Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration.
Platelet-rich plasma has been used to treat articular cartilage defects, with the expectations of anabolic and anti-inflammatory effects. However, its role on cellular chondrogenic or fibrogenic commitment is still a controversy. Herein, the role of platelet-rich plasma releasate, the product obtained following platelet-rich plasma activation, on cellular commitment toward the chondrogenic lineage was evaluated in vitro. Human nasoseptal chondrogenic cells and human bone marrow mesenchymal stromal cells were used as cell types already committed to the chondrogenic lineage and undifferentiated cells, respectively, as different concentrations of platelet-rich plasma releasate were tested in comparison to commonly used fetal bovine serum. Low concentration of platelet-rich plasma releasate (2.5%) presented similar effects on cellular growth compared to 10% fetal bovine serum, for both cell types. In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation. Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum. Histological analyses and measurements of pellet area demonstrated that even low concentrations of platelet-rich plasma releasate were enough to prevent nasoseptal chondrogenic cells from losing their chondrogenic potential due to in vitro expansion thereby promoting their recommitment. Low concentration of platelet-rich plasma releasate supplemented in chondrogenic medium also increased the chondrogenic potential of mesenchymal stromal cells seeded on collagen-hyaluronic acid scaffolds, as observed by an increase in chondrogenic-related gene expression, sulfated glycosaminoglycan production, and compressive modulus following in vitro culture. On the contrary, higher concentration of platelet-rich plasma releasate (10%) hampered some of these features. In conclusion, platelet-rich plasma releasate was able to prevent cellular chondrogenic capacity loss, inducing regain of their phenotype, and modulate cell commitment. Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.


Bolus arginine supplementation affects neither muscle blood flow nor muscle protein synthesis in young men at rest or after resistance exercise.
The aim of this study was to investigate the ergogenic potential of arginine on NO synthesis, muscle blood flow, and skeletal muscle protein synthesis (MPS). Eight healthy young men (22.1 ± 2.6 y, 1.79 ± 0.06 m, 76.6 ± 6.2 kg; mean ± SD) participated in 2 trials where they performed a bout of unilateral leg resistance exercise and ingested a drink containing either 10 g essential amino acids with 10 g l-arginine (ARG) or an isonitrogenous control (CON). Femoral artery blood flow of both the nonexercised and exercised leg was measured continuously using pulsed-wave Doppler ultrasound, while rates of mixed and myofibrillar MPS were determined using a primed continuous infusion of L-[ring-(13)C(6)] or L-[ring-(2)H(5)]phenylalanine. The plasma arginine concentration increased 300% during the ARG trial but not during the CON trial (P < 0.001). Plasma nitrate, nitrite, and endothelin-1, all markers of NO synthesis, did not change during either the ARG or CON trial. Plasma growth hormone increased to a greater degree after exercise in the ARG trial than CON trial (P < 0.05). Femoral artery blood flow increased 270% above basal in the exercised leg (P < 0.001) but not in the nonexercised leg, with no differences between the ARG and CON trials. Mixed and myofibrillar MPS were both greater in the exercised leg compared with the nonexercised leg (P < 0.001), but did not differ between the ARG and CON treatments. We conclude that an oral bolus (10 g) of arginine does not increase NO synthesis or muscle blood flow. Furthermore, arginine does not enhance mixed or myofibrillar MPS either at rest or after resistance exercise beyond that achieved by feeding alone.


Long-term effects on symptoms by reducing electric fields from visual display units.
The purpose of the study was to see whether the results of an earlier study [ie, that skin symptoms were reduced by reducing electric fields from visual display units (VDU)] could be reproduced or not. In addition, an attempt was made to determine whether eye symptoms and symptoms from the nervous system could be reduced by reducing VDU electric fields. The study was designed as a controlled double-blind intervention. The electric fields were reduced by using electric-conducting screen filters. Forty-two persons completed the study while working at their ordinary job, first 1 week with no filter, then 3 months with an inactive filter and then 3 months with an active filter (or in reverse order). The inactive filters were identical to the active ones, except that their ground cables were replaced by empty plastic insulation. The inactive filters did not reduce the fields from the VDU. The fields were significantly lower with active filters than with inactive filters. Most of the symptoms were statistically significantly less pronounced in the periods with the filters when compared with the period with no filter. This finding can be explained by visual effects and psychological effects. No statistically significant difference in symptom severeness was observed between the period with an inactive filter and the one with an active filter. The study does not support the hypothesis that skin, eye, or nervous system symptoms can be reduced by reducing VDU electric fields.


[Educational issues and strategies to improve APN education].
This study was aimed at exploring the current status of graduate programs for an advanced practice nurse(APN) to recommend future directions of APN education. A total of 142 students enrolled in seven APN specialty programs, 67 professors who were involved in APN education, and nine nurse administrators participated in the study. Data was collected by questionnaires and focus group interviews. The current definition of APN was found not to be specific enough to represent expected roles of APN in regards to knowledge, attitudes, roles, and skills. Standard curricula employed regardless of the area of APN specialty, lack of qualified clinical practice settings, as well as prepared instructors were found to be problematic. The following needs to be addressed: 1. redefining of APN roles, 2. tailoring specialty areas of APN, 3. consolidating educational programs, and 4. ensuring APN role models and faculty. Suggesting a CNS role in Korean APN, areas of APN should be rearranged to clarify their roles and educational programs need to be further developed to meet the expectations and quality of APNs. It is necessary to ensure APN's employment in the health care system by laws and policies to perform advanced nursing roles.


Patterns and trends of transcatheter aortic valve implantation in Italy: insights from RISPEVA.
Clinical trials have shown that transcatheter aortic valve implantation for aortic stenosis compares favorably to surgical replacement in high-risk patients and is superior to medical therapy in those at prohibitive risk. There is uncertainty however on patterns and trends in transcatheter aortic valve implantation, especially focusing on Italy. The RISPEVA study is a prospective Italian registry including 21 institutions. Patients have been enrolled since late 2012, and data collection includes several baseline, procedural, in-hospital, and follow-up details. For the present analysis on patterns and trends, we focused on patients enrolled between 2012 and 2015, and as primary variable on the prevalence of high versus prohibitive surgical risk, limiting our scope to procedural outcomes. A total of 1157 patients were included. The temporal breakdown was 376 (33%) patients enrolled in 2013, 408 (35%) in 2014, and 373 (32%) in 2015. Several patient features differed over time, including risk score, peripheral artery disease, end-stage pulmonary disease, and prior valvuloplasty (all P < 0.05). Several procedural features differed significantly over time, including sheath size, use of general anesthesia, Prostar closure device, predilation, antiembolic device, new TAVI device, and multiple prostheses (all P < 0.05). No significant temporal differences were found for major clinical outcomes, whereas the occurrence of moderate or severe postprocedural regurgitation and pacemaker dependency decreased over the years (both P < 0.05). According to the RISPEVA results, the Italian uptake of TAVI is steady, with evident trends toward less invasive approaches and fitter patients.


Crystalluria and its possible significance. A patient-control study.
The significance of crystalluria in the diagnosis and prognosis of urolithiasis remains a controversial subject in the current urological literature. In this study, in addition to the standard urolithiasis clinical and biochemical work-up, routine urine microscopy was performed to study crystals in 1 fresh and 2 stored morning urine samples from 140 urinary stone patients and 42 controls. Crystalluria was more frequently detected in patients (9.3% of the fresh samples) than in controls (2%). Storing the samples for 6 hours did not increase the frequency percent of detected crystalluria either in patients or controls. However, in the samples stored for 24 hours, the frequency of crystalluria increased to 27.1% in patients and only to 12% in controls, though the pH did not change from that of the fresh sample. In addition, while calcium oxalate crystals in patients formed aggregates whether in fresh or 24 hour samples, those of controls did not. This denotes a characteristic change in the physico-chemical properties of the urine of stone formers from that of controls. Accordingly, the study of crystalluria in patients with urolithiasis seems to help in the proper evaluation and, maybe, treatment of the disease.


Non-random nature of 2-acetylaminofluorene-induced alterations of DNA template capacity.
Male rats were fed a diet containing 0.03% (w/w) 2-acetylaminofluorene (AAF) and their hepatic DNA was isolated and transcribed with E. coli RNA polymerase. Ingestion of the carcinogen-containing diet for 4 days substantially reduced the template capacity of the isolated DNA. This reduction in template capacity was due to an apparent decreased RNA chain size (up to 50%), with no significant changes in initiation or re-initiation of RNA synthesis. This premature termination of RNA synthesis was accompanied, in some instances, by a reduced rate of RNA chain elongation. When the rats were returned to a basal diet for 7 days following 4 days of AAF ingestion, template capacity and RNA chain size returned to control values. Fractionation of hepatic chromatin on a glycerol gradient revealed that inhibition of DNA template capacity occurs on portions exhibiting characteristics of expressed, as well as those with characteristics of repressed, segments of the genome. In contrast, the DNA isolated from a small, highly condensed chromatin fraction (15% of total chromatin-DNA) showed no significant reduction in total template capacity. Analysis of the fidelity of RNA synthesis on this DNA template was performed by determining the rate of addition of individual nucleotide triphosphates to a growing RNA chain. Large reductions in the rates of adenosine and uridine polymerization were observed while no changes in guanosine or cytidine polymerization were found. This suggests the presence of functionally significant carcinogen-induced modifications of adenine. The inhibition in the rate of adenosine and uridine polymerization was reversed when the animals were placed on a basal diet after AAF ingestion.


Diagnostic heterogeneity of diabetes in lean young adults: classification based on immunological and genetic parameters.
The aim of our study was to investigate the relative prevalence of the different forms of diabetes in young adults and their respective clinical characteristics. Included were 51 nonobese patients (BMI < 27 kg/m2) with diabetes diagnosed before age 40, excluding typical IDDM. Each patient was subjected to screening for glucokinase gene (MODY2) and mitochondrial DNA (at nucleotide 3243) mutations, to HLA class II genotyping, and screening for the presence of islet cell antibodies (ICAs) and anti-GAD antibodies. Informative families were analyzed for linkage of diabetes to chromosome 12q (MODY3). Based on clinical criteria, patients were subdivided into MODY (n = 19) and non-MODY (n = 32). In the MODY group, we identified three patients with MODY2, one with the 3243 mitochondrial mutation, and another with autoimmune diabetes. One of the five MODY families available for linkage study was shown to have MODY3. In the non-MODY group, we found five patients with autoimmune diabetes and one with MODY2. No clinical parameter was helpful to classify patients in one of these subclasses of diabetes; however, the glucagon-stimulated C-peptide was useful to discriminate between MODY2 patients and the others. In conclusion, young and lean non-insulin-dependent diabetic patients constitute a very heterogeneous group, although they present similar clinical characteristics. The clinical distinction of MODY and non-MODY patients allows correct classification in, at most, 75% of the patients and thus is not sufficient to predict clinical course. However, immunological and genetic parameters allowed us to classify only 25% of the patients in specific diagnostic classes.


