin, Madison). Effect of amino acids and oxygen on chemotaxis in Escherichia coli. J. Bacteriol. 92:121-129. 1966.-Motile cells of Escherichia coli placed at one end of a capillary tube containing a mixture of the 20 amino acids commonly found in proteins migrate out into the tube in two bands. The bands are clearly visible to the naked eye, and they can also be demonstrated by microscopy, photography, and densitometry, and by assaying for bacteria throughout the tube. The occurrence of more than one band is not due to heterogeneity among the bacteria, since each band can be used over to give rise to two again. The first band uses all the oxygen to oxidize portions of one or more of the amino acids, including serine, and the second band consumes the residual serine anaerobically. The results are interpreted to mean that E. coli shows chemotaxis toward oxygen and serine. When no energy source is added to the medium, a band of bacteria still appears. It consumes all the oxygen to oxidize an endogenous energy source. The addition of any one of 10 oxidizable amino acids stimulates the rate of travel of this band. Alanine, an example that was studied in detail, supports such a band that consumes all the oxygen to oxidize a portion of the alanine. Serine, the only amino acid that this strain can use either aerobically or anaerobically when grown under the conditions used here, gives rise to two bands.


Neuroendocrine cells in prostate cancer.
Neuroendocrine (NE) cells are found in prostate tumors, and their incidence is considered a promising prognostic indicator for the development of androgen-independent disease. NE cells are derived from non-NE prostate cancer cells and secrete factors that can act in a paracrine manner to stimulate the survival, growth, motility, and metastatic potential of prostatic carcinoma cells. Factors such as IL-6, epinephrine, and forskolin induce NE differentiation in prostate cancer cells; the mechanisms involve increases in intracellular cAMP, protein kinase A (PKA) activation and reduced intracellular calcium levels. Transcription factors implicated in the acquisition of NE characteristics by prostate cancer cells include STAT3, CREB, EGR1, c-fos, and NF-kappaB. Expression of Chromogranin A, neuron-specific enolase, bcl-2, and the androgen receptor are modulated during NE differentiation and serve as molecular markers for NE cells. Most importantly, NE cells secrete neuropeptides, such as bombesin, neurotensin, PTHrP, serotonin, and calcitonin, which trigger growth and survival responses in androgen-independent prostate cancer cells. Prostate cancer cell receptors that play a role in these processes include the gastrin-releasing peptide (GRP) receptor, neurotensin receptors, and the epidermal growth-factor receptor (EGFR). Signal-transduction molecules activated by these neuropeptides include Src, focal adhesion kinase (FAK), ERK, and PI3K/Akt, with subsequent activation of Elk-1, NF-kappaB, and c-myc transcription factors. A multitude of genes are then expressed by prostate cancer cells, which are involved in proliferation, anti-apoptosis, migration, metastasis, and angiogenesis. Targeting of these pathways at multiple levels can be exploited to inhibit the process by which NE cells contribute to the progression of androgen-independent, treatment-refractory prostate cancer.


Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors.
Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear. Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA. Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors. Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.


Grape Seed Proanthocyanidin Inhibits Mucin Synthesis and Viral Replication by Suppression of AP-1 and NF-κB via p38 MAPKs/JNK Signaling Pathways in Respiratory Syncytial Virus-Infected A549 Cells.
Airway epithelial cells are often infected by respiratory syncytial virus (RSV), one of the most common causes of asthma, bronchiolitis, chronic obstructive pulmonary disease, and pneumonia. During the infection process, excessive mucins instigate airway inflammation. However, the mechanism underlying RSV-induced airway hyper-responsiveness and inflammation is poorly understood. Furthermore, no reliable vaccines or drugs for antiviral therapy are available. In this study, the effect of the natural compound grape seed proanthocyanidin (GSP) on RSV-infected human airway epithelial cells A549 was evaluated. After pretreatment of the cells with or without exposure to RSV with 5-10 μg GSP/mL, the expression of various mucins (MUC1, MUC2, MUC5AC, MUC5B, and MUC8) was evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting, as well as confocal microscopy. We found that GSP significantly decreased RSV-induced mucin synthesis at the mRNA and protein levels. In addition, GSP suppressed the RSV-induced signaling pathways, including extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38, together with nuclear factor kappa B (NF-κB) and activating protein-1 family members (c-Jun and c-Fos). Concomitantly, GSP inhibited the replication of RSV within A549 cells. Taken together, all our results suggest that GSP could be a potent therapeutic agent to suppress excessive mucus production and viral replication in RSV-induced airway inflammatory disorders.


