Peptide-pulsed dendritic cells induce the hepatitis C viral epitope-specific responses of naïve human T cells.
Hepatitis C virus (HCV) is a major cause of liver disease. Spontaneous resolution of infection is associated with broad, MHC class I- (CD8(+)) and class II-restricted (CD4(+)) T cell responses to multiple viral epitopes. Only 20% of patients clear infection spontaneously, however, most develop chronic disease. The response to chemotherapy varies; therapeutic vaccination offers an additional treatment strategy. To date, therapeutic vaccines have demonstrated only limited success in clinical trials. Vector-mediated vaccination with multi-epitope-expressing DNA constructs provides an improved approach. Highly-conserved, HLA-A2-restricted HCV epitopes and HLA-DRB1-restricted immunogenic consensus sequences (ICS, each composed of multiple overlapping and highly conserved epitopes) were predicted using bioinformatics tools and synthesized as peptides. HLA binding activity was determined in competitive binding assays. Immunogenicity and the ability of each peptide to stimulate naïve human T cell recognition and IFN-γ production were assessed in cultures of total PBMCs and in co-cultures composed of peptide-pulsed dendritic cells (DCs) and purified T lymphocytes, cell populations derived from normal blood donors. Essentially all predicted HLA-A2-restricted epitopes and HLA-DRB1-restricted ICS exhibited HLA binding activity and the ability to elicit immune recognition and IFN-γ production by naïve human T cells. The ability of DCs pulsed with these highly-conserved HLA-A2- and -DRB1-restricted peptides to induce naïve human T cell reactivity and IFN-γ production ex vivo demonstrates the potential efficacy of a multi-epitope-based HCV vaccine targeted to dendritic cells.


Unexpected consequences of administering bacteriocinogenic probiotic strains for Salmonella populations, revealed by an in vitro colonic model of the child gut.
New biological strategies for the treatment of Salmonella infection are needed in response to the increase in antibiotic-resistant strains. Escherichia coli L1000 and Bifidobacterium thermophilum RBL67 were previously shown to produce antimicrobial proteinaceous compounds (microcin B17 and thermophilicin B67, respectively) active in vitro against a panel of Salmonella strains recently isolated from clinical cases in Switzerland. In this study, two three-stage intestinal continuous fermentation models of Salmonella colonization inoculated with immobilized faeces of a two-year-old child were implemented to study the effects of the two bacteriocinogenic strains compared with a bacteriocin-negative mutant of strain L1000 on Salmonella growth, as well as gut microbiota composition and metabolic activity. Immobilized E. coli L1000 added to the proximal colon reactor showed a low colonization, and developed preferentially in the distal colon reactor independent of the presence of genetic determinants for microcin B17 production. Surprisingly, E. coli L1000 addition strongly stimulated Salmonella growth in all three reactors. In contrast, B. thermophilum RBL67 added in a second phase stabilized at high levels in all reactors, but could not inhibit Salmonella already present at a high level (>10(7) c.f.u. ml(-1)) when the probiotic was added. Inulin added at the end of fermentation induced a strong bifidogenic effect in all three colon reactors and a significant increase of Salmonella counts in the distal colon reactor. Our data show that under the simulated child colonic conditions, the microcin B17 production phenotype does not correlate with inhibition of Salmonella but leads to a better colonization of E. coli L1000 in the distal colon reactor. We conclude that in vitro models with complex and complete gut microbiota are required to accurately assess the potential and efficacy of probiotics with respect to Salmonella colonization in the gut.


Effect of differing levels of tobacco-specific nitrosamines in cigarette smoke on the levels of biomarkers in smokers.
Smokers are exposed to significant doses of carcinogens, including tobacco-specific nitrosamines (TSNA). Previous studies have shown significant global differences in the levels of TSNAs in cigarette smoke because of the variation in tobacco blending and curing practices around the world. Mouth-level exposure to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) measured in cigarette butts and urinary concentrations of its major metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) were examined among 126 daily smokers in four countries over a 24-hour study period. As mouth-level exposure of NNK increased, the urinary NNAL increased even after adjustment for other covariates (beta = 0.46, P = 0.004). The relationship between mouth-level exposure to nicotine and its salivary metabolite, cotinine, was not statistically significant (beta = 0.29, P = 0.057), likely because of the very limited range of differences in mouth-level nicotine exposure in this population. We have shown a direct association between the 24-hour mouth-level exposure of NNK resulting from cigarette smoking and the concentration of its primary metabolite, NNAL, in the urine of smokers. Internal dose concentrations of urinary NNAL are significantly lower in smokers in countries that have lower TSNA levels in cigarettes such as Canada and Australia in contrast to countries that have high levels of these carcinogens in cigarettes, such as the United States. Lowering the levels of NNK in the mainstream smoke of cigarettes through the use of specific tobacco types and known curing practices can significantly affect the exposure of smokers to this known carcinogen.


GAP domains responsible for ras p21-dependent inhibition of muscarinic atrial K+ channel currents.
The interaction between the low molecular weight G protein ras p21 and a guanosine triphosphatase activating protein (GAP) uncouples a heterotrimeric G protein (Gk) from muscarinic receptors. Through the use of isolated atrial cell membranes and genetically engineered GAP deletion mutants, the src homology regions (SH2-SH3) at the amino terminus of GAP have been identified as the domains responsible for this effect. Deletion of the domain required to stimulate the guanosine triphosphatase activity of ras p21 relieves the requirement for ras p21 in this system. A model is presented that suggests that ras p21 induces a conformational change in GAP, which allows the SH2-SH3 regions of GAP to function.


Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene nonessential for growth in culture.
The gene designated gamma 134.5 maps in the inverted repeats flanking the long unique sequence of herpes simplex virus-1 (HSV-1) DNA, and therefore it is present in two copies per genome. This gene is not essential for viral growth in cell culture. Four recombinant viruses were genetically engineered to test the function of this gene. These were (i) a virus from which both copies of the gene were deleted, (ii) a virus containing a stop codon in both copies of the gene, (iii) a virus containing after the first codon an insert encoding a 16-amino acid epitope known to react with a specific monoclonal antibody, and (iv) a virus in which the deleted sequences were restored. The viruses from which the gene was deleted or which carried stop codons were avirulent on intracerebral inoculation of mice. The virus with the gene tagged by the sequence encoding the epitope was moderately virulent, whereas the restored virus reacquired the phenotype of the parent virus. Significant amounts of virus were recovered only from brains of animals inoculated with virulent viruses. Inasmuch as the product of the gamma 134.5 gene extended the host range of the virus by enabling it to replicate and destroy brain cells, it is a viral neurovirulence factor.


Housekeeping genes for quantitative expression studies in the three-spined stickleback Gasterosteus aculeatus.
During the last years the quantification of immune response under immunological challenges, e.g. parasitation, has been a major focus of research. In this context, the expression of immune response genes in teleost fish has been surveyed for scientific and commercial purposes. Despite the fact that it was shown in teleostei and other taxa that the gene for beta-actin is not the most stably expressed housekeeping gene (HKG), depending on the tissue and experimental treatment, the gene has been used as a reference gene in such studies. In the three-spined stickleback, Gasterosteus aculeatus, other HKG than the one for beta-actin have not been established so far. To establish a reliable method for the measurement of immune gene expression in Gasterosteus aculeatus, sequences from the now available genome database and an EST library of the same species were used to select oligonucleotide primers for HKG, in order to perform quantitative reverse-transcription (RT) PCR. The expression stability of ten candidate reference genes was evaluated in three different tissues, and in five parasite treatment groups, using the three algorithms BestKeeper, geNorm and NormFinder. Our results showed that in most of the tissues and treatments HKG that could not be used so far due to unknown sequences, proved to be more stably expressed than the one for beta-actin. As they were the most stably expressed genes in all tissues examined, we suggest using the genes for the L13a ribosomal binding protein and ubiquitin as alternative or additional reference genes in expression analysis in Gasterosteus aculeatus.


Somatic symptoms and panic attacks: a retrospective study of learning experiences.
Individuals with panic attacks evaluate physical anxiety symptoms as dangerous and tend to respond to them with fear. In a retrospective questionnaire study, we explored childhood and adolescent learning experiences with respect to somatic symptoms of panickers. Compared to normal controls (N = 61), patients with panic disorder (N = 121), infrequent panickers (N = 86) and patients with other anxiety disorders (N = 38) reported more frequent instances prior to age 18 when they had experienced symptoms like dizziness, shortness of breath, palpitations or nausea, accompanied by special attention from their parents and instructions to restrain from strenuous or social activities. The differences were due to higher symptom frequencies in the anxiety groups. All anxiety groups reported more frequent uncontrolled behavior of their parents than controls. Patients with panic disorder and infrequent panickers reported that their parents had suffered more frequently from physical symptoms typical of anxiety than patients with other anxiety disorders or normal controls. Panickers, but not patients with other anxiety disorders, had observed sick-role behavior related to panic symptoms in their parents more often than controls. Panic attack Ss reported a higher number of household members suffering from chronic illnesses than controls and patients with other anxiety disorders. No group differences were found in the reported behavior of parents when Ss had colds. Overall, the results point to the role of severe illnesses and physical symptoms typical of anxiety in significant others in the history of Ss with panic attacks. These experiences during childhood and adolescence may contribute to their belief that physical symptoms are dangerous. In contrast, there was no specificity for panic with respect to the Ss' own physical symptoms or cold-related symptoms.


The Ides of March. The match 1993 in perspective.
The total number of US medical school graduates who selected categorical programs in internal medicine in this year's match fell by 7.9%. Consequently, the program fill rate for US graduates declined for the eighth consecutive year, from 55.9% to 53.1%. Preliminary positions and international graduates continue to increase, though questions remain whether these residents serve as good role models to encourage student interest in internal medicine. We reviewed the 1993 National Resident Matching Program data and several curricular reforms implemented in our internal medicine clerkship and residency program to evaluate whether such reform might enhance interest in general internal medicine. Reform in our internal medicine clerkship curriculum included increased emphasis on problem-based learning and exposure to generalist role models. This resulted in substantial increases on the average miniboard examination and a 22% increase in the number of students pursuing residency in internal medicine from the previous year. Residency curricular reform centered around enhanced ambulatory care teaching along with improved exposure to generalist role models. This resulted in a gradual increase in the number of residency graduates who were staying in general internal medicine from 21% in 1991 to 40% in 1993. Innovative curricular changes in our internal medicine clerkship and residency programs have led to enhanced interest in general internal medicine. Although our results are preliminary, such change is necessary, not only to continue program excellence, but for simple survival.


