Radioimmunotherapy of lymphoma: a UC Davis experience.
B-cell malignancies, such as malignant lymphoma and chronic lymphocytic leukemia, commonly present with advanced disease and multiple sites of involvement. Consequently, systemic combination chemotherapy is the standard therapeutic approach and cures about one half of these patients. Development of novel therapies is required if the remaining patients are to be cured of their malignancy. Lym-1, a mouse monoclonal antibody that is reactive with these malignancies, has been coupled with 131I or 67Cu and used to treat 55 patients with advanced B-cell malignancies that had proven resistant to standard therapy. The majority of the patients responded to this therapy and the survival of responders was longer than that of non-responders. Similar results have recently been reported by others. These results represent a remarkable achievement for single agent therapy because these trials were exploratory in nature and most of the patients had failed many chemotherapy regimens. The toxicities were in general mild and readily manageable. It appears that enhancing strategies are likely to improve upon these results by increasing the therapeutic index.


Melanocortin-4 receptor (MC4R) genotypes have no major effect on fatness in a Large White x Wild Boar intercross.
The melanocortin-4 receptor (MC4R), a G-protein coupled receptor, is implicated in mediating the effect of leptin on food intake and energy balance. A previous candidate gene study reported an association between an MC4R missense mutation (Asp298Asn) and fatness, growth and feed intake in pigs. To assess this association further, we analysed the segregation of this missense mutation in relation to variation in fatness traits using a Wild Boar x Large White intercross. The Wild Boar and Large White founders were homozygous for different MC4R alleles. The MC4R was assigned to the expected region on pig chromosome 1. The statistical evaluation did not reveal any indication of a significant effect on fatness related traits in this pedigree.


Estimated gain in life expectancy. A simple tool to select optimal reperfusion treatment in individual patients with evolving myocardial infarction.
Currently several modes of reperfusion therapy for acute myocardial infarction are available. Streptokinase, accelerated alteplase and direct angioplasty are the most frequently used. These options are increasingly effective, but are also increasingly complex and costly. Since, unfortunately, physicians are often restricted by budget limitations, choices must be made in clinical practice to provide optimal therapy to individual patients. In order to guide such decision making, we developed a model to predict the expected benefit of therapy in terms of gain in life expectancy. Patients' life expectancy will decrease after infarction. Part of this loss can be prevented by early reperfusion therapy. The clinical benefit of therapy ranges from negligible gain in patients with small infarcts treated relatively late to an expected gain of more than 2 years in patients with extensive infarction treated within 3 h of onset of symptoms. The expected benefits are presented in a set of tables and depend on age, previous infarction, estimated infarct size, treatment delay and intracranial bleeding risk. With the help of these table, resources will be allocated in such a manner that patients who will benefit the most will receive the most effective therapy. Patients with similar expected treatment benefit will be offered the same mode of therapy. Future life years were discounted at 5% per year. The arbitrary thresholds currently applied for decision making at the Thoraxcenter are: no reperfusion therapy when the estimated gain in discounted life expectancy was < 1 month, streptokinase for 1-4 months and accelerated alteplase for a gain > or = 5 months. Direct angioplasty is recommended in patients with an estimated gain > or = 12 months, and in patients with an increased risk of intracranial bleeding. In this way, approximately 80% of our patients will be treated with thrombolytics (40% streptokinase and 40% accelerated alteplase), while in 10% direct angioplasty will be initiated. Patients with small infarcts presenting late will not receive reperfusion therapy. These threshold values have been chosen arbitrarily, and different thresholds may be selected in other centres. However, the developed model would guarantee that treatment decisions are made in a consistent manner, to provide optimal therapy for patients with evolving myocardial infarction, in spite of limited resources.


Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes.
An accelerated rate of glucose transport is among the most characteristic biochemical markers of cellular transformation. To study the molecular mechanism by which transporter activity is altered, cultured rodent fibroblasts transfected with activated myc, ras, or src oncogenes were used. In myc-transfected cells, the rate of 2-deoxy-D-glucose uptake was unchanged. However, in cells transfected with activated ras and src oncogenes, the rate of glucose uptake was markedly increased. The increased transport rate in ras- and src-transfected cells was paralleled by a marked increase in the amount of glucose transporter protein, as assessed by immunoblots, as well as by a markedly increased abundance of glucose transporter messenger RNA. Exposure of control cells to the tumor-promoting phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) for 18 hours had a similar effect of increasing the rate of glucose transport and the abundance of transporter messenger RNA. For ras, src, and TPA, the predominant mechanism responsible for activation of the transport system is increased expression of the structural gene encoding the glucose transport protein.


Threshold dose distributions for 5 major allergenic foods in children.
For most allergenic foods, insufficient threshold dose information within the population restricts the advice on levels of unintended allergenic foods which should trigger precautionary labeling on prepackaged foods. We wanted to derive threshold dose distributions for major allergenic foods and to elaborate the protein doses at which a proportion of the allergic population is likely to respond. For 7 allergenic foods double-blind, placebo-controlled food challenges (DBPCFCs) with a positive outcome for allergic reactions were selected from the clinical database of children routinely tested to diagnose food allergy at the University Medical Center Groningen. For each allergen 2 population threshold distributions were determined with the individual minimal eliciting dose and the preceding dose of each DBPCFC for objective symptoms and any symptom (either subjective or objective). Individual positive DBPCFCs were available for peanut (n = 135), cow's milk (n = 93), hen's egg (n = 53), hazelnut (n = 28), and cashew nut (n = 31). Fewer children were challenged with soy (n = 10) or walnut (n = 13). Threshold dose distributions showed a good statistical and visual fit. The protein dose at which 5% of the allergic population is likely to respond with objective reactions was 1.6 mg for peanut, 1.1 mg for cow's milk, 1.5 mg for hen's egg, 7.4 mg for cashew nut, and 0.29 mg for hazelnut. Thresholds for any symptom were on average 2 to 6 times lower than for objective symptoms. The 95% upper and lower confidence intervals of the threshold distributions were overlapping. The peanut threshold distribution on objective symptoms was similar to the distribution of another European center. Threshold distribution curves and eliciting doses are a powerful tool to compare different allergenic foods and for informing policy on precautionary labeling.


Oral prenylation inhibition with lonafarnib in chronic hepatitis D infection: a proof-of-concept randomised, double-blind, placebo-controlled phase 2A trial.
Therapies for chronic hepatitis delta virus (HDV) infection are unsatisfactory. Prenylation is essential for HDV and inhibition abrogates HDV production in experimental models. In a proof-of-concept study, we aimed to assess the effect on HDV RNA levels, safety, and tolerability of the prenylation inhibitor lonafarnib in patients with chronic delta hepatitis. In this phase 2A double-blind, randomised, placebo-controlled study, patients aged 18 years or older with chronic HDV infection were randomly assigned (3:1 in group 1 and 2:1 in group 2) to receive lonafarnib 100 mg (group 1) or lonafarnib 200 mg (group 2) twice daily for 28 days with 6 months' follow-up. Participants were randomised by random-number tables blocked in groups of four without stratification. Both groups enrolled six treatment participants and two placebo participants. Group 1 placebo patients received open-label lonafarnib as group 2 participants. The primary therapeutic endpoint was a decrease in HDV RNA viral titre in serum and the primary safety endpoint was the ability to tolerate the drug at the prescribed dose for the full 4-week duration, defined as drug discontinuation due to intolerance or grade 3/4 adverse events. This trial is registered with ClinicalTrials.gov, number NCT01495585. Between Jan 19, 2012, and April 28, 2014, 14 patients were enrolled, of whom eight were assigned to group 1 and six were assigned to group 2. At day 28, compared with placebo, mean log HDV RNA declines from baseline were -0·73 log IU/mL in group 1 (95% CI 0·17-1·31; p=0·03) and -1·54 log IU/mL in group 2 (1·21-1·93; p<0·0001). Lonafarnib serum concentrations correlated with HDV RNA change (r(2)=0·78, p<0·0001). Model fits show that hepatitis B surface antigen (HBsAg) remained stable after a short pharmacological delay (0·75 days [SE 0·24]), lonafarnib effectiveness in blocking HDV production was greater in group 2 than in group 1 (0·952 [SE 0·06] vs 0·739 [0·05], p<0·001), and the HDV half-life was 1·62 days (0·07). There was no evidence of virological resistance. Adverse events were mainly mild to moderate with group 1 patients experiencing diarrhoea in three patients (50%) and nausea in two patients (33%) and in group 2 with all patients (100%) experiencing nausea, diarrhoea, abdominal bloating, and weight loss greater than 2 kg (mean of 4 kg). No treatment discontinuations occurred in any treatment groups. Treatment of chronic HDV with lonafarnib significantly reduces virus levels. The decline in virus levels significantly correlated with serum drug levels, providing further evidence for the efficacy of prenylation inhibition in chronic HDV. National Institute of Diabetes and Digestive and Kidney Diseases and National Cancer Institute, National Institutes of Health, and Eiger Biopharmaceuticals Inc.


Real-time increased detection of neoplastic tissue in Barrett's esophagus with probe-based confocal laser endomicroscopy: final results of an international multicenter, prospective, randomized, controlled trial.
Probe-based confocal laser endomicroscopy (pCLE) allows real-time detection of neoplastic Barrett's esophagus (BE) tissue. However, the accuracy of pCLE in real time has not yet been extensively evaluated. To compare the sensitivity and specificity of pCLE in addition to high-definition white-light endoscopy (HD-WLE) with HD-WLE alone for the detection of high-grade dysplasia (HGD) and early carcinoma (EC) in BE. International, prospective, multicenter, randomized, controlled trial. Five tertiary referral centers. A total of 101 consecutive BE patients presenting for surveillance or endoscopic treatment of HGD/EC. All patients were examined by HD-WLE, narrow-band imaging (NBI), and pCLE, and the findings were recorded before biopsy samples were obtained. The order of HD-WLE and NBI was randomized and performed by 2 independent, blinded endoscopists. All suspicious lesions on HD-WLE or NBI and 4-quadrant random locations were documented. These locations were examined by pCLE, and a presumptive diagnosis of benign or neoplastic (HGD/EC) tissue was made in real time. Finally, biopsies were taken from all locations and were reviewed by a central pathologist, blinded to endoscopic and pCLE data. Diagnostic characteristics of pCLE. The sensitivity and specificity for HD-WLE were 34.2% and 92.7%, respectively, compared with 68.3% and 87.8%, respectively, for HD-WLE or pCLE (P = .002 and P < .001, respectively). The sensitivity and specificity for HD-WLE or NBI were 45.0% and 88.2%, respectively, compared with 75.8% and 84.2%, respectively, for HD-WLE, NBI, or pCLE (P = .01 and P = .02, respectively). Use of pCLE in conjunction with HD-WLE and NBI enabled the identification of 2 and 1 additional HGD/EC patients compared with HD-WLE and HD-WLE or NBI, respectively, resulting in detection of all HGD/EC patients, although not statistically significant. Academic centers with enriched population. pCLE combined with HD-WLE significantly improved the ability to detect neoplasia in BE patients compared with HD-WLE. This may allow better informed decisions to be made for the management and subsequent treatment of BE patients. ( NCT00795184.).