Internet-Based Direct-to-Consumer Genetic Testing: A Systematic Review.
Direct-to-consumer genetic tests (DTC-GT) are easily purchased through the Internet, independent of a physician referral or approval for testing, allowing the retrieval of genetic information outside the clinical context. There is a broad debate about the testing validity, their impact on individuals, and what people know and perceive about them. The aim of this review was to collect evidence on DTC-GT from a comprehensive perspective that unravels the complexity of the phenomenon. A systematic search was carried out through PubMed, Web of Knowledge, and Embase, in addition to Google Scholar according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist with the key term "Direct-to-consumer genetic test." In the final sample, 118 articles were identified. Articles were summarized in five categories according to their focus on (1) knowledge of, attitude toward use of, and perception of DTC-GT (n=37), (2) the impact of genetic risk information on users (n=37), (3) the opinion of health professionals (n=20), (4) the content of websites selling DTC-GT (n=16), and (5) the scientific evidence and clinical utility of the tests (n=14). Most of the articles analyzed the attitude, knowledge, and perception of DTC-GT, highlighting an interest in using DTC-GT, along with the need for a health care professional to help interpret the results. The articles investigating the content analysis of the websites selling these tests are in agreement that the information provided by the companies about genetic testing is not completely comprehensive for the consumer. Given that risk information can modify consumers' health behavior, there are surprisingly few studies carried out on actual consumers and they do not confirm the overall concerns on the possible impact of DTC-GT. Data from studies that investigate the quality of the tests offered confirm that they are not informative, have little predictive power, and do not measure genetic risk appropriately. The impact of DTC-GT on consumers' health perceptions and behaviors is an emerging concern. However, negative effects on consumers or health benefits have yet to be observed. Nevertheless, since the online market of DTC-GT is expected to grow, it is important to remain aware of a possible impact.


Microbiology of diabetic foot infections in a teaching hospital in Malaysia: a retrospective study of 194 cases.
Diabetes mellitus is a progressive disease with chronic complications. Foot infections are a major complication of diabetes and eventually lead to development of gangrene and lower extremity amputation. The microbiological characteristics of diabetic foot infections have not been extensively studied in Malaysia. This study investigated the microbiology of diabetic foot infections and their resistance to antibiotics in patients with diabetic foot infections treated at University of Malaya Medical Centre in Kuala Lumpur, Malaysia. A retrospective analysis was conducted of clinical specimens taken from patients with diabetic foot infections over a 12-month period from July 1, 2004 to June 30, 2005. A total of 194 patients with positive clinical specimens were identified. The clinical specimens were cultured using standard aerobic and anaerobic microbiological techniques. Antibiotic sensitivity testing to different antimicrobial agents was carried out using the disk diffusion method. 287 pathogens were isolated from 194 patients, an average of 1.47 organisms per lesion. The most frequently isolated pathogens were Gram-negative bacteria (52%), including Proteus spp. (28%), Pseudomonas aeruginosa (25%), Klebsiella pneumoniae (15%) and Escherichia coli (9%). Gram-positive bacteria accounted for 45% of all bacterial isolates. Staphylococcus aureus was predominant (44%) among Gram-positive bacteria, followed by Group B streptococci (25%) and Enterococcus spp. (9%). Antimicrobial susceptibility results showed that Gram-negative bacterial isolates were sensitive to imipenem and amikacin while vancomycin showed good activity against Gram-positive bacteria. The antibiogram results of this study suggest that pathogens remain sensitive to a number of widely used agents. Imipenem was equally effective against Gram-negative bacilli and Gram-positive cocci.


Bipolar mixed states: still mixed up?
To provide an update regarding the nosological status of, and treatments for, bipolar mixed states. Mixed states are increasingly recognized as relatively common. The nosological approach in DSM-5 reflects a more liberal application of a mixed states specifier to either the depressed or manic pole of bipolar disorder. The presence of mixed features is associated with a worse clinical course and higher rates of comorbidities including substance abuse. There is ongoing debate about the role of antidepressants in the evolution of such states, but there is increasing consensus that the presence of mixed features should preclude the use of antidepressants. Clinical vigilance and careful evaluation are required to ensure mixed states are not missed in the clinical context. Atypical antipsychotics are emerging as the medications of choice in the pharmacological management of mixed states. Furthermore, research is required into the aetiogenesis and effective treatment of mixed states.


