[Prognostic impact of plasma Epstein-Barr virus DNA concentration on distant metastasis in nasopharyngeal carcinoma].
Distant metastasis is one of the main causes of treatment failure in nasopharyngeal carcinoma (NPC). Trials of combined modality treatment could not demonstrate obvious decrease of the rate of distant metastasis and survival rate, therefore eligible individualized and best combined modality treatment has not been established. Plasma Epstein-Barr virus (EBV) DNA is a sensitive and specific molecular marker, which may reflect the stage, response to treatments and prognosis of NPC. This prospective study was to investigate whether the plasma EBV DNA concentration could be used to predict distant metastasis in NPC, and thus to provide us a molecular marker for individualized combined modality treatment. Blood samples from 69 patients with primary NPC were collected before and after radiotherapy. The content of EBV DNA in the plasma samples was detected by real-time quantitative PCR. All patients received consequent follow up and long-term efficacy and survival assessment. The correlation of pre/post treatment EBV DNA concentrations to survival was analyzed by Kaplan-Meier method. The prognostic factors were evaluated by Cox proportional hazards model. Both pretreatment EBV DNA median concentration (27,000 copies/ml) and post-treatment EBV DNA detecting rate (55.56%) in patients with distant metastasis were higher than those with continuous remission (4,000 copies/ml, 5.56%) and those with local relapse (3,850 copies/ml, 0%)(P = 0.039 and 0.001, respectively). With a cut-off value of 20,000 copies/ml and 0 copies/ml respectively for pretreatment and post-treatment EBV DNA concentration, patients with lower EBV DNA concentration had statistically preferable progression-free survival (PFS), metastasis-free survival(MFS) and overall survival (OS) than those with higher EBV DNA concentration. Cox regression analysis demonstrated that both pretreatment EBV DNA (P = 0.050; RR = 3.95) and post-treatment EBV DNA (P = 0.001; RR = 11.74) were risk factors for MFS. Further analysis of pretreatment and post-treatment EBV DNA concentration revealed that whether EBV DNA concentration could be decreased to 0 after treatment dominated metastasis-free survival (P = 0.000). Plasma EBV DNA concentration before and after treatment, especially whether post-treatment concentration could be decreased to 0 may predict distant metastasis, which helps to select patients with high risks and determine the combined modality treatment.


Oxidative stress biomarker monitoring in elite women volleyball athletes during a 6-week training period.
The objectives of this study were to determine (a) if reactive oxygen metabolites (ROMs) are a reliable parameter for monitoring oxidative stress in athletes alone or in association with other parameters of oxidative stress and depending on whether antioxidant supplements are taken or not; (b) the level of oxidative stress in athletes before the competition season; and (c) if oxidative status could be improved in volleyball athletes. Sixteen women athletes (supplemented group) received an antioxidant cocktail containing vitamin E, vitamin C, zinc gluconate, and selenium as a dietary supplement during a 6-week training period, whereas 12 of them (control group) received no dietary supplement. Blood samples were taken before and after the training period. The following parameters were measured: ROMs, superoxide anion (O2⁻₂), malondialdehyde (MDA), advanced oxidation protein products (AOPP), lipid hydroperoxide (LOOH), biological antioxidative potential (BAP), paraoxonase activity toward paraoxon (POase) and diazoxon (DZOase), superoxide dismutase(SOD), total sulfydryl group concentration (SH groups), and lipid status. Reactive oxygen metabolites were taken as the dependent variable and MDA, O2⁻₂, AOPP, and LOOH as independent variables. In the group of athletes who have received supplementation, linear regression analysis revealed that the implemented model had a lower influence on dROMs (70.4 vs. 27.9%) after the training period. The general linear model showed significant differences between parameters before and after training/supplementation (Wilks' lambda = 0.074, F = 11.76, p < 0.01). At the partial level, significant increases in ROM levels (p <0.05, 95% confidence interval [CI]: 286-337), SOD activity (CI: 113-144), and BAP (CI: 2,388-2,580) (p < 0.01) were observed. The association between ROMs and other parameters of oxidative stress was reduced in athletes who received supplements. During the precompetition training period, treatment with dietary supplements prevented the depletion of antioxidative defense in volleyball athletes.


Rotation, scale, and translation resilient watermarking for images.
Many electronic watermarks for still images and video content are sensitive to geometric distortions. For example, simple rotation, scaling, and/or translation (RST) of an image can prevent blind detection of a public watermark. In this paper, we propose a watermarking algorithm that is robust to RST distortions. The watermark is embedded into a one-dimensional (1-D) signal obtained by taking the Fourier transform of the image, resampling the Fourier magnitudes into log-polar coordinates, and then summing a function of those magnitudes along the log-radius axis. Rotation of the image results in a cyclical shift of the extracted signal. Scaling of the image results in amplification of the extracted signal, and translation of the image has no effect on the extracted signal. We can therefore compensate for rotation with a simple search, and compensate for scaling by using the correlation coefficient as the detection measure. False positive results on a database of 10,000 images are reported. Robustness results on a database of 2000 images are described. It is shown that the watermark is robust to rotation, scale, and translation. In addition, we describe tests examining the watermarks resistance to cropping and JPEG compression.