Quantitative Thermal Imaging Biomarkers to Detect Acute Skin Toxicity From Breast Radiation Therapy Using Supervised Machine Learning.
Radiation-induced dermatitis is a common side effect of breast radiation therapy (RT). Current methods to evaluate breast skin toxicity include clinical examination, visual inspection, and patient-reported symptoms. Physiological changes associated with radiation-induced dermatitis, such as inflammation, may also increase body-surface temperature, which can be detected by thermal imaging. Quantitative thermal imaging markers were identified and used in supervised machine learning to develop a predictive model for radiation dermatitis. Ninety patients treated for adjuvant whole-breast RT (4250 cGy/fx = 16) were recruited for the study. Thermal images of the treated breast were taken at 4 intervals: before RT, then weekly at fx = 5, fx = 10, and fx = 15. Parametric thermograms were analyzed and yielded 26 thermal-based features that included surface temperature (°C) and texture parameters obtained from (1) gray-level co-occurrence matrix, (2) gray-level run-length matrix, and (3) neighborhood gray-tone difference matrix. Skin toxicity was evaluated at the end of RT using the Common Terminology Criteria for Adverse Events (CTCAE) guidelines (Ver.5). Binary group classes were labeled according to a CTCAE cut-off score of ≥2, and thermal features obtained at fx = 5 were used for supervised machine learning to predict skin toxicity. The data set was partitioned for model training, independent testing, and validation. Fifteen patients (∼17% of the whole data set) were randomly selected as an unseen test data set, and 75 patients (∼83% of the whole data set) were used for training and validation of the model. A random forest classifier with leave-1-patient-out cross-validation was employed for modeling single and hybrid parameters. The model performance was reported using receiver operating characteristic analysis on patients from an independent test set. Thirty-seven patients presented with adverse skin effects, denoted by a CTCAE score ≥2, and had significantly higher local increases in skin temperature, reaching 36.06°C at fx = 10 (P = .029). However, machine-learning models demonstrated early thermal signals associated with skin toxicity after the fifth RT fraction. The cross-validated model showed high prediction accuracy on the independent test data (test accuracy = 0.87) at fx = 5 for predicting skin toxicity at the end of RT. Early thermal markers after 5 fractions of RT are predictive of radiation-induced skin toxicity in breast RT.


Relationship of prevalence of non-insulin-dependent diabetes mellitus to Amerindian admixture in the Mexican Americans of San Antonio, Texas.
A genetic and epidemiological survey of non-insulin-dependent diabetes mellitus (NIDDM) was conducted among the Mexican Americans residing in three socioeconomically distinct areas of San Antonio, Texas: a low socioeconomic (SES) traditional area (barrio), a middle SES, ethnically balanced area (transitional), and a high SES, predominantly Anglo area (suburb). Seventeen polymorphic markers were used to relate the prevalences of NIDDM with the extent of Amerindian ancestry of 1,237 Mexican Americans of these three residential areas. While only the RH and haptoglobin loci showed evidence of association with NIDDM, an admixture analysis of the combined allele frequency data revealed a pattern of decreasing NIDDM prevalence with increasing socioeconomic status (as approximated by neighborhood of residence) and a parallel decrease in Amerindian ancestry. The rank-order correlation between NIDDM prevalence and Amerindian admixture is 0.943 (P less than .001) for the crude prevalence rate and 0.829 (P less than .02) for the age-adjusted rate. Nested gene diversity analysis revealed that the heterogeneity of allele frequencies is more pronounced when individuals were classified by their NIDDM disease status as compared to the classification by neighborhood. Estimation of Amerindian ancestry of each individual did not reveal any significant change in the shape of the distributions of individual admixture proportions in diabetics as compared to the controls. Nevertheless, the results suggest that genetic factors partially explain the differences in NIDDM prevalence observed between the Mexican American and Anglo populations in the southwestern United States.


Pretreatment elevated fibrinogen level predicts worse oncologic outcomes in upper tract urothelial carcinoma.
This study aimed to further validate the prognostic role of fibrinogen in upper tract urothelial carcinoma (UTUC) in a large Chinese cohort. A total of 703 patients who underwent radical nephroureterectomy were retrospectively identified. Fibrinogen levels of ≥4.025 g l-1 were defined as elevated. Logistic regression analysis was performed to determine the association between fibrinogen and adverse pathological features. Kaplan-Meier analysis and Cox regression models were used to assess the associations of fibrinogen with cancer-specific survival (CSS), disease recurrence-free survival (RFS), and overall survival (OS). Harrell c-index and decision curve analysis were used to assess the clinical utility of multivariate models. The median follow-up duration was 42 (range: 1-168) months. Logistic regression analysis revealed that elevated fibrinogen was associated with higher tumor stage and grade, lymph node involvement, lymphovascular invasion, sessile carcinoma, concomitant variant histology, and positive surgical margins (all P < 0.05). Multivariate Cox regression analysis demonstrated that elevated fibrinogen was independently associated with decreased CSS (hazard ratio [HR]: 2.33; P < 0.001), RFS (HR: 2.09; P < 0.001), and OS (HR: 2.09; P < 0.001). The predictive accuracies of the multivariate models were improved by 3.2%, 2.0%, and 2.8% for CSS, RFS, and OS, respectively, when fibrinogen was added. Decision curve analysis showed an added benefit for CSS prediction when fibrinogen was added to the model. Preoperative fibrinogen may be a strong independent predictor of worse oncologic outcomes in UTUC; therefore, it may be valuable to apply this marker to the current risk stratification in UTUC.


Abnormal brain response to cholinergic challenge in chronic encephalopathy from the 1991 Gulf War.
Several case definitions of chronic illness in veterans of the 1991 Persian Gulf War have been linked epidemiologically with environmental exposure to cholinesterase-inhibiting chemicals, which cause chronic changes in cholinergic receptors in animal models. Twenty-one chronically ill Gulf War veterans (5 with symptom complex 1, 11 with complex 2, and 5 with complex 3) and 17 age-, sex- and education-matched controls, underwent an 99mTc-HMPAO-SPECT brain scan following infusion of saline and >48 h later a second scan following infusion of physostigmine in saline. From each SPECT image mean normalized regional cerebral blood flow (nrCBF) from 39 small blocks of correlated voxels were extracted with geostatistical spatial modeling from eight deep gray matter structures in each hemisphere. Baseline nrCBF in symptom complex 2 was lower than controls throughout deep structures. The change in nrCBF after physostigmine (challenge minus baseline) was negative in complexes 1 and 3 and controls but positive in complex 2 in some structures. Since effects were opposite in different groups, no finding typified the entire patient sample. A hold-out discriminant model of nrCBF from 17 deep brain blocks predicted membership in the clinical groups with sensitivity of 0.95 and specificity of 0.82. Gulf War-associated chronic encephalopathy in a subset of veterans may be due to neuronal dysfunction, including abnormal cholinergic response, in deep brain structures.


Antipsychotic Medication Treatment Patterns in Adult Depression.
To characterize the role of antipsychotic medications in the community treatment of adult depression. We identified adults (aged 18-64 years) with new episodes of depression treatment (ICD-9-CM 296.2, 296.3, 300.4, or 311) in US national Medicaid data (2001-2010). Patients with alternative ICD-9-CM antipsychotic indications, such as schizophrenia or bipolar disorder, were excluded. Each patient was followed for at least 1 year to characterize antipsychotic and antidepressant treatment and emerging alternative antipsychotic indications. For patients without alternative indications through day 45 following start of antipsychotic treatment, antipsychotics were considered to be intended for treatment of depression. Among this group, we determined whether antipsychotic initiation was preceded by minimally adequate treatment with antidepressants, defined as active antidepressant treatment for ≥ 31 days prior to and including the day of antipsychotic initiation. Within 1 year following onset, 14.0% of patients started an antipsychotic medication. A total of 41.3% of antipsychotic initiators developed an antipsychotic indication other than depression through day 45 following antipsychotic initiation, most often bipolar disorder or depression with psychotic features. The remaining 58.7% of antipsychotic initiators presumably started antipsychotics for nonpsychotic depression. Of these, 71.3% did not have minimally adequate antidepressant treatment prior to starting the antipsychotic medication. Antipsychotic medications are used in approximately 1 in 7 patients with a new episode of depression. For 1 in 12 patients, the antipsychotic was considered to be intended for nonpsychotic depression. Almost three-quarters of these patients did not receive minimally adequate treatment with antidepressants prior to antipsychotic initiation. This pattern suggests potentially inappropriate and premature initiation of a drug class with substantial adverse effects and medical risks.


Why does the Secchi disk disappear? An imaging perspective.
The widely-used Secchi disk method is re-examined from the modulation transfer aspect. Namely, by assuming a volume scattering function and applying small angle scattering approximation, we show that the Secchi depth and horizontal visibility can be determined using the water modulation transfer function and the corresponding spatial frequencies associated with the disk. A basic equation of Secchi disk is reached that is comparable to the radiative transfer approach, in that the Secchi depth is inversely proportional to the attenuation coefficient (c). With typical values for parameters applied, we demonstrate that the modulation transfer technique produces a horizontal visibility range of about 4.8/c, which is inline with previous studies. The improvement lies in the fact that the current approach correctly addresses the response of all spatial frequencies according to the modeled optical transfer function of the water. In terms of Secchi disk theory, the current approach helps to understand the effect of disk size as well as the role of scattering on the Secchi disk depth. The approach presented provides an understanding of Secchi disk disappearance by showing that as the disk is moved away from the observer, the spatial frequencies corresponding to the disk size increase, while the modulation transfer dampens contrast at an increased rate.