Asian dust: seasonal transport to the hawaiian islands.
Analyses of atmospheric particles collected at Mauna Loa Observatory in Hawaii from February 1979 through September 1982 reveal strong influxes of Asian dust in the spring of each year. Concentrations of a typical crustal element, aluminum, are more than an order of magnitude greater between February and June than during the remainder of the year (71 +/- 51 versus 6.7 +/- 2.3 nanograms per cubic meter). The mass of crustal material transported during the relatively short dust episodes accounts for an average of 80 percent of the total yearly mass of atmospheric particles at 3400 meters on Mauna Loa.


Cardiorespiratory function is significantly improved following corrective surgery for severe pectus excavatum. Proposed treatment guidelines.
Pectus excavatum is a congenital chest abnormality which may become more marked during childhood and teenage growth. Young teenagers with severe PE deformities often become short of breath during strenuous exercise and complain of becoming easily fatigued. The pathophysiologic effects of PE remain controversial because they have been difficult to measure, especially in young children. Symptomatic patients often have significant subjective improvement during exercise following PE repair and there is a major cosmetic impact on teenagers with severe deformities. This study was designed to measure pulmonary and cardiac function with exercise before and after corrective surgery. Pulmonary function testing and incremental exercise testing were performed in 36 adolescents with pectus excavatum (PE) and 10 age-matched, healthy control subjects. Six months after corrective surgery was performed, 15 of these PE patients and 6 control subjects were re-evaluated for pulmonary function. Before surgery, PE subjects had a lower forced vital capacity (FVC) than controls; there was no change in FVC after surgery. Before surgery, 58% of PE patients had subjective complaints of exercise limitation; 66% of the patients were significantly improved after surgery. PE subjects exercised at a similar workload to controls. Respiratory parameters during exercise were similar between the two groups before surgery, indicating that exercise was not limited by restrictive lung disease. After surgery, PE subjects exercised longer and had a higher oxygen pulse than before surgery, whereas controls showed no such changes. Although some PE subjects showed mild restrictive lung function, surgical repair did not influence this mild degree of restriction. After corrective surgery, PE patients have increased exercise tolerance and a higher oxygen pulse. Oxygen pulse is a measure of cardiac output. Results suggest that PE repair improves cardiopulmonary function during vigorous exercise. Based upon these studies, and our experience in the treatment of more than 700 surgical patients with pectus excavatum over a 40-year period, guidelines for the diagnosis and management of children with pectus excavatum are proposed.


Differential vibratory characteristics of adductor spasmodic dysphonia and muscle tension dysphonia on high-speed digital imaging.
The purpose of this study was to quantify disorder-specific signature kinematic disturbances of vibratory motion in adductor spasmodic dysphonia (AdSD) and muscle tension dysphonia (MTD), in voice disturbances of a severe nature, with the use of high-speed digital imaging (HSDI). A secondary hypothesis of the study was to investigate the sensitivity and specificity of the signature kinematic features obtained from HSDI, in differentiating between AdSD and MTD. We used vibratory features from automated extraction of vocal fold motion waveforms and glottal cycle montage analysis from HSDI for differential kinematic profiling of AdSD and MTD. Novel features of motion irregularities and micromotions (as small as 27 ms) were greater in number for AdSD, whereas reduced motion irregularities, absence of oscillatory breaks, absence of micromotions, and increased hyperfunction characterized the MTD group. Oscillatory breaks (as small as 8 ms), although present only in the AdSD group, were not statistically significant because of their reduced number of occurrences compared to the other features. Further montage analysis of successive glottal cycles of oscillatory breaks in the AdSD group revealed 3 different kinematic patterns within the AdSD group, indicative of likely AdSD with: 1) possible predominant thyroarytenoid muscle involvement, 2) possible predominant cricothyroid muscle involvement, and 3) possible combined involvements of the thyroarytenoid and lateral cricoarytenoid muscles. Four consistent but unique kinematic patterns were identified within the MTD group: 1) diplophonia, 2) vocal fry, 3) breathy phonation, and 4) pressed phonation. Sensitivity and specificity analysis revealed that only motion irregularity was a significant predictor of the presence of AdSD. Fine kinematic analysis from HSDI can be used to aid detailed clinical profiling of the source characteristics of AdSD and MTD.


Risk communication during COVID-19: A descriptive study on familiarity with, adherence to and trust in the WHO preventive measures.
Risk communication is a key component of public health interventions during an outbreak. As the coronavirus pandemic unfolded in late 2019, the World Health Organization (WHO) was at the forefront in the development of risk communication strategies. The WHO introduced a range of activities with the purpose of enabling the public to avail verified and timely information on COVID-19 prevention behaviors. Given the various WHO activities to protect the public health during COVID-19, it is important to investigate the extent of familiarity and uptake of the WHO recommendations among the public during the first wave of the pandemic. To do this, we conducted a large-scale Pan-European survey covering around 7500 individuals that are representative of populations from seven European countries, collected online during April 2-April 15, 2020. We use descriptive statistics including proportions and correlations and graphical representations such as bar charts to analyze and display the data. Our findings suggest that information from the WHO in the context of COVID-19 is well trusted and acted upon by the public. Overall familiarity and adherence were quite high in most countries. Adherence was higher for social distancing recommendations compared to hygiene measures. Familiarity and adherence were higher among older, female, and highly educated respondents. However, country level heterogeneities were observed in the level of trust in information from the WHO, with countries severely affected by the pandemic reporting lower levels of trust. Our findings call for efforts from health authorities to get regular feedback from the public on their familiarity and compliance with recommendations for preventive measures at all stages of the pandemic, to further develop and adapt risk communication as the pandemic evolves.


Annual air pollution caused by the Hungry Ghost Festival.
Burning of joss paper and incense is still a very common traditional custom in countries with a majority Chinese population. The Hungry Ghost Festival which is celebrated in the 7 month of the Chinese calendar is one of the events where joss paper and incense are burned as offerings. This study investigates the impact of the Ghost Month Festival (open burning event) on air quality by analysis of the chemical composition of particulate matter (PM) and rainwater samples collected during this event, compared with data collected throughout the year, as well as bottom ash samples from burning the original joss paper and incense. The results showed that the change in the chemical composition of the rainwater and PM2.5 (PM ≤ 2.5 μm) atmospheric samples could be correlated directly with burning events during this festival, with many elements increasing between 18% and 60% during August and September compared to the yearly mean concentrations. The order of percentage increase in elemental composition (in rain water and PM2.5) during the Hungry Ghost Festival is as follows: Zn > Ca > K > Mg > Fe > Al > Na ∼ Mn ∼ Ti ∼ V > Cu > As > Ni > Co > Cd > Cr > Pb. The chemical composition of the original source materials (joss paper and incense for combustion) and their associated bottom ash were analysed to explain the impact of burning on air quality.


Epidemic and Non-Epidemic Hot Spots of Malaria Transmission Occur in Indigenous Comarcas of Panama.
From 2002-2005, Panama experienced a malaria epidemic that has been associated with El Niño Southern Oscillation weather patterns, decreased funding for malaria control, and landscape modification. Case numbers quickly decreased afterward, and Panama is now in the pre-elimination stage of malaria eradication. To achieve this new goal, the characterization of epidemiological risk factors, foci of transmission, and important anopheline vectors is needed. Of the 24,681 reported cases in these analyses (2000-2014), ~62% occurred in epidemic years and ~44% in indigenous comarcas (5.9% of Panama's population). Sub-analyses comparing overall numbers of cases in epidemic and non-epidemic years identified females, comarcas and some 5-year age categories as those disproportionately affected by malaria during epidemic years. Annual parasites indices (APIs; number of cases per 1,000 persons) for Plasmodium vivax were higher in comarcas compared to provinces for all study years, though P. falciparum APIs were only higher in comarcas during epidemic years. Interestingly, two comarcas report increasing numbers of cases annually, despite national annual decreases. Inclusion of these comarcas within identified foci of malaria transmission confirmed their roles in continued transmission. Comparison of species distribution models for two important anophelines with Plasmodium case distribution suggest An. albimanus is the primary malaria vector in Panama, confirmed by identification of nine P. vivax-infected specimen pools. Future malaria eradication strategies in Panama should focus on indigenous comarcas and include both active surveillance for cases and comprehensive anopheline vector surveys.


[Profile of hospital admissions due to acute poisoning among children under 6 years of age in the metropolitan region of Rio de Janeiro, Brazil].
To describe the profile of poisoning cases among children attended during three years at emergency hospitals in the metropolitan area of Rio de Janeiro, Brazil. All confirmed or suspected poisoning cases, due to drugs and biological substances; organic solvents and hydrocarbons; chemical products; carbon monoxide and other gases; and pesticides, were collected. 1,574 cases of poisoning in children up to 5 years of age were detected. Around 40% of the cases involved chemical products of domestic use, 35% were caused by drugs, and 15% by pesticides. More than half of pesticide poisonings involved the 'chumbinho', an illegal product sold as a rodenticide, and usually including in its formulation a carbamate. Distribution of agents varied significantly by gender, the relative frequency of poisonings due to drugs and 'chumbinho' being higher among females than males. Participation of chemical products of domestic use decreased with age, but the role of drugs increased as agents of poisoning. During the observation period there was no significant reduction in the total number of annual cases. Supporting the expansion of the network of Poison Control Centers, developing intervention for the production of safer packaging devices for drugs chemical products, suppressing the illegal commerce of 'chumbinho', and fostering health education activities might contribute to modify this situation.