Edward Forbes (1815-1854) and the exhibition of natural order in Edinburgh.
The roles, affordances and social agency of natural history museums are discussed in relation to the writings of Edward Forbes. These signal a motivation, in the mid-nineteenth-century, to naturalize the established social order through the systematic arrangement and display of natural history specimens. The perceived importance of the embodied messages of social order, as an antidote to radicalism and revolution, overrode concerns about temperance and abstinence and immediate fears for the physical safety of collections. The tensions between temperance, and the broader concerns about social order, were played out over the matter of the museums themselves being licensed premises.


Monocyte superoxide anion production in rheumatoid arthritis: preliminary evidence for enhanced rates of superoxide anion production by monocytes from patients receiving penicillamine, sodium aurothiomalate and corticosteroids.
In-vitro studies of superoxide (O-2) anion production by blood monocytes after stimulation with either serum treated zymosan (STZ), IgG treated zymosan (IgGTZ), or fluoride ion (F-) were performed on cells from normal controls (n = 22) and patients with classical or definite rheumatoid arthritis (RA) (n = 35). Twenty-two of the patients were on nonsteroidal anti-inflammatory drugs (NSAID) alone and 13 were on either sodium aurothiomalate, penicillamine, corticosteroids, or a combination. Monocytes from RA patients on 'second-line therapy' showed significantly increased rates of O-2 release in response to STZ compared with normal controls, but no increase was seen in monocytes from patients on NSAID alone. With IgGTZ as the stimulus, rates of O-2 release were increased in monocytes from patients on NSAID alone compared with normal controls (p less than 0.02), but were increased to a greater extent in monocytes from patients on second-line therapy (p less than 0.01). There were no differences in basal unstimulated O-2 production and no differences after stimulation with F-. The enhanced release of O-2 by monocytes from patients on second-line therapy could not be attributed to increased disease activity and may be an effect of therapy.


Central-cue discriminability modulates object-based attention by influencing spatial attention.
The role of central-cue discriminability in modulating object-based effects was examined using Egly, Driver, and Rafal's (1994) "double-rectangle" spatial cueing paradigm. Based on the attentional focusing hypothesis (Goldsmith & Yeari, 2003), we hypothesized that highly discriminable central-arrow cues would be processed with attention spread across the two rectangles (potential target locations), thereby strengthening the perceptual representation of these objects so that they influence the subsequent endogenous deployment of attention, yielding object-based effects. By contrast, less discriminable central-arrow cues should induce a more narrow attentional focus to the center of the display, thereby weakening the rectangle object representations so that they no longer influence the subsequent attentional deployment. Central-arrow-cue discriminability was manipulated by size and luminance contrast. The results supported the predictions, reinforcing the attentional focusing hypothesis and highlighting the need to consider central-cue discriminability when designing experiments and in comparing experimental results.


Using accelerometry to classify physical activity intensity in older adults: What is the optimal wear-site?
Abstract This study aimed to determine the optimal accelerometer wear-site specific cut-points for discrimination of the sedentary time, light physical activity and moderate-to-vigorous physical activity (MVPA) in older adults. Twenty-three adults (14 females) aged 55-77 years wore a GENEActiv accelerometer on their non-dominant wrist, dominant wrist, waist and dominant ankle whilst undertaking eight, five-minute bouts of activity: lay supine, seated reading, slow walking, medium walking, fast walking, folding laundry, sweeping and stationary cycling. VO2 was assessed concurrently using indirect calorimetry. Receiver-operating-characteristic (ROC) analyses were used to derive wear-site specific cut-points for classifying intensity. Indirect calorimetry indicated that being lay supine and seated reading were classified as sedentary (<1.5 METs), laundry as light (1.51-2.99 METs) and sweeping, slow, medium and fast walking and cycling all classified as moderate intensity (>3 METs). Areas under ROC curves indicated that the classification of sedentary activity was good for the non-dominant wrist and excellent for all other wear sites. Classification of MVPA was excellent for the waist and ankle, good for the waist and poor for the dominant and non-dominant wrists. Overall, the ankle location performed better than in other locations. Ankle-worn accelerometry appears to provide the most suitable wear-site to discriminate between sedentary time and MVPA in older adults.