Metabolic and behavioral responses to high-fat feeding in mice selectively bred for high wheel-running activity.
Increased dietary fat intake is a precipitating factor for the development of obesity and associated metabolic disturbances. Physically active individuals generally have a reduced risk of developing these unhealthy states, but the underlying mechanisms are poorly understood. In the present study, we investigated the effects of feeding a high-fat diet (HFD) on obesity development and fuel homeostasis in male and female mice with a trait for increased physical activity and in their controls. Male and female mice selectively bred for a high level of wheel running behavior over 30 generations and nonselected controls (background strain Hsd:ICR) were maintained on a standard lab chow high-carbohydrate diet (HCD) or on an HFD (60% fat). Food intake, body weight, indirect calorimetry parameters, spontaneous locomotor activity and several hormones relevant to metabolism and energy balance were measured. On HFD, mice reduced food intake and increased body fat mass and plasma leptin levels, with the notable exception of the selected females, which increased their ingested calories without any effects on body mass or plasma leptin levels. In addition, they had an elevated daily energy expenditure (DEE), increased spontaneous cage activity ( approximately 700% relative to controls) and higher resting metabolic rate (RMR) on the HFD compared with feeding the HCD. The selected males also had a higher DEE compared with controls, but no interaction with diet was observed. On HCD, adiponectin levels were higher in selected male, but not female, mice relative to controls. A marked increase in the level of plasma adiponectin was observed on the HFD in selected females, an effect of diet that was not observed in selected males. Genetically based high locomotor activity renders female, but not male, mice resistant to HFD-induced obesity by alterations in behavioral, endocrine and metabolic traits that facilitate fat utilization rather than limiting HFD intake.


Contact urticaria syndrome. Contact urticaria to diethyltoluamide (immediate-type hypersensitivity).
Contact urticaria refers to a wheal-and-flare response occurring on the application of chemicals to intact skin. Standard closed patch tests read at 48 hours after application yield misleading information; observations should be made instead using open patch tests 15 to 30 minutes after application. There are three major subdivisions of the syndrome: nonimmunologic cause (such as application of histamine), immunologic cause (immediate hypersensitivity), and uncertain cause (such as application of ammonium persulfate). Our patient had contact urticaria due to the insect repellent, diethyltoluamide. The experimental data suggest that this case was due to an immunologic response (immediate hypersensitivity) and demarcates the specificity of response. The immunologically mediated cases cover a broad spectrum of manifestations from contact urticaria only to local urticaria plus asthma and, in extreme sensitivity, includes anaphylactoid responses.


Ketamine Anesthesia Does Not Improve Depression Scores in Electroconvulsive Therapy: A Randomized Clinical Trial.
Although interest in ketamine use during electroconvulsive therapy (ECT) has increased, studies have been equivocal with regard to its efficacy. The aims of this clinical trial were to evaluate ketamine's antidepressive effects in ECT as a primary anesthetic, determine ketamine's tolerability when compared with standard anesthesia, and determine if plasma brain-derived neurotrophic factor (BDNF) is necessary for treatment response. Adults meeting criteria for treatment-resistant depression undergoing index course ECT received either methohexital (1 to 2 mg/kg) or ketamine (1 to 2 mg/kg) anesthesia in this dual-arm double-blinded randomized clinical trial (NCT02752724). The primary outcome of this study is change in depression questionnaire scores before and after ECT. Seizure data, depression severity using self-reported and clinician-assessed questionnaires, cognitive scoring, and plasma BDNF concentrations were obtained before and after completion of ECT. There were no differences in seizure lengths, hemodynamics, or seizure stimuli between the ketamine (n=23;138 ECTs) and methohexital (n=27;159 ECTs) groups. Depression scores improved similarly after ECT in both groups. In the methohexital group, 15% of patients failed to achieve adequate seizures and were switched to ketamine and 26% were converted to bilateral ECT stimulus, whereas all ketamine patients achieved adequate seizures and only 4% required bilateral stimulus. Plasma BDNF increased after ECT only in the ketamine group. Our data show that ketamine does not significantly improve depression when compared with methohexital as a single induction agent for ECT, increases serum BDNF and does not increase rates of post-ECT agitation. Ketamine use in ECT may have some benefits for some patients that are not captured through standard depression assessment questionnaires alone.


Mutational analysis of the Arabidopsis RPS2 disease resistance gene and the corresponding pseudomonas syringae avrRpt2 avirulence gene.
Plants have evolved a large number of disease resistance genes that encode proteins containing conserved structural motifs that function to recognize pathogen signals and to initiate defense responses. The Arabidopsis RPS2 gene encodes a protein representative of the nucleotide-binding site-leucine-rich repeat (NBS-LRR) class of plant resistance proteins. RPS2 specifically recognizes Pseudomonas syringae pv. tomato strains expressing the avrRpt2 gene and initiates defense responses to bacteria carrying avrRpt2, including a hypersensitive cell death response (HR). We present an in planta mutagenesis experiment that resulted in the isolation of a series of rps2 and avrRpt2 alleles that disrupt the RPS2-avrRpt2 gene-for-gene interaction. Seven novel avrRpt2 alleles incapable of eliciting an RPS2-dependent HR all encode proteins with lesions in the C-terminal portion of AvrRpt2 previously shown to be sufficient for RPS2 recognition. Ten novel rps2 alleles were characterized with mutations in the NBS and the LRR. Several of these alleles code for point mutations in motifs that are conserved among NBS-LRR resistance genes, including the third LRR, which suggests the importance of these motifs for resistance gene function.


Integrity of white matter in the corpus callosum correlates with bimanual co-ordination skills.
Variation in brain structure may reflect variation in functional properties of specific brain systems. Structural variation may therefore reflect variation in behavioural performance. Here, we use diffusion-weighted magnetic resonance imaging to show that variation in white matter integrity in a specific region in the body of the corpus callosum is associated with variation in performance of a bimanual co-ordination task. When the callosal region showing this association is used as a seed for probabilistic tractography, inter-hemispheric pathways are generated to the supplementary motor area and caudal cingulate motor area. This provides further evidence for the role of medial wall motor areas in bimanual co-ordination and supports the idea that variation in brain structure reflects inter-individual differences in skilled performance.


The effects of daily consumption of grapefruit on body weight, lipids, and blood pressure in healthy, overweight adults.
Folklore has suggested that consuming grapefruit may promote weight control. Sparse data exist to support this hypothesis, although there is some evidence of health promotion effects with regard to blood pressure control and modulation of circulating lipids. The aim of this randomized controlled trial was to prospectively evaluate the role of grapefruit in reducing body weight and blood pressure and in promoting improvements in the lipid profile in overweight adults (N = 74). Following a 3-week washout diet low in bioactive-rich fruits and vegetables, participants were randomized to either the control diet (n = 32) or daily grapefruit (n = 42) in the amount of one half of a fresh Rio-Red grapefruit with each meal (3× daily) for 6 weeks. No differences between group in weight, blood pressure, or lipids were demonstrated. Grapefruit consumption was associated with modest weight loss (-0.61 ± 2.23 kg, P = .097), a significant reduction in waist circumference (-2.45 ± 0.60 cm, P = .0002), and a significant reduction in systolic blood pressure (-3.21 ± 10.13 mm Hg, P = .03) compared with baseline values. Improvements were observed in circulating lipids of those consuming grapefruit, with total cholesterol and low-density lipoprotein significantly decreasing by -11.7 mg/dL (P = .002) and -18.7 mg/dL (P < .001), respectively, compared with baseline values. This study suggests that consumption of grapefruit daily for 6 weeks does not significantly decrease body weight, lipids, or blood pressure as compared with the control condition. However, the improvements in blood pressure and lipids demonstrated in the intervention group suggest that grapefruit should be further evaluated in the context of obesity and cardiovascular disease prevention.


Hepatic mitochondrial inner membrane properties and carnitine palmitoyltransferase A and B. Effect of diabetes and starvation.
Intact mitochondria and inverted submitochondrial vesicles were prepared from the liver of fed, starved (48 h) and streptozotocin-diabetic rats in order to characterize carnitine palmitoyltransferase kinetics and malonyl-CoA sensitivity in situ. In intact mitochondria, both starved and diabetic rats exhibited increased Vmax., increased Km for palmitoyl-CoA, and decreased sensitivity to malonyl-CoA inhibition. Inverted submitochondrial vesicles also showed increased Vmax. with starvation and diabetes, with no change in Km for either palmitoyl-CoA or carnitine. Inverted vesicles were uniformly less sensitive to malonyl-CoA regardless of treatment, and diabetes resulted in a further decrease in sensitivity. In part, differences in the response of carnitine palmitoyltransferase to starvation and diabetes may reside in differences in the membrane environment, as observed with Arrhenius plots, and the relation of enzyme activity and membrane fluidity. In all cases, whether rats were fed, starved or diabetic, and whether intact or inverted vesicles were examined, increasing membrane fluidity was associated with increasing activity. Malonyl-CoA was found to produce a decrease in intact mitochondrial membrane fluidity in the fed state, particularly at pH 7.0 or less. No effect was observed in intact mitochondria from starved or diabetic rats, or in inverted vesicles from any of the treatment groups. Through its effect on membrane fluidity, malonyl-CoA could regulate carnitine palmitoyltransferase activity on both surfaces of the inner membrane through an interaction with only the outer surface.