DNA-induced size-selective separation of mixtures of gold nanoparticles.
We present a novel method for size-selectively separating mixtures of nanoparticles in aqueous media utilizing the inherent chemical recognition properties of DNA and the cooperative binding properties of DNA-functionalized gold nanoparticles. We have determined that the melting temperatures (T(m)s) of aggregates formed from nanoparticles interconnected by duplex DNA are dependent upon particle size. This effect is proposed to derive from larger contact areas between the larger particles and therefore increased cooperativity, leading to higher T(m)s. The separation protocol involves taking two aliquots of a mixture of particles that vary in size and functionalizing them with complementary DNA. These aliquots are mixed at a temperature above the T(m) for aggregates formed from the smaller particles but below the T(m) for aggregates formed from the larger particles. Therefore, the aggregates that form consist almost exclusively of the larger particles and can be easily separated by sedimentation and centrifugation from the smaller dispersed particles. This unusual size-dependent behavior and separation protocol are demonstrated for three binary mixtures of particles and one ternary mixture.


The in vivo rate of somatic adenomatous polyposis coli mutation.
Cancer arises through successive somatic mutations/epimutations of oncogenes and tumor-suppressor genes. Accurate estimates of the rates at which these (epi)mutations occur are a vital but missing link in our emerging quantitative understanding of tumorigenesis. Their absence has hindered arguments concerning the importance of genetic instability in tumorigenesis and the number of mutations that precede malignant conversion of healthy cell lineages. Herein, a novel method for calculating the in vivo mutation rate of the adenomatous polyposis coli (APC) tumor-suppressor gene is presented. The large majority of bowel cancers are thought to be initiated by a partial loss of APC function, with the age-onset pattern dramatically altered for the worse in familial adenomatous polyposis (FAP) because these patients harbor selected germline APC mutations. Colon cancer in the context of FAP can be thought of as occurring "one hit quicker" than in the sporadic setting. We were able to isolate and estimate the rate of the initiating APC mutation in sporadic cases using the age incidence of FAP to approximate the time taken for a cell lineage in a sporadic patient with one APC mutation to present clinically as a cancer. Our result of approximately 10(-5) mutations per allele per year, although higher than previous estimates, appears to be consistent with the mutational spectrum of APC. The quality of fit provided by this method supports the theory that FAP and sporadic bowel cancer follow the same genetic pathway and are separated by only one mutation.


Bacterial pyrogenic exotoxins as superantigens.
The recent discovery of the mode of interaction between a group of microbial proteins known as superantigens and the immune system has opened a wide area of investigation into the possible role of these molecules in human diseases. Superantigens produced by certain viruses and bacteria, including Mycoplasma species, are either secreted or membrane-bound proteins. A unique feature of these proteins is that they can interact simultaneously with distinct receptors on different types of cells, resulting in enhanced cell-cell interaction and triggering a series of biochemical reactions that can lead to excessive cell proliferation and the release of inflammatory cytokines. However, although superantigens share many features, they can have very different biological effects that are potentiated by host genetic and environmental factors. This review focuses on a group of secreted pyrogenic toxins that belong to the superantigen family and highlights some of their structural-functional features and their roles in diseases such as toxic shock and autoimmunity. Deciphering the biological activities of the various superantigens and understanding their role in the pathogenesis of microbial infections and their sequelae will enable us to devise means by which we can intervene with their activity and/or manipulate them to our advantage.


Influenza vaccination in patients with lung cancer receiving anti-programmed death receptor 1 immunotherapy does not induce immune-related adverse events.
Influenza vaccination is recommended in patients with cancer to reduce influenza-related complications. Recently, more immune-related adverse events (irAEs) were demonstrated in patients with lung cancer who were vaccinated with the trivalent seasonal influenza vaccine during anti-programmed death receptor 1 (PD-1) immunotherapy. Confirmation of these findings is essential before recommendations on influenza vaccination may be revoked. In this cohort study in patients with lung cancer receiving nivolumab 3 mg/kg every 2 weeks during two influenza seasons (2015/16-2016/17), irAEs have been monitored. Incidence, timing and severity of irAEs were compared between vaccinated patients and non-vaccinated patients. In a compassionate use programme, 127 patients with lung cancer had been treated with at least one dose of nivolumab during two national influenza vaccination campaigns from September until December of 2015 and 2016. Forty-two patients had received the influenza vaccine, and 85 patients were not vaccinated. Median follow-up period was 118 days (interquartile range 106-119). Mean age was 64 years (range 46-83). In vaccinated and non-vaccinated patients, the incidence of irAEs was 26% and 22%, respectively, rate ratio 1.20 (95% confidence interval [CI] 0.51-2.65). The incidence of serious irAEs was 7% and 4%, respectively, rate ratio 2.07 (95% CI 0.28-15.43). Influenza vaccination while receiving nivolumab did not result in significant differences in the rates of discontinuation, death, clinical deterioration or tumour response between the groups. Influenza vaccination in patients with lung cancer receiving anti-PD-1 immunotherapy does not induce irAEs in our cohort. With this result, influenza vaccination should not be deterred from this group of patients.


High-resolution maps of Swiss apiaries and their applicability to study spatial distribution of bacterial honey bee brood diseases.
Honey bees directly affect and are influenced by their local environment, in terms of food sources, pollinator densities, pathogen and toxin exposure and climate. Currently, there is a lack of studies analyzing these data with Geographic Information Systems (GIS) to investigate spatial relationships with the environment. Particularly for inter-colonial pathogen transmission, it is known that the likelihood of a healthy colony to become infested (e.g., Varroosis) or infected (e.g., American foulbrood-AFB, European foulbrood-EFB) increases with higher colony density. Whether these transmission paths can actually be asserted at apiary level is largely unknown. Here, we unraveled spatial distribution and high-resolution density of apiaries and bacterial honey bee brood diseases in Switzerland based on available GIS data. Switzerland as 'model country' offers the unique opportunity to get apiary data since 2010 owing to compulsory registration for every beekeeper. Further, both destructive bee brood diseases (AFB and EFB) are legally notifiable in Switzerland, and EFB has an epizootic character for the last decades. As governmental data sets have to be ameliorated, raw data from the cantonal agricultural or veterinary offices have been included. We found a mean density of 0.56 apiaries per km2, and high resolution spatial analyzes showed strong correlation between density of apiaries and human population density as well as agricultural landscape type. Concerning two bacterial bee brood diseases (AFB, EFB), no significant correlation was detectable with density of apiaries on cantonal level, though a high correlation of EFB cases and apiary density became obvious on higher resolution (district level). Hence, Swiss EFB epizootics seem to have benefited from high apiary densities, promoting the transmission of pathogens by adult bees. The GIS-based method presented here, might also be useful for other bee diseases, anthropogenic or environmental factors affecting bee colonies.


Gastrointestinal bleeding after high intake of omega-3 fatty acids, cortisone and antibiotic therapy: a case study.
Omega-3 fatty acids exert a plethora of physiological actions including triglycerides lowering, reduction of inflammatory indices, immunomodulation, anti- thrombotic effects and possibly promotion of exercise performance. Their use is widespread and for commonly ingested doses their side- effects are minimal. We report a case of a 60 y amateur athlete who consumed about 20 g omega-3 fatty acids daily from supplements and natural sources for a year. After the intake of cortisone and antibiotics he presented duodenum ulcer and bleeding although he had no previous history of gastrointestinal problems. Although several animal data support gastro-protective effects of omega-3 fatty acids in the present case they were not able to prevent ulcer generation. The present observation may be explained by (i) the high dose of omega-3 fatty acids and their effect on bleeding, (ii) the fact that cortisone increases their oxidation and may render them proinflammatory, (iii) other antithrombotic microconstituents included in the consumed cod-oil and/or the diet of the subject and (iv) the differences in the coagulation and fibrinolytic systems of well- trained subjects. Further studies are needed to substantiate any possible interaction of cortisone and omega-3 fatty acids in wide ranges of intake.


Combined organic biomarker and use-wear analyses of stone artefacts from Liang Bua, Flores, Indonesia.
Organic biomarker and lithic use-wear analyses of archaeological implements manufactured and/or used by hominins in the past offers a means of assessing how prehistoric peoples utilised natural resources. Currently, most studies focus on one of these techniques, rather than using both in sequence. This study aims to assess the potential of combining both methods to analyse stone artefacts, using a set of 69 stones excavated from the cave site of Liang Bua (Flores, Indonesia). Prior to chemical analysis, an initial inspection of the artefacts revealed potential use-wear traces but no visible residues. Gas chromatography mass spectrometry (GC-MS) analysis, including the targeting of 86 lipids, terpenes, terpenoids, alkanes and their analogues, found compounds with plant or animal origin on 27 of the 69 stones. The artefacts were subsequently cleaned, and use-wear analysis identified traces of use on 43 artefacts. Use-wear analysis confirmed traces of use on 23 of the 27 artefacts with potential use-residues that were determined by GC-MS. The GC-MS results were broadly consistent with the functional classes identified in the later use-wear analysis. This inclusive approach for stone artefact analysis strengthens the identifications made through multiple lines of enquiry. There remain conflicts and uncertainties in specific cases, suggesting the need for further refinement and analyses of the relationships between use-wear and residues.


Changes in mortality rates and causes of death in a population-based cohort of persons living with and without HIV from 1996 to 2012.
Non-HIV/AIDS-related diseases are gaining prominence as important causes of morbidity and mortality among people living with HIV. The purpose of this study was to characterize and compare changes over time in mortality rates and causes of death among a population-based cohort of persons living with and without HIV in British Columbia (BC), Canada. We analysed data from the Comparative Outcomes And Service Utilization Trends (COAST) study; a retrospective population-based study created via linkage between the BC Centre for Excellence in HIV/AIDS and Population Data BC, and containing data for HIV-infected individuals and the general population of BC, respectively. Our analysis included all known HIV-infected adults (≥ 20 years) in BC and a random 10% sample of uninfected BC adults followed from 1996 to 2012. Deaths were identified through Population Data BC - which contains information on all registered deaths in BC (BC Vital Statistics Agency dataset) and classified into cause of death categories using International Classification of Diseases (ICD) 9/10 codes. Age-standardized mortality rates (ASMR) and mortality rate ratios were calculated. Trend test were performed. 3401 (25%), and 47,647 (9%) individuals died during the 5,620,150 person-years of follow-up among 13,729 HIV-infected and 510,313 uninfected individuals, respectively. All-cause and cause-specific mortality rates were consistently higher among HIV-infected compared to HIV-negative individuals, except for neurological disorders. All-cause ASMR decreased from 126.75 (95% CI: 84.92-168.57) per 1000 population in 1996 to 21.29 (95% CI: 17.79-24.79) in 2011-2012 (83% decline; p < 0.001 for trend), compared to a change from 7.97 (95% CI: 7.61-8.33) to 6.87 (95% CI: 6.70-7.04) among uninfected individuals (14% decline; p < 0.001). Mortality rates from HIV/AIDS-related causes decreased by 94% from 103.85 per 1000 population in 1996 to 6.72 by the 2011-2012 era (p < 0.001). Significant ASMR reductions were also observed for hepatic/liver disease and drug abuse/overdose deaths. ASMRs for neurological disorders increased significantly over time. Non-AIDS-defining cancers are currently the leading non-HIV/AIDS-related cause of death in both HIV-infected and uninfected individuals. Despite the significant mortality rate reductions observed among HIV-infected individuals from 1996 to 2012, they still have excess mortality risk compared to uninfected individuals. Additional efforts are needed to promote effective risk factor management and appropriate screening measures among people living with HIV.