Peroral infection of suckling mice with milk-borne mouse mammary tumour virus: uptake of the main viral antigens by the gut.
Persistence of mouse mammary tumour virus (MMTV) components in the digestive tract of suckling mice was investigated by immunoperoxidase staining of the main viral antigens and micro-immunoenzyme assays of gp52 and p28; these latter assays were also performed after ingestion of milk enriched in viral antigens using Cr2O3 as a marker for the alimentary bolus migration. When compared to the ingested antigens, the amounts of gp52 and p28 decreased during transit, p28 being more rapidly digested than gp52. The antigens were, however, destroyed to a much larger extent in the gut of the adult than in that of the newborn mouse. A fraction of the marker remained for a long time in the stomach; a prolonged retention was also observed with gp52 and especially with p28. Significant amounts of viral antigens were detected in the intestinal walls: both p28 and gp52 were found in the duodenum and small intestine. Moreover, the four viral antigens gp52, gp36, p28 and p8 were clearly observed in very large supranuclear vacuoles inside the epithelial cells of the distal part of the gut. Total particles can reach the intestine; the viral material could then be either destroyed or taken up in the epithelial cells by endocytosis, so that the intestinal epithelium might serve as a portal of entry for MMTV in the suckling mouse.


Overall Survival of Patients With Locoregional and Metastatic Breast Cancer: Is the Influence of Baseline Characteristics the Same?
To compare the overall survival (OS) of patients with locoregional and metastatic breast cancer (BC) considering baseline demographic, clinical and contextual characteristics. A retrospective analysis of a cancer registry was conducted, using the Kaplan-Meier and Mantel-Cox analyses for the calculation of median OS and cumulative survival. The median OS was 112 months, being longer in patients with locoregional versus those with metastatic BC at diagnosis (115 vs. 31 months, p<0.001). The cumulative survival at 1, 3 and 5 years were 94.9%, 85.6% and 76.5%, respectively. More recent year of diagnosis [hazard ratio (HR)=1.09] and age at diagnosis (≥65 vs. 40 years, HR=2.79) and presence of metastatic disease (HR=5.69) were associated with a shorter OS. The region of residence, morphology and topography of the tumor were also associated with survival in patients with BC. Rurality was only associated with lower survival in patients with metastatic BC. This study identified significant differences in the median OS of patients with locoregional and those with metastatic BC considering their baseline characteristics.


DNA-Mediated Patterning of Single Quantum Dot Nanoarrays: A Reusable Platform for Single-Molecule Control.
We present a facile strategy of general applicability for the assembly of individual nanoscale moieties in array configurations with single-molecule control. Combining the programming ability of DNA as a scaffolding material with a one-step lithographic process, we demonstrate the patterning of single quantum dots (QDs) at predefined locations on silicon and transparent glass surfaces: as proof of concept, clusters of either one, two, or three QDs were assembled in highly uniform arrays with a 60 nm interdot spacing within each cluster. Notably, the platform developed is reusable after a simple cleaning process and can be designed to exhibit different geometrical arrangements.


Neural correlates of self-injurious behavior in Prader-Willi syndrome.
Individuals with Prader-Willi syndrome (PWS), a genetic disorder caused by mutations to the q11-13 region on chromosome 15, commonly show severe skin-picking behaviors that can cause open wounds and sores on the body. To our knowledge, however, no studies have examined the potential neural mechanisms underlying these behaviors. Seventeen individuals with PWS, aged 6-25 years, who showed severe skin-picking behaviors, were recruited and scanned on a 3T scanner. We used functional magnetic resonance imaging (fMRI) while episodes of skin picking were recorded on an MRI-safe video camera. Three participants displayed skin picking continuously throughout the scan, three participants did not display skin picking, and the data for one participant evidenced significant B0 inhomogeneity that could not be corrected. The data for the remaining 10 participants (six male, four female) who displayed a sufficient number of picking and nonpicking episodes were subjected to fMRI analysis. Results showed that regions involved in interoceptive, motor, attention, and somatosensory processing were activated during episodes of skin-picking behavior compared with nonpicking episodes. Scores obtained on the Self-Injury Trauma scale were significantly negatively correlated with mean activation within the right insula and left precentral gyrus. These data indicate that itch and pain processes appear to underlie skin-picking behaviors in PWS, suggesting that interoceptive disturbance may contribute to the severity and maintenance of abnormal skin-picking behaviors in PWS. Implications for treatments are discussed.


Deuterium kinetic isotope effects and the mechanism of the bacterial luciferase reaction.
A combined experimental and theoretical investigation of the deuterium isotope effects on the bacterial luciferase reaction is described. The experimental studies focus on determining if the unusual aldehydic deuterium isotope effect of approximately 1.5 observed in these reactions is an intrinsic isotope effect resulting from a single rate-limiting step or is a composite of multiple rate-limiting steps. The isotope effect observed is not significantly affected by variation in the aldehyde chain length, changes in the pH over a range of 6-9, use of alphaC106A and alphaC106S site-directed mutants, or chloride substitution at the 8-position of the reduced flavin, though the isotope effect is decreased when the 8-methoxy-substituted flavin is used as a substrate. From these observations it is concluded that the aldehydic isotope effect arises from the change in rate of a single kinetic step. A stopped-flow kinetic analysis of the microscopic rate constants for the reactions of 1-[1H]decanal and 1-[2H]decanal in the bacterial luciferase reaction was carried out, and aldehyde hydration isotope effects were determined. From the results it is estimated that the aldehydic deuterium isotope effect is approximately 1.9 after formation of an intermediate flavin C4a-hydroperoxy hemiacetal. Ab initio calculations were used to examine the transformation of the aldehyde into a carboxylic acid and to predict isotope effects for possible mechanisms. These calculations indicate that the mechanism involving rate-limiting electron transfer from the flavin C4a-hydroxide to an intermediate dioxirane is consistent with the enigmatic aldehydic isotope effect and that the intermediacy of a dioxirane is energetically plausible.