Multicenter Evaluation of Circulating Cell-Free DNA Extraction and Downstream Analyses for the Development of Standardized (Pre)analytical Work Flows.
In cancer patients, circulating cell-free DNA (ccfDNA) can contain tumor-derived DNA (ctDNA), which enables noninvasive diagnosis, real-time monitoring, and treatment susceptibility testing. However, ctDNA fractions are highly variable, which challenges downstream applications. Therefore, established preanalytical work flows in combination with cost-efficient and reproducible reference materials for ccfDNA analyses are crucial for analytical validity and subsequently for clinical decision-making. We describe the efforts of the Innovative Medicines Initiative consortium CANCER-ID (http://www.cancer-id.eu) for comparing different technologies for ccfDNA purification, quantification, and characterization in a multicenter setting. To this end, in-house generated mononucleosomal DNA (mnDNA) from lung cancer cell lines carrying known TP53 mutations was spiked in pools of plasma from healthy donors generated from 2 different blood collection tubes (BCTs). ccfDNA extraction was performed at 15 partner sites according to their respective routine practice. Downstream analysis of ccfDNA with respect to recovery, integrity, and mutation analysis was performed centralized at 4 different sites. We demonstrate suitability of mnDNA as a surrogate for ccfDNA as a process quality control from nucleic acid extraction to mutation detection. Although automated extraction protocols and quantitative PCR-based quantification methods yielded the most consistent and precise results, some kits preferentially recovered spiked mnDNA over endogenous ccfDNA. Mutated TP53 fragments derived from mnDNA were consistently detected using both next-generation sequencing-based deep sequencing and droplet digital PCR independently of BCT. This comprehensive multicenter comparison of ccfDNA preanalytical and analytical work flows is an important contribution to establishing evidence-based guidelines for clinically feasible (pre)analytical work flows.


Development of injectable, resorbable drug-releasing copolymer scaffolds for minimally invasive sustained ophthalmic therapeutics.
Copolymers based on N-isopropylacrylamide (NIPAAm), acrylic acid N-hydroxysuccinimide (NAS) and varying concentrations of acrylic acid (AA) and acryloyloxy dimethyl-γ-butyrolactone (DBA) were synthesized to create thermoresponsive, resorbable copolymers for minimally invasive drug and/or cell delivery to the posterior segment of the eye to combat retinal degenerative diseases. Increasing DBA content was found to decrease both copolymer water content and lower critical solution temperature. The incorporation of NAS provided an amine-reactive site, which can be exploited for facile conjugation of bioactive agents. Proton nuclear magnetic resonance analysis revealed the onset of hydrolysis-dependent opening of the DBA lactone ring, which successfully eradicated copolymer phase transition properties and should allow the gelled polymer to re-hydrate, enter systemic circulation and be cleared from the body without the production of degradation byproducts. Hydrolytic ring opening occurs slowly, with over 85% copolymer mass remaining after 130 days of incubation in 37°C phosphate buffered saline. These slow-degrading copolymers are hypothesized to be ideal delivery vehicles to provide minimally invasive, sustained, localized release of pharmaceuticals within the posterior segment of the eye to combat retinal degenerative diseases.


Constant, cycling, hot and cold thermal environments: strong effects on mean viability but not on genetic estimates.
It has frequently been suggested that trait heritabilities are environmentally sensitive, and there are genetic trade-offs between tolerating different environments such as hot and cold or constant and fluctuating temperatures. Future climate predictions suggest an increase in both temperatures and their fluctuations. How species will respond to these changes is uncertain, particularly as there is a lack of studies which compare genetic performances in constant vs. fluctuating environments. In this study, we used a nested full-sib/half-sib breeding design to examine how the genetic variances and heritabilities of egg-to-adult viability differ at high and low temperatures with and without daily fluctuations in temperatures using Drosophila melanogaster as a model organism. Although egg-to-adult viability was clearly sensitive to developmental temperatures, heritabilities were not particularly sensitive to developmental temperatures. Moreover, we found that egg-to-adult viabilities at different developmental temperatures were positively correlated, suggesting a common genetic background for egg-to-adult viability at different temperatures. Finding both a uniform genetic background coupled with rather low heritabilities insensitive to temperatures, our results suggest evolutionary responses are unlikely to be limited by temperature effects on genetic parameters or negative genetic correlations, but by the direct effects of stressful temperatures on egg-to-adult viability accompanied with low heritabilities.


Skin biopsy rates and incidence of melanoma: population based ecological study.
To describe changes in skin biopsy rates and to determine their relation with changes in the incidence of melanoma. Population based ecological study. Nine geographical areas of the United States. Participants of the Surveillance Epidemiology and End Results (SEER) programme aged 65 and older. For the period 1986 to 2001, annual skin biopsy rates for each surveillance area from Medicare claims and incidence rates for melanoma for the same population. Between 1986 and 2001 the average biopsy rate across the nine participating areas increased 2.5-fold among people aged 65 and older (2847 to 7222 per 100,000 population). Over the same period the average incidence of melanoma increased 2.4-fold (45 to 108 per 100,000 population). Assuming that the occurrence of true disease was constant, the extra number of melanoma cases that were diagnosed after carrying out 1000 additional biopsies was 12.6 (95% confidence interval 11.2 to 14.0). After controlling for a potential increase in the true occurrence of disease, 1000 additional biopsies were still associated with 6.9 (3.1 to 10.8) extra melanoma cases diagnosed. Stage specific analyses suggested that 1000 biopsies were associated with 4.4 (2.1 to 6.8) extra cases of in situ melanoma diagnosed and 2.3 (0.0 to 4.6) extra cases of local melanoma, but not with the incidence of advanced melanoma. Mortality from melanoma changed little during the period. The incidence of melanoma is associated with biopsy rates. That the extra cases diagnosed were confined to early stage cancer while mortality remained stable suggests overdiagnosis-the increased incidence being largely the result of increased diagnostic scrutiny and not an increase in the incidence of disease.


Predicting global scale exposure of humans to PCB 153 from historical emissions.
Predicting human exposure to an environmental contaminant based on its emissions is one of the great challenges of environmental chemistry. It has been done successfully on a local or regional scale for some persistent organic pollutants. Here we assess whether it can be done at a global scale, using PCB 153 as a test chemical. The global multimedia fate model BETR Global and the human exposure model ACC-HUMAN were employed to predict the concentration of PCB 153 in human milk for 56 countries around the world from a global historical emissions scenario. The modeled concentrations were compared with measurements in pooled human milk samples from the UNEP/WHO Global Monitoring Plan. The modeled and measured concentrations were highly correlated (r = 0.76, p < 0.0001), and the concentrations were predicted within a factor of 4 for 49 of 78 observations. Modeled concentrations of PCB 153 in human milk were higher than measurements for some European countries, which may reflect weaknesses in the assumptions made for food sourcing and an underestimation of the rate of decrease of concentrations in air during the last decades. Conversely, modeled concentrations were lower than measurements in West African countries, and more work is needed to characterize exposure vectors in this region.


The multi-track angiography catheter: a new tool for complex catheterisation in congenital heart disease.
To develop a simple and versatile catheter system for complex cardiac catheterisation because angiography and pressure measurements during diagnostic and interventional cardiac catheterisation are often unsatisfactory. The Multi-Track Angio catheter system is a single lumen side-hole catheter with a short distal extension containing a lumen for a standard guidewire. The catheter is introduced over a previously placed guidewire running through this distal extension. It can then be manipulated within the heart by sliding along the guidewire. The tip of the catheter is always stabilised by the guidewire. This stability enhances angiography and pressure recordings. The Multi-Track Angio catheter system was used in 84 patients (age 1 day-20 years). Thirty one procedures were diagnostic and 53 interventional. The decision to use the Multi-Track Angio catheter was based on three criteria: firstly, unsatisfactory angiography obtained with conventional equipment; secondly, difficult catheter course requiring use of a guidewire; and thirdly, requirement for angiography and pressure recordings during interventional procedures. No complications were encountered. High quality angiography could be performed in all cases without catheter recoil. The Multi-Track Angio catheter system allows for high quality angiography and pressure recordings during diagnostic and interventional cardiac catheterisation. The advantage of the system is that both angiography and pressure recordings can be performed repeatedly from stable catheter positions using a previously placed guidewire. This reduces the need for guidewire manipulations or catheter exchanges and decreases procedure time and the risk of complications.


Integrating PET and PET/CT into the risk-adapted therapy of lymphoma.
Imaging with 18F-FDG PET is increasingly accepted as a valuable tool for lymphoma management. A recent shift in the use of PET and PET/CT in medical practice has become evident. We selected aggressive lymphomas as a platform for the discussion of these imaging modalities in oncology patients and the resulting management questions. On the basis of our clinical experience and a review of the literature, we evaluated the emerging role of 18F-FDG PET in staging, response assessment, risk stratification, and tailored therapy. We explored the biologic meaning of true-positive or true-negative PET results in assessing tumor killing and the implications for risk-adapted therapy of lymphoma. PET/CT improves the accuracy of staging and response assessment over that of conventional anatomic imaging. The strong prognostic value of PET for aggressive lymphomas is established, whether the imaging is performed at the end of therapy or after only a few cycles of chemotherapy. How to modify therapy on the basis of PET results is not yet established, although it is clear that high-risk patient subsets can be reliably identified. PET/CT improves the accuracy of staging and response assessment over that of CT alone. A negative midtreatment PET result does not indicate the absence of a viable tumor or that therapy can be abbreviated or reduced in intensity. Similarly, a positive PET result does not necessarily indicate a viable tumor or that extending or intensifying treatment will benefit the patient. In assessing response, it is possible that prognosis rests not only on whether the PET result is positive or negative but also on the intensity of the signal. Although the prognostic value of PET for lymphoma is now clear, how to tailor therapy accordingly is a separate matter that requires further investigation.