Awareness and prevention of osteoporosis among South Asian women.
We examined awareness of osteoporosis prevention among peri- and post-menopausal South Asian women attending two community centers in the Dallas/Fort-Worth Metroplex. We conducted a quasi-experimental study (final N = 61) assessing knowledge about osteoporosis among South Asian women (>or=40 years). The mean age was 52.3 years (SD = 8.72). Over 50% were college educated and 64% had no health insurance. We administered a baseline knowledge test, followed by a health education intervention and, 2 weeks later, by a post-test. Participants received one point for each correct answer and scores were added (<or=14). Participants showed a significant increase in osteoporosis knowledge post intervention (paired t (60) = -9.5, P < .01). For example, women reported highest knowledge gains on the following: adequate calcium intake is achievable from two glasses of milk a day; very thin women are at risk for developing osteoporosis, and family history of osteoporosis is a risk factor. Intervention completers were better prepared to prevent and manage osteoporosis. Results indicate the efficacy of educational intervention in improving osteoporosis awareness; and point to the potential for knowledge acquisition aimed at developing community-based prevention strategies at the community level.


Effects of chronic acetazolamide administration on gas exchange and acid-base control in pulmonary circulation in exercising horses.
Carbonic anhydrase (CA) catalyses the hydration/dehydration reaction of CO(2) and increases the rate of Cl(-) and HCO(3)(-) exchange between the erythrocytes and plasma. Therefore, chronic inhibition of CA has a potential to attenuate CO(2) output and induce greater metabolic and respiratory acidosis in exercising horses. To determine the effects of Carbonic anhydrase inhibition on CO(2) output and ionic exchange between erythrocytes and plasma and their influence on acid-base balance in the pulmonary circulation (across the lung) in exercising horses with and without CA inhibition. Six horses were exercised to exhaustion on a treadmill without (Con) and with CA inhibition (AczTr). CA inhibition was achieved with administration of acetazolamide (10 mg/kg bwt t.i.d. for 3 days and 30 mg/kg bwt before exercise). Arterial, mixed venous blood and CO(2) output were sampled at rest and during exercise. An integrated physicochemical systems approach was used to describe acid base changes. AczTr decreased the duration of exercise by 45% (P < 0.0001). During the transition from rest to exercise CO(2) output was lower in AczTr (P < 0.0001). Arterial PCO(2) (P < 0.0001; mean ± s.e. 71 ± 2 mmHg AczTr, 46 ± 2 mmHg Con) was higher, whereas hydrogen ion (P = 0.01; 12.8 ± 0.6 nEq/l AczTr, 15.5 ± 0.6 nEq/l Con) and bicarbonate (P = 0.007; 5.5 ± 0.7 mEq/l AczTr, 10.1 ± 1.3 mEq/l Con) differences across the lung were lower in AczTr compared to Con. No difference was observed in weak electrolytes across the lung. Strong ion difference across the lung was lower in AczTr (P = 0.0003; 4.9 ± 0.8 mEq AczTr, 7.5 ± 1.2 mEq Con), which was affected by strong ion changes across the lung with exception of lactate. CO(2) and chloride changes in erythrocytes across the lung seem to be the major contributors to acid-base and ions balance in pulmonary circulation in exercising horses.


A comparison of the predictive accuracy of three screening models for pulmonary arterial hypertension in systemic sclerosis.
There is evidence that early screening for pulmonary arterial hypertension (PAH) in systemic sclerosis (SSc) improves outcomes. We compared the predictive accuracy of two recently published screening algorithms (DETECT 2013 and Australian Scleroderma Interest Group (ASIG) 2012) for SSc-associated PAH (SSc-PAH) with the commonly used European Society of Cardiology/European Respiratory Society (ESC/ERS 2009) guidelines. We included 73 consecutive SSc patients with suspected PAH undergoing right heart catheterization (RHC). The three screening models were applied to each patient. For each model, contingency table analysis was used to determine sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values for PAH. These properties were also evaluated in an 'alternate scenario analysis' in which the prevalence of PAH was set at 10%. RHC revealed PAH in 27 (36.9%) patients. DETECT and ASIG algorithms performed equally in predicting PAH with sensitivity and NPV of 100%. The ESC/ERS guidelines had sensitivity of 96.3% and NPV of only 91%, missing one case of PAH; these guidelines could not be applied to three patients who had absent tricuspid regurgitant (TR) jet. The ASIG algorithm had the highest specificity (54.5%). With PAH prevalence set at 10%, the NPV of the models was unchanged, but the PPV dropped to less than 20%. In this cohort, the DETECT and ASIG algorithms out-perform the ESC/ERS guidelines, detecting all patients with PAH. The ESC/ERS guidelines have limitations in the absence of a TR jet. Ultimately, the choice of SSc-PAH screening algorithm will also depend on cost and ease of application.


Electrostatic screening and charge correlation effects in micellization of ionic surfactants.
We have used atomistic simulations to study the role of electrostatic screening and charge correlation effects in self-assembly processes of ionic surfactants into micelles. Specifically, we employed grand canonical Monte Carlo simulations to investigate the critical micelle concentration (cmc), aggregation number, and micellar shape in the presence of explicit sodium chloride (NaCl). The two systems investigated are cationic dodecyltrimethylammonium chloride (DTAC) and anionic sodium dodecyl sulfate (SDS) surfactants. Our explicit-salt results, obtained from a previously developed potential model with no further adjustment of its parameters, are in good agreement with experimental data for structural and thermodynamic micellar properties. We illustrate the importance of ion correlation effects by comparing these results with a Yukawa-type surfactant model that incorporates electrostatic screening implicitly. While the effect of salt on the cmc is well-reproduced even with the implicit Yukawa model, the aggregate size predictions deviate significantly from experimental observations at low salt concentrations. We attribute this discrepancy to the neglect of ion correlations in the implicit-salt model. At higher salt concentrations, we find reasonable agreement of the Yukawa model with experimental data. The crossover from low to high salt concentrations is reached when the electrostatic screening length becomes comparable to the headgroup size.


Arsenic response of AtPCS1- and CePCS-expressing plants - effects of external As(V) concentration on As-accumulation pattern and NPT metabolism.
Phytochelatins (PCs) are small, cysteine-rich peptides, known to play a major role in detoxification of both cadmium and arsenic. The aim of this study was to determine whether overexpression of either of two PC synthase (PCS) genes, AtPCS1 and CePCS in Nicotiana tabacum (previously shown to cause decrease and increase, respectively, of cadmium tolerance of tobacco - Wojas et al., 2008) also contributes to such contrasting phenotypes with respect to arsenic (As) tolerance and accumulation, and how observed responses relate to non-protein thiol (NPT) metabolism. The expression of both genes resulted in an increase of As-tolerance, with CePCS plants most tolerant. We showed for the first time that the response of PCS overexpressing plants to As qualitatively depends on the external As(V) concentration. At the less toxic 50muM As(V), AtPCS1 and CePCS transformants accumulated more As in roots and leaves than WT. An increase in PC production and the level of PC2 species was detected in leaves of AtPCS1 and CePCS plants, which might explain their enhanced As-accumulation and tolerance. In contrast, at the highly toxic 200muM As(V), several disturbances in thiol metabolism of PCS overexpressing plants were found, surprisingly, including decrease of PC levels both in roots and leaves of transgenic plants relative to WT. The increase in As-tolerance and accumulation due to AtPCS1 and CePCS overexpression, observed at the As(V) concentrations similar to those found in As-contaminated soils, makes these genes promising candidates for plant engineering for phytoremediation.


Droplet-based microfluidics at the femtolitre scale.
We have built a toolbox of modules for droplet-based microfluidic operations on femtolitre volume droplets. We have demonstrated monodisperse production, sorting, coalescence, splitting, mixing, off-chip incubation and re-injection at high frequencies (up to 3 kHz). We describe the constraints and limitations under which satisfactory performances are obtained, and discuss the physics that controls each operation. For some operations, such as internal mixing, we obtained outstanding performances: for instance, in 75 fL droplets the mixing time was 45 μs, 35-fold faster than previously reported for a droplet microreactor. In practice, in all cases, a level of control comparable to nanolitre or picolitre droplet manipulation was obtained despite the 3 to 6 order of magnitude reduction in droplet volume. Remarkably, all the operations were performed using devices made using standard soft-lithography techniques and PDMS rapid prototyping. We show that femtolitre droplets can be used as microreactors for molecular biology with volumes one billion times smaller than conventional microtitre plate wells: in particular, the Polymerase Chain Reaction (PCR) was shown to work efficiently in 20 fL droplets.


Psychophysics with children: Investigating the effects of attentional lapses on threshold estimates.
When assessing the perceptual abilities of children, researchers tend to use psychophysical techniques designed for use with adults. However, children's poorer attentiveness might bias the threshold estimates obtained by these methods. Here, we obtained speed discrimination threshold estimates in 6- to 7-year-old children in UK Key Stage 1 (KS1), 7- to 9-year-old children in Key Stage 2 (KS2), and adults using three psychophysical procedures: QUEST, a 1-up 2-down Levitt staircase, and Method of Constant Stimuli (MCS). We estimated inattentiveness using responses to "easy" catch trials. As expected, children had higher threshold estimates and made more errors on catch trials than adults. Lower threshold estimates were obtained from psychometric functions fit to the data in the QUEST condition than the MCS and Levitt staircases, and the threshold estimates obtained when fitting a psychometric function to the QUEST data were also lower than when using the QUEST mode. This suggests that threshold estimates cannot be compared directly across methods. Differences between the procedures did not vary significantly with age group. Simulations indicated that inattentiveness biased threshold estimates particularly when threshold estimates were computed as the QUEST mode or the average of staircase reversals. In contrast, thresholds estimated by post-hoc psychometric function fitting were less biased by attentional lapses. Our results suggest that some psychophysical methods are more robust to attentiveness, which has important implications for assessing the perception of children and clinical groups.


Total aberrations compensation in digital holographic microscopy with a reference conjugated hologram.
In this paper we present a new method to achieve quantitative phase contrast imaging in Digital Holographic Microscopy (DHM) that allows to compensate for phase aberrations and image distortion by recording of a single reference hologram.We demonstrate that in particular cases in which the studied specimen does not have abrupt edges, the specimen's hologram itself can be used as reference hologram. We show that image distortion and phase aberrations introduced by a lens ball used as microscope objective are completely suppressed with our method. Finally the concept of self-conjugated reference hologram is applied on a biological sample (Trypanosoma Brucei) to maintain a spatial phase noise level under 3 degrees.