The influence of asthma on neuroinflammation and neurodevelopment: From epidemiology to basic models.
Asthma is a highly heterogeneous inflammatory disease that can have a significant effect on both the respiratory system and central nervous system. Population based studies and animal models have found asthma to be comorbid with a number of neurological conditions, including depression, anxiety, and neurodevelopmental disorders. In addition, maternal asthma during pregnancy has been associated with neurodevelopmental disorders in the offspring, such as autism spectrum disorders and attention deficit hyperactivity disorder. In this article, we review the most current epidemiological studies of asthma that identify links to neurological conditions, both as it relates to individuals that suffer from asthma and the impacts asthma during pregnancy may have on offspring neurodevelopment. We also discuss the relevant animal models investigating these links, address the gaps in knowledge, and explore the potential future directions in this field.


Absent nasal bone at 11-14 weeks of gestation and chromosomal defects.
To examine the association between absence of the nasal bone at the 11-14-week ultrasound scan and chromosomal defects. Ultrasound examination was carried out in 3829 fetuses at 11-14 weeks' gestation immediately before fetal karyotyping. At the scan the fetal crown-rump length (CRL) and nuchal translucency (NT) thickness were measured and the fetal profile was examined for the presence or absence of the nasal bone. Maternal characteristics including ethnic origin were also recorded. The fetal profile was successfully examined in 3788 (98.9%) cases. In 3358/3788 cases the fetal karyotype was normal and in 430 it was abnormal. In the chromosomally normal group the incidence of absent nasal bone was related firstly to the ethnic origin of the mother (2.8% for Caucasians, 10.4% for Afro-Caribbeans and 6.8% for Asians), secondly to fetal CRL (4.6% for CRL of 45-54 mm, 3.9% for CRL of 55-64 mm, 1.5% for CRL of 65-74 mm and 1.0% for CRL of 75-84 mm) and thirdly, to NT thickness, (1.8% for NT < 2.5 mm, 3.4% for NT 2.5-3.4 mm, 5.0% for NT 3.5-4.4 mm and 11.8% for NT > or = 4.5 mm. In the chromosomally abnormal group the nasal bone was absent in 161/242 (66.9%) with trisomy 21, in 48/84 (57.1%) with trisomy 18, in 7/22 (31.8%) with trisomy 13, in 3/34 (8.8%) with Turner syndrome and in 4/48 (8.3%) with other defects. At the 11-14-week scan the incidence of absent nasal bone is related to the presence or absence of chromosomal defects, CRL, NT thickness and ethnic origin.


Insulin receptors in hepatocytes: postreceptor events mediate down regulation.
Down regulation of the insulin receptor of primary cultures of rat hepatocytes occurs in the presence of insulin and several agents with insulin-like activity, which act through or distal to the insulin receptor. These findings indicate that the interaction of insulin with its specific binding site is not in itself sufficient to down-regulate this receptor and that one or more steps subsequent to this interaction are necessary. Thus, down regulation may be a complex biological response to insulin, and if a cell were resistant to this effect of insulin, our data may explain how target cells from a patient or animal can have a normal number of receptors in the presence of increased concentrations of circulating insulin.


Variability in immunization guidelines in children before and after lung transplantation.
Lung transplant candidates and recipients are at high risk of infections from vaccine-preventable diseases. However, well-established guidelines neither exist for pre- and post-transplant vaccination nor do monitoring guidelines for pediatric lung transplant recipients. To ascertain the current vaccination and monitoring practices of pediatric lung transplant centers, a self-administered questionnaire was distributed to the 18 pediatric lung transplant centers within the International Pediatric Lung Transplant Collaborative in April 2006. Sixteen of 18 centers (89%) surveyed responded. Pretransplant, national vaccination guidelines are followed. Eleven centers reported following standardized vaccination guidelines post-transplant. Vaccines were more commonly provided by the primary-care physician pretransplant (69%) rather than post-transplant (38%). Post-transplant, 50% of the centers recommend live vaccines for household contacts but not for the transplant recipient. Pretransplant monitoring of response to prior vaccination was performed inconsistently except for varicella (88%). Only 44% of the transplant centers measure for response to vaccination post-transplant, mostly hepatitis B. Current vaccination practices of pediatric lung transplant centers are heterogeneous. The lung transplant community would be well served by studies designed to evaluate the efficacy of vaccinations in this population.