Comparison of endovascular and open surgical repairs for abdominal aortic aneurysm.
Evaluate treatment options for nonruptured abdominal aortic aneurysms (AAA); the relationship of hospital and physician volume to outcomes for endovascular repair (EVAR); affect of patient and AAA factors on outcomes; cost-benefits of treatments. PubMed, Cochrane Library, FDA, and other electronic websites until May 2006. Reference lists and content experts were used to identify additional reports. Randomized controlled trials (RCT) of open surgical repair (OSR), EVAR, or active surveillance, systematic reviews, nonrandomized U.S. trials, and national registries were used to assess clinical outcomes. Volume-outcome articles published after 2000 were reviewed if they reported the relationship between U.S. hospital or physician volume and outcomes, were population-based, and the analysis was adjusted for risk factors. Cost studies included at least 50 EVAR and provided data on costs or charges, and cost-effectiveness analyses. Initial or attained diameter is the strongest known predictor of rupture. The annual risk of rupture is below 1 percent for AAA <5.5 cm in diameter. Among medically ill patients unfit for OSR with AAA >/=5.5 cm, the risk of rupture may be as high as 10 percent per year. Early/immediate OSR of AAA <5.5 cm (two trials n=2,226) did not reduce all-cause mortality compared with surveillance and delayed OSR. Results did not differ according to age, gender, baseline AAA diameter or creatinine concentration. Two RCT with followup of at least 2 years compared EVAR to OSR for AAA >/=5.5 cm. EVAR reduced postoperative 30-day mortality compared to OSR (1.6 percent EVAR vs. 4.7 percent OSR, RR = 0.34 [0.17 to 0.65]). Early reduction in all-cause mortality with EVAR disappeared before 2 years. Post-operative complications and reinterventions were higher with EVAR. Quality of life differences were small and disappeared after 3-6 months. One RCT of patients with AAA >/=5.5 cm judged medically unfit for OSR (n=338), reported no difference in all-cause mortality or AAA mortality between EVAR and no intervention (HR = 1.21; 95 percent CI 0.87 to 1.69). Forty-eight nonrandomized reports evaluated EVAR. Patient, AAA characteristics, and outcomes were similar to RCT comparing EVAR to OSR. A volume outcome relationship has been shown for OSR, but there are no data adequate to estimate the effect of hospital or physician volume on EVAR outcomes or to identify a volume threshold for policymakers. Immediate OSR for AAA <5.5 cm costs more and does not improve long-term survival compared to active surveillance and delayed OSR. The cost effectiveness of EVAR relative to OSR is difficult to determine. However, compared to OSR for AAA >/=5.5 cm, EVAR has greater in-hospital costs primarily due to the cost of the prosthesis. EVAR has shorter length of stay, lower 30-day morbidity and mortality but does not improve quality of life beyond 3 months or survival beyond 2 years, and is associated with complications, need for reintervention, long-term monitoring, and higher long-term costs. Compared to no intervention in patients medically unfit for OSR, EVAR costs more and does not improve survival or quality of life. For AAA <5.5 cm in diameter, active surveillance with delayed OSR results in equivalent mortality but lesser morbidity and operative costs due to fewer interventions compared to immediate OSR. For AAA >/=5.5 cm, EVAR has not been shown to improve long-term survival or health status over OSR though peri-operative outcomes are improved. EVAR does not improve survival in patients who are medically unfit for OSR. EVAR is associated with more complications, need for reintervention, monitoring, and costs compared to OSR or no intervention. U.S. RCT are needed using approved EVAR devices to evaluate patient outcomes.


Survey Breakoffs in a Computer-Assisted Telephone Interview.
Nearly 23% of all telephone interviews in the most recently completed wave of the Panel Study of Income Dynamics break off at least once, requiring multiple sessions to complete the interview. Given this high rate, a study was undertaken to better understand the causes and consequences of temporary breakoffs in a computer-assisted telephone interview setting. The majority of studies examining breakoffs have been conducted in the context of self-administered web surveys. The present study uses new paradata collected on telephone interview breakoffs to describe their prevalence, associated field effort, the instrument sections and questions on which they occur, their source - whether respondent-initiated, interviewer-initiated, or related to telephone problems - and associations with respondent and interviewer characteristics. The results provide information about the survey response process and suggest a set of recommendations for instrument design and interviewer training, as well as additional paradata that should be collected to provide more insight into the breakoff phenomenon.


Chemoattractant-induced phosphatidylinositol 3,4,5-trisphosphate accumulation is spatially amplified and adapts, independent of the actin cytoskeleton.
Experiments in amoebae and neutrophils have shown that local accumulations of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P(3)] mediate the ability of cells to migrate during gradient sensing. To define the nature of this response, we subjected Dictyostelium discoideum cells to measurable temporal and spatial chemotactic inputs and analyzed the accumulation of PI(3,4,5)P(3) on the membrane, as well as the recruitment of the enzymes phosphoinositide 3-kinase and PTEN. In latrunculin-treated cells, spatial gradients elicited a PI(3,4,5)P(3) response only on the front portion of the cell where the response increased more steeply than the gradient and did not depend on its absolute concentration. Phosphoinositide 3-kinase bound to the membrane only at the front, although it was less sharply localized than PI(3,4,5)P(3). Membrane-bound PTEN was highest at the rear and varied inversely with receptor occupancy. The localization of PI(3,4,5)P(3) was enhanced further in untreated polarized cells containing an intact cytoskeleton. Interestingly, the treated cells could respond to two independent gradients simultaneously, demonstrating that a response at the front does not necessarily inhibit the back. Combinations of temporal and spatial stimuli provided evidence of an inhibitory process and showed that a gradient generates a persistent steady-state response independent of a previous history of exposure to chemoattractant. These results support a local excitation/global inhibition model and argue against other schemes proposed to explain directional sensing.


Evaluation of high-perimeter electrode designs for deep brain stimulation.
Deep brain stimulation (DBS) is an effective treatment for movement disorders and a promising therapy for treating epilepsy and psychiatric disorders. Despite its clinical success, complications including infections and mis-programing following surgical replacement of the battery-powered implantable pulse generator adversely impact the safety profile of this therapy. We sought to decrease power consumption and extend battery life by modifying the electrode geometry to increase stimulation efficiency. The specific goal of this study was to determine whether electrode contact perimeter or area had a greater effect on increasing stimulation efficiency. Finite-element method (FEM) models of eight prototype electrode designs were used to calculate the electrode access resistance, and the FEM models were coupled with cable models of passing axons to quantify stimulation efficiency. We also measured in vitro the electrical properties of the prototype electrode designs and measured in vivo the stimulation efficiency following acute implantation in anesthetized cats. Area had a greater effect than perimeter on altering the electrode access resistance; electrode (access or dynamic) resistance alone did not predict stimulation efficiency because efficiency was dependent on the shape of the potential distribution in the tissue; and, quantitative assessment of stimulation efficiency required consideration of the effects of the electrode-tissue interface impedance. These results advance understanding of the features of electrode geometry that are important for designing the next generation of efficient DBS electrodes.


Echinococcus granulosus and other intestinal helminths: current status of prevalence and management in rural dogs of eastern Australia.
Ascertain the prevalence of intestinal helminths in rural dogs from eastern Australia and Tasmania. Identify farm management practices contributing to the perpetuation and transmission of Echinococcus granulosus. Helminth infection in dogs was determined microscopically through faecal flotation. Infection with E. granulosus was determined via faecal antigen-capture ELISA and coproPCR. Taeniid eggs were identified using molecular methods. Data on dog management and owner understanding of hydatid disease were collected via questionnaire. Faeces were collected from 1425 Australian rural dogs (1119 mainland; 306 Tasmania). Eggs of hookworms were most prevalent, up to 40.2%, followed by whipworms (Trichuris vulpis), up to 21.2%. Roundworms (Toxocara canis and Toxascaris leonine) were least common, up to 6.1%. Taeniid eggs were found in 11 dogs (5 Taenia pisiformis; 2 T. serialis; 4 T. hydatigena); 2 of the T. hydatigena-infected dogs were also E. granulosus coproantigen-positive. Of the 45 dogs found to be E. granulosus coproantigen-positive, 24 were in Tasmania, 16 in NSW, 3 in Victoria and 2 in Queensland. Three Tasmanian coproantigen ELISA-positive dogs were also coproPCR-positive. The most common dog ration was commercial dry food, but half the owners fed raw meat to their dogs and some fed offal of lambs (8.9%) or mutton (7.8%). More than half (69%) of owners weighed their dogs before deworming. Few dewormed their dogs often enough to ensure they remained cestode-free and owners hunting wildlife usually left carcases where they were shot. E. granulosus is still present in Australian rural dogs, including Tasmania, but at low levels. Owner behaviour perpetuates transmission of cestodes.


Arachidonic acid and docosahexaenoic acid supplementation increases coronary flow velocity reserve in Japanese elderly individuals.
Arachidonic acid (ARA) and docosahexaenoic acid (DHA) are important components of phospholipids and cell membranes. There has, however, been no clinical report on the direct effects of ARA and DHA on coronary circulation. To evaluate the effects of ARA and DHA on coronary circulation using the measurement of coronary flow velocity reserve (CFVR) by transthoracic Doppler echocardiography (TTDE). A double-blind, placebo-matched study of 28 Japanese elderly individuals (19 men, mean age 65 years) conducted to compare the effects of polyunsaturated fatty acids (PUFA; ARA 240 mg/day, DHA 240 mg/day) and placebo on CFVR. Coronary flow velocity (CFV) of the left anterior descending coronary artery was measured at rest and during hyperaemia by TTDE to determine CFVR. There were no significant differences in CFV at rest or during hyperaemia in CFVR at baseline in the two groups (PUFA versus placebo 17 (7 SD) versus 16 (6), 62 (20) versus 59 (12), and 3.85 (1.04) versus 3.98 (0.83) cm/s, respectively). After three months' supplementation, CFV during hyperaemia was significantly higher in the PUFA than in the placebo group (73 (19) versus 64 (12) cm/s, p<0.01) although no significant difference was found between the two groups in CFV at rest (17 (7) versus 16 (4) cm/s). CFVR thus significantly increased after PUFA consumption (3.85 (1.04) versus 4.46 (0.95), p = 0.0023). Three months' supplementation of PUFA increased CFVR in Japanese elderly individuals, which suggests beneficial effects of PUFA on the coronary microcirculation.


From undocumented to lawfully present: Do changes to legal status impact psychological wellbeing among latino immigrant young adults?
Exclusionary immigration policies, as a form of structural racism, have led to a sizeable undocumented population that is largely barred from access to resources in the United States. Existing research suggests that undocumented immigration status detrimentally impacts mobility, yet few studies have tested the impacts of legal status on psychological wellbeing. Most importantly, we know little about how changes to legal status impact wellbeing. Announced in 2012, the Deferred Action for Childhood Arrivals (DACA) program allows eligible undocumented youth to apply for temporary lawful status. Drawing on cross-sectional survey data from 487 Latino immigrant young adults in California collected in 2014 and 2015, we analyze the predictors of three specialized outcomes related to immigrants' psychological wellbeing-distress, negative emotions, and deportation worry before and after a transition from undocumented to lawfully present status. Results show that retrospective reports of past psychological wellness, when all respondents were undocumented, are predicted primarily by socioeconomic status. However, reports of current psychological wellness are predicted by DACA status. Our results demonstrate, for the first time, the positive emotional consequences of transitioning out of undocumented status for immigrant young adults.