The interaction between systemic inflammation and psychosocial stress in the association with cardiac troponin elevation: A new approach to risk assessment and disease prevention.
We have previously shown that there is a complex and dynamic biological interaction between acute mental stress and acute release of inflammatory factors into the blood stream in relation to heart disease. We now hypothesize that the presence of chronic psychosocial stress may modify the weight of single test results for inflammation as a predictor of heart disease. Using a cross-sectional design, 500 participants free from heart disease drawn from the Whitehall II study in UK in 2006-2008 were tested for plasma fibrinogen as an inflammatory factor, financial strain as a marker of chronic psychosocial stress, coronary calcification measured using computed tomography, and for plasma high-sensitivity cardiac troponin T (HS-CTnT) as a marker of cardiac risk. Fibrinogen concentration levels above the average were associated with a 5-fold increase in the odds of HS-CTnT positivity only among individuals with financial strain (N=208, OR=4.73, 95%CI=1.67 to 13.40, P=0.003). Fibrinogen was in fact not associated with HS-CTnT positivity in people without financial strain despite the larger size of that subsample (n=292, OR=0.84, 95%CI=0.42 to 1.67, P=0.622). A test for interaction on the full sample (N=500) showed a P value of 0.010 after adjusting for a range of demographics, health behaviours, traditional cardiovascular risk factors, psychosocial stressors, inflammatory cytokines, and coronary calcification. In conclusion, elevated fibrinogen seems to be cardio-toxic only when is combined with financial strain. Chronic psychosocial stress may modify the meaning that we should give to single test results for inflammation. Further research is needed to confirm our results.


Increased Body Weight and Fat Mass After Subchronic GIP Receptor Antagonist, but Not GLP-2 Receptor Antagonist, Administration in Rats.
Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-2 (GLP-2) are hormones secreted from the enteroendocrine cells after a meal. They exert their actions through activation of G protein-coupled receptors (R), the GIPR and GLP-2R, respectively. Both have been reported to influence metabolism. The purpose of the study was to investigate the role of the hormones in the regulation of lipid and bone homeostasis by subchronic treatment with novel GIPR and GLP-2R antagonists. Rats were injected once daily with vehicle, GIPR, or GLP-2R antagonists for 3 weeks. Body weight, food intake, body composition, plasma lipoprotein lipase (LPL), adipokines, triglycerides and the marker of bone resorption carboxy-terminal collagen crosslinks (CTX), were examined. In rats, subchronic treatment with GIPR antagonist, rat GIP (3-30)NH2, did not modify food intake and bone resorption, but significantly increased body weight, body fat mass, triglycerides, LPL, and leptin levels compared with vehicle treated rats. Subchronic (Pro3)GIP (a partial GIPR agonist), GLP-2(11-33), and GLP-2(3-33) (GLP-2R antagonists) treatment did not affect any parameter. The present results would be consistent with a role for GIP, but not GLP-2, in the maintenance of lipid homeostasis in rats, while neither GIPR nor GLP-2R antagonism appeared to influence bone resorption in rats.


Starvation-induced activation of ATM/Chk2/p53 signaling sensitizes cancer cells to cisplatin.
Optimizing the safety and efficacy of standard chemotherapeutic agents such as cisplatin (CDDP) is of clinical relevance. Serum starvation in vitro and short-term food starvation in vivo both stress cells by the sudden depletion of paracrine growth stimulation. The effects of serum starvation on CDDP toxicity were investigated in normal and cancer cells by assessing proliferation, cell cycle distribution and activation of DNA-damage response and of AMPK, and were compared to effects observed in cells grown in serum-containing medium. The effects of short-term food starvation on CDDP chemotherapy were assessed in xenografts-bearing mice and were compared to effects on tumor growth and/or regression determined in mice with no diet alteration. We observed that serum starvation in vitro sensitizes cancer cells to CDDP while protecting normal cells. In detail, in normal cells, serum starvation resulted in a complete arrest of cellular proliferation, i.e. depletion of BrdU-incorporation during S-phase and accumulation of the cells in the G0/G1-phase of the cell cycle. Further analysis revealed that proliferation arrest in normal cells is due to p53/p21 activation, which is AMPK-dependent and ATM-independent. In cancer cells, serum starvation also decreased the fraction of S-phase cells but to a minor extent. In contrast to normal cells, serum starvation-induced p53 activation in cancer cells is both AMPK- and ATM-dependent. Combination of CDDP with serum starvation in vitro increased the activation of ATM/Chk2/p53 signaling pathway compared to either treatment alone resulting in an enhanced sensitization of cancer cells to CDDP. Finally, short-term food starvation dramatically increased the sensitivity of human tumor xenografts to cisplatin as indicated not only by a significant growth delay, but also by the induction of complete remission in 60% of the animals bearing mesothelioma xenografts, and in 40% of the animals with lung carcinoma xenografts. In normal cells, serum starvation in vitro induces a cell cycle arrest and protects from CDDP induced toxicity. In contrast, proliferation of cancer cells is only moderately reduced by serum starvation whereas CDDP toxicity is enhanced. The combination of CDDP treatment with short term food starvation improved outcome in vivo. Therefore, starvation has the potential to enhance the therapeutic index of cisplatin-based therapy.


Local-scale genetic structure in the peatmoss Sphagnum fuscum.
Sphagnum (peatmoss) dominates huge areas of the Northern Hemisphere and acts as a significant carbon sink on a global scale, yet little is known about the genetic structure of Sphagnum populations. We investigated genetic structure within a population of the common peatmoss Sphagnum fuscum, to assess local patterns of genetic diversity and the spatial extent of clones. One hundred seventeen shoots were sampled from five transects in Fuglmyra, central Norway, and sequenced for three anonymous DNA regions. Five neighbourhood patches were marked along each transect, and from each patch, five stems were sampled for molecular analyses. Seventeen haplotypes could be distinguished and two major groups of haplotypes differed by 12 mutational steps. The two major haplotype groups differed significantly in microhabitat association along the distance to groundwater table and the pH gradients, indicating microhabitat differentiation. The haplotypes within these groups were all genetically similar, differing by one or two mutations. The most common haplotype occurred in four transects separated by 250-m distance. Most of the molecular variation in the population was found among transects, and within patches. Large dominating clones within each transect resulted in low variation explained by the among-patch-within-transect component of spatial structure. Mutation appears to account for a larger proportion of the population variation than recombination. Within the population, vegetative growth and asexual reproduction from gametophyte fragments dominate as the main reproductive mode.


Control of T helper cell differentiation--in search of master genes.
Naïve T helper (T(H)0) cells can differentiate into one of two distinct populations: T(H)1 and T(H)2. Each population is characterized by the expression of specific cytokines and their ability to participate in cell-mediated or humoral immune responses. Recent efforts at identifying the molecular mechanisms through which T(H)0 cells become T(H)1 or T(H)2 cells have been promising. A number of transcription factors, including GATA-3 and T-bet, have been identified that promote the differentiation of T(H)0 cells and the maintenance of the differentiated cell phenotype. Dong and Flavell review recent findings on proteins that control the fate of T(H)0 differentiation, whether by promotion or inhibition, and discuss the role of epigenesis in the differentiation process.


Evidenced-based practice: priorities and implementation strategies.
Evidenced-based practice (EBP) should be a driving force behind establishing optimal clinical practices. Recently, clinicians and hospitals have started efforts to introduce key EBP. These efforts hold the potential to improve patient outcomes and reduce costs. However, many practices need updating with EBP. Which ones should be chosen? While practices often vary in terms of importance for each hospital, some changes in practice are likely to have a better return on the investment (ROI). Two key areas affecting most hospitals are practices associated with high costs and increased mortality. In critical care areas, these two key areas often involve addressing outlier management and severe sepsis. In addition, the recognition of the need for the change is only one step in ensuring EBP. To ensure EBP is implemented, clinical leaders who will ensure that the new practice standards are being utilized are necessary. Fortunately, many hospitals have strong leaders. The advanced practice nurse (APN) is one such leader. The APN is often in a unique position to help recognize, prioritize, and implement EBP into the hospitals culture. This article illustrates steps in making EBP a reality by highlighting the management of outliers and severe sepsis and the implementation strategies for these conditions.


Cytotoxic effects of air freshener biocides in lung epithelial cells.
This study evaluated the cytotoxicity of mixtures of citral (CTR) and either benzisothiazolinone (BIT, Mix-CTR-BIT) or triclosan (TCS, Mix-CTR-TCS) in human A549 lung epithelial cells. We investigated the effects of various mix ratios of these common air freshener ingredients on cell viability, cell proliferation, reactive oxygen species (ROS) generation, and DNA damage. Mix-CTR-BIT and Mix-CTR-TCS significantly decreased the viability of lung epithelial cells and inhibited cell growth in a dose-dependent manner. In addition, both mixtures increased ROS generation, compared to that observed in control cells. In particular, cell viability, growth, and morphology were affected upon increase in the proportion of BIT or TCS in the mixture. However, comet analysis showed that treatment of cells with Mix-CTR-BIT or Mix-CTR-TCS did not increase DNA damage. Taken together, these data suggested that increasing the content of biocides in air fresheners might induce cytotoxicity, and that screening these compounds using lung epithelial cells may contribute to hazard assessment.


Street choice logit model for visitors in shopping districts.
In this study, we propose two models for predicting people's activity. The first model is the pedestrian distribution prediction (or postdiction) model by multiple regression analysis using space syntax indices of urban fabric and people distribution data obtained from a field survey. The second model is a street choice model for visitors using multinomial logit model. We performed a questionnaire survey on the field to investigate the strolling routes of 46 visitors and obtained a total of 1211 street choices in their routes. We proposed a utility function, sum of weighted space syntax indices, and other indices, and estimated the parameters for weights on the basis of maximum likelihood. These models consider both street networks, distance from destination, direction of the street choice and other spatial compositions (numbers of pedestrians, cars, shops, and elevation). The first model explains the characteristics of the street where many people tend to walk or stay. The second model explains the mechanism underlying the street choice of visitors and clarifies the differences in the weights of street choice parameters among the various attributes, such as gender, existence of destinations, number of people, etc. For all the attributes considered, the influences of DISTANCE and DIRECTION are strong. On the other hand, the influences of Int.V, SHOPS, CARS, ELEVATION, and WIDTH are different for each attribute. People with defined destinations tend to choose streets that "have more shops, and are wider and lower". In contrast, people with undefined destinations tend to choose streets of high Int.V. The choice of males is affected by Int.V, SHOPS, WIDTH (positive) and CARS (negative). Females prefer streets that have many shops, and couples tend to choose downhill streets. The behavior of individual persons is affected by all variables. The behavior of people visiting in groups is affected by SHOP and WIDTH (positive).