The microRNA 424/503 cluster reduces CDC25A expression during cell cycle arrest imposed by transforming growth factor β in mammary epithelial cells.
Recently, we demonstrated that the microRNA 424(322)/503 [miR-424(322)/503] cluster is transcriptionally controlled by transforming growth factor β (TGF-β) in the mammary epithelium. Induction of this microRNA cluster impacts mammary epithelium fate by regulating apoptosis and insulin-like growth factor 1 (IGF1) signaling. Here, we expanded our finding to demonstrate that miR-424(322)/503 is an integral component of the cell cycle arrest mediated by TGF-β. Mechanistically, we showed that after TGF-β exposure, increased levels of miR-424(322)/503 reduce the expression of the cell cycle regulator CDC25A. miR-424(322)/503-dependent posttranscriptional downregulation of CDC25A cooperates with previously described transcriptional repression of the CDC25A promoter and proteasome-mediated degradation to reduce the levels of CDC25A expression and to induce cell cycle arrest. We also provide evidence that the TGF-β/miR-424(322)/503 axis is part of the mechanism that regulates the proliferation of hormone receptor-positive (HR(+)) mammary epithelial cells in vivo.


[Bran in the treatment of irritable bowel syndrome].
The paper deals with the problem of using wheat bran in the treatment of patients with irritable colon syndrome expressed as spastic constipation. The effectiveness of the treatment with the wheat bran only (in a dose of 30-35 g/day), and in combination with drugs was comparatively studied in 105 patients. The bran fractions differing in the particle size, in the content of cellulose, starch and vitamins were used in the treatment. The combined therapy proved to be advantageous only in the rate of the clinical effect, while the acceleration of the movement along the large intestine did not depend on the treatment type. A long-term (during one year) follow-up of the patients showed that the bran intake led to the cessation of the disease relapse; when the bran was abolished the symptoms of the disease appeared in 11 out of 12 cases. The highest effect was recorded with the bran fraction containing 55.3% cellulose, 18.3% lignin, 157 micrograms tocopherol and the lowest amount of starch--18.0%. A conclusion has been made that the wheat bran are effective in the treatment and prevention of intestinal diseases, the effectiveness of the treatment depends on the summary content of food fibers in the nutrition.


The arthritis of inflammatory childhood myositis syndromes.
Arthritis has been an associated finding in juvenile dermatomyositis (JDM), but its prevalence, course, and response to therapy has not been well described. We investigated the frequency, course, and clinical and radiographic features in a large cohort of patients with JDM. The charts of 94 patients with idiopathic myositis (1984-99) were reviewed: 80 JDM, 3 juvenile polymyositis (JPM), 5 amyopathic JDM, and 6 overlap myositis syndromes. Compiled data included demographics, clinical features, a detailed description of the arthritis, investigations (radiographs, autoantibodies), course, and response to therapy. All radiographs were independently reviewed by a single radiologist. Sixty-one percent (95% CI 50-72%) of patients with JDM had arthritis. The arthritis was reported a median 4.5 mo (range -73.6 to 76.6 mo) after the JDM onset. When compared to patients with no arthritis, the occurrence of arthritis was not significantly related to sex, race, positive antinuclear antibody or rheumatoid factor, calcinosis, nodules, vasculitis, or Raynaud's phenomenon. The initial involvement was pauciarticular in 67% and polyarticular in 33%. In the pauci group, asymptomatic knee effusions were the predominant finding (n = 19, 58%), and in 18 patients may have been the result of steroid therapy. Two patients evolved from a pauci onset to a polyarticular course. All responded to therapy (corticosteroids; 47 were taking other medications) with remission of the arthritis within a median of 2.0 mo (range 0.1-64.5 mo). However, the arthritis recurred in 39% as the corticosteroids were tapered. Four patients with JDM eventually required corticosteroid wrist injections, with resolution of the arthritis. The arthritis was nonerosive in all cases. No patient with JPM had arthritis. Three of 5 patients with amyopathic JDM and 4 of 6 with overlap myositis syndrome had a nonerosive polyarthritis. Nonerosive arthritis involving the knees, wrists, elbows, and fingers is a frequent manifestation of JDM and other idiopathic childhood myositis. The arthritis is seen early in the course of JDM and often responds to treatment. However, the arthritis may recur with tapering of corticosteroids despite remission of the JDM. In a significant proportion of JDM cases, arthritis is the major sequela and may warrant further medical therapy or intraarticular corticosteroid injections.


5-HIAA in cerebrospinal fluid and deficit schizophrenic characteristics.
Higher CSF 5-HIAA concentrations and lower CSF HVA concentrations have been associated with various measures of slowed motor behaviour and communication in schizophrenic patients. To derive a single, reliable measure of deficit characteristics in schizophrenic patients, we entered four items of the BPRS reflecting negative symptoms, a work history measure derived from the Strauss-Carpenter scale, and three subscale scores of the WAIS-R into a principal-components analysis to derive a single factor score. The CSF 5-HIAA concentrations were within the normal range of values, and correlated directly with this factor score, but CSF HVA concentrations were not associated with the deficit factor score. These findings add support to the hypothesis that brain serotonin function is associated with deficit schizophrenic characteristics.