Effect of addition of 2% chlorhexidine or 10% doxycycline on antimicrobial activity of biodentine.
The purpose of this in vitro study was to determine whether the addition of 2% chlorhexidine gluconate or 10% doxycycline would enhance the antimicrobial activity of Biodentine against Staphylococcus aureus (ATCC-25923), Enterococcus faecalis (ATCC-29212), Candida albicans (ATCC-90028), and Streptococcus mutans (MTCC-497). Three wells of 4 mm diameter and 4 mm depth on each plate were prepared on the agar medium with standardized suspensions of each microorganism. Biodentine powder mixed with 2% chlorhexidine (0.06 g) or 10% doxycycline (0.30 g) in its liquid or liquid alone was placed to fill each well. Plates were incubated at 37°C as required for microbial growth. A blinded, independent observer measured zones of inhibition. The data were analyzed using independent "t" test to compare the differences among the three cement preparations for different micro-organisms. All Biodentine samples inhibited microbial growth. The highest mean diameters of zone of inhibition for all the micro-organisms were found around Biodentine/chlorhexidine (13.417) followed by Biodentine alone (12.236) and Biodentine/doxycycline (11.25). In conclusion, adding 2% chlorhexidine gluconate in liquid of Biodentine enhanced the antimicrobial activity of Biodentine against all the tested micro-organisms except Candida albicans, while addition of 10% doxycycline decreased the antimicrobial activity of Biodentine. The differences were significant statistically (P < 0.05).


Differential effects of neuromuscular electrical stimulation parameters on submental motor-evoked potentials.
Neuromuscular electrical stimulation (NMES) of the muscles underlying the pharynx and faucial pillars affects the excitability of corticobulbar projections in a frequency- and duration-specific manner. The anterior hyomandibular (submental) muscles are primary targets for the clinical application of NMES to improve disordered swallowing, but the optimal NMES parameters for this application are unknown. To determine the influence of NMES parameters on the excitability of corticobulbar projections to the submental musculature. Transcranial magnetic stimulation (TMS) was used in event-related protocols, triggered by either volitional contraction of the submental muscles or pharyngeal swallowing, to assess corticobulbar excitability prior to, immediately following, and 30, 60, and 90 minutes post-NMES in 25 healthy volunteers. In the first 2 experiments, 4 stimulus frequencies (5, 20, 40, and 80 Hz) and 3 NMES dosages, manipulated through stimulus train durations or number of repetitions, were evaluated. The optimal excitatory NMES triggered by volitional swallowing (event-related NMES) was then replicated in a new sample and contrasted with non-event-related NMES (either discrete events or continuously for 1 hour). It was found that 80Hz NMES increased motor-evoked potential (MEP) amplitude at 30 minutes and 60 minutes poststimulation only after 60 repetitions of 4-s event-related NMES trains. Non-event-related and continuous NMES did not affect MEP amplitudes. No changes in MEP onset latencies were observed. Changes in corticobulbar excitability induced by NMES of the submental muscle group are frequency and dose dependent and only occur after NMES triggered by volitional swallowing. Underlying neural mechanisms are discussed.


Cortical activity during walking and balance tasks in older adults and in people with Parkinson's disease: A structured review.
An emerging body of literature has examined cortical activity during walking and balance tasks in older adults and in people with Parkinson's disease, specifically using functional near infrared spectroscopy (fNIRS) or electroencephalography (EEG). This review provides an overview of this developing area, and examines the disease-specific mechanisms underlying walking or balance deficits. Medline, PubMed, PsychInfo and Scopus databases were searched. Articles that described cortical activity during walking and balance tasks in older adults and in those with PD were screened by the reviewers. Thirty-seven full-text articles were included for review, following an initial yield of 566 studies. This review summarizes study findings, where increased cortical activity appears to be required for older adults and further for participants with PD to perform walking and balance tasks, but specific activation patterns vary with the demands of the particular task. Studies attributed cortical activation to compensatory mechanisms for underlying age- or PD-related deficits in automatic movement control. However, a lack of standardization within the reviewed studies was evident from the wide range of study protocols, instruments, regions of interest, outcomes and interpretation of outcomes that were reported. Unstandardized data collection, processing and reporting limited the clinical relevance and interpretation of study findings. Future work to standardize approaches to the measurement of cortical activity during walking and balance tasks in older adults and people with PD with fNIRS and EEG systems is needed, which will allow direct comparison of results and ensure robust data collection/reporting. Based on the reviewed articles we provide clinical and future research recommendations.