How were new medicines discovered?
Preclinical strategies that are used to identify potential drug candidates include target-based screening, phenotypic screening, modification of natural substances and biologic-based approaches. To investigate whether some strategies have been more successful than others in the discovery of new drugs, we analysed the discovery strategies and the molecular mechanism of action (MMOA) for new molecular entities and new biologics that were approved by the US Food and Drug Administration between 1999 and 2008. Out of the 259 agents that were approved, 75 were first-in-class drugs with new MMOAs, and out of these, 50 (67%) were small molecules and 25 (33%) were biologics. The results also show that the contribution of phenotypic screening to the discovery of first-in-class small-molecule drugs exceeded that of target-based approaches - with 28 and 17 of these drugs coming from the two approaches, respectively - in an era in which the major focus was on target-based approaches. We postulate that a target-centric approach for first-in-class drugs, without consideration of an optimal MMOA, may contribute to the current high attrition rates and low productivity in pharmaceutical research and development.


Inhibition of AP-1 binding and transcription by gold and selenium involving conserved cysteine residues in Jun and Fos.
Gold(I) salts and selenite, which have diverse therapeutic and biological effects, are noted for their reactivity with thiols. Since the binding of Jun-Jun and Jun-Fos dimers to the AP-1 DNA binding site is regulated in vitro by a redox process involving conserved cysteine residues, we hypothesized that some of the biological actions of gold and selenium are mediated via these residues. In electrophoretic mobility-shift analyses, AP-1 DNA binding was inhibited by gold(I) thiolates and selenite, with 50% inhibition occurring at approximately 5 microM and 1 microM, respectively. Thiomalic acid had no effect in the absence of gold(I), and other metal ions inhibited at higher concentrations, in a rank order correlating with their thiol binding affinities. Cysteine-to-serine mutants demonstrated that these effects of gold(I) and selenite require Cys272 and Cys154 in the DNA-binding domains of Jun and Fos, respectively. Gold(I) thiolates and selenite did not inhibit nonspecific protein binding to the AP-1 site and were at least an order of magnitude less potent as inhibitors of sequence-specific binding to the AP-2, TFIID, or NF1 sites compared with the AP-1 site. In addition, 10 microM gold(I) or 10 microM selenite inhibited expression of an AP-1-dependent reporter gene, but not an AP-2-dependent reporter gene. These data suggest a mechanism regulating transcription factor activity by inorganic ions which may contribute to the known antiarthritic action of gold and cancer chemoprevention by selenium.


Clinical, radiologic, and manometric characteristics of chronic intestinal dysmotility: the Stanford experience.
The clinical spectrum of chronic intestinal dysmotility (CID) is not well known. We determined the spectrum of motor abnormalities, underlying pathology, clinical course, and response to treatment of adults with CID at a tertiary referral center. This was a descriptive retrospective analysis of a CID cohort conducted at a tertiary referral gastrointestinal (GI) motility center. A total of 113 referred patients underwent gastroduodenal manometry, other motility studies as appropriate, and radiologic and/or endoscopic assessment to exclude mechanical intestinal obstruction. Common symptoms included abdominal distention, abdominal pain, nausea, and constipation. The course was chronic with intermittent symptoms. Gastroduodenal manometry was abnormal in all patients; a pattern suggestive of a neuropathic process was the most common. Other GI motility studies showed delayed gastric, gallbladder, and colonic transit, nonspecific esophageal dysmotility, sphincter of Oddi hypertonicity, and poor rectal balloon sensation/expulsion. Treatment involved nutritional support, prokinetics, analgesics, antinausea agents, and laxatives, with variable response and high morbidity, multiple emergency admissions, need for nutritional support, and poor response to surgery. Nearly 40% of the patients underwent abdominal surgery. Patients with CID have a chronic course and high morbidity. Because any segment of the GI tract may be involved in CID, functional assessment of the entire GI tract is recommended. CID presents several unmet clinical needs even in tertiary centers with expertise.


Phage display peptide probes for imaging early response to bevacizumab treatment.
Early evaluation of cancer response to a therapeutic regimen can help increase the effectiveness of treatment schemes and, by enabling early termination of ineffective treatments, minimize toxicity, and reduce expenses. Biomarkers that provide early indication of tumor therapy response are urgently needed. Solid tumors require blood vessels for growth, and new anti-angiogenic agents can act by preventing the development of a suitable blood supply to sustain tumor growth. The purpose of this study is to develop a class of novel molecular imaging probes that will predict tumor early response to an anti-angiogenic regimen with the humanized vascular endothelial growth factor antibody bevacizumab. Using a bevacizumab-sensitive LS174T colorectal cancer model and a 12-mer bacteriophage (phage) display peptide library, a bevacizumab-responsive peptide (BRP) was identified after six rounds of biopanning and tested in vitro and in vivo. This 12-mer peptide was metabolically stable and had low toxicity to both endothelial cells and tumor cells. Near-infrared dye IRDye800-labeled BRP phage showed strong binding to bevacizumab-treated tumors, but not to untreated control LS174T tumors. In addition, both IRDye800- and (18)F-labeled BRP peptide had significantly higher uptake in tumors treated with bevacizumab than in controls treated with phosphate-buffered saline. Ex vivo histopathology confirmed the specificity of the BRP peptide to bevacizumab-treated tumor vasculature. In summary, a novel 12-mer peptide BRP selected using phage display techniques allowed non-invasive visualization of early responses to anti-angiogenic treatment. Suitably labeled BRP peptide may be potentially useful pre-clinically and clinically for monitoring treatment response.


Amyloid goiter due to familial mediterranean Fever in a patient with byler syndrome: a case report.
Familial Mediterranean Fever (FMF), also inherited with autosomal recessive trait, is characterized by recurrent episodes of fever, arthritis, and serositis. Congenital Byler Syndrome (Progressive Familial Intrahepatic Cholestasis) inherited with autosomal recessive trait and characterized by defective secretion of bile acids. FMF associated Amyloid A deposition occurs in many tissues and organs, but amyloid goiter is a rare entity that leads to enlargement and dysfunction of the thyroid. We present a rare case of 24 year old male patient who had liver and kidney transplantation due to Byler Syndrome and secondary amyloidosis related to FMF, diagnosed as rapidly growing large amyloid goiter. Deposits of extracellular amyloid and dense adipose metaplasia diagnostic for amyloid goiter are determined upon histopathological examination of thyroidectomy material. When goiter was detected in cases with history of systemic amyloidosis and rapidly growing goitre, amyloid goiter should be remembered at first. This case is unique since two autosomal genetic disorders are together in the same patient and important as it emphasizes the consequences of consanguineous marriage, early diagnosis and treatment compliance of FMF and the awareness of amyloid goiter in patients followed by primary care physicians and healthcare professionals.


Modeling amino acid side chains in proteins: 15N NMR spectra of guanidino groups in nonpolar environments.
Natural-abundance 15N NMR spectroscopy on dodecylguanidine reveals solvent and protonation effects that model those that could occur for the arginine side chain in proteins. Our results demonstrate that the 15N chemical shifts of the terminal guanine nitrogens strongly depend on the solvent chosen for measurements. A polar H-bond-donating solvent like water has strongly deshielding effects on the neutral guanidine group (with the latter acting predominantly as an H-bond acceptor). As a result, a substantial upfield shift occurs when neutral guanidine is dissolved instead in a non-H-bonding solvent (chloroform). These solvent effects can be as large as those induced by protonation changes. This limits the ability of 15N chemical shifts to distinguish the protonation state of the arginine side chain, at least without specific knowledge of its environment. These results help to reconcile previous interpretations about the protonation state arg-82 in the M state of bacteriorhodopsin based on FTIR and 15N NMR spectroscopy. That is, contrary to earlier conclusions from solid-state NMR, the side chain of arg-82 could undergo a deprotonation between the bR and M states, but only if it also experienced a significant decrease in the H-bonding character and polarity of its environment. In fact, the average 15N chemical shift of the two Neta of arg-82 in bacteriorhodopsin's M intermediate (from the previous NMR measurements) is 17 ppm upfield from the corresponding value for the deprotonated arginine side chain in aqueous solution at pH >14, but only 3 ppm upfield from the value for deprotonated dodecylguanidine in chloroform.


Definitions and Assessment Approaches for Emergency Medical Services for Children.
Pediatric Life Support (PLS) courses and instructional programs are educational tools developed to teach resuscitation and stabilization of children who are critically ill or injured. A number of PLS courses have been developed by national professional organizations for different health care providers (eg, pediatricians, emergency physicians, other physicians, prehospital professionals, pediatric and emergency advanced practice nurses, physician assistants). PLS courses and programs have attempted to clarify and standardize assessment and treatment approaches for clinical practice in emergency, trauma, and critical care. Although the effectiveness of PLS education has not yet been scientifically validated, the courses and programs have significantly expanded pediatric resuscitation training throughout the United States and internationally. Variability in terminology and in assessment components used in education and training among PLS courses has the potential to create confusion among target groups and in how experts train educators and learners to teach and practice pediatric emergency, trauma, and critical care. It is critical that all educators use standard terminology and patient assessment to address potential or actual conflicts regarding patient evaluation and treatment. This article provides a consensus of several organizations as to the proper order and terminology for pediatric patient assessment. The Supplemental Information provides definitions for terms and nomenclature used in pediatric resuscitation and life support courses.


Application of surface roughness analysis on micro-computed tomographic images of bone erosion: examples using a rodent model of rheumatoid arthritis.
Quantifying the bone erosion in preclinical models of rheumatoid arthritis is valuable for the evaluation of drug treatments. This study introduces a three-dimensional method for bone surface roughness measurement from micro-computed tomographic data obtained from rats subjected to collagen-induced arthritis (CIA), in which the degree of bone erosion is related to the severity and the duration of the disease. In two studies of rat CIA, the surface roughness of the talus bone following 21 days of disease increased 559% and 486% from the control group. At 41 days following disease induction, the roughness of the bone surface increased 857% above baseline. The roughness of the control samples was similar from each study (less than 4% different), demonstrating the robustness of the algorithm. Treatment with methotrexate at 0.1 mg/kg daily demonstrated significant protection from bone erosion, whereas the 0.05 mg/kg daily dose was not efficacious (98% versus 22% inhibition of roughness-measured bone erosion). The main advantage of such an algorithm is demonstrated in the preclinical drug study of rat CIA with methotrexate treatment, indicating the immediate utility of this approach in drug development studies.