Body cues, not facial expressions, discriminate between intense positive and negative emotions.
The distinction between positive and negative emotions is fundamental in emotion models. Intriguingly, neurobiological work suggests shared mechanisms across positive and negative emotions. We tested whether similar overlap occurs in real-life facial expressions. During peak intensities of emotion, positive and negative situations were successfully discriminated from isolated bodies but not faces. Nevertheless, viewers perceived illusory positivity or negativity in the nondiagnostic faces when seen with bodies. To reveal the underlying mechanisms, we created compounds of intense negative faces combined with positive bodies, and vice versa. Perceived affect and mimicry of the faces shifted systematically as a function of their contextual body emotion. These findings challenge standard models of emotion expression and highlight the role of the body in expressing and perceiving emotions.


Evidence for apoptosis of murine macrophages by Actinobacillus actinomycetemcomitans infection.
The gram-negative bacterium Actinobacillus actinomycetemcomitans is considered an important etiological agent in periodontal diseases. In this study, we show that A. actinomycetemcomitans strains are cytotoxic for the murine macrophage cell line J774.1. On the other hand, Porphyromonas gingivalis strains, other gram-negative oral species implicated in adult periodontitis, showed weak cytotoxic effects. For this to occur, A. actinomycetemcomitans had to gain entry into the macrophages, since cytotoxicity was prevented by cytochalasin D. We demonstrate that cell death induced by A. actinomycetemcomitans Y4 occurs through apoptosis, as shown by changes in nuclear morphology, an increase in the proportion of fragmented DNA, and the typical ladder pattern of DNA fragmentation indicative of apoptosis. We further sought to determine whether the cytotoxicity induced by A. actinomycetemcomitans Y4 could be modulated by the protein kinase inhibitors H7 and HA1004. Apoptotic cell death induced by A. actinomycetemcomitans Y4 was suppressed by H7 but was relatively unaffected by HA1004. These findings suggest that the signals of protein kinases may regulate apoptosis induced by A. actinomycetemcomitans Y4. The ability of A. actinomycetemcomitans to promote the apoptosis of macrophages may be important for the initiation of infection and the development of periodontal diseases.


Adipocyte iron regulates leptin and food intake.
Dietary iron supplementation is associated with increased appetite. Here, we investigated the effect of iron on the hormone leptin, which regulates food intake and energy homeostasis. Serum ferritin was negatively associated with serum leptin in a cohort of patients with metabolic syndrome. Moreover, the same inverse correlation was observed in mice fed a high-iron diet. Adipocyte-specific loss of the iron exporter ferroportin resulted in iron loading and decreased leptin, while decreased levels of hepcidin in a murine hereditary hemochromatosis (HH) model increased adipocyte ferroportin expression, decreased adipocyte iron, and increased leptin. Treatment of 3T3-L1 adipocytes with iron decreased leptin mRNA in a dose-dependent manner. We found that iron negatively regulates leptin transcription via cAMP-responsive element binding protein activation (CREB activation) and identified 2 potential CREB-binding sites in the mouse leptin promoter region. Mutation of both sites completely blocked the effect of iron on promoter activity. ChIP analysis revealed that binding of phosphorylated CREB is enriched at these two sites in iron-treated 3T3-L1 adipocytes compared with untreated cells. Consistent with the changes in leptin, dietary iron content was also directly related to food intake, independently of weight. These findings indicate that levels of dietary iron play an important role in regulation of appetite and metabolism through CREB-dependent modulation of leptin expression.


All fiber-based Yb-doped high energy, high power femtosecond fiber lasers.
Two all fiber-based laser systems are demonstrated to achieve high energy and high average power femtosecond pulsed outputs at wavelength of 1 µm. In the high energy laser system, a pulse energy of 1.05 mJ (0.85 mJ after pulse compressor) at 100 kHz repetition rate has been realized by a Yb-doped ultra large-core single-mode photonic crystal fiber (PCF) rod amplifier, seeded with a 50 µJ fiber laser. The pulse duration is 705 fs. In the high average power experiment, a large mode area (LMA) fiber has been used in the final stage amplifier, seeded with a 50 W mode locked fiber laser. The system is running at a repetition rate of 69 MHz producing 1052 W of average power before compressor. After pulse compression, a pulse duration of 800 fs was measured.


Detoxification of Neuromodulation-Eligible Patients by a Standardized Protocol: A Retrospective Pilot Study.
Patients eligible for spinal cord stimulation (SCS) generally experience excruciating pain, requiring more opioid consumption, which is usually an indication for SCS implantation. After final implantation, SCS has the ability to stabilize or decrease opioid usage in half of the patients. In this study, opioids were actively eliminated prior to implantation of any neuromodulation device with a standardized detoxification protocol. This pilot study aims to explore the feasibility, effectiveness, and safety of this opioid detoxification protocol prior to neuromodulation techniques. In this retrospective pilot study, 70 patients who were taking opioids and who were eligible for neuromodulation techniques underwent the detoxification program. A combined in- and out-patient clinic protocol was applied, whereby clonidine was the main component of both parts of the program. A multidisciplinary team with pain physicians and psychologists was responsible for performing this detoxification program. Safety and feasibility were systematically recorded during the hospitalization. No serious safety issues were reported. At the start of the program, patients reported a mild sedative effect of clonidine. Additionally, most patients presented mild symptoms of opioid withdrawal, which were partially countered by the sedative effect of clonidine. Both patients and the medical staff found this protocol feasible in clinical practice. Concerning the effectiveness, a statistically significant decrease in median morphine milligram equivalents (MMEs) was found with an MME of 175 (Q1-Q3: 118.1-240) at baseline, and at the last available follow-up visit the MME was 0 (Q1-Q3: 0-16.88). This standardized detoxification program has proven its effectiveness, safety, and feasibility in this single-center experience pilot study in patients eligible for neuromodulation techniques.


Determinants of government HIV/AIDS financing: a 10-year trend analysis from 125 low- and middle-income countries.
Trends and predictors of domestic spending from public sources provide national authorities and international donors with a better understanding of the HIV financing architecture, the fulfillment of governments' commitments and potential for long-term sustainability. We analyzed government financing of HIV using evidence from country reports on domestic spending. Panel data from 2000 to 2010 included information from 647 country-years amongst 125 countries. A random-effects model was used to analyze ten year trends and identify independent predictors of public HIV spending. Low- and middle-income countries spent US$ 2.1 billion from government sources in 2000, growing to US$ 6.6 billion in 2010, a three-fold increase. Per capita spending in 2010 ranged from 5 cents in low-level HIV epidemics in the Middle East to US$ 32 in upper-middle income countries with generalized HIV epidemics in Southern Africa. The average domestic public spending per capita was US$ 2.55. The analysis found that GDP per capita and HIV prevalence are positively associated with increasing levels of HIV-spending from public sources; a 10 percent increase in HIV prevalence is associated with a 2.5 percent increase in domestic funding for HIV. Additionally, a 10 percent increase in GDP per capita is associated with an 11.49 percent increase in public spending for HIV and these associations were highly significant. Domestic resources in low- and middle-income countries showed a threefold increase between 2000 and 2010 and currently support 50 percent of the global response with 41 percent coming from sub-Saharan Africa. Domestic spending in LMICs was associated with increased economic growth and an increased burden of HIV. Sustained increases in funding for HIV from public sources were observed in all regions and emphasize the increasing importance of government financing.


The Multiple-Slit Technique (MUST) for Penile Length and Girth Restoration.
Patients with severe erectile dysfunction (ED) and penile size issues, especially seen in Peyronie's disease (PD), are candidates for more invasive penile prosthesis insertion techniques that aim for penile length and girth reconstruction. To present the feasibility and safety of penile length and girth restoration based on the so-called multiple-slit technique (MUST) for patients with severe ED and significant penile shortening with or without PD. From July 2013 through January 2016, 138 patients underwent the MUST. The International Index of Erectile Function (IIEF) and the Erectile Dysfunction Inventory of Treatment Satisfaction were completed. Outcome analysis was focused on penile length restoration, penile curvature correction, intra- and postoperative complications, and patient satisfaction. 138 patients underwent the procedure (103 malleable and 35 inflatable devices). Etiologies of penile shortening and narrowing were PD, severe ED, post-radical prostatectomy, and androgen-deprivation therapy with or without brachytherapy or external radiotherapy for prostate cancer, and post-penile fracture in 60.1%, 24.6%, 10.1%, 3.6%, and 2.2%, respectively. In PD cases, the mean deviation of the penile axis was 55° (range = 0-90°). Mean subjective penile length loss reported was 3.2 cm (range = 1-5 cm), and shaft constriction was present in 44.9%. Median follow-up was 15.2 months (range = 6-36 months). Mean penile length gain was 3.1 cm (range = 2-5 cm). No penile prosthesis infection caused device explantation. One glans necrosis was encountered. The average IIEF score increased from 22 points at baseline to 66 points at 6-month follow-up. The MUST helps address penile size issues in cases of severe ED with concomitant conditions that impair penile length or girth. The strength of the study is its applicability to provide surgeons with a solution for cases in which patients have severe ED and penile size impairment owing to underlying conditions such as PD. The study is limited by the relatively short follow-up. The MUST is an effective, safe, and viable treatment option for a selected patient cohort. Because of the potential complications, proper counseling should take place and only experienced surgeons should perform this type of surgery. Egydio PH, Kuehhas FE. The Multiple-Slit Technique (MUST) for Penile Length and Girth Restoration. J Sex Med 2018;15:261-269.