Comparing hiking, mountain biking and horse riding impacts on vegetation and soils in Australia and the United States of America.
Hiking, horse riding and mountain biking are popular in protected areas in Australia and the United States of America. To help inform the often contentious deliberations about use of protected areas for these three types of activities, we review recreation ecology research in both countries. Many impacts on vegetation, soils and trails are similar for the three activities, although there can be differences in severity. Impacts include damage to existing trails, soil erosion, compaction and nutrification, changes in hydrology, trail widening, exposure of roots, rocks and bedrock. There can be damage to plants including reduction in vegetation height and biomass, changes in species composition, creation of informal trails and the spread of weeds and plant pathogens. Due to differences in evolutionary history, impacts on soil and vegetation can be greater in Australia than in the USA. There are specific social and biophysical impacts of horses such as those associated with manure and urine, grazing and the construction and use of tethering yards and fences. Mountain bike specific impacts include soil and vegetation damage from skidding and the construction of unauthorised trails, jumps, bridges and other trail technical features. There are gaps in the current research that should be filled by additional research: (1) on horse and mountain bike impacts to complement those on hiking. The methods used need to reflect patterns of actual usage and be suitable for robust statistical analysis; (2) that directly compares types and severity of impacts among activities; and (3) on the potential for each activity to contribute to the spread of weeds and plant pathogens. Additional research will assist managers and users of protected areas in understanding the relative impacts of these activities, and better ways to manage them. It may not quell the debates among users, managers and conservationists, but it will help put it on a more scientific footing.


First Report of Prune dwarf virus and Prunus necrotic ringspot virus on Peach in Montenegro.
In September and October 2011, samples were collected from mature peach trees (~17 years old) exhibiting symptoms of chlorotic rings and spots, vein clearing, mosaic, necrosis, leaf distortion, stunting, and rosette formation in a major commercial orchard (~80 ha) near Podgorica, Montenegro. Samples were collected from nine different peach varieties (cvs. Adriana, Caldesi, Gloria, Maria Marta, May Crest, Morsiani, Rita Star, Spring Belle, and Spring Crest). Samples (n = 58) were tested using DAS-ELISA for the presence of Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV). Commercial positive and negative controls were included in each ELISA (antisera and controls supplied by BIOREBA AG, Reinach, Switzerland). Only one symptomatic sample from cv. Gloria tested positive for PDV (sample reference: 399/11), a further 11 samples (cvs. Rita Star [six], May Crest [four] and Spring Crest [one]) were positive for PNRSV. Samples were also tested for Plum pox virus (PPV) by real-time RT-PCR (1). The PDV positive sample (399/11) showing mosaic was in mixed infection with PPV, as were 6 of the 11 PNRSV samples, including sample 373/11 with yellow mottling and leaf distortion symptoms. On single-infected PNRSV, sample 368/11 chlorotic line patterns and leaf deformations were observed. To confirm the presence of PDV and PNRSV, positive samples were also tested by RT-PCR. Total RNA was extracted using RNeasy Plant Mini kit (Qiagen, Hilden, Germany). RT-PCR was performed with primer pairs PDV2F/PDV1R (3) and MG1/MG2 (2) specific for PDV and PNRSV, respectively. Amplicons of the expected size, 173 bp for PDV and 675 bp for PNRSV, were obtained from corresponding ELISA-positive samples. Amplified products from three samples (PDV 399/11 and PNRSV 368/11 and 373/11) were cloned into pGEM-T Easy Vector (Promega, Madison, WI) then sent for sequence analysis (MWG-Biotech AG, Edersberg, Germany). Sequence data was compared to sequences published in GenBank. Analysis of sequence obtained from isolate 399/11 (cv. Gloria) corresponded to partial CP gene of PDV, with a high degree of similarity to isolates reported from other parts of the world ranging from 94.2 to 95.9%, showing highest similarity with isolate Ch 137 (L28145). Sequence analyses of CP gene from PNRSV isolates 368/11 (JX569825) and 373/11 (JX569826) proved to be 89.3 to 99.7% identical with corresponding sequences of isolates previously described. In particular, the Montenegrin PNRSV isolates were most closely related to Chilean NctCl.augl isolate from nectarine (EF565253). To demonstrate that the virus was infectious, seedlings of peach cv. GF305 were side grafted with bud-woods from PDV (sample 399/11) and PNRSV-positive samples (samples 368/11 and 373/11) and a healthy control sample. Grafted seedlings were kept in a greenhouse with a under 16-h light regime at 22 to 24°C and observed for symptom development. No symptoms were observed in grafted plants with the healthy control. All plants inoculated with virus-positive samples exhibited stunted vegetation and mild mottle with no difference in symptoms between the two viruses. Indicator plants of peach cv. GF305 inoculated with PPV dual-infected samples (399/11 and 373/11) were subsequently shown to be positive for PPV by real-time RT-PCR. Subsequent DAS-ELISA test on samples from experimentally inoculated trees using specific antisera as described above confirmed PDV and PNRSV infections as expected. These viruses have recently been reported from sour cherry (Prunus cerasus L.) in Serbia (4), ~600 km to the northeast. However, to our knowledge, this is the first report on the occurrence of PDV and PNRSV in Montenegro. References: (1) N. Capote et al. Int. Microbiol. 12:1, 2009. (2) M. Glasa et al. Ann. Appl. Biol. 140:279, 2002. (3) D. R. Parakh et al. Acta Hortic. 386:421, 1996. (4) S. Radičević et al. Genetika 44:285, 2012.


Developing a tool for evaluating community-based rehabilitation in Uganda.
The overall purpose of this study is to use the conceptual criteria of the 'Joint Position Statement (JPS)' (see http://www.ilo.org/public/english/employment/skills/download/jointpaper.pdf )on Community-based rehabilitation (CBR), together with the data collected from a CBR programme in Uganda (Tororo) to generate a prototype CBR evaluation tool. A case study approach is used to examine the Tororo CBR programme. Project documentation was examined to generate base line data which was used to inform sampling for interviews and focus group discussions. These were conducted with a wide range of stakeholders. The interview and focus group data are recorded, translated, transcribed and imported into a computer-assisted analysis programme, NVivo. The data was analysed in two stages: (i) data led development of themes (ii) allocation of the generated themes to the conceptual structure of the 'JPS'. A third stage uses the data to generate an outcome measurement tool for evaluating CBR programmes. Fifty different themes were identified during the analysis and used to construct a questionnaire framed around the concepts of the 'JPS'. In addition, the questionnaire reflected the words and phrases used by participants. The outcome is a short and feasible tool for evaluating CBR programmes. The 'JPS' has been used as a framework for the analysis of qualitative data to generate the basis of a prototype evaluative questionnaire which can now be piloted, amended and then used for the evaluation of other CBR programmes.


Assessment of the clinical relevance of 17q25.3 copy number and three-dimensional telomere organization in non-small lung cancer patients.
To identify potential biomarkers that may provide new therapeutic targets or prognostic indicators for non-small cell lung cancer (NSCLC), we investigated the three-dimensional (3D) organization of telomeres and cytoband 17q25.3 copy number in NSCLC tissues. NSCLC paraffin-embedded tissue specimens from 18 patients were assessed for 3D telomere organization by 3D nuclear telomere imaging followed by quantitative analysis. Patients were stratified by smoking, histology, and EGFR status. Cytoband 17q25.3 was examined by fluorescent in situ hybridization. Data from comparative genomic hybridization and/or single nucleotide polymorphism arrays for cytoband 17q25.3 were obtained and correlated with Q-FISH and 3D telomere results. 3D telomeric profiling demonstrated that the smokers, EGFR-negative, and squamous cell carcinoma subgroups tended to have higher numbers of lower-intensity telomeres, indicative of shorter telomeres, as well as higher numbers of telomeric aggregations compared to non-smokers, EGFR-positive, and adenocarcinomas, respectively. Gains of cytoband 17q25.3 in conjunction with an increase in the control region 17p11.2 were observed in 7 of 18 (38.9 %) patients, reflecting a gain of chromosome 17. Clonal gains of cytoband 17q25.3 were observed in 11 of 18 (61 %) patients, highlighting a potential biological significance for the genes in this region in NSCLC tumourigenesis. The 3D telomere profiles may differentiate NSCLC patients with different histologies, EGFR, and smoking statuses, rendering them a potential biomarker for distinguishing these clinically relevant histological and molecular subtypes of lung cancer. Highly frequent clonal gain of cytoband 17q25.3 was also demonstrated, suggesting an important biological role for the genes in this region.