Ecstasy use and its association with sexual behaviors among drug users in New York City.
In the past two decades, recreational use of ecstasy has become a growing concern in the United States, although most studies assessing ecstasy use have focused on white, middle-class adolescents who use ecstasy during raves and in clubs. We assessed the prevalence of recent ecstasy use among predominantly minority heroin, cocaine, and crack users in New York City and the association between ecstasy and sexual risk above and beyond that of the other drugs. Between 2002 and 2004, injection and non-injection heroin, crack and cocaine users (N= 534) completed a risk behavior questionnaire that included items on ecstasy use. Logistic regression was used to investigate the relation between current ecstasy use and sexual behaviors. Of 534 illicit drug users, 69.7% were aged 25 years or older, 65.2% were Hispanic, 27.9% Black and 77.4% male; 36.7% were injectors. 17.2% of respondents reported recent (last six months) ecstasy use. In a multivariable logistic regression model, current ecstasy use was associated both with initiating sex before age 14 (adjusted odds ratio (AOR) = 1.51) and having two or more partners in the past two months (AOR = 1.86) after adjusting for age at study entry, current cocaine and marijuana use and being an injection drug user. This study suggests that ecstasy use may be more prevalent among urban drug users. Ecstasy use in urban settings, beyond clubs and raves, should continue to be monitored.


Optimum Temperature of Oxygen for Transpulmonary Hypothermia with Cooled Oxygen Inhalation: A Preliminary Study in a Rat Model.
Cooled oxygen inhalation was hypothesized as a novel hypothermia technique in a previous study. In the current study, we aimed to determine the optimal temperature of oxygen for this method. This is a prospective, randomized, controlled, examiner-blinded experimental study conducted with 45 healthy, adult, Wistar Hannover male rats. Rats were randomly divided into five groups; group 1: +4°C intubated group (n = 7), group 2: +4°C nonintubated group (n = 9), group 3: +8°C intubated group (n = 9), group 4: +8°C nonintubated group (n = 9), and group 5: control group (n = 9). The control group received only a standardized anesthesia protocol, and no hypothermia technique was administered. Cooled oxygen was administered in the four study groups until the rectal temperature reached 34°C. The target temperature was maintained between 32°C and 34°C for 2 hours. Then, hypothermia protocols were terminated and rats were rewarmed externally with a blanket. Main outcomes were the speed (°C/minute) of temperature decrease (S) and the time required to reach the target body temperature (T). All study groups had better results than the control group in T and S values (p < 0.001) for both parameters. Group 1 had a better T value than group 4 (p = 0.01), but no difference in S value (p = 0.223). Comparison of group 2 and group 4 showed that group 4 had better results in T and S (p = 0.04 and 0.001, respectively). No pathologic changes in histologic examination were observed in any group. Our study showed that the optimal temperature of oxygen for the cooled oxygen technique was +4°C through an intubation tube.


Social tipping points in global groundwater management.
Groundwater is critical to global food security, environmental flows, and millions of rural livelihoods in the face of climate change 1 . Although a third of Earth's largest groundwater basins are being depleted by irrigated agriculture 2 , little is known about the conditions that lead resource users to comply with conservation policies. Here we developed an agent-based model 3,4 of irrigated agriculture rooted in principles of cooperation 5,6 and collective action 7 and grounded on the World Values Survey Wave 6 (n = 90,350). Simulations of three major aquifer systems facing unsustainable demands reveal tipping points where social norms towards groundwater conservation shift abruptly with small changes in cultural values and monitoring and enforcement provisions. These tipping points are amplified by group size and best invoked by engaging a minority of rule followers. Overall, we present a powerful tool for evaluating the contingency of regulatory compliance upon cultural, socioeconomic, institutional and physical conditions, and its susceptibility to change beyond thresholds. Managing these thresholds may help to avoid unsustainable groundwater development, reduce enforcement costs, better account for cultural diversity in transboundary aquifer management and increase community resilience to changes in regional climate. Although we focus on groundwater, our methods and findings apply broadly to other resource management issues.


Radiographic assessment of the thickness lost from polyethylene tibial inserts that had been sterilized differently.
Archived serial radiographs of knee replacements provide indirect evidence of the clinical performance of ultra-high molecular weight polyethylene tibial bearings. Our purpose was to determine the loss of thickness in polyethylene tibial inserts that were of the same design but had been sterilized differently. Four hundred and sixteen knees with an Anatomic Modular Knee primary total knee replacement had five to eighteen years of follow-up and a posterior cruciate ligament-retaining polyethylene tibial insert that had been sterilized with either gamma radiation in air (from 1987 to 1993) or gamma radiation in an inert gas (from 1993 to 1995), or by means of a nonirradiation gas-plasma method (from 1995 to 2001). Readings were taken from 1975 weight-bearing anteroposterior radiographs of the knee on which the top surface of the metal tibial tray had nearly projected as a line. In each tibiofemoral compartment, loss of polyethylene thickness was calculated as the standard thickness of the insert minus the measured magnification-corrected thickness. For each sterilization method, thickness loss was plotted versus the corresponding follow-up time, and a mixed linear regression model was used to quantify the change in thickness over time. Multiple linear regression analysis was used to determine whether loss of thickness was associated with implant factors, patient variables, and early postoperative limb alignment. According to the mixed model regression coefficients, loss of medial compartment thickness was greatest for inserts that had been sterilized with gamma radiation in air (0.15 mm/yr), least for those sterilized with gamma radiation in an inert gas (0.02 mm/yr), and of an intermediate magnitude for those sterilized with a nonirradiation method (0.06 mm/year); loss of lateral compartment thickness ranged from 0.03 to 0.06 mm/yr. Notably, no insert irradiated in an inert gas showed a loss of thickness of >1 mm, and no nonirradiated insert showed a loss of >2 mm. Loss of medial compartment thickness in inserts sterilized with radiation in air increased significantly with a greater polyethylene shelf age, lesser patient age, and more varus alignment of the limb in the early postoperative period (p < 0.01). In this total knee design, tibial polyethylene performance improved markedly after discontinuation of the gamma radiation in air sterilization technique. Future concerns are that bearings sterilized with radiation in an inert gas may oxidize in vivo and develop fatigue wear because of free radicals generated during sterilization with radiation and that nonirradiated bearings may undergo greater losses in thickness from routine burnishing since they lack the cross-linking that accompanies sterilization with radiation.


The use of herbal medicine in Alzheimer's disease-a systematic review.
The treatments of choice in Alzheimer's disease (AD) are cholinesterase inhibitors and NMDA-receptor antagonists, although doubts remain about the therapeutic effectiveness of these drugs. Herbal medicine products have been used in the treatment of Behavioral and Psychological Symptoms of Dementia (BPSD) but with various responses. The objective of this article was to review evidences from controlled studies in order to determine whether herbs can be useful in the treatment of cognitive disorders in the elderly. Randomized controlled studies assessing AD in individuals older than 65 years were identified through searches of MEDLINE, LILACS, Cochrane Library, dissertation Abstract (USA), ADEAR (Alzheimer's Disease Clinical Trials Database), National Research Register, Current Controlled trials, Centerwatch Trials Database and PsychINFO Journal Articles. The search combined the terms Alzheimer disease, dementia, cognition disorders, Herbal, Phytotherapy. The crossover results were evaluated by the Jadad's measurement scale. The systematic review identified two herbs and herbal formulations with therapeutic effects for the treatment of AD: Melissa officinalis, Salvia officinalis and Yi-Gan San and BDW (Ba Wei Di Huang Wan). Ginkgo biloba was identified in a meta-analysis study. All five herbs are useful for cognitive impairment of AD. M. officinalis and Yi-Gan San are also useful in agitation, for they have sedative effects. These herbs and formulations have demonstrated good therapeutic effectiveness but these results need to be compared with those of traditional drugs. Further large multicenter studies should be conducted in order to test the cost-effectiveness of these herbs for AD and the impact in the control of cognitive deterioration.


Transmission of calicivirus by a foodhandler in the pre-symptomatic phase of illness.
After a Christmas party in a restaurant, 48 (68%) of the 82 guests contracted calicivirus gastroenteritis. The epidemiological investigation showed that salad was strongly associated with the disease episode (RR = 2.43, P = 0.0005). Similar symptoms occurred among other customers who had had a meal at the same restaurant on the same evening. A foodhandler who had only prepared salad and appetizers became sick about 30 min after the end of his shift. He had been free of symptoms while preparing food. Few outbreak investigations have shown calicivirus transmission by foodhandlers some hours before becoming symptomatic.


Air transport of patients with pneumothorax: is tube thoracostomy required before flight?
It is conventionally thought that patients with pneumothorax (PTX) require tube thoracostomy (TT) before air medical transport (AMT), especially in unpressurized rotor-wing (RW) aircraft, to prevent deterioration from expansion of the PTX or development of tension PTX. We hypothesize that patients with PTX transported without TT tolerate RW AMT without serious deterioration, as defined by hypotension, hypoxemia, respiratory distress, intubation, bag valve mask ventilation, needle thoracostomy (NT), or cardiac arrest during transport. We conducted a retrospective review of a case-series of trauma patients transported to a single Level 1 trauma center via RW with confirmed PTX and no TT. Using standardized abstraction forms, we reviewed charts for signs of deterioration. Those patients identified as having clinical deterioration were independently reviewed for the likelihood that the clinical deterioration was a direct consequence of PTX. During the study period, 66 patients with confirmed PTX underwent RW AMT with an average altitude gain of 1890 feet, an average barometric pressure 586-600 mmHg, and average flight duration of 28 minutes. All patients received oxygen therapy; 14/66 patients (21%) were supported with positive pressure ventilation. Eleven of 66 patients (17%) had NT placed before flight and 4/66 (6%) had NT placed during flight. Four of 66 patients (6% CI0.3-11.7) may have deteriorated during AMT as a result of PTX; all were successfully managed with NT. In this series, 6% of patients with PTX deteriorated as result of AMT without TT, yet all patients were managed successfully with NT. Routine placement of TT in patients with PTX before RW AMT may not be necessary. Further prospective evaluation is warranted.