Nanocrystalline materials: recent advances in crystallographic characterization techniques.
Most properties of nanocrystalline materials are shape-dependent, providing their exquisite tunability in optical, mechanical, electronic and catalytic properties. An example of the former is localized surface plasmon resonance (LSPR), the coherent oscillation of conduction electrons in metals that can be excited by the electric field of light; this resonance frequency is highly dependent on both the size and shape of a nanocrystal. An example of the latter is the marked difference in catalytic activity observed for different Pd nanoparticles. Such examples highlight the importance of particle shape in nanocrystalline materials and their practical applications. However, one may ask 'how are nanoshapes created?', 'how does the shape relate to the atomic packing and crystallography of the material?', 'how can we control and characterize the external shape and crystal structure of such small nanocrystals?'. This feature article aims to give the reader an overview of important techniques, concepts and recent advances related to these questions. Nucleation, growth and how seed crystallography influences the final synthesis product are discussed, followed by shape prediction models based on seed crystallography and thermodynamic or kinetic parameters. The crystallographic implications of epitaxy and orientation in multilayered, core-shell nanoparticles are overviewed, and, finally, the development and implications of novel, spatially resolved analysis tools are discussed.


Dynamics of fluctuations in smectic membranes.
We present a comprehensive account of the dynamics of layer-displacement fluctuations in smectic liquid-crystal membranes as studied by x-ray photon correlation spectroscopy (XPCS) and neutron-spin echo (NSE). Combining these two techniques at fast relaxation times, three distinct relaxation regimes can be distinguished. For thin membranes, at the specular Bragg position oscillatory relaxation occurs, which transforms for thicker samples into exponential decay. Above a critical off-specular angle, in XPCS exponential relaxation is observed that does not depend on the scattering angle. This indicates relaxation times that are independent of the wavelength of the fluctuations. In this regime the relaxation of the fluctuations is dominated by the surface tension. Using NSE larger off-specular angles can be reached than by XPCS, for which the relaxation time decreases with the scattering angle. This regime is dominated by the bulk elasticity of the smectic membrane. The results are compared with theoretical models for the fluctuation behavior of smectic membranes, in which effects of the mosaic distribution and of the center of mass movement of the smectic membranes must be incorporated.


Marker-free plasmids for biotechnological applications - implications and perspectives.
Nonviral gene therapy and DNA vaccines have become the first promising approaches to treat, cure, or ultimately prevent disease by providing genetic information encoded on a plasmid. Since 1989, more than 1800 clinical trials have been approved worldwide, and approximately 20% of them are using plasmid DNA (pDNA) as a vector system. Although much safer than viral approaches, DNA vectors generally do encode antibiotic resistance genes in the plasmid backbone. These antibiotic resistance markers constitute a possible safety risk, and they are associated with structural plasmid instabilities and decreased gene delivery efficiency. These drawbacks have initiated the development of various antibiotic marker-free selection approaches. We provide an overview on the potential implications of marker-free plasmids and perspectives for their successful biotechnological use in the future.


Soluble recombinant HTLV-1 surface glycoprotein competitively inhibits syncytia formation and viral infection of cells.
Efficient entry into, and infection of, human cells by human T-cell leukaemia virus type-1 (HTLV-1) is mediated by the viral envelope glycoproteins, gp46 and gp21. The gp46 surface glycoprotein binds to an as yet unidentified cell surface receptor, thereby, allowing the gp21 transmembrane glycoprotein to initiate fusion of the viral and cellular membranes. In the absence of membrane fusion viral penetration and entry into the host cell cannot occur. The envelope glycoproteins are also a major target for neutralising antibodies and cytotoxic T lymphocytes following a protective immune response, and represent ideal constituents for a recombinant HTLV-1 vaccine. Given the importance of the envelope proteins in HTLV-1 pathogenesis there is increasing interest in obtaining sufficient quantities of these proteins for biochemical, biophysical and biological analyses. We have now developed a system for production of large amounts of a glycosylated and functional form of soluble recombinant gp46 (sRgp46), and have used this recombinant material for analysis of envelope function and receptor binding activity. We find that, the sRgp46 molecules expressed in our system are immunologically indistinguishable from the native virally expressed surface glycoproteins; that sRgp46 binds to T-cells in a dose dependent and saturable manner; and that cell surface binding by sRgp46 can be inhibited by neutralising antibodies. Importantly, we demonstrate that these sRgp46 molecules potently inhibit syncytia formation and viral infection of target cells, and that regions outwith the SU domain of envelope are not required for binding to target cells or for inhibiting membrane fusion. The sRgp46 produced in our study will provide new opportunities to investigate envelope-receptor interactions, and will be of utility in defining the conformationally sensitive antigenic determinants of the HTLV-1 surface glycoprotein.


Health Beliefs of Wearing Facemasks for Influenza A/H1N1 Prevention: A Qualitative Investigation of Hong Kong Older Adults.
Although vaccination is the first-line strategy controlling and preventing influenza A/H1N1 for older adults, personal protective measures, such as wearing facemasks, are also important preventive behaviors to reduce the risk of becoming infected with influenza A/H1N1 during a pandemic. In the current study, we aimed to explore the belief-related psychological factors of wearing facemasks for influenza A/H1N1 prevention in a sample of Hong Kong older adults. Community-dwelling Chinese adults (N = 137) aged between 65 and 80 years (Mage = 75.10; SDage = 6.49) participated in semistructured interviews. Data were analyzed using theoretical thematic analysis and identified themes were matched deductively within the belief-based processes of the health belief model. Results revealed beliefs that underpinned 4 general dimensions of facemask wearing: (1) perceived susceptibility and seriousness of influenza A/H1N1 pandemic (e.g., influenza A/H1N1 as contagious and lethal), (2) modifying factors (e.g., social responsibility to prevent influenza), (3) cues to action (e.g., seeing others doing it), and (4) perceived benefits and barriers (e.g., protects oneself and others, difficult to breathe). Future interventions can target these beliefs to improve facemask wearing of older adults and, thus, curb preventable infection rates during an influenza A/H1N1 pandemic.


Translating clinical trial results into practice: the effect of an AIDS clinical trial on prescribed antiretroviral therapy for HIV-infected pregnant women.
The success of Pediatric AIDS Clinical Trials Group (PACTG) Protocol 076 in preventing vertical HIV transmission prompted intensive efforts to inform lay-persons and professionals about the trial's results. To explore community responsiveness to these efforts by assessing temporal, maternal, and health care factors associated with prescribed antiretroviral therapy before and after PACTG Protocol 076. Retrospective cohort study. New York State Medicaid program. 2607 HIV-infected women who delivered a living child between January 1993 and September 1996. Adjusted odds of being prescribed antiretroviral treatment in the second or third trimester for women who delivered in period 1 (during the trial [January 1993 to February 1994]), period 2 (after the trial's end and announcement of the results to publication of the results [March 1994 to November 1994]), and period 3 (after publication of the trial results [December 1994 to September 1996]). The adjusted odds of being prescribed antiretroviral therapy increased 21% per month in period 2 and decreased to 3% per month in period 3. In all time periods, the adjusted odds of being prescribed antiretroviral therapy were at least 60% greater (P < 0.05) for women who were treated at an institution that performed HIV clinical trials, received HIV-focused ambulatory care, or had adequate prenatal care visits. After the trial, women receiving methadone treatment had at least twofold (95% CI, 1.5- to 4.3-fold) greater adjusted odds of being prescribed antiretroviral therapy than women who did not take any illicit drugs. Latin-American women, older women, and women born in the United States had greater adjusted odds (P < 0.05) of being prescribed treatment in period 3. Community practice responded rapidly to efforts to disseminate the results of PACTG Protocol 076; however, the absolute increase in prescribed therapy was greatest for women who had adequate prenatal visits or were receiving HIV-focused care, care at a site that performed clinical trials, or methadone therapy.


Psychosocial aspects of systemic sclerosis.
In 2010, an international consortium of researchers published a consensus agenda for research on psychosocial aspects of systemic sclerosis (SSc; scleroderma). The present review summarizes recent research on SSc-related depression and anxiety, fatigue and sleep, pain, pruritus, body image distress, sexual function, work disability, healthcare needs, psychosocial interventions, and psychoneuroimmunology. Researchers have used structured interviews to establish prevalence rates for clinical mood disorders in SSc, although anxiety remains understudied and distress may be a useful outcome to consider. Longitudinal research has identified predictors of fatigue. Research on body image distress suggests the importance of changes in the facial skin and hands. Studies have identified sexual function problems in women and men with SSc. A prospective study found that breathing problems and fatigue predicted workplace disability. A randomized controlled trial evaluating multidisciplinary care showed benefits for health-related quality of life. There has been a recent expansion in studies of psychosocial aspects of SSc, and in the validation of psychosocial measures that can be used in clinics to identify problems and track outcomes; however, prospective studies remain scarce. To better address the psychosocial needs of persons with SSc, interventions need to be developed and tested via randomized controlled trials with power to detect clinically meaningful changes.


Design features of the Diabetes and Periodontal Therapy Trial (DPTT): a multicenter randomized single-masked clinical trial testing the effect of nonsurgical periodontal therapy on glycosylated hemoglobin (HbA1c) levels in subjects with type 2 diabetes and chronic periodontitis.
Evidence suggests that periodontitis is associated with prevalent and incident type 2 diabetes mellitus (T2DM), raising the question of whether periodontitis treatment may improve glycemic control in patients with T2DM. Meta-analyses of mostly small clinical trials suggest that periodontitis treatment results in a modest reduction in glycosylated hemoglobin (Hb) A1c. The purpose of the Diabetes and Periodontal Therapy Trial (DPTT) was to determine if periodontal treatment reduces HbA1c in patients with T2DM and periodontitis. DPTT was a phase-III, single-masked, multi-center, randomized trial with a planned enrollment of 600 participants. Participants were randomly assigned to receive periodontal treatment immediately (Treatment Group) or after 6 months (Control Group). HbA1c values and clinical periodontal measures were determined at baseline and 3 and 6 months following randomization. Medication usage and dosing were assessed at each visit. Periodontal treatment consisted of scaling and root planing for a minimum of two 90-minute sessions, plus the use of an antibacterial mouth rinse for at least 32 days afterwards. The primary outcome was change in HbA1c from baseline to 6 months and the trial was powered to detect a between-group difference of 0.6%. Secondary outcomes included changes in periodontal clinical measures, fasting plasma glucose, the Homeostasis Model Assessment (HOMA2) and the need for rescue diabetes or periodontal therapy. Dental and medical researchers collaborated to recruit, treat and monitor participants with two chronic diseases to determine if treatment of one condition affects the status of the other.