MicroRNA let-7c Inhibits Cell Proliferation and Induces Cell Cycle Arrest by Targeting CDC25A in Human Hepatocellular Carcinoma.
Down-regulation of the microRNA let-7c plays an important role in the pathogenesis of human hepatocellular carcinoma (HCC). The aim of the present study was to determine whether the cell cycle regulator CDC25A is involved in the antitumor effect of let-7c in HCC. The expression levels of let-7c in HCC cell lines were examined by quantitative real-time PCR, and a let-7c agomir was transfected into HCC cells to overexpress let-7c. The effects of let-7c on HCC proliferation, apoptosis and cell cycle were analyzed. The in vivo tumor-inhibitory efficacy of let-7c was evaluated in a xenograft mouse model of HCC. Luciferase reporter assays and western blotting were conducted to identify the targets of let-7c and to determine the effects of let-7c on CDC25A, CyclinD1, CDK6, pRb and E2F2 expression. The results showed that the expression levels of let-7c were significantly decreased in HCC cell lines. Overexpression of let-7c repressed cell growth, induced cell apoptosis, led to G1 cell cycle arrest in vitro, and suppressed tumor growth in a HepG2 xenograft model in vivo. The luciferase reporter assay showed that CDC25A was a direct target of let-7c, and that let-7c inhibited the expression of CDC25A protein by directly targeting its 3' UTR. Restoration of CDC25A induced a let-7c-mediated G1-to-S phase transition. Western blot analysis demonstrated that overexpression of let-7c decreased CyclinD1, CDK6, pRb and E2F2 protein levels. In conclusion, this study indicates that let-7c suppresses HCC progression, possibly by directly targeting the cell cycle regulator CDC25A and indirectly affecting its downstream target molecules. Let-7c may therefore be an effective therapeutic target for HCC.


AtREV1, a Y-family DNA polymerase in Arabidopsis, has deoxynucleotidyl transferase activity in vitro.
To clarify the functions of the Arabidopsis thaliana REV1 (AtREV1) protein, we expressed it in Escherichia coli and purified it to near homogeneity. The deoxynucleotidyl transferase activity of the recombinant AtREV1 was examined in vitro using a primer extension assay. The recombinant AtREV1 transferred one or two nucleotides to the primer end. It efficiently inserted dCMP regardless of the opposite base. AtREV1 also inserted a dCMP opposite an apurinic/apyrimidinic site, which is physiologically generated or induced by various DNA-damaging agents. In contrast, AtREV1 had no insertion activities against UV-inducible DNA lesions as reported in yeast or mammalian system. Although the substrate specificity of AtREV1 was rather narrow in the presence of magnesium ion, it widened in the presence of manganese ion. These results suggest that AtREV1 serves as a deoxycytidyl transferase in plant cells.


Corneal endothelial cell dysfunction: etiologies and management.
A transparent cornea is essential for the formation of a clear image on the retina. The human cornea is arranged into well-organized layers, and each layer plays a significant role in maintaining the transparency and viability of the tissue. The endothelium has both barrier and pump functions, which are important for the maintenance of corneal clarity. Many etiologies, including Fuchs' endothelial corneal dystrophy, surgical trauma, and congenital hereditary endothelial dystrophy, lead to endothelial cell dysfunction. The main treatment for corneal decompensation is replacement of the abnormal corneal layers with normal donor tissue. Nowadays, the trend is to perform selective endothelial keratoplasty, including Descemet stripping automated endothelial keratoplasty and Descemet's membrane endothelial keratoplasty, to manage corneal endothelial dysfunction. This selective approach has several advantages over penetrating keratoplasty, including rapid recovery of visual acuity, less likelihood of graft rejection, and better patient satisfaction. However, the global limitation in the supply of donor corneas is becoming an increasing challenge, necessitating alternatives to reduce this demand. Consequently, in vitro expansion of human corneal endothelial cells is evolving as a sustainable choice. This method is intended to prepare corneal endothelial cells in vitro that can be transferred to the eye. Herein, we describe the etiologies and manifestations of human corneal endothelial cell dysfunction. We also summarize the available options for as well as recent developments in the management of corneal endothelial dysfunction.