Single delayed rectifier channels in the membrane of rabbit ventricular myocytes.
In rabbit ventricular cells, the delayed rectifier current (IK) has not been extensively studied, and properties of single IK channels still need to be determined. In this study, we present data on a voltage-dependent channel in rabbit ventricular cells; the properties indicate that it is an IK channel. Patch-clamp experiments were carried out on cell-attached and inside-out patches of rabbit ventricular cells. Single-channel currents were recorded at negative potentials as inward currents with 150 mM K+ in the pipette. Voltage-dependent channel activity was only present after the return from a depolarizing test pulse, indicating activation on depolarization. Single-channel conductance calculated from the current-voltage relation was 13.1 pS (pooled data, n = 8). The shift in reversal potential of the unitary currents, determined at 150 and 300 mM K+ at the intracellular side of the membrane, showed that the channels were highly permeable to potassium ions. Increase of the duration or the amplitude of the depolarizing test pulse increased channel activity. The time constant for activation at +30 mV was 187 msec (pooled data, n = 4). Half-activation potential was -4.9 +/- 3.8 mV (mean +/- SD), and the slope factor was 7.2 +/- 3.7 mV (mean +/- SD). Current tails, reconstructed from averaged single-channel currents, revealed that the time course of deactivation decreased from 694 +/- 73 msec at -80 mV to 136 +/- 39 msec at -110 mV. Additional evidence that the channel was indeed an IK channel was provided by the observation that the channel was blocked by 10(-7) M E-4031, a class III antiarrhythmic agent that has been shown to block a component of the macroscopic IK in guinea pig heart.


Hemoglobin is associated with retinal vascular fractals in type 1 diabetes patients.
Retinal vascular fractal dimension, a measure of the density of the retinal vasculature, has been suggested as a marker of systemic microvascular disorders in diabetes. As hemoglobin concentration is tightly related to vascular physiology and hypoxia, the hypothesis was that hemoglobin concentration would be associated with retinal vascular fractals in a relevant population. In a cross-sectional study of 204 long-term type 1 diabetes patients from a population-based cohort, retinal digital photos were captured and graded for fractal dimension (Df) by International Retinal Imaging Software - Fractal (IRIS-Fractal). Df was calculated from a disc-centered retinal photo from the right eye. Hemoglobin concentrations were measured using routine equipment. Of 175 patients with gradable images, median age was 57.7 years and median duration of diabetes was 42 years. Median retinal Df was 1.4606 (inter-quartile range 0.0264). A positive correlation was found between hemoglobin concentration and retinal vascular Df (r = 0.23, p = 0.0018). In a multiple linear regression model, Df was associated with hemoglobin (coefficient 0.0054 per 1.0 mmol/L increase in hemoglobin, p = 0.01) and age (coefficient -0.0046 for each 10-year increase in age, p = 0.04). Hemoglobin correlated independently with retinal vascular fractals indicating a relationship between hemoglobin availability and retinal vascular structure.


Investigating the potential role of TRPA1 in locomotion and cardiovascular control during hypertension.
Radiotelemetry was used to investigate the in vivo cardiovascular and activity phenotype of both TRPA1 (transient receptor potential ankyrin 1) wild-type (WT) and TRPA1 knockout (KO) mice. After baseline recording, experimental hypertension was induced using angiotensin II infusion (1.1 mg(-1) kg(-1) a day, for 14 days). TRPA1 WT and KO mice showed similar morphological and functional cardiovascular parameters, including similar basal blood pressure (BP), heart rate, size, and function. Similar hypertension was also displayed in response to angiotensin II (156 ± 7 and 165 ± 11 mmHg, systolic BP ± SEM, n = 5-6). TRPA1 KO mice showed increased hypertensive hypertrophy (heart weight:tibia length: 7.3 ± 1.6 mg mm(-1) vs. 8.8 ± 1.7 mg mm(-1)) and presented with blunted interleukin 6 (IL-6) production compared with hypertensive WT mice (151 ± 24 vs. 89 ± 16 pg mL(-1)). TRPA1 expression in dorsal root ganglion (DRG) neurones was upregulated during hypertension (163% of baseline expression). Investigations utilizing the TRPA1 agonist cinnamaldehyde (CA) on mesenteric arterioles isolated from näive mice suggested a lack of TRPA1-dependent vasoreactivity in this vascular bed; a site with notable ability to alter total peripheral resistance. However, mesenteric arterioles isolated from TRPA1 KO hypertensive mice displayed significantly reduced ability to relax in response to nitric oxide (NO) (P < 0.05). Unexpectedly, naïve TRPA1 KO mice also displayed physical hyperactivity traits at baseline, which was exacerbated during hypertension. In conclusion, our study provides a novel cardiovascular characterization of TRPA1 KO mice in a model of hypertension. Results suggest that TRPA1 has a limited role in global cardiovascular control, but we demonstrate an unexpected capacity for TRPA1 to regulate physical activity.


Detection of human papillomavirus DNA in cytologically normal women and subsequent cervical squamous intraepithelial lesions.
Human papillomavirus (HPV) infection has been strongly associated with cervical carcinoma and its cytologic precursors, squamous intraepithelial lesions (SIL). We investigated the risk of SIL prospectively following polymerase chain reaction (PCR)-based DNA testing for a wide range of genital HPV types in a cohort of initially cytologically normal women, to clarify the role of HPV in the etiology of SIL. Starting in April 1989, 17,654 women who were receiving routine cytologic screening at Kaiser Permanente (Portland, OR) were followed for the development of incident SIL. During follow-up, 380 incident case patients and 1037 matched control subjects were eligible for this nested case-control study. Cervical lavages collected at enrollment and, later, at the time of case diagnosis (or the corresponding time for selection of control subjects) were tested for HPV DNA using a PCR-based method. The data were analyzed as contingency tables with two-sided P values or, for multivariable analyses, using odds ratios (ORs) with 95% confidence intervals (CIs). In comparison with initially HPV-negative women, women who tested positive for HPV DNA at enrollment were 3.8 times (95% CI = 2.6-5.5) more likely to have low-grade SIL subsequently diagnosed for the first time during follow-up and 12.7 times more likely (95% CI = 6.2-25.9) to develop high-grade SIL. At the time of diagnosis, the cross-sectional association of HPV DNA and SIL was extremely strong (OR = 44.4 and 95% CI = 24.2-81.5 for low-grade SIL and OR = 67.1 and 95% CI = 19.3-233.7 for high-grade SIL). HPV16 was the virus type most predictive of SIL, even low-grade SIL. These findings are consistent with the hypothesis that HPV infection is the primary cause of cervical neoplasia. Furthermore, they support HPV vaccine research to prevent cervical cancer and efforts to develop HPV DNA diagnostic tests.


Mango Supplementation Modulates Gut Microbial Dysbiosis and Short-Chain Fatty Acid Production Independent of Body Weight Reduction in C57BL/6 Mice Fed a High-Fat Diet.
High-fat (HF) diet-induced obesity is associated with changes in the gut microbiota. Fiber and other bioactive compounds in plant-based foods are suggested to prevent gut dysbiosis brought on by HF feeding. Mango is high in fiber and has been reported to have anti-obesogenic, hypoglycemic, and immunomodulatory properties. We investigated the effects of freeze-dried mango pulp combined with an HF diet on the cecal microbial population and its relation to body composition, lipids, glucose parameters, short-chain fatty acid (SCFA) production, and gut inflammatory markers in a mouse model of diet-induced obesity. Six-wk-old male C57BL/6 mice were randomly assigned to 1 of 4 dietary treatment groups: control (AIN-93M, 10% fat kcal), HF (60% fat kcal), and HF + 1% or 10% mango (HF+1%M or HF+10%M, wt:wt) for 12 wk. The cecal microbial population was assessed by use of 16S rDNA sequencing. Body composition, plasma glucose and lipids, cecal and fecal SCFAs, and mRNA abundance of inflammatory markers in the ileum and colonic lamina propria were assessed. Compared with the control group, HF feeding significantly reduced (P < 0.05) 1 operational taxonomic unit (OTU) of the genus Bifidobacteria (64-fold) and 5 OTUs of the genus Akkermansia (≥16-fold). This reduction was prevented in the HF+10%M group, members of which had 10% higher final body weight compared with the HF group (P = 0.01) and similar fasting blood glucose concentrations (P = 0.24). The HF+10%M group had 135% (P = 0.004) and 133% (P < 0.0001) greater fecal acetic and n-butyric acids concentrations than the HF group, suggesting greater microbial fermentation. Furthermore, a 59% greater colonic interleukin 10 (Il10) gene expression was observed in the HF+10%M group than in the HF group (P = 0.048), indicating modulation of gut inflammation. The HF+1%M group generally did not differ from the HF group. The addition of mango to an HF diet modulated the gut microbiota and production of SCFAs in C57BL/6 mice; these changes may improve gut tolerance to the insult of an HF diet.


Examining the short-term anxiolytic and antidepressant effect of Floatation-REST.
Floatation-REST (Reduced Environmental Stimulation Therapy) reduces sensory input to the nervous system through the act of floating supine in a pool of water saturated with Epsom salt. The float experience is calibrated so that sensory signals from visual, auditory, olfactory, gustatory, thermal, tactile, vestibular, gravitational and proprioceptive channels are minimized, as is most movement and speech. This open-label study aimed to examine whether Floatation-REST would attenuate symptoms of anxiety, stress, and depression in a clinical sample. Fifty participants were recruited across a spectrum of anxiety and stress-related disorders (posttraumatic stress, generalized anxiety, panic, agoraphobia, and social anxiety), most (n = 46) with comorbid unipolar depression. Measures of self-reported affect were collected immediately before and after a 1-hour float session, with the primary outcome measure being the pre- to post-float change score on the Spielberger State Anxiety Inventory. Irrespective of diagnosis, Floatation-REST substantially reduced state anxiety (estimated Cohen's d > 2). Moreover, participants reported significant reductions in stress, muscle tension, pain, depression and negative affect, accompanied by a significant improvement in mood characterized by increases in serenity, relaxation, happiness and overall well-being (p < .0001 for all variables). In reference to a group of 30 non-anxious participants, the effects were found to be more robust in the anxious sample and approaching non-anxious levels during the post-float period. Further analysis revealed that the most severely anxious participants reported the largest effects. Overall, the procedure was well-tolerated, with no major safety concerns stemming from this single session. The findings from this initial study need to be replicated in larger controlled trials, but suggest that Floatation-REST may be a promising technique for transiently reducing the suffering in those with anxiety and depression. ClinicalTrials.gov NCT03051074.