Postoperative chemoradiotherapy improves survival in esophageal squamous cell cancer with extracapsular lymph node extension.
In esophageal squamous cell carcinoma (ESCC), extracapsular extention (ECE) in metastatic lymph nodes portends high rate of recurrence and poor prognosis. To our knowledge, the effectiveness of postoperative chemoradiotherapy (CRT) in these patients has never been investigated. In this retrospective study, we compared the outcomes of surgery with or without postoperative chemoradiotherapy in ESCC patients with ECE. From 2008 to 2009, 90 ECSS patients with ECE were included. Among those patients, 47 only received curative surgery alone, and 43 received additional postoperative concurrent CRT which consisted of radiotherapy (median dose 50 Gy) and chemotherapy (5-fluorouracil 1000 mg/m(2), days 1-4 and 29-32; cisplatinum 25 mg/m(2), days 1-3 and 29-31). Patients treated with postoperative CRT had significantly more T3/4 tumors (p=0.023). Based on log-rank stratified by T stage, postoperative adjuvant CRT significantly improved the overall survival (p=0.017) and progression free survival (p=0.002). In multivariate analysis, adjuvant CRT was identified as an independent prognostic factor (HR=0.494, CI 0.290-0.844, p=0.010). Compared with surgery alone, the CRT group had significantly fewer cases of regional recurrence (P=0.048) and overall recurrence (P=0.024). However, there was no significant difference in distant metastasis between two groups (P=0.755). In conclusion, our data suggest that the postoperative adjuvant CRT might be beneficial in selected subgroups of ESCC patients with ECE. To further verify these results, a prospective trial with a large sample size is needed.


Targeting the polyadenylation factor EhCFIm25 with RNA aptamers controls survival in Entamoeba histolytica.
Messenger RNA 3'-end polyadenylation is an important regulator of gene expression in eukaryotic cells. In our search for new ways of treating parasitic infectious diseases, we looked at whether or not alterations in polyadenylation might control the survival of Entamoeba histolytica (the agent of amoebiasis in humans). We used molecular biology and computational tools to characterize the mRNA cleavage factor EhCFIm25, which is essential for polyadenylation in E. histolytica. By using a strategy based on the systematic evolution of ligands by exponential enrichment, we identified single-stranded RNA aptamers that target EhCFIm25. The results of RNA-protein binding assays showed that EhCFIm25 binds to the GUUG motif in vitro, which differs from the UGUA motif bound by the homologous human protein. Accordingly, docking experiments and molecular dynamic simulations confirmed that interaction with GUUG stabilizes EhCFIm25. Incubating E. histolytica trophozoites with selected aptamers inhibited parasite proliferation and rapidly led to cell death. Overall, our data indicate that targeting EhCFIm25 is an effective way of limiting the growth of E. histolytica in vitro. The present study is the first to have highlighted the potential value of RNA aptamers for controlling this human pathogen.


Effects of long-term blood pressure lowering and dual antiplatelet treatment on cognitive function in patients with recent lacunar stroke: a secondary analysis from the SPS3 randomised trial.
The primary outcome results for the SPS3 trial suggested that a lower systolic target blood pressure (<130 mm Hg) might be beneficial for reducing the risk of recurrent stroke compared with a higher target (130-149 mm Hg), but that the addition of clopidogrel to aspirin was not beneficial compared with aspirin plus placebo. In this prespecified secondary outcome analysis of the SPS3 trial, we aimed to assess whether blood pressure reduction and dual antiplatelet treatment affect changes in cognitive function over time in patients with cerebral small vessel disease. In the SPS3 trial, patients with recent (within 6 months) symptomatic lacunar infarcts from 81 centres in North America, Latin America, and Spain were randomly assigned, in a two-by-two factorial design, to target levels of systolic blood pressure (1:1; 130-149 mm Hg vs <130 mm Hg; open-label) and to a once-daily antiplatelet treatment (1:1; aspirin 325 mg plus clopidogrel 75 mg vs aspirin 325 mg plus placebo; double-blind). For this analysis, the main cognitive outcome was change in Cognitive Abilities Screening Instrument (CASI) during follow-up. Patients were tested annually for up to 5 years, during which time the mean difference in systolic blood pressure was 11 mm Hg (SD 16) between the two targets (138 mm Hg vs 127 mm Hg at 1 year). We used linear mixed models to compare changes in CASI Z scores over time. The SPS3 trial is registered with ClinicalTrials.gov, number NCT00059306. The study took place between March 23, 2003, and April 30, 2012. 2916 of 3020 SPS3 participants (mean age 63 years [SD 11]) with CASI scores at study entry were included in the analysis, with a median follow-up of 3·0 years (IQR 1·0-4·9). Mean changes in CASI Z scores from study entry to assessment at years 1 (n=2472), 2 (n=1968), 3 (n=1521), 4 (n=1135), and 5 (n=803) were 0·12 (SD 0·83), 0·15 (0·84), 0·16 (0·95), 0·19 (0·99), and 0·14 (1·09), respectively. Changes in CASI Z scores over time did not differ between assigned antiplatelet groups (p=0·858) or between assigned blood pressure target groups (p=0·520). There was no interaction between assigned antiplatelet groups and assigned blood pressure target groups and change over time (p=0·196). Cognitive function is not affected by short-term dual antiplatelet treatment or blood pressure reduction in fairly young patients with recent lacunar stroke. Future studies of cognitive function after stroke should be of longer duration or focus on patients with higher rates of cognitive decline. US National Institute of Neurological Disorders and Stroke.


The thioredoxin reductase-1 inhibitor aurothioglucose attenuates lung injury and improves survival in a murine model of acute respiratory distress syndrome.
Inflammation and oxygen toxicity increase free radical production and contribute to the development of acute respiratory distress syndrome (ARDS), which is a significant cause of morbidity and mortality in intensive care patients. We have previously reported increased glutathione (GSH) levels in lung epithelial cells in vitro and attenuated adult murine hyperoxic lung injury in vivo after pharmacological thioredoxin reductase-1 (TrxR1) inhibition. Using a murine ARDS model, we tested the hypothesis that aurothioglucose (ATG) treatment increases pulmonary GSH levels, attenuates lung injury, and decreases mortality in a GSH-dependent manner. Adult mice received a single intratracheal dose of 0.375 μg/g lipopolysaccharide (LPS) 12 h before a single intraperitoneal injection of 25 mg/kg ATG. Control mice received intratracheal and/or intraperitoneal saline. Mice were then exposed to room air or hyperoxia (>95% O2). Lung injury was assessed by bronchoalveolar lavage protein concentrations. Expression of glutamate-cysteine ligase modifier subunit (GCLM), GSH, cytokines, and chemokines was determined. Exposure to LPS/hyperoxia induced inflammation and lung injury. ATG treatment significantly attenuated lung injury, increased lung GCLM expression and GSH levels, and decreased mortality. GSH depletion completely prevented the protective effects of ATG in LPS/hyperoxia-exposed mice. ATG treatment significantly attenuates lung injury and enhances survival in a clinically relevant murine model of ARDS. The protective effects of ATG are GSH dependent. Augmentation of GSH systems by TrxR1 inhibition could represent a promising therapeutic approach to attenuate oxidant-mediated lung injury and improve patient outcomes.


Repetitive DNA in the pea (Pisum sativum L.) genome: comprehensive characterization using 454 sequencing and comparison to soybean and Medicago truncatula.
Extraordinary size variation of higher plant nuclear genomes is in large part caused by differences in accumulation of repetitive DNA. This makes repetitive DNA of great interest for studying the molecular mechanisms shaping architecture and function of complex plant genomes. However, due to methodological constraints of conventional cloning and sequencing, a global description of repeat composition is available for only a very limited number of higher plants. In order to provide further data required for investigating evolutionary patterns of repeated DNA within and between species, we used a novel approach based on massive parallel sequencing which allowed a comprehensive repeat characterization in our model species, garden pea (Pisum sativum). Analysis of 33.3 Mb sequence data resulted in quantification and partial sequence reconstruction of major repeat families occurring in the pea genome with at least thousands of copies. Our results showed that the pea genome is dominated by LTR-retrotransposons, estimated at 140,000 copies/1C. Ty3/gypsy elements are less diverse and accumulated to higher copy numbers than Ty1/copia. This is in part due to a large population of Ogre-like retrotransposons which alone make up over 20% of the genome. In addition to numerous types of mobile elements, we have discovered a set of novel satellite repeats and two additional variants of telomeric sequences. Comparative genome analysis revealed that there are only a few repeat sequences conserved between pea and soybean genomes. On the other hand, all major families of pea mobile elements are well represented in M. truncatula. We have demonstrated that even in a species with a relatively large genome like pea, where a single 454-sequencing run provided only 0.77% coverage, the generated sequences were sufficient to reconstruct and analyze major repeat families corresponding to a total of 35-48% of the genome. These data provide a starting point for further investigations of legume plant genomes based on their global comparative analysis and for the development of more sophisticated approaches for data mining.


[Dietary habits and cardiovascular disease: the experience of EPIC Italian collaboration].
to report and evaluate the evidence produced by the EPIC Italian collaboration (EPICOR Project) on the dietary determinants of cardiovascular diseases in Italy. prospective study carried out in a large Italian population, composed by cohorts recruited in Northern, Central and Southern Italy. data on dietary habits collected at the baseline observation through standardised questionnaires on 47,749 free-living adults at the time of the recruitment of the study (1993-1998). major coronary and cerebrovascular events (acute coronary syndrome, PTCA, CABG, ischemic and haemorrhagic stroke, TEA of supraortic vessels) identified at follow-up. The longitudinal analyses here reported have measured risks through the use of multivariate Cox regression models, adjusted for potential confounders. the longitudinal analyses of EPICOR indicate that Mediterranean-oriented dietary habits, measured through specific indicators and the consumption of various typical food, are able to reduce coronary and cerebrovascular risks, and that this protection is possible even nowadays, although many changes in diet have occurred in the last decades in Italy. Habitual consumption of plant origin products, including all foods with low glycemic index, is an advantage for cardiovascular risk. the EPICOR Project is the largest, long-lasting Italian study on the relationship between diet and cardiovascular diseases. It is also the study with the greater number of observed variables. Its results point out the importance to support preventive programmes and industrial policies able to favour a dietary style inspired to the Italian Mediterranean tradition.