Molecular dynamics simulations of the adipocyte lipid binding protein reveal a novel entry site for the ligand.
The adipocyte lipid binding protein (ALBP) binds fatty acids (FA) in a cavity that is inaccessible from the bulk. Therefore, the penetration of the FA necessitates conformational changes whose nature is still unknown. It was suggested that the lipid first enters through a "portal region" which consists of the alphaII helix and the adjacent tight turns. The initial event in the ligand binding must be the interaction of the lipid with the protein surface. To analyze this interaction, we have carried out three molecular dynamics simulations of the apo-ALBP, with a palmitate ion initially located at different positions near the protein surface. The simulation indicated that the ligand could adsorb to the protein in more than one location. Yet, in one case, the ligand managed to penetrate the protein by entering a newly formed cavity some 10 A deep. The entry site is located near the N-terminus, at the junction between the loops connecting the beta-strands. Further progression of the penetration seems to be arrested by the need for desolvation of the COOH end of the headgroup. Evolutionary analysis showed that amino acids in this entry site are well conserved. On the basis of these observations, we suggest that the ligand may enter the protein from a site other than the portal region. Furthermore, the rate-limiting step is proposed to be the desolvation of the FA polar headgroup.


Altering Visual Perception Abnormalities: A Marker for Body Image Concern.
The body image concern (BIC) continuum ranges from a healthy and positive body image, to clinical diagnoses of abnormal body image, like body dysmorphic disorder (BDD). BDD and non-clinical, yet high-BIC participants have demonstrated a local visual processing bias, characterised by reduced inversion effects. To examine whether this bias is a potential marker of BDD, the visual processing of individuals across the entire BIC continuum was examined. Dysmorphic Concern Questionnaire (DCQ; quantified BIC) scores were expected to correlate with higher discrimination accuracy and faster reaction times of inverted stimuli, indicating reduced inversion effects (occurring due to increased local visual processing). Additionally, an induced global or local processing bias via Navon stimulus presentation was expected to alter these associations. Seventy-four participants completed the DCQ and upright-inverted face and body stimulus discrimination task. Moderate positive associations were revealed between DCQ scores and accuracy rates for inverted face and body stimuli, indicating a graded local bias accompanying increases in BIC. This relationship supports a local processing bias as a marker for BDD, which has significant assessment implications. Furthermore, a moderate negative relationship was found between DCQ score and inverted face accuracy after inducing global processing, indicating the processing bias can temporarily be reversed in high BIC individuals. Navon stimuli were successfully able to alter the visual processing of individuals across the BIC continuum, which has important implications for treating BDD.


Evidence-based development of school-based and family-involved prevention of overweight across Europe: the ENERGY-project's design and conceptual framework.
There is an urgent need for more carefully developed public health measures in order to curb the obesity epidemic among youth. The overall aim of the "EuropeaN Energy balance Research to prevent excessive weight Gain among Youth" (ENERGY)-project is the development and formative evaluation of a theory-informed and evidence-based multi-component school-based and family-involved intervention program ready to be implemented and evaluated for effectiveness across Europe. This program aims at promoting the adoption or continuation of health behaviors that contribute to a healthy energy balance among school-aged children. Earlier studies have indicated that school and family environments are key determinants of energy-balance behaviors in schoolchildren. Schools are an important setting for health promotion in this age group, but school-based interventions mostly fail to target and involve the family environment. Led by a multidisciplinary team of researchers from eleven European countries and supported by a team of Australian experts, the ENERGY-project is informed by the Environmental Research Framework for Weight gain Prevention, and comprises a comprehensive epidemiological analysis including 1) systematic reviews of the literature, 2) secondary analyses of existing data, 3) focus group research, and 4) a cross European school-based survey. The theoretical framework and the epidemiological analysis will subsequently inform stepwise intervention development targeting the most relevant energy balance-related behaviors and their personal, family-environmental and school-environmental determinants applying the Intervention Mapping protocol. The intervention scheme will undergo formative and pilot evaluation in five countries. The results of ENERGY will be disseminated among key stakeholders including researchers, policy makers and the general population. The ENERGY-project is an international, multidisciplinary effort to develop and test an evidence-based and theory-informed intervention program for obesity prevention among school-aged children.


Influence of Architecture on the Interfacial Properties of Polymers: Linear Chains, Stars, and Microgels.
Surface-active polymers have important applications as effective and responsive emulsifiers, foaming agents, and coatings. In this contribution, we explore the impact of the polymer architecture on the behavior at oil-water interfaces by comparing different poly(N-isopropylacrylamide) (pNIPAM)-based systems, namely, monolayers of linear and star-shaped macromolecules, ultralow cross-linked, regular cross-linked, and hollow microgels. Compression isotherms were determined experimentally as well as by computer simulations. The latter provides information about the conformational changes of the individual macromolecules as well as the interfacial properties of the monolayer, including the surface structure and the density distribution of an ensemble of interacting macromolecules near an interface. Surprisingly, the isotherms of the linear polymer, of the star polymer, and of the ultralow cross-linked microgel have an identical shape that differs from the isotherms of regular and hollow microgels. We introduced the mass fraction of adsorbed polymer, which gives a measure of the polymer segments contributing to the isotherm in relation to the most flexible architecture, i.e., the linear polymer, and allows a comparison of polymers with different architectures. The data demonstrate that increasing the number of cross-links leads to a significantly lower amount of polymer in the proximity of the interface as the increase in cross-linker reduces the deformability or softness of the polymers at the interface. The volume fraction profiles along the normal to the interface are essentially different in the microgel monolayers as compared to those in the linear and star polymer. The profiles through the microgel contact line and their growth upon initial compression are similar to those of the linear chains. Herewith, the profiles through the center of mass practically do not change upon compression. Therefore, the initial growth in the microgel surface pressure reveals the polymer-like behavior and is related to the deformation of the peripheral part of the microgel. Further compression of the microgel monolayer leads to 3D interactions of the microgels within the aqueous side of the interface and soft colloid-like behavior.