The CD85J/leukocyte inhibitory receptor-1 distinguishes between conformed and beta 2-microglobulin-free HLA-G molecules.
For a proper development of the placenta, maternal NK cells should not attack the fetal extravillous cytotrophoblast cells. This inhibition of maternal NK cells is partially mediated via the nonclassical MHC class I molecule HLA-G. Recently, we demonstrated that HLA-G forms disulfide-linked high molecular complexes on the surface of transfected cells. In the present study, we demonstrate that HLA-G must associate with beta(2)m for its interaction with CD85J/leukocyte Ig-like receptor-1 (LIR-1). Although HLA-G free H chain complexes are expressed on the surface, they are not recognized and possibly interfere with CD85J/LIR-1 and HLA-G interaction. The formation of these complexes on the cell surface might represent a novel mechanism developed specifically by the HLA-G protein aimed to control the efficiency of the CD85J/LIR-1-mediated inhibition. We also show that endogenous HLA-G complexes are expressed on the cell surface. These findings provide novel insights into the delicate interaction between extravillous cytotrophoblast cells and NK cells in the decidua.


A tidal ventilation model for oxygenation in respiratory failure.
We develop tidal-ventilation pulmonary gas-exchange equations that allow pulmonary shunt to have different values during expiration and inspiration, in accordance with lung collapse and recruitment during lung dysfunction (Am. J. Respir. Crit. Care Med. 158 (1998) 1636). Their solutions are tested against published animal data from intravascular oxygen tension and saturation sensors. These equations provide one explanation for (i) observed physiological phenomena, such as within-breath fluctuations in arterial oxygen saturation and blood-gas tension; and (ii) conventional (time averaged) blood-gas sample oxygen tensions. We suggest that tidal-ventilation models are needed to describe within-breath fluctuations in arterial oxygen saturation and blood-gas tension in acute respiratory distress syndrome (ARDS) subjects. Both the amplitude of these oxygen saturation and tension fluctuations, and the mean oxygen blood-gas values, are affected by physiological variables such as inspired oxygen concentration, lung volume, and the inspiratory:expiratory (I:E) ratio, as well as by changes in pulmonary shunt during the respiratory cycle.


Riboregulation in the Major Gastric Pathogen Helicobacter pylori.
Helicobacter pylori is a Gram-negative bacterial pathogen that colonizes the stomach of about half of the human population worldwide. Infection by H. pylori is generally acquired during childhood and this bacterium rapidly establishes a persistent colonization. H. pylori causes chronic gastritis that, in some cases, progresses into peptic ulcer disease or adenocarcinoma that is responsible for about 800,000 deaths in the world every year. H. pylori has evolved efficient adaptive strategies to colonize the stomach, a particularly hostile acidic environment. Few transcriptional regulators are encoded by the small H. pylori genome and post-transcriptional regulation has been proposed as a major level of control of gene expression in this pathogen. The transcriptome and transcription start sites (TSSs) of H. pylori strain 26695 have been defined at the genome level. This revealed the existence of a total of 1,907 TSSs among which more than 900 TSSs for non-coding RNAs (ncRNAs) including 60 validated small RNAs (sRNAs) and abundant anti-sense RNAs, few of which have been experimentally validated. An RNA degradosome was shown to play a central role in the control of mRNA and antisense RNA decay in H. pylori. Riboregulation, genetic regulation by RNA, has also been revealed and depends both on antisense RNAs and small RNAs. Known examples will be presented in this review. Antisense RNA regulation was reported for some virulence factors and for several type I toxin antitoxin systems, one of which controls the morphological transition of H. pylori spiral shape to round coccoids. Interestingly, the few documented cases of small RNA-based regulation suggest that their mechanisms do not follow the same rules that were well established in the model organism Escherichia coli. First, the genome of H. pylori encodes none of the two well-described RNA chaperones, Hfq and ProQ that are important for riboregulation in several organisms. Second, some of the reported small RNAs target, through "rheostat"-like mechanisms, repeat-rich stretches in the 5'-untranslated region of genes encoding important virulence factors. In conclusion, there are still many unanswered questions about the extent and underlying mechanisms of riboregulation in H. pylori but recent publications highlighted original mechanisms making this important pathogen an interesting study model.


The incidence and distribution of stress fractures in elite tennis players.
Modern professional tennis involves powerful movements repeatedly subjecting the musculoskeletal system to heavy mechanical load. Thus tennis players are exposed to high risk of overuse injuries including stress fractures. To determine the incidence and distribution of stress fractures in elite tennis players. Retrospective cohort study. The cohort study population consisted of 139 elite players (mean (SD) age, 20.0 (5.0) years; 48 female, 91 male). Stress fractures were identified and confirmed radiologically from medical records during a two year period. Injuries were analysed according to age, sex, site, severity, delay in diagnosis, and time needed to return to sports. 15 players sustained 18 stress fractures, corresponding to an overall case incidence of 12.9% (95% confidence interval (CI), 8.1 to 20.0). The tarsal navicular was most affected (n = 5; 27%), followed by the pars interarticularis (n = 3; 16%), the metatarsals (n = 3; 16%), the tibia (n = 2; 11%) and the lunate (n = 2; 11%). Magnetic resonance imaging showed a greater incidence of "high grade" lesions (94.4%). Stress fracture incidence was significantly higher in juniors (20.3% (95% CI, 11.4 to 33.2)) than in professional players (7.5% (2.8 to 15.6)) (p = 0.045). There was a high absolute risk (12.9%) of stress fractures in elite tennis players over a two year period. Junior players were at highest risk. The lesions are a major cause of disruption both of training and of competition. Risk factors should therefore be identified and prevention emphasised.


Insulin resistance and normal thyroid hormone levels: prospective study and metabolomic analysis.
While hyperthyroidism and hypothyroidism cause dysglycemia, the relationship between thyroid hormone levels within the normal range and insulin resistance (IR) is unclear. In 940 participants with strictly normal serum concentrations of free triiodothyronine (fT3), free thyroxine (fT4), and thyroid-stimulating hormone (TSH) followed up for 3 yr, we measured insulin sensitivity (by the insulin clamp technique) and 35 circulating metabolites. At baseline, across quartiles of increasing fT3 levels (or fT3/fT4 ratio) most features of IR emerged [i.e., male sex, greater body mass index (BMI), waist circumference, heart rate, blood pressure, fatty liver index, free fatty acids, and triglycerides; reduced insulin-mediated glucose disposal; and β-cell glucose sensitivity). In multiadjusted analyses, fT3 was reciprocally related to insulin sensitivity and, in a subset of 303 subjects, directly related to endogenous glucose production. In multiple regression models adjusting for sex, age, BMI, and baseline value of insulin sensitivity, higher baseline fT3 levels were significant predictors of decreases in insulin sensitivity. Moreover, baseline fT3 predicted follow-up increases in glycemia independently of sex, age, BMI, insulin sensitivity, β-cell glucose sensitivity, and baseline glycemia. Serum tyrosine levels were higher with IR and were directly associated with fT3; higher α-hydroxybutyrate levels signaled enhanced oxidative stress, thereby impairing tyrosine degradation. In 25 patients with morbid obesity, surgery-induced weight loss improved IR and consensually lowered fT3 levels. High-normal fT3 levels are associated with IR both cross-sectionally and longitudinally, and predict deterioration of glucose tolerance. This association is supported by a metabolite pattern that points at increased oxidative stress as part of the IR syndrome.


Vascular action of cocoa flavanols in humans: the roots of the story.
Diet patterns are widely recognized as contributors to hypertension. Widely studied potential contributors include intake of sodium, potassium, magnesium, calcium, soluble fiber, omega-3 fatty acids, alcohol, protein, and calories. We add to that list the effect of dietary flavanols present in certain cocoas, which have sufficient activity on vascular nitric oxide to influence blood pressure control. Kuna Indians who live on islands near Panama have little age-related rise in blood pressure or hypertension. On migration to Panama City, blood pressure rises with age, and the frequency of essential hypertension matches urban levels elsewhere. We have identified a specific food that probably makes an important contribution to cardiovascular status. Island-dwelling Kuna drink more than 5 cups of flavanol-rich cocoa per day and incorporate that cocoa into many recipes. Mainland Kuna ingest little cocoa, and what they take is commercially available and flavanol-poor. The flavanol-rich cocoa activates nitric oxide synthase in vitro and in intact humans in the doses that the Kuna employ. Vasodilator responses to flavonoid-rich cocoa are prevented or reversed by the arginine analog, N-nitro-L-arginine methyl ester. Island-dwelling Kuna have a 3-fold larger urinary nitrate:nitrite than do Mainland dwellers. As endothelial dysfunction is central to current thinking on cardiovascular pathophysiology, a food that enhances endothelial function could have broad implications. The list of candidate conditions that might be influenced is impressive, ranging from atherosclerosis and diabetes mellitus to hypertension and preeclampsia, to vascular dementias and end-stage renal disease. The next decade will be interesting.


Mechanism of the diastolic dysfunction induced by glycolytic inhibition. Does adenosine triphosphate derived from glycolysis play a favored role in cellular Ca2+ homeostasis in ferret myocardium?
Several lines of evidence indicate that glycolysis is especially important for normal diastolic relaxation and for the maintenance of cellular ion homeostasis in myocardium. To elucidate whether the glycolytic flux of ATP contributes to diastolic tone and to the regulation of intracellular Ca2+, myocardial content of sugar phosphates ([SP]) and intracellular Ca2+ concentration ([Ca2+]i) were measured in isolated, perfused ferret hearts using nuclear magnetic resonance. Glucose and acetate were used as substrates for glycolysis and oxidative phosphorylation, respectively. Glycogen was effectively depleted after 15-min perfusion with glucagon (2 mg/liter), as verified by the lack of rise in [SP] during exposure to iodoacetate (100 microM) in substrate-free perfusate. Despite the fact that glycolytic flux had been blocked both by iodoacetate and by absence of substrate, end-diastolic left ventricular pressure (EDP) remained unchanged (P > 0.15, n = 6). The subsequent addition of glucose to the perfusate led to SP accumulation and a marked rise in EDP, with a significant correlation between EDP and [SP] (r = 0.86 +/- 0.04, P < 0.01, n = 6). A similar correlation was observed when glucose in the perfusate was replaced by 2-deoxyglucose (r = 0.78 +/- 0.09, P < 0.01, n = 3). Fluorine nuclear magnetic resonance measurements of [Ca2+]i verified that EDP faithfully reports changes in diastolic [Ca2+]i under the present experimental conditions. Thus, intracellular Ca2+ overload is caused by the accumulation of SP rather than by the inhibition of glycolysis per se. Glycolysis may appear to be important because its by-products are deleterious, and not necessarily because glycolytically derived ATP plays a favored role in ion homeostasis.