Evaluation of a novel syndromic surveillance system for the detection of the 2007 melamine-related nephrotoxicosis foodborne outbreak in dogs and cats in the United States.
The 2007 nephrotoxicosis outbreak associated with melamine and cyanuric acid adulteration of pet foods in the United States sparked an urgent need for a nationwide companion animal surveillance program. In 2016, we introduced a syndromic surveillance system based on a novel epidemiological algorithm, the proportionate diagnostic outcome ratio (PDOR). The PDOR procedure was validated using simulated outbreaks of foodborne illness (i.e., aflatoxicosis and gastrointestinal illness) in dogs and cats. In this study, we further evaluated the PDOR procedure using the 2007 melamine-related outbreak of nephrotoxicosis. The performance of the PDOR procedure was assessed by the time to alert and positive predictive value (PPV). Electronic medical records of dogs and cats seen at networked primary care veterinary hospitals across the United States were retrieved from a centralized database. The data of four relevant syndromic components: elevated serum creatinine concentration, vomiting, anorexia, and lethargy from July 28, 2006 to May 31, 2007 were prospectively analyzed using the PDOR algorithm. The results showed that the alerts generated from the analysis of elevated serum creatinine concentration could have led to an early detection of this nephrotoxicosis foodborne outbreak and were well matched to the reported timeline of the outbreak. Additionally, we also observed variations in the performance of the PDOR procedure across age of animals and syndromic components, with the PPVs ranged from 0.61 to 1.0. Combined with the findings from previous evaluations using simulated outbreak scenarios, this study provided additional evidence that the PDOR procedure can be applied in syndromic surveillance to effectively and accurately detect various types of foodborne illness outbreaks in companion animals. However, the interpretations of and responses to alerts require an understanding of clinical veterinary medicine and relevant syndromic knowledge, and should not be based solely on quantitative measures.


Bombesin stimulates mucosal growth in jejunal and ileal Thiry-Vella fistulas.
The authors determined whether the trophic effects of bombesin (BBS) on the small bowel mucosa are mediated by either nonluminal factors or endogenous luminal secretion. The gut hormone bombesin stimulates growth of small bowel mucosa. The mechanisms responsible for this trophic effect are not known. Rats underwent construction of a Thiry-Vella fistula (TVF) of either the jejunum or ileum. On postoperative day 10, the two groups were subdivided to receive either saline (control) or bombesin (10 micrograms/kg, subcutaneously, three times a day). After 14 days, rats were killed and the TVF was removed. The mucosa was scraped and weighed, and DNA and protein content was determined. Bombesin significantly increased mucosal weight and DNA and protein content of both the jejunal and ileal TVF compared with the control rats. Bombesin-mediated stimulation of small bowel mucosal growth is mediated by factors that are independent of luminal contents and pancreaticobiliary secretion. Bombesin may prove to be an important enterotrophic factor for gut mucosal proliferation.


Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus.
A real-time polymerase chain reaction (PCR) assay for the rapid detection of African swine fever virus (ASFV), multiplexed for simultaneous detection of swine beta-actin as an endogenous control, has been developed and validated by four National Reference Laboratories of the European Union for African swine fever (ASF) including the European Union Reference Laboratory. Primers and a TaqMan(®) probe specific for ASFV were selected from conserved regions of the p72 gene. The limit of detection of the new real-time PCR assay is 5.7-57 copies of the ASFV genome. High accuracy, reproducibility and robustness of the PCR assay (CV ranging from 0.7 to 5.4%) were demonstrated both within and between laboratories using different real-time PCR equipments. The specificity of virus detection was validated using a panel of 44 isolates collected over many years in various geographical locations in Europe, Africa and America, including recent isolates from the Caucasus region, Sardinia, East and West Africa. Compared to the OIE-prescribed conventional and real-time PCR assays, the sensitivity of the new assay with internal control was improved, as demonstrated by testing 281 field samples collected in recent outbreaks and surveillance areas in Europe and Africa (170 samples) together with samples obtained through experimental infections (111 samples). This is particularly evident in the early days following experimental infection and during the course of the disease in pigs sub-clinically infected with strains of low virulence (from 35 up to 70dpi). The specificity of the assay was also confirmed on 150 samples from uninfected pigs and wild boar from ASF-free areas. Measured on the total of 431 tested samples, the positive deviation of the new assay reaches 21% or 26% compared to PCR and real-time PCR methods recommended by OIE. This improved and rigorously validated real-time PCR assay with internal control will provide a rapid, sensitive and reliable molecular tool for ASFV detection in pigs in newly infected areas, control in endemic areas and surveillance in ASF-free areas.


Validation of the comprehensive ICF Core Set for patients in geriatric post-acute rehabilitation facilities.
To examine the relevance and completeness of the comprehensive International Classification of Functioning, Disability and Health (ICF) Core Set for patients in geriatric post-acute rehabilitation facilities. Multi-centre cohort study. A total of 209 patients (67% female, mean age 80.4 years) in geriatric wards of 2 Austrian and 3 German hospitals. Data on functioning were collected using the respective comprehensive ICF Core Set. Data were extracted from patients' medical record sheets and interviews with health professionals and patients. Most of the categories of the comprehensive ICF Core Set describing impairments, limitations or restrictions occurred in a considerable proportion of the study population. The most outstanding limitations and restrictions of the patients were problems with walking and moving around, and difficulties with self-care. Fourteen aspects of functioning not previously covered by the comprehensive ICF Core Set were reported as relevant. Most categories of the comprehensive ICF Core Set could be confirmed. Limitations in categories of intellectual and seeing functions appeared less frequently than might have been expected for a population of older hospitalized people. Some additional categories not covered by the present version of the comprehensive ICF Core Set emerged from the interviews and should be considered for inclusion in the final version.


Lysosomal localization of GLUT8 in the testis--the EXXXLL motif of GLUT8 is sufficient for its intracellular sorting via AP1- and AP2-mediated interaction.
The class III sugar transport facilitator GLUT8 co-localizes with the lysosomal protein LAMP1 in heterologous expression systems. GLUT8 carries a [D/E]XXXL[L/I]-type dileucine sorting signal that has been postulated to retain the protein in an endosomal/lysosomal compartment via interactions with clathrin adaptor protein (AP) complexes. However, contradictory findings have been described regarding the subcellular localization of the endogenous GLUT8 and the adaptor proteins that interact with its dileucine motif. Here we demonstrate that endogenous GLUT8 is localized in a late endosomal/lysosomal compartment of spermatocytes and spermatids, and that the adaptor complexes AP1 and AP2, but not AP3 or AP4, interact with its N-terminal intracellular domain (NICD). In addition, fusion of the GLUT8 NICD to the tailless lumenal domain of the IL-2 receptor alpha chain (TAC) protein (interleukin-2 receptor a chain) targeted the protein to intracellular membranes, indicating that its N-terminal dileucine signal is sufficient for endosomal/lysosomal targeting of the transporter. The localization and targeting of GLUT8 show striking similarities to sorting mechanisms reported for lysosomal proteins. Therefore, we suggest a potential role for GLUT8 in the so far unexplored substrate transport across intracellular membranes.


Parents' early life stressful experiences, their present well-being, and that of their children.
Parents' early life stressful experiences have lifelong consequences, not only for themselves but also for their children. The current study utilized a sample of military families (n = 266) including data from both active-duty and civilian parents and their adolescent children. Hypotheses reflecting principles of persistence, transmission, and proximity as pertaining to parents and their children were examined. The impact of parents' childhood experiences on their functioning later in life and, consequently, their adolescent children's well-being were examined. Adults who encountered more stressful childhood experiences, including relatively prevalent and less severe adversities (e.g., verbal conflict between parents) experienced poorer functioning than adults who encountered little early stress. Civilian parents' current functioning was related to adolescent children's well-being, whereas the functioning of active-duty parents was generally not related to children's well-being. Persistence, transmission, and proximity hypotheses were generally supported but with variations attributable to whether an adult was a military member. (PsycINFO Database Record


Benchmarking DFT approaches for the calculation of polarizability inputs for refractive index predictions in organic polymers.
In a previous study, we introduced a new computational protocol to accurately predict the index of refraction (RI) of organic polymers using a combination of first-principles and data modeling. This protocol is based on the Lorentz-Lorenz equation and involves the calculation of static polarizabilities and number densities of oligomer sequences, which are extrapolated to the polymer limit. We chose to compute the polarizabilities within the density functional theory (DFT) framework using the PBE0/def2-TZVP-D3 model chemistry. While this ad hoc choice proved remarkably successful, it is also relatively expensive from a computational perspective. It represents the bottleneck step in the overall RI modeling protocol, thus limiting its utility for virtual high-throughput screening studies, in which efficiency is essential. For polymers that exhibit late-onset extensivity, the employed linear extrapolation scheme can require demanding calculations on long-oligomer sequences, thus becoming another bottleneck. In the work presented here, we benchmark DFT model chemistries to identify approaches that optimize the balance between accuracy and efficiency for this application domain. We compare results for conjugated and non-conjugated polymers, augment our original extrapolation approach with a non-linear option, analyze how the polarizability errors propagate into the RI predictions, and offer guidance for method selection.


Culture at the centre of community based aged care in a remote Australian Indigenous setting: a case study of the development of Yuendumu Old People's Programme.
Yuendumu is a Warlpiri Aboriginal community 300 km north west of Alice Springs in Central Australia. Since emerging from the welfare period in the early 1970s, a range of services have evolved with the aim of developing a comprehensive community based aged care service. In 2000 Mampu Maninja-kurlangu Jarlu Patu-ku Aboriginal Corporation (Yuendumu Old Peoples Programme; YOPP) commenced operation to manage the developing services. This case study aims to describe, from the analytic standpoint of community control and cultural comfort, the main features of the 'Family Model of Care', which underpins the operations of the service and YOPP management processes. Data were mostly generated from participant observation by the authors in the development and management of YOPP between 1993 and 2009. A literature review of Indigenous history and public health in Central Australia was also undertaken, which was supplemented by a review of Programme documentation, including evaluations, needs assessments and annual reports. The design and operations of YOPP are embodied in a documented 'Family Model of Care' which provides important lessons for the provision of aged care in a cross-cultural context. According to the concepts 'community control' and 'cultural comfort' outlined in this article, mainstream services can function in a complementary and supportive manner with professional services being accountable and responsive to a local management system that is governed by the structures and norms of community tradition. The notions of 'cultural comfort' and 'community control' as operating principles have enabled YOPP to continue under the management of local people, sustain core cultural strengths and values, and meet the needs for increased quality of care for the aged in Yuendumu. This model of care emphasizes and recognizes paradigms of mutual competence between traditional and mainstream human service culture, and offers important lessons for improvement to the quality of aged care in remote Indigenous communities in Australia and elsewhere.