Amended safety assessment of sodium picramate and picramic acid.
Sodium picramate is the sodium salt of picramic acid, a substituted phenolic compound. Sodium picramate and picramic acid function as hair colorants; they are reportedly used in 31 and 3 hair-dye products, respectively. No concentration-of-use data were available for sodium picramate, but picramic acid was reported to be used at 0.6%. The Cosmetic Ingredient Review Expert Panel recognized that adding picramic acid to a hair-dye formulation likely results in formation of a salt such as sodium picramate, which suggested that safety test data for one ingredient would be applicable to the other. Hair dyes containing these ingredients bear a caution statement and patch test instructions for determining whether the product causes skin irritation. The panel finds that the available data support the safety of these colorants in hair dyes and expects that sodium picramate would be used at concentrations comparable to those reported for picramic acid.


Major Improvements to the Heliconius melpomene Genome Assembly Used to Confirm 10 Chromosome Fusion Events in 6 Million Years of Butterfly Evolution.
The Heliconius butterflies are a widely studied adaptive radiation of 46 species spread across Central and South America, several of which are known to hybridize in the wild. Here, we present a substantially improved assembly of the Heliconius melpomene genome, developed using novel methods that should be applicable to improving other genome assemblies produced using short read sequencing. First, we whole-genome-sequenced a pedigree to produce a linkage map incorporating 99% of the genome. Second, we incorporated haplotype scaffolds extensively to produce a more complete haploid version of the draft genome. Third, we incorporated ∼20x coverage of Pacific Biosciences sequencing, and scaffolded the haploid genome using an assembly of this long-read sequence. These improvements result in a genome of 795 scaffolds, 275 Mb in length, with an N50 length of 2.1 Mb, an N50 number of 34, and with 99% of the genome placed, and 84% anchored on chromosomes. We use the new genome assembly to confirm that the Heliconius genome underwent 10 chromosome fusions since the split with its sister genus Eueides, over a period of about 6 million yr.


Expression of the heparin-binding cytokines, midkine (MK) and HB-GAM (pleiotrophin) is associated with epithelial-mesenchymal interactions during fetal development and organogenesis.
Midkine (MK) and heparin binding-growth associated molecule (HB-GAM or pleiotrophin), constitute a new family of heparin-binding proteins implicated in the regulation of growth and differentiation (T. Muramatsu (1993) Int. J. Dev. Biol. 37, 183-188). We used affinity-purified antibodies against MK and HB-GAM to analyze their distribution during mouse embryonic development. From 9 to 14.5 day post-coitum (dpc), both proteins were detected in central and peripheral nervous systems, facial processes, limb buds, sense organs, respiratory, digestive, urogenital, and skeletal systems. MK and HB-GAM were often localized on the surface of differentiating cells and in basement membranes of organs undergoing epithelial-mesenchymal interactions. The level of MK protein decreased considerably in the 16.5 dpc embryo, whereas HB-GAM staining persisted in many tissues. Our in situ hybridization results revealed a widespread expression of MK transcripts that was not always consistent with the distribution of MK protein in developing tissues. In many epithelio-mesenchymal organs MK and HB-GAM were codistributed with syndecan-1, a cell surface proteoglycan. In limb buds and facial processes, MK, HB-GAM, and syndecan-1 were localized to the apical epithelium and the adjacent proliferating mesenchyme. Both MK and HB-GAM bound syndecan-1 in solid-phase assays in a heparan sulfate-dependent manner. The biological effects of MK and HB-GAM on limb and facial mesenchyme were studied in vitro by application of beads preloaded with the proteins. Neither MK nor HB-GAM stimulated mesenchymal cell proliferation or induced syndecan-1 expression. Taken together these results indicate that MK and HB-GAM may play regulatory roles in differentiation and morphogenesis of the vertebrate embryo, particularly in epithelio-mesenchymal organs, and suggest molecular interactions with syndecan-1.


Nintedanib in Idiopathic Pulmonary Fibrosis: Practical Management Recommendations for Potential Adverse Events.
Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease with a dismal survival rate of only 3 years and no curative pharmacological therapy. The recent approval of 2 anti-fibrotic drugs (nintedanib and pirfenidone) that slow disease progression has provided some hope for patients. However, effectively managing anti-fibrotic treatment can be a challenge due to tolerability issues, the presence of pulmonary and extra-pulmonary comorbidities, and the need for concomitant medications in many patients. In general, making clear evidence-based decisions can be difficult for physicians because patients with comorbidities are often excluded from clinical trials. Since currently anti-fibrotic drugs are the only effective therapeutics capable of slowing disease progression, it is imperative that all treatment options are thoroughly evaluated and exhausted in each individual, irrespective of complicating factors, to permit the best outcome for the patient. In this review, we present data from clinical trials, post hoc analyses, post-marketing surveillance, and real-world studies that are relevant to the management of nintedanib treatment. In addition, we also provide practical recommendations developed by a multidisciplinary panel of experts for the management of nintedanib treatment in patients with IPF associated complications and those experiencing gastrointestinal side effects.