Constrained tibial vibration in mice: a method for studying the effects of vibrational loading of bone.
Vibrational loading can stimulate the formation of new trabecular bone or maintain bone mass. Studies investigating vibrational loading have often used whole-body vibration (WBV) as their loading method. However, WBV has limitations in small animal studies because transmissibility of vibration is dependent on posture. In this study, we propose constrained tibial vibration (CTV) as an experimental method for vibrational loading of mice under controlled conditions. In CTV, the lower leg of an anesthetized mouse is subjected to vertical vibrational loading while supporting a mass. The setup approximates a one degree-of-freedom vibrational system. Accelerometers were used to measure transmissibility of vibration through the lower leg in CTV at frequencies from 20 Hz to 150 Hz. First, the frequency response of transmissibility was quantified in vivo, and dissections were performed to remove one component of the mouse leg (the knee joint, foot, or soft tissue) to investigate the contribution of each component to the frequency response of the intact leg. Next, a finite element (FE) model of a mouse tibia-fibula was used to estimate the deformation of the bone during CTV. Finally, strain gages were used to determine the dependence of bone strain on loading frequency. The in vivo mouse leg in the CTV system had a resonant frequency of 60 Hz for +/-0.5 G vibration (1.0 G peak to peak). Removing the foot caused the natural frequency of the system to shift from 60 Hz to 70 Hz, removing the soft tissue caused no change in natural frequency, and removing the knee changed the natural frequency from 60 Hz to 90 Hz. By using the FE model, maximum tensile and compressive strains during CTV were estimated to be on the cranial-medial and caudolateral surfaces of the tibia, respectively, and the peak transmissibility and peak cortical strain occurred at the same frequency. Strain gage data confirmed the relationship between peak transmissibility and peak bone strain indicated by the FE model, and showed that the maximum cyclic tibial strain during CTV of the intact leg was 330+/-82microepsilon and occurred at 60-70 Hz. This study presents a comprehensive mechanical analysis of CTV, a loading method for studying vibrational loading under controlled conditions. This model will be used in future in vivo studies and will potentially become an important tool for understanding the response of bone to vibrational loading.


Sudden onset of proteinuria in chronic serum sickness in rats.
1. Daily intravenous administration of bovine serum albumin (BSA) to rats produces chronic serum sickness glomerulonephritis, an immune-complex-mediated renal disease that is eventually always fatal. We have performed a detailed study of the onset of proteinuria in chronic serum sickness in order to assess the long-term consequences of discontinuing daily BSA injections precisely at that very early and well-defined stage of disease. 2. Urine and plasma samples from rats receiving daily BSA injections were collected and analysed daily before the onset of proteinuria, at which time the rats were divided into three groups. Group 1 continued to receive daily BSA injections, in group 2 injections were stopped on the first, and in group 3 on the third, day of proteinuria. Proteinuria began suddenly and was not preceded either by microalbuminuria or abnormalities of plasma composition. The sudden expression of proteinuria was accompanied by an equally rapid development of hypoalbuminaemia and hypercholesterolaemia. Development of the characteristic glomerular histopathology of serum sickness coincided with, but did not precede, the onset of proteinuria. Despite the discontinuation of antigen injections at the onset of proteinuria, basement membrane thickening was evident 8 weeks later; proteinuria persisted and hypercholesterolaemia increased. 3. In this model of immune complex glomerulonephritis, changes in kidney function and immunopathology were abrupt and closely linked, precluding the use of those criteria to predict when proteinuria would begin. Furthermore, extremely brief periods of active immunological injury to the peripheral capillary wall were sufficient to produce persistent abnormalities of glomerular structure and function.


Suppression of Fusarium Wilt of Watermelon by Soil Amendment with Hairy Vetch.
Hairy vetch (Vicia villosa Roth) as a soil amendment was evaluated for suppression of Fusarium wilt of watermelon and soil populations of Fusarium oxysporum f. sp. niveum in greenhouse, microplot, and field studies. When mixed at 1 or 5% (wt/wt) in a loamy sand soil that was artificially or naturally infested with race 2 of F. oxysporum f. sp. niveum, pulverized dry hairy vetch, crab shell, and urea provided the best suppression (53 to 87% reduction) of Fusarium wilt on watermelon seedlings among 13 plant and animal residues screened. Soil amended with hairy vetch at 0.25 or 0.5% (wt/wt) in microplots resulted in 54 to 69% decreased wilt incidence and 100 to 220% increase of watermelon plant biomass. Hairy vetch winter cover crop incorporated into field plots under black plastic provided 42 to 48% reduction of wilt incidence, 64 to 100% increase of plant biomass, and a 34 to 68% increase in weight of fruit, comparable to improvements achieved by the soil fumigants methyl bromide or 1,3-dichloropropene plus 35% chloropicrin. Soil amendment with hairy vetch also increased the sugar content of watermelon fruit 10 to 15%. Significant reductions in the populations of F. oxysporum f. sp. niveum were not observed in hairy vetch-amended soil in microplots and field plots, but were observed in greenhouse pot soil amended with 5% (wt/wt) hairy vetch, which was attributed primarily to increased levels of fungicidal ammonia produced during decomposition. Incorporating hairy vetch into mulched soil can be an alternative or supplement to cultivar resistance and crop rotation for management of Fusarium wilt of watermelon.


Replicator dynamics in public goods games with reward funds.
Which punishment or rewards are most effective at maintaining cooperation in public goods interactions and deterring defectors who are willing to freeload on others' contribution? The sanction system is itself a public good and can cause problematic "second-order free riders" who do not contribute to the provisions of the sanctions and thus may subvert the cooperation supported by sanctioning. Recent studies have shown that public goods games with punishment can lead to a coercion-based regime if participation in the game is optional. Here, we reveal that even with compulsory participation, rewards can maintain cooperation within an infinitely large population. We consider three strategies for players in a standard public goods game: to be a cooperator or a defector in a standard public goods game, or to be a rewarder who contributes to the public good and to a fund that rewards players who contribute during the game. Cooperators do not contribute to the reward fund and are therefore classified as second-order free riders. The replicator dynamics for the three strategies exhibit a rock-scissors-paper cycle, and can be analyzed fully, despite the fact that the expected payoffs are nonlinear. The model does not require repeated interaction, spatial structure, group selection, or reputation. We also discuss a simple method for second-order sanctions, which can lead to a globally stable state where 100% of the population are rewarders.


Comparison of ureteric stone size, on bone window versus standard soft-tissue window settings, on multi-detector non-contrast computed tomography.
To compare the difference in mean stone size, as measured on bone window vs standard soft-tissue window setting using multi-detector computed tomography (MDCT) in patients with a solitary ureteric stone. In all, 60 patients presenting to the emergency and outpatient departments of a University Hospital from May 2015 to October 2015 and fulfilling the inclusion criteria were included in the study. A 64-slice MDCT was used to assess the locations and size of the ureteric stones. A consultant radiologist independently analysed the MDCT scans of all the patients. The mean difference in stone size was calculated between both window settings in axial and coronal planes. The mean (SD) age of the patients was 37.13 (11.9) years. Males constituted ∼68% of the cohort and 32% were female. In all, 85% of the patients had left ureteric stones and 15% had right ureteric stones. The mean (SD) stone size, as measured on the soft-tissue window setting was 6.68 (2.01) mm, and on the bone window setting was 4.8 (1.9) mm. The mean (SD) difference in stone size between the two window settings was +1.85 (0.55) mm. The two means were compared using Student's t-test, and the difference was found to be statistically significant (P < 0.05). The stone size measured using the soft-tissue window setting on a MDCT is significantly different from the measurement on the bone window setting.


Reconstructing Bronze Age diets and farming strategies at the early Bronze Age sites of La Bastida and Gatas (southeast Iberia) using stable isotope analysis.
The El Argar society of the Bronze Age in the southeast of the Iberian Peninsula (2200-1550 cal BCE) was among the first complex societies in Europe. Its economy was based on cereal cultivation and metallurgy, it was organized hierarchically, and successively expanded its territory. Most of the monumentally fortified settlements lay on steeply sloped mountains, separated by fertile plains, and allowed optimal control of the area. Here, we explore El Argar human diets, animal husbandry strategies, and food webs using stable carbon and nitrogen isotope analysis of charred cereal grains as well as human and animal bone collagen. The sample comprised 75 human individuals from the sites of La Bastida (n = 52) and Gatas (n = 23), 32 domesticated and wild animals as well as 76 barley and 29 wheat grains from two chronological phases of a total time span of ca. 650 years. The grains indicate extensive cereal cultivation under rain-fed conditions with little to moderate application of manure. Especially at La Bastida, crops and their by-products contributed significantly to the forage of the domesticated animals, which attests to a strong interrelation of cultivation and animal husbandry. Trophic level spacing and Bayesian modelling confirm that human diets were largely based on barley with some contribution of meat or dairy products. A cross-sectional analysis of bone collagen suggests that children were breastfed until about 1.5-2 years old, and infants from Gatas may have suffered from more metabolic stress than those at La Bastida. Adults of both sexes consumed similar diets that reflect social and chronological variation to some extent. Despite significantly higher δ13C and δ15N values at La Bastida than at Gatas, the isotopic data of the staple crops and domestic animals from both sites indicate that such differences do not necessarily correspond to different average human diets, but to agricultural strategies. These results urge for a reassessment of previous isotope studies in which only human remains have been taken into account. The study highlights that disentangling the complex influences on human isotope compositions requires a firm set of comparative data.