Psychedelic Treatment for Trauma-Related Psychological and Cognitive Impairment Among US Special Operations Forces Veterans.
U.S. Special Operations Forces Veterans are at increased risk for a variety of mental health problems and cognitive impairment associated with military service. Current treatments are lacking in effectiveness and adherence. Therefore, this study examined psychedelic treatment with ibogaine and 5-methoxy-N,N-dimethyltryptamine for trauma-related psychological and cognitive impairment among U.S. Special Operations Forces Veterans. We conducted a survey of Veterans who completed a specific psychedelic clinical program in Mexico between 2017 and 2019. Questions probed retrospective reports of mental health and cognitive functioning during the 30 days before and 30 days after treatment. A total of 65 people completed treatment during this time frame and were eligible for contact. Of these, 51 (78%) completed the survey and were included in data analyses (mean age = 40; male = 96%; married = 55%; Caucasian/White = 92%; Operation Enduring Freedom/Operation Iraqi Freedom Service = 96%). Results indicated significant and very large reductions in retrospective report of suicidal ideation (p < .001; d = -1.9), cognitive impairment (p < .001; d = -2.8), and symptoms of posttraumatic stress disorder (p < .001; d = -3.6), depression (p < .001; d = -3.7), and anxiety (p < .001; d = -3.1). Results also showed a significant and large increase in retrospective report of psychological flexibility (p < .001; d = 2.9) from before-to-after the psychedelic treatment. Increases in the retrospective report of psychological flexibility were strongly associated with retrospective report of reductions in cognitive impairment, and symptoms of posttraumatic stress disorder, depression, and anxiety (rs range -0.61 to -0.75; p < .001). Additionally, most participants rated the psychedelic experiences as one of the top five personally meaningful (84%), spiritually significant (88%), and psychologically insightful (86%) experiences of their lives.Limitations: Several limitations should be considered including the retrospective, self-report, survey design of the study, and the lack of randomization and blinding, thus making these finding preliminary. U.S. Special Operations Forces Veterans may have unique treatment needs because of the sequela of problems associated with repeated trauma exposure and the nature of the exposure. Psychedelic-assisted therapy with these under-researched psychedelics may hold unique promise for this population. However, controlled studies are needed to determine whether this treatment is efficacious in relieving mental health and cognitive impairment among U.S. Special Operations Forces Veterans.


Dilated Cardiomyopathy: Phosphorus 31 MR Spectroscopy at 7 T.
Purpose To test whether the increased signal-to-noise ratio of phosphorus 31 (31P) magnetic resonance (MR) spectroscopy at 7 T improves precision in cardiac metabolite quantification in patients with dilated cardiomyopathy (DCM) compared with that at 3 T. Materials and Methods Ethical approval was obtained, and participants provided written informe consent. In a prospective study, 31P MR spectroscopy was performed at 3 T and 7 T in 25 patients with DCM. Ten healthy matched control subjects underwent 31P MR spectroscopy at 7 T. Paired Student t tests were performed to compare results between the 3-T and 7-T studies. Results The phosphocreatine (PCr) signal-to-noise ratio increased 2.5 times at 7 T compared with that at 3 T. The PCr to adenosine triphosphate (ATP) concentration ratio (PCr/ATP) was similar at both field strengths (mean ± standard deviation, 1.48 ± 0.44 at 3 T vs 1.54 ± 0.39 at 7 T, P = .49), as expected. The Cramér-Rao lower bounds in PCr concentration (a measure of uncertainty in the measured ratio) were 45% lower at 7 T than at 3 T, reflecting the higher quality of 7-T 31P spectra. Patients with dilated cardioyopathy had a significantly lower PCr/ATP than did healthy control subjects at 7 T (1.54 ± 0.39 vs 1.95 ± 0.25, P = .005), which is consistent with previous findings. Conclusion 7-T cardiac 31P MR spectroscopy is feasible in patients with DCM and gives higher signal-to-noise ratios and more precise quantification of the PCr/ATP than that at 3 T. PCr/ATP was significantly lower in patients with DCM than in control subjects at 7 T, which is consistent with previous findings at lower field strengths.


Natural selection of parental ability to vary the sex ratio of offspring.
Theory and data suggest that a male in good condition at the end of the period of parental investment is expected to outreproduce a sister in similar condition, while she is expected to outreproduce him if both are in poor condition. Accordingly, natural selection should favor parental ability to adjust the sex ratio of offspring produced according to parental ability to invest. Data from mammals support the model: As maternal condition declines, the adult female tends to produce a lower ratio of males to females.


HIV-1 specific IgA detected in vaginal secretions of HIV uninfected women participating in a microbicide trial in Southern Africa are primarily directed toward gp120 and gp140 specificities.
Many participants in microbicide trials remain uninfected despite ongoing exposure to HIV-1. Determining the emergence and nature of mucosal HIV-specific immune responses in such women is important, since these responses may contribute to protection and could provide insight for the rational design of HIV-1 vaccines. We first conducted a pilot study to compare three sampling devices (Dacron swabs, flocked nylon swabs and Merocel sponges) for detection of HIV-1-specific IgG and IgA antibodies in vaginal secretions. IgG antibodies from HIV-1-positive women reacted broadly across the full panel of eight HIV-1 envelope (Env) antigens tested, whereas IgA antibodies only reacted to the gp41 subunit. No Env-reactive antibodies were detected in the HIV-negative women. The three sampling devices yielded equal HIV-1-specific antibody titers, as well as total IgG and IgA concentrations. We then tested vaginal Dacron swabs archived from 57 HIV seronegative women who participated in a microbicide efficacy trial in Southern Africa (HPTN 035). We detected vaginal IgA antibodies directed at HIV-1 Env gp120/gp140 in six of these women, and at gp41 in another three women, but did not detect Env-specific IgG antibodies in any women. Vaginal secretions of HIV-1 infected women contained IgG reactivity to a broad range of Env antigens and IgA reactivity to gp41. In contrast, Env-binding antibodies in the vaginal secretions of HIV-1 uninfected women participating in the microbicide trial were restricted to the IgA subtype and were mostly directed at HIV-1 gp120/gp140.


Anharmonic and Quantum Fluctuations in Molecular Crystals: A First-Principles Study of the Stability of Paracetamol.
Molecular crystals often exist in multiple competing polymorphs, showing significantly different physicochemical properties. Computational crystal structure prediction is key to interpret and guide the search for the most stable or useful form, a real challenge due to the combinatorial search space, and the complex interplay of subtle effects that work together to determine the relative stability of different structures. Here we take a comprehensive approach based on different flavors of thermodynamic integration in order to estimate all contributions to the free energies of these systems with density-functional theory, including the oft-neglected anharmonic contributions and nuclear quantum effects. We take the two main stable forms of paracetamol as a paradigmatic example. We find that anharmonic contributions, different descriptions of van der Waals interactions, and nuclear quantum effects all matter to quantitatively determine the stability of different phases. Our analysis highlights the many challenges inherent in the development of a quantitative and predictive framework to model molecular crystals. However, it also indicates which of the components of the free energy can benefit from a cancellation of errors that can redeem the predictive power of approximate models, and suggests simple steps that could be taken to improve the reliability of ab initio crystal structure prediction.


Factors determining sensitivity and resistance of tumor cells to arsenic trioxide.
Previously, arsenic trioxide showed impressive regression rates of acute promyelocytic leukemia. Here, we investigated molecular determinants of sensitivity and resistance of cell lines of different tumor types towards arsenic trioxide. Arsenic trioxide was the most cytotoxic compound among 8 arsenicals investigated in the NCI cell line panel. We correlated transcriptome-wide microarray-based mRNA expression to the IC(50) values for arsenic trioxide by bioinformatic approaches (COMPARE and hierarchical cluster analyses, Ingenuity signaling pathway analysis). Among the identified pathways were signaling routes for p53, integrin-linked kinase, and actin cytoskeleton. Genes from these pathways significantly predicted cellular response to arsenic trioxide. Then, we analyzed whether classical drug resistance factors may also play a role for arsenic trioxide. Cell lines transfected with cDNAs for catalase, thioredoxin, or the anti-apoptotic bcl-2 gene were more resistant to arsenic trioxide than mock vector transfected cells. Multidrug-resistant cells overexpressing the MDR1, MRP1 or BCRP genes were not cross-resistant to arsenic trioxide. Our approach revealed that response of tumor cells towards arsenic trioxide is multi-factorial.


Energy Metabolism Disorder as a Contributing Factor of Rheumatoid Arthritis: A Comparative Proteomic and Metabolomic Study.
To explore the pathogenesis of rheumatoid arthritis (RA), the different metabolites were screened in synovial fluid by metabolomics. Synovial fluid from 25 RA patients and 10 normal subjects were analyzed by GC/TOF MS analysis so as to give a broad overview of synovial fluid metabolites. The metabolic profiles of RA patients and normal subjects were compared using multivariate statistical analysis. Different proteins were verified by qPCR and western blot. Different metabolites were verified by colorimetric assay kit in 25 inactive RA patients, 25 active RA patients and 20 normal subjects. The influence of hypoxia-inducible factor (HIF)-1α pathway on catabolism was detected by HIF-1α knockdown. A subset of 58 metabolites was identified, in which the concentrations of 7 metabolites related to energy metabolism were significantly different as shown by importance in the projection (VIP) (VIP ≥ 1) and Student's t-test (p<0.05). In the 7 metabolites, the concentration of glucose was decreased, and the concentration of lactic acid was increased in the synovial fluid of RA patients than normal subjects verified by colorimetric assay Kit. Receiver operator characteristic (ROC) analysis shows that the concentration of glucose and lactic acid in synovial fluid could be used as dependable biomarkers for the diagnosis of active RA, provided an AUC of 0.906 and 0.922. Sensitivity and specificity, which were determined by cut-off points, reached 84% and 96% in sensitivity and 95% and 85% in specificity, respectively. The verification of different proteins identified in our previous proteomic study shows that the enzymes of anaerobic catabolism were up-regulated (PFKP and LDHA), and the enzymes of aerobic oxidation and fatty acid oxidation were down-regulated (CS, DLST, PGD, ACSL4, ACADVL and HADHA) in RA patients. The expression of HIF-1α and the enzymes of aerobic oxidation and fatty acid oxidation were decreased and the enzymes of anaerobic catabolism were increased in FLS cells after HIF-1α knockdown. It was found that enhanced anaerobic catabolism and reduced aerobic oxidation regulated by HIF pathway are newly recognized factors contributing to the progression of RA, and low glucose and high lactic acid concentration in synovial fluid may be the potential biomarker of RA.