Mirror aneurysms: a reflection on natural history.
Investigators conducting the International Study of Unruptured Intracranial Aneurysms, sponsored by the National Institutes of Health, sought to evaluate predictors of future hemorrhage in patients who had unruptured mirror aneurysms. These paired aneurysms in bilateral arterial positions mirror each other; their natural history is unknown. Centers in the US, Canada, and Europe enrolled patients for prospective assessment of unruptured intracranial aneurysms. Central radiological review confirmed the presence or absence of mirror aneurysms in patients without a history of prior subarachnoid hemorrhage (SAH) (Group 1). Outcome at 1 and 5 years and aneurysm characteristics are compared. Of 3120 patients with aneurysms treated in 61 centers, 376 (12%) had mirror aneurysms, which are more common in women than men (82% [n = 308] vs 73% [n = 1992], respectively; p <0.001) and in patients with a family history of aneurysm or SAH (p <0.001). Compared with patients with nonmirror saccular aneurysms, a greater percentage of patients with mirror aneurysms had larger (>10 mm) aneurysms (mean maximum diameter 11.7 vs 10.4 mm, respectively; p <0.001). The most common distribution for mirror aneurysms was the middle cerebral artery (34% [126 patients]) followed by noncavernous internal carotid artery (32% [121]), posterior communicating artery (16% [60]), cavernous internal carotid artery (13% [48]), anterior cerebral artery/anterior communicating artery (3% [13]), and vertebrobasilar circulation (2% [8]). When these patients were compared with patients without mirror aneurysms, no statistically significant differences were found in age (mean age 54 years in both groups), blood pressure, smoking history, or cardiac disease. Aneurysm rupture rates were similar (3.0% for patients with mirror aneurysms vs 2.8% for those without). Overall, patients with mirror aneurysms were more likely to be women, to report a family history of aneurysmal SAH, and to have larger aneurysms. The presence of a mirror aneurysm was not an independent predictor of future SAHs.


Assessing the Impact of the Pediatric Orthopaedic Society of North America (POSNA) Visiting Scholar Program.
The primary objective of the Pediatric Orthopaedic Society of North America (POSNA)-Children's Orthopaedics in Underserved Regions (COUR) Visiting Scholar Program is to engage emerging leaders from low-income and middle-income countries (LMICs) in POSNA educational offerings. This study aims to outline the educational and leadership activities pursued by the alumni of the POSNA-COUR Visiting Scholars Program. We hypothesized that there may be a discrepancy between visiting scholar interest level and actual participation in key follow-up educational and leadership activities. A 32-question online survey developed by the POSNA-COUR committee was electronically delivered to the POSNA-COUR visiting scholarship recipients from 2007 to 2019. The survey inquired about scholars' educational background, clinical practice, and academic interests. Respondents also reported educational, research, and leadership activities that occurred following program participation. Fifty-seven percent (44/77) of the previous scholars participated in the survey. Sixty-eight percent reported that >75% of their practice was comprised of pediatric patients. The COUR scholarship afforded 15 scholars the opportunity to attend their first educational conference in North America. The majority (86%) of respondents consulted POSNA members on care-related issues, 52% organized a POSNA member's visit to their country, and 13% have become members of POSNA. Scholars shared their knowledge with colleagues through lectures (73%), demonstration of surgical skills (63%), mentorship (59%), hosting local courses (43%), and research (14%). There was a significant discrepancy between interest and actual participation in research (98% vs. 16%, P<0.0001), hosting a regional course (98% vs. 43%, P<0.0001), and becoming a member of POSNA (96% vs. 13%, P<0.0001). The POSNA-COUR Visiting Scholar Program is effective at providing beneficial educational opportunities to orthopaedic surgeons from LMICs. There is, however, a significant discrepancy between reported scholar interest and actual engagement in leadership endeavors including: research, organization of regional courses, and POSNA membership. The impact of the POSNA-COUR visiting scholar program can be enhanced through collaboration with scholarship alumni in these key areas. Level IV-case series.