Influenza surveillance among outpatients and inpatients in Morocco, 1996-2009.
There is limited information about the epidemiology of influenza in Africa. We describe the epidemiology and seasonality of influenza in Morocco from 1996 to 2009 with particular emphasis on the 2007-2008 and 2008-2009 influenza seasons. Successes and challenges of the enhanced surveillance system introduced in 2007 are also discussed. Virologic sentinel surveillance for influenza virus was initiated in Morocco in 1996 using a network of private practitioners that collected oro-pharyngeal and naso-pharyngeal swabs from outpatients presenting with influenza-like-illness (ILI). The surveillance network expanded over the years to include inpatients presenting with severe acute respiratory illness (SARI) at hospitals and syndromic surveillance for ILI and acute respiratory infection (ARI). Respiratory samples and structured questionnaires were collected from eligible patients, and samples were tested by immunofluorescence assays and by viral isolation for influenza viruses. We obtained a total of 6465 respiratory specimens during 1996 to 2009, of which, 3102 were collected during 2007-2009. Of those, 2249 (72%) were from patients with ILI, and 853 (27%) were from patients with SARI. Among the 3,102 patients, 98 (3%) had laboratory-confirmed influenza, of whom, 85 (87%) had ILI and 13 (13%) had SARI. Among ILI patients, the highest proportion of laboratory-confirmed influenza occurred in children less than 5 years of age (3/169; 2% during 2007-2008 and 23/271; 9% during 2008-2009) and patients 25-59 years of age (8/440; 2% during 2007-2009 and 21/483; 4% during 2008-2009). All SARI patients with influenza were less than 14 years of age. During all surveillance years, influenza virus circulation was seasonal with peak circulation during the winter months of October through April. Influenza results in both mild and severe respiratory infections in Morocco, and accounted for a large proportion of all hospitalizations for severe respiratory illness among children 5 years of age and younger.


Sepsis in Acute Mediastinitis - A Severe Complication after Oesophageal Perforations. A Review of the Literature.
Despite recent advancements in antibiotic therapy and the progress made in critical care and modern diagnostic methods, acute mediastinitis continues to be a severe condition. Acute mediastinitis can occur in the context of cardio-thoracic surgery, oesophageal perforations and oropharyngeal infections condition. Forty-five percent of oesophageal perforations occurs during simple endoscopy. Spontaneous perforation (Boerhaave syndrome) accounts for 15% of perforations, and twelve percent are due to the ingestion of foreign bodies. Other causes include blind or penetrating trauma, and circa 9% to intraoperative lesions. CT scan is the standard investigation that reveals direct signs of mediastinitis.The oral administration of contrast substances can underscore the level of oesophageal perforation. Conservative treatment is the first-choice treatment and surgical treatment is reserved only for specific situations.The principles of surgical treatment consist of drainage, primary suture, oesophageal exclusion with or without the application of oesophagectomy, endoscopic vacuum wound assisted therapy of the perforation and associated paraoesophageal mediastinal drainage and endoscopic stenting associated with drainage. The lowest mortality rate is recorded in patients with perforations diagnosed less than twenty-four hours after the onset of symptoms. Surgical treatment remains the gold standard especially in cases of thoracic and abdominal perforations while further investigations are mandatory before endoscopic stenting is carried out.


CHK1-targeted therapy to deplete DNA replication-stressed, p53-deficient, hyperdiploid colorectal cancer stem cells.
Cancer stem cells (CSCs) are responsible for tumour formation and spreading, and their targeting is required for tumour eradication. There are limited therapeutic options for advanced colorectal cancer (CRC), particularly for tumours carrying RAS-activating mutations. The aim of this study was to identify novel CSC-targeting strategies. To discover potential therapeutics to be clinically investigated as single agent, we performed a screening with a panel of FDA-approved or investigational drugs on primary CRC cells enriched for CSCs (CRC-SCs) isolated from 27 patients. Candidate predictive biomarkers of efficacy were identified by integrating genomic, reverse-phase protein microarray (RPPA) and cytogenetic analyses, and validated by immunostainings. DNA replication stress (RS) was increased by employing DNA replication-perturbing or polyploidising agents. The drug-library screening led to the identification of LY2606368 as a potent anti-CSC agent acting in vitro and in vivo in tumour cells from a considerable number of patients (∼36%). By inhibiting checkpoint kinase (CHK)1, LY2606368 affected DNA replication in most CRC-SCs, including RAS-mutated ones, forcing them into premature, lethal mitoses. Parallel genomic, RPPA and cytogenetic analyses indicated that CRC-SCs sensitive to LY2606368 displayed signs of ongoing RS response, including the phosphorylation of RPA32 and ataxia telangiectasia mutated serine/threonine kinase (ATM). This was associated with mutation(s) in TP53 and hyperdiploidy, and made these CRC-SCs exquisitely dependent on CHK1 function. Accordingly, experimental increase of RS sensitised resistant CRC-SCs to LY2606368. LY2606368 selectively eliminates replication-stressed, p53-deficient and hyperdiploid CRC-SCs independently of RAS mutational status. These results provide a strong rationale for biomarker-driven clinical trials with LY2606368 in patients with CRC.


Deconstructing risk: separable encoding of variance and skewness in the brain.
Risky choice entails a need to appraise all possible outcomes and integrate this information with individual risk preference. Risk is frequently quantified solely by statistical variance of outcomes, but here we provide evidence that individuals' choice behaviour is sensitive to both dispersion (variance) and asymmetry (skewness) of outcomes. Using a novel behavioural paradigm in humans, we independently manipulated these 'summary statistics' while scanning subjects with fMRI. We show that a behavioural sensitivity to variance and skewness is mirrored in neuroanatomically dissociable representations of these quantities, with parietal cortex showing sensitivity to the former and prefrontal cortex and ventral striatum to the latter. Furthermore, integration of these objective risk metrics with subjective risk preference is expressed in a subject-specific coupling between neural activity and choice behaviour in anterior insula. Our findings show that risk is neither monolithic from a behavioural nor neural perspective and its decomposition is evident both in distinct behavioural preferences and in segregated underlying brain representations.


SAGES's advanced GI/MIS fellowship curriculum pilot project.
The American health care system faces deficits in quality and quantity of surgeons. SAGES is a major stakeholder in surgical fellowship training and is responsible for defining the curriculum for the Advanced GI/MIS fellowship. SAGES leadership is actively adapting this curriculum. The process of reform began in 2014 through a series of iterative meetings and discussions. A working group within the Resident and Fellow Training Committee reviewed case log data from 2012 to 2015. These data were used to propose new criteria designed to provide adequate exposure to core content. The working group also proposed using video assessment of an MIS case to provide objective assessment of competency. Case log data were available for 326 fellows with a total of 85,154 cases logged (median 227 per fellow). The working group proposed new criteria starting with minimum case volumes for five defined categories including foregut (20), bariatrics (25), inguinal hernia (10), ventral hernia (10), and solid organ/colon/thoracic (10). Fellows are expected to perform an additional 75 complex MIS cases of any category for a total of 150 required cases overall. The proposal also included a minimum volume of flexible endoscopy (50) and submission of an MIS foregut case for video assessment. The new criteria more clearly defined which surgeon roles count for major credit within individual categories. Fourteen fellowships volunteered to pilot these new criteria for the 2017-2018 academic year. The new SAGES Advanced GI/MIS fellowship has been crafted to better define the core content that should be contained in these fellowships, while still allowing sufficient heterogeneity so that individual learners can tailor their training to specific areas of interest. The criteria also introduce innovative, evidence-based methods for assessing competency. Pending the results of the pilot program, SAGES will consider broad implementation of the new fellowship criteria.


Effect of rhBMP-2 Immobilized Anorganic Bovine Bone Matrix on Bone Regeneration.
Anorganic bovine bone matrix (Bio-Oss®) has been used for a long time for bone graft regeneration, but has poor osteoinductive capability. The use of recombinant human bone morphogenetic protein-2 (rhBMP-2) has been suggested to overcome this limitation of Bio-Oss®. In the present study, heparin-mediated rhBMP-2 was combined with Bio-Oss® in animal experiments to investigate bone formation performance; heparin was used to control rhBMP-2 release. Two calvarial defects (8 mm diameter) were formed in a white rabbit model and then implanted or not (controls) with Bio-Oss® or BMP-2/Bio-Oss®. The Bio-Oss® and BMP-2/Bio-Oss® groups had significantly greater new bone areas (expressed as percentages of augmented areas) than the non-implanted controls at four and eight weeks after surgery, and the BMP-2/Bio-Oss® group (16.50 ± 2.87 (n = 6)) had significantly greater new bone areas than the Bio-Oss® group (9.43 ± 3.73 (n = 6)) at four weeks. These findings suggest that rhBMP-2 treated heparinized Bio-Oss® markedly enhances bone regeneration.


Brief historical background and principal nature of process.
After a brief outline of the historical background, the biological principle of structural tissue adaptation to alterations in load is discussed with special reference to the cardiovascular system. It is stressed that adaptive changes of cardiac and vascular design to local load are rapid, precise, and, on the local level, highly appropriate. However, in hypertension, when they affect the entire systemic resistance section (structural autoregulation), as well as some major "barostats," an upward structural resetting of the entire pressure equilibrium occurs. Further, by way of the consequent vascular resistance vessel hyperreactivity, a positive feedback interaction is established with functional pressure effects and thus tends to further escalate the pressure rise. For these reasons the "normal" structural factor becomes of key importance for the initiation, maintenance, and gradual aggravation of chronic high pressure states.


Correlating viral phenotypes with phylogeny: accounting for phylogenetic uncertainty.
Many recent studies have sought to quantify the degree to which viral phenotypic characters (such as epidemiological risk group, geographic location, cell tropism, drug resistance state, etc.) are correlated with shared ancestry, as represented by a viral phylogenetic tree. Here, we present a new Bayesian Markov-Chain Monte Carlo approach to the investigation of such phylogeny-trait correlations. This method accounts for uncertainty arising from phylogenetic error and provides a statistical significance test of the null hypothesis that traits are associated randomly with phylogeny tips. We perform extensive simulations to explore and compare the behaviour of three statistics of phylogeny-trait correlation. Finally, we re-analyse two existing published data sets as case studies. Our framework aims to provide an improvement over existing methods for this problem.


Relationship of race-, sexual orientation-, and HIV-related discrimination with adherence to HIV treatment: a pilot study.
Adherence to highly active antiretroviral therapy (HAART) must be close to perfect in order to maintain suppression of HIV viral load, and to prevent the development of drug resistant strains of HIV. People living with HIV (PLWH) often report low levels of adherence. One variable that has been linked to poor adherence is perceived discrimination; however, research has generally not considered the possible unique effects of different types of discrimination on adherence. The present pilot study aimed to examine the association of three types of discrimination (due to HIV+ status, race, or sexual orientation) with adherence among 57 PLWH. Logistic regression analyses were conducted to demonstrate the relationships between each type of discrimination and self-reported adherence. Racial discrimination significantly predicted lower adherence levels, whereas sexual orientation- and HIV-related discrimination did not. Results underscore the importance of addressing discrimination issues, specifically racial, when designing interventions to improve adherence to HAART.