Systemic chemotherapy plus cetuximab after complete surgery in the treatment of isolated colorectal peritoneal carcinoma: COCHISE phase II clinical trial.
The primary objective of this non-randomised phase II study was to evaluate the combination of systemic chemotherapy plus cetuximab after complete cytoreductive surgery (CCS) for treatment of isolated colorectal peritoneal carcinoma (CRPC). This multicentre, prospective phase II clinical trial was conducted in seven national cancer referral centres, however research published during study recruitment indicated cetuximab treatment as ineffective in patients with mutated KRAS genes, leading to an additional exclusion criterion to the current protocol, excluding patients with mutated KRAS genes. This significantly impacted recruitment and the study did not achieve the necessary recruitment of 46 patients. Fourteen patients underwent CCS and were included in the study, however one did not provide informed consent and another received only one cycle of chemotherapy leading to 12 patients in the per protocol population for analysis. Adjuvant Folfox Cetuximab was administered when CCS was achieved for patients > 18 years with histologically proven CRPC and no other metastatic disease (liver, lungs, lymphadenopathy, etc.). CRPC median index was 5.00 (range: 1-17). Median PFS was 12.3 months [95% CI (3.7-28.2)] with 8.3% [95% CI (0.5-31.1)] and 0% PFS at 3 and 5 years respectively. Median OS was 43.4 months [95% CI (16.8-60)]. Trial registration Clinical Trials NCT00766142, October 3, 2008. Retrospectively registered.


Restoration of Somatosensory Function by Pairing Vagus Nerve Stimulation with Tactile Rehabilitation.
Sensory dysfunction is a common consequence of many forms of neurological injury, including stroke and nerve damage. Rehabilitative paradigms that incorporate sensory retraining can provide modest benefits, but the majority of patients are left with lasting sensory loss. We have developed a novel strategy that uses closed-loop vagus nerve stimulation (VNS) paired with tactile rehabilitation to enhance synaptic plasticity and facilitate recovery of sensory function. A clinical case report provides initial evidence that a similar implementation of closed-loop VNS paired with a tactile rehabilitation regimen could improve recovery of somatosensory function. Here, we sought to build on these promising initial clinical data and rigorously evaluate the ability of VNS paired with tactile rehabilitation to improve recovery in an animal model of chronic sensory loss. The study design, including planned sample size, assessments, and statistical comparisons, was preregistered prior to beginning data collection (https://osf.io/xsnj5/). VNS paired with tactile rehabilitation resulted in a significant and nearly complete recovery of mechanosensory withdrawal thresholds. Equivalent tactile rehabilitation without VNS failed to improve sensory function. This VNS-dependent restoration of sensory thresholds was maintained for several months after the cessation of stimulation, illustrating long-term benefits. Moreover, VNS paired with tactile rehabilitation resulted in significant generalized improvements in other measures of sensorimotor forepaw function. Given the safety and tolerability of VNS therapy, these findings suggest that incorporating VNS paired with sensory retraining into rehabilitative regimens may represent a fundamentally new method to increase recovery of sensory function after neurological injury. ANN NEUROL 2020;87:194-205.


Elaiomycins K and L, new azoxy antibiotics from Streptomyces sp. Tü 6399*.
Elaiomycins K and L, two new azoxy-type antibiotics, were detected by HPLC-diode array screening in the culture filtrate extract of Streptomyces sp. Tü 6399. The structures were determined by high-resolution MS and 2-dimensional (1)H and (13)C correlated NMR spectroscopy including (15)N-NMR experiments and established these compounds as new members of the elaiomycin family. Both metabolites show a weak antibacterial activity against Bacillus subtilis and Staphylococcus lentus as well as against the phytophathogenic strain Xanthomonas campestris.


Search for invisible decays of the Higgs boson produced in association with a hadronically decaying vector boson in pp collisions at [Formula: see text] TeV with the ATLAS detector.
A search for Higgs boson decays to invisible particles is performed using 20.3 [Formula: see text] of pp collision data at a centre-of-mass energy of 8 TeV recorded by the ATLAS detector at the Large Hadron Collider. The process considered is Higgs boson production in association with a vector boson ([Formula: see text] or Z) that decays hadronically, resulting in events with two or more jets and large missing transverse momentum. No excess of candidates is observed in the data over the background expectation. The results are used to constrain VH production followed by H decaying to invisible particles for the Higgs boson mass range [Formula: see text] GeV. The 95 % confidence-level observed upper limit on [Formula: see text] varies from 1.6 pb at 115 GeV to 0.13 pb at 300 GeV. Assuming Standard Model production and including the [Formula: see text] contribution as signal, the results also lead to an observed upper limit of 78 % at 95 % confidence level on the branching ratio of Higgs bosons decays to invisible particles at a mass of 125 GeV.


Glucose in dry and moist ionic liquid: vibrational circular dichroism, IR, and possible mechanisms.
Ionic liquids and their mixtures with water show remarkable features in cellulose processing. For this reason, understanding the behavior of carbohydrates in ionic liquids is important. In the present study, we investigated three d-glucose isomers (α, β and open-chain) in 1-ethyl-3-methylimidazolium acetate in the presence and absence of water, through ab initio molecular dynamics simulations. In the complex hydrogen bonding network of these mixtures, the most interesting observation is that upon water addition every hydrogen bond elongates, except the glucose-glucose hydrogen bond for the open-chain and the α-form which shortens, clearly showing the beginning of the crystallization process. The ring glucose rearranges from on-top to in-plane and the open form changes from a coiled to a more linear arrangement when adding water which explains the contradiction that the center of mass distances of the glucose molecules with other glucose molecules grow while the hydrogen bonds shorten. The appearance of coiled open forms indicates that the previously suggested isomerization between these forms is possible and might play a role in the solubility of the related carbohydrates. The calculated IR and VCD spectra reveal insight into the intermolecular interactions, with good to excellent agreements with experimental spectra. Investigating the role of the cation, distances between the acidic carbon atom of the cation and the glucose carbon atom where ring closure and opening occurs are found, which are way shorter than dispersion-like interactions between aliphatic hydrocarbons.


Telomere length maintenance, shortening, and lengthening.
Telomeres maintain chromosome stability and cell replicative capacity. Telomere shortening occurs concomitant with aging. Short telomeres are associated with some diseases, such as dyskeratosis congenita, idiopathic pulmonary fibrosis, and aplastic anemia. Telomeres are longer in pluripotent stem cells than in somatic cells and lengthen significantly during preimplantation development. Furthermore, telomere elongation during somatic cell reprogramming is of great importance in the acquisition of authentic pluripotency. This review focuses primarily on regulatory mechanisms of telomere length maintenance in pluripotent cells, telomere length extension in early embryo development, and also telomere rejuvenation in somatic cell reprogramming. Telomere related diseases are also discussed in this review.


Development and application of a database of food ingredient fraud and economically motivated adulteration from 1980 to 2010.
Food ingredient fraud and economically motivated adulteration are emerging risks, but a comprehensive compilation of information about known problematic ingredients and detection methods does not currently exist. The objectives of this research were to collect such information from publicly available articles in scholarly journals and general media, organize into a database, and review and analyze the data to identify trends. The results summarized are a database that will be published in the US Pharmacopeial Convention's Food Chemicals Codex, 8th edition, and includes 1305 records, including 1000 records with analytical methods collected from 677 references. Olive oil, milk, honey, and saffron were the most common targets for adulteration reported in scholarly journals, and potentially harmful issues identified include spices diluted with lead chromate and lead tetraoxide, substitution of Chinese star anise with toxic Japanese star anise, and melamine adulteration of high protein content foods. High-performance liquid chromatography and infrared spectroscopy were the most common analytical detection procedures, and chemometrics data analysis was used in a large number of reports. Future expansion of this database will include additional publically available articles published before 1980 and in other languages, as well as data outside the public domain. The authors recommend in-depth analyses of individual incidents. This report describes the development and application of a database of food ingredient fraud issues from publicly available references. The database provides baseline information and data useful to governments, agencies, and individual companies assessing the risks of specific products produced in specific regions as well as products distributed and sold in other regions. In addition, the report describes current analytical technologies for detecting food fraud and identifies trends and developments.


Freezing tolerance in plants requires lipid remodeling at the outer chloroplast membrane.
Plants show complex adaptations to freezing that prevent cell damage caused by cellular dehydration. Lipid remodeling of cell membranes during dehydration is one critical mechanism countering loss of membrane integrity and cell death. SENSITIVE TO FREEZING 2 (SFR2), a gene essential for freezing tolerance in Arabidopsis, encodes a galactolipid remodeling enzyme of the outer chloroplast envelope membrane. SFR2 processively transfers galactosyl residues from the abundant monogalactolipid to different galactolipid acceptors, forming oligogalactolipids and diacylglycerol, which is further converted to triacylglycerol. The combined activity of SFR2 and triacylglycerol-biosynthetic enzymes leads to the removal of monogalactolipids from the envelope membrane, changing the ratio of bilayer- to non-bilayer-forming membrane lipids. This SFR2-based mechanism compensates for changes in organelle volume and stabilizes membranes during freezing.


Estrogen plus progestin and risk of benign proliferative breast disease.
Women with benign proliferative breast disease are at increased risk of subsequent breast cancer. Estrogens and progesterone exert proliferative effects on mammary epithelium, and combined hormone replacement therapy has been associated with increased breast cancer risk. We tested the effect of conjugated equine estrogen plus progestin on the risk of benign proliferative breast disease in the Women's Health Initiative (WHI) randomized controlled trial. In the WHI trial of estrogen plus progestin, 16,608 postmenopausal women were randomly assigned either to 0.625 mg/day of conjugated equine estrogen plus 2.5 mg/day of medroxyprogesterone acetate or to placebo. Baseline and annual breast exams and mammograms were required. The trial was terminated early (average follow-up, 5.5 years). We identified women who had had a biopsy for benign breast disease, and subjected histologic sections from the biopsies to standardized review. Overall, 178 incident cases of benign proliferative breast disease were ascertained in the estrogen plus progestin group and 99 in the placebo group. The use of estrogen plus progestin was associated with a 74% increase in the risk of benign proliferative breast disease [hazard ratio, 1.74; 95% confidence interval (CI), 1.35-2.25]. For benign proliferative breast disease without atypia the hazard ratio was 2.00 (95% CI, 1.50-2.66), while for atypical hyperplasia it was 0.76 (95% CI, 0.38-1.52). The risk varied little by levels of baseline characteristics. The results of this study suggest that the use of estrogen plus progestin may increase the risk of benign proliferative breast disease.