Cerebrospinal fluid abnormalities and rate of decline in everyday function across the dementia spectrum: normal aging, mild cognitive impairment, and Alzheimer disease.
To investigate the effect of cerebrospinal fluid (CSF) abnormalities on the rate of decline in everyday function in normal aging, mild cognitive impairment (MCI), and mild Alzheimer disease (AD). Immunoassays of total tau (t-tau), tau phosphorylated at threonine 181 (p-tau(181)), and beta-amyloid 1-42 (Abeta(42)) concentrations were performed in CSF obtained from participants in the Alzheimer's Disease Neuroimaging Initiative. Random effects regressions were used to examine the relationship among CSF abnormalities, cognitive impairment (assessed with the Alzheimer Disease Assessment Scale-cognitive subscale [ADAS-Cog]), and functional decline (assessed with the Pfeffer Functional Activities Questionnaire) and to determine whether the impact of CSF abnormalities on functional decline is mediated by cognitive impairment. Fifty-eight sites in the United States and Canada. One hundred fourteen cognitively intact adults, 195 patients with MCI, and 100 patients with mild AD. Main Outcome Measure Decline in the Pfeffer Functional Activities Questionnaire score. Abnormalities in all CSF analytes were associated with functional decline in MCI, and all but the t-tau:Abeta(42) ratio were associated with functional decline in controls. No abnormal CSF analyte was associated with functional decline in AD. Among controls, p-tau(181) concentration was the most sensitive to functional decline, whereas in MCI it was Abeta(42) concentration. Cerebrospinal fluid biomarkers were uniformly more sensitive to functional decline than the ADAS-Cog score among controls and variably so in MCI, whereas the ADAS-Cog score was unequivocally more sensitive than CSF biomarkers in AD. The impact of CSF abnormalities on functional decline in MCI was partially mediated by their effect on cognitive status. Across all diagnostic groups, persons with both tau and Abeta(42) abnormalities exhibited the steepest rate of functional decline. Abnormalities in CSF are associated with functional decline and thus with future development of AD in controls and patients with MCI. However, they do not predict further functional degradation in patients with AD. Persons with comorbid tau and Abeta(42) abnormalities are at greatest risk of functional loss.


Ultrasonographic evaluation of abdominal wall fat index, carotid intima-media thickness and plaque score in obstructive sleep apnea syndrome.
Obstructive sleep apnea syndrome (OSAS) is strongly related with increased risk of cardiovascular diseases andvisceral obesity. Abdominal wall fat index (AFI) is an indicator of visceral fat accumulation determined by ultrasonography(US). Carotid intima-media thickness (C-IMT) and carotid plaque score (C-PS) are the indicators of cardiovascular risk. Theaim of this study was to investigate the relation between OSAS and AFI, C-IMT or C-PS. One-hundred and four subjects (31 females, 73 males) between 23-73year-old, candidate for polysomnography (PSG) with suspect of OSAS and without other atherosclerotic risk factor, were evaluated by US. AFI, C-IMTmean and C-PS values were determined and the subjects were grouped according to their apnea-hypopnea index (AHI) values as follows: no OSAS (<5), mild OSAS (5-15), moderate OSAS (15-30) and severe OSAS (>30). There was a statistically significant correlation between AFI and AHI (p=0.019). The C-IMTmean values of subjects with OSAS (AHI >5) were significantly higher than those without OSAS (AHI <5) (p=0.035). C-PS was not correlated with AHI (p=0.345) and also there was not a statistically significant difference between OSAS groups in terms of C-PS (p=0.775). This study revealed that AFI correlates with AHI and C-IMT increases in OSAS. The two parameters could be used as indicators of risk of metabolic disorders and atheroscleroticdiseases in subjects with sleep apnea in the future.


Effect of Thuja occidentalis and its polysaccharide on cell-mediated immune responses and cytokine levels of metastatic tumor-bearing animals.
Tumor microenvironment induces an active immune tolerance and escapes immune surveillance. In order to achieve an effective antitumor immune response, appropriately activated immune cells should maintain their antitumor activity to overcome the immune suppressive tumor microenvironment. This study focuses on the effect of Thuja occidentalis L. (Cupressaceae) extract and its polysaccharide (TPS) on cell-mediated immune response (CMI) in metastasis bearing mice. Metastasis was induced by injecting B16F-10 melanoma cells in mice through the tail vein and effector mechanisms of CMI was studied by analyzing cytotoxic T-lymphocyte (CTL) activity, natural killer (NK) cell activity, antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent complement-mediated cytotoxicity (ACC). The effect of T. occidentalis and TPS on pro-inflammatory cytokines and tissue inhibitor matrix metalloproteinases (TIMP) levels were also analyzed. Administration of T. occidentalis and TPS enhanced the NK cell activity, ADCC and ACC much earlier than the control tumor-bearing animals. T. occidentalis and TPS were also found to decrease the elevated level of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, GM-CSF and tumor necrosis factor (TNF)-α in the serum of metastatic tumor-bearing animals. The level of antitumor factors such as IL-2 and TIMP was elevated by the treatment with T. occidentalis and TPS in the serum, which was lowered in the untreated tumor-bearing animals. This study clearly suggests that T. occidentalis and TPS effectively stimulate cell-mediated immune system and decrease pro-inflammatory cytokines, thereby inhibiting metastasis of tumor cells.