Understanding the intracellular effect of enhanced nutrient feeding toward high titer antibody production process.
One of the major goals in cell culture process development for therapeutic antibody production is to develop methods to reach high titer in classical fed-batch processes. This goal is often achieved through the optimizations of expression vector, cell line, media and cell culture process controls to increase cell specific productivity, viable cell density, and culture longevity. During process optimization for a selected production cell line, cell specific productivity (qP) can vary significantly with culture conditions. Therefore, identifying strategies to maintain maximal specific productivity throughout the entire fed-batch culture and to eliminate cellular/process bottlenecks that prevent high levels of antibody production would be crucial for further advancements in this area. In this work, specific productivity was increased and maintained at high level throughout the course of the culture by the optimization of feed media and feeding strategy. Through the enhancement of nutrient feeding, final titer was increased by 2.5-fold from the platform fed-batch process and reached 7.5 g/L. In addition, further insight upon possible cellular bottlenecks in high yield antibody production was obtained by comparing the levels of heavy chain (HC) and light chain (LC) mRNA and the levels of intracellular antibody between the non-optimized and optimized feeding processes. The mRNA levels of the two processes were measured and exhibited no significant difference suggesting that transcription is not the bottleneck. When intracellular antibody level was studied, the relatively constant level of HC, LC, and intact antibody between days 9 and 14 suggested that translation could be the rate-limiting step under the non-optimized nutrient feeding condition due to the dramatic drop of qP to roughly zero which correlated with the depletion of tyrosine as one of the key amino acids for protein synthesis. Finally, accumulation of unassembled HC but not intact antibody was observed at days 14-18 under the enhanced feeding condition, implying that folding and assembly may be the bottleneck toward the end of the culture.


Comparative effectiveness of traditional chemoembolization with or without sorafenib for hepatocellular carcinoma.
To compare the overall survival (OS) and progression-free survival (PFS) with associated adverse events (AE) in patients with unresectable hepatocellular carcinoma (HCC) treated with transarterial chemoembolization (TACE) + sorafenib vs TACE alone. In this retrospective cohort study we collected data on all consecutive patients with a diagnosis of unresectable HCC between 2007 and 2011 who had been treated with TACE + sorafenib or TACE alone. We hypothesized that the combination therapy is superior to TACE alone in improving the survival in these patients. Data extracted included patient's demographics, etiology of liver disease, histology of HCC, stage of liver disease with respect to model of end stage liver disease score and Child-Turcotte-Pugh (CTP) classification and Barcelona Clinic Liver Cancer (BCLC) staging for HCC. Computed tomography scan findings, alpha fetoprotein levels, number of treatments and related AE were also recorded and analyzed. Of the 43 patients who met inclusion criteria, 13 were treated with TACE + sorafenib and 30 with TACE alone. There was no significant difference in median survival: 20.6 mo (95%CI: 13.4-38.4) for the TACE + sorafenib and 18.3 mo (95%CI: 11.8-32.9) for the TACE alone (P = 0.72). There were also no statistically significant differences between groups in OS (HR = 0.82, 95%CI: 0.38-1.77; P = 0.61), PFS (HR = 0.93, 95%CI: 0.45-1.89; P = 0.83), and treatment-related toxicities (P = 0.554). CTP classification and BCLC staging for HCC were statistically significant (P = 0.001, P = 0.04 respectively) in predicting the survival in patients with HCC. The common AE observed were abdominal pain, nausea, vomiting and mild elevation of liver enzymes. Combination therapy with TACE + sorafenib is safe and equally effective as TACE alone in patients with unresectable HCC. CTP classification and BCLC staging were the significant predictors of survival. Future trials with large number of patients are needed to further validate this observation.


Modulation of transient receptor vanilloid 1 activity by transient receptor potential ankyrin 1.
Transient receptor potential vanilloid 1 (TRPV1) is a nonselective ligand-gated cation channel responding to noxious heat, protons, and chemicals such as capsaicin. TRPV1 is expressed in sensory neurons and plays a critical role in pain associated with tissue injury, inflammation, and nerve lesions. Transient receptor potential ankyrin 1 (TRPA1) is coexpressed with TRPV1. It is activated by compounds that cause a burning sensation (e.g., mustard oil) and, indirectly, by components of the inflammatory milieu eliciting nociceptor excitation and pain hypersensitivity. Previous studies indicate an interaction of TRPV1 and TRPA1 signaling pathways. Here we sought to examine the molecular mechanisms