Molecular Characterization and Event-Specific Real-Time PCR Detection of Two Dissimilar Groups of Genetically Modified Petunia (Petunia x hybrida) Sold on the Market.
Petunia plants with unusual orange flowers were noticed on the European market and confirmed to be genetically modified (GM) by the Finnish authorities in spring 2017. Later in 2017, inspections and controls performed by several official laboratories of national competent authorities in the European Union detected several GM petunia varieties with orange flowers, but also another group of unusually colored flowers. In the latter group, a so far undetected gene coding for a flavonoid 3'5' hydroxylase (F3'5'H) responsible for the purple color was identified by German and Dutch authorities, suggesting that the petunias found on the markets contain different genetic constructs. Here, a strategy is described for the identification of GM petunia varieties. It is based on an initial GMO screening for known elements using (real-time) PCR and subsequent identification of the insertion sites by a gene walking-like approach called ALF (amplification of linearly-enriched fragments) in combination with Sanger and MinION sequencing. The results indicate that the positively identified GM petunias can be traced back to two dissimilar GM events used for breeding of the different varieties. The test results also confirm that the transgenic petunia event RL01-17 used in the first German field trial in 1991 is not the origin of the GM petunias sold on the market. On basis of the obtained sequence data, event-specific real-time PCR confirmatory methods were developed and validated. These methods are applicable for the rapid detection and identification of GM petunias in routine analysis. In addition, a decision support system was developed for revealing the most likely origin of the GM petunia.


Therapies for depression in Parkinson's disease.
Depression is one of the most common neuropsychiatric disturbances in Parkinson's disease. 40% of observed variation in quality of life is due to depression. However, there is little hard evidence of the efficacy and safety of antidepressant therapies in Parkinson's disease. To assess the efficacy and safety of antidepressant therapies in idiopathic Parkinson's disease. Safety refers to both the direct side-effects of the therapy and also the therapy's interactions with the symptoms of Parkinson's disease and with the antiparkinsonian medications. Relevant clinical trials were identified by electronic searches the Cochrane Controlled Trials Register (the Cochrane Library Issue 3, 2001), MEDLINE(1996-2001), EMBASE (1974-2001), PsychLit (1800's-2001), CINAHL (1982-2001) databases. The reference list of all trial reports and reviews were examined. Queries were sent out to all manufacturers and distributors of antidepressants within the UK requesting information on any relevant clinical trials. Randomised controlled trials (RCT) examining licensed oral antidepressants, electroconvulsive therapy (ECT) or behavioural therapy in the treatment of depression in idiopathic Parkinson's disease. Data was extracted and assessed independently by three of the authors. Disagreements were resolved by discussion. Three randomised controlled trials were found examining oral antidepressant medications in Parkinson's disease in a total of 106 patients. No eligible trials of ECT or behavioural therapy were found. In the first arm of the crossover trial by Andersen 1980 (n=22) patients in the nortriptyline group showed a larger improvement than placebo group in median depression score in a self-made 31-item depression rating scale after 16 weeks of treatment but statistical significance was not calculated. A parallel group trial by Wermuth 1998 (n=37) did not show any statistically significant difference between the citalopram and placebo groups in the Hamilton Depression Scale after 52 weeks of treatment. The third study by Rabey 1996 (n=47) was a randomised open-label trial to compare fluvoxamine versus amitriptyline. Similar numbers of patients in amitriptyline and fluvoxamine groups (60% vs 55%) had a 50% reduction of Hamilton score after 16 months of treatment. However, further assessment of this trial was not possible because only summary results were available from an abstract and attempts to contact the authors failed. Visual hallucinations or confusion had been reported in patients with fluvoxamine and amitriptyline. Otherwise, no other major side effects were found in the other two trials. Insufficient data on the effectiveness and safety of any antidepressants therapies in Parkinson's disease are available on which to make recommendations for their use. Further large scale randomised controlled trials are urgently required in this area.


Combination therapy in a xenograft model of glioblastoma: enhancement of the antitumor activity of temozolomide by an MDM2 antagonist.
OBJECTIVE Improvement in treatment outcome for patients with glioblastoma multiforme (GBM) requires a multifaceted approach due to dysregulation of numerous signaling pathways. The murine double minute 2 (MDM2) protein may fulfill this requirement because it is involved in the regulation of growth, survival, and invasion. The objective of this study was to investigate the impact of modulating MDM2 function in combination with front-line temozolomide (TMZ) therapy in GBM. METHODS The combination of TMZ with the MDM2 protein-protein interaction inhibitor nutlin3a was evaluated for effects on cell growth, p53 pathway activation, expression of DNA repair proteins, and invasive properties. In vivo efficacy was assessed in xenograft models of human GBM. RESULTS In combination, TMZ/nutlin3a was additive to synergistic in decreasing growth of wild-type p53 GBM cells. Pharmacodynamic studies demonstrated that inhibition of cell growth following exposure to TMZ/nutlin3a correlated with: 1) activation of the p53 pathway, 2) downregulation of DNA repair proteins, 3) persistence of DNA damage, and 4) decreased invasion. Pharmacokinetic studies indicated that nutlin3a was detected in human intracranial tumor xenografts. To assess therapeutic potential, efficacy studies were conducted in a xenograft model of intracranial GBM by using GBM cells derived from a recurrent wild-type p53 GBM that is highly TMZ resistant (GBM10). Three 5-day cycles of TMZ/nutlin3a resulted in a significant increase in the survival of mice with GBM10 intracranial tumors compared with single-agent therapy. CONCLUSIONS Modulation of MDM2/p53-associated signaling pathways is a novel approach for decreasing TMZ resistance in GBM. To the authors' knowledge, this is the first study in a humanized intracranial patient-derived xenograft model to demonstrate the efficacy of combining front-line TMZ therapy and an inhibitor of MDM2 protein-protein interactions.


Central venous catheter-related bloodstream infection and colonization: the impact of insertion site and distribution of multidrug-resistant pathogens.
Placement of central-venous catheters (CVCs) is an essential practice in the management of hospitalized patients, however, insertion at the commonly used sites has often the potential of inducing major complications. Neverthelss, the impact of specific site central line catheter insertion on catheter-associated bloodstream infections (CLABSIs) has not been clarified yet in the literature. The aim of the study was to compare CLABSIs and catheter colonization rates among the three catheter insertion sites: subclavian (SC), internal jugular (IJ) and femoral (FEM) in hospitalized patients. Moreover, to analyze the distribution of pathogens and their antimicrobial resistance profiles at these three sites, concurrently. We performed a retrospective analysis of data collected prospectively from all catheterized patients at a tertiary care Greek hospital from May 2016 to May 2018. Data was collected on 1414 CVCs and 13,054 CVC-days. Τhe incidence of CLABSIs among the three sites was as follows: SC:5.1/1000 catheter/days, IJ: 3.73/1000 catheter/days and FEM: 6.93/1000 catheter/days (p = 0.37). The incidence of colonization was as follows: SC:13.39/1000 catheter/days; IJ:7.34/ 1000 catheter/days; FEM:22.91/1000 catheter/days (p = 0.009). MDROs predominated in both CLABSIs and tip colonizations (59.3 and 61%, respectively) with Acinetobacter baumanii being the predominant pathogen (16/59, 27.1% and 44/144, 30.5%, respectively). The incidence of CLABSIs due to multidrug-resistant organisms (MDROs) was as follows: SC:3.83/1000 catheter days; IJ:1.49/1000 catheter days; FEM:5.86/1000 catheter days (p = 0.04). The incidence of tip colonization by MDROs among the 3 sites was as follows: SC:8.93/1000 catheter/days; IJ:4.48/1000 catheter/days; FEM:12.79/1000 catheter/days (p = 0.06). There was no significant difference in the type of pathogen isolated among site groups for both CLABSIs and tip colonizations. FEM site of catheter insertion was associated with a higher rate of bloodstream infection and catheters' colonization compared to IJ and SC sites. Furthermore, this survey highlights the changing trend of the distribution of frequent pathogens and resistance patterns towards MDR Gram-negative pathogens, underscoring the need for consistent monitoring of antimicrobial resistance patterns of these specific infections.


Highly dispersive micro-ring resonator based on one dimensional photonic crystal waveguide design and analysis.
We propose and analyze a novel design of a hybrid micro-ring resonator and photonic crystal device. The proposed device is based on a micro-ring resonator with the addition of a series of periodic defects that are introduced to the microring. When the wavelength of operation approaches the band-gap of the periodic structure, the modal dispersion is significantly increased. The huge dispersion leads to narrowing of the spectral linewidth of the resonator. We predict an order of magnitude linewidth narrowing for a microring radius of the order of 10mum. The proposed hybrid device is analyzed theoretically and numerically using finite-elements calculations and finite-difference-time-domain calculations. We also present as well as the design and analysis of add-drop and notch filters based on the highly dispersive ring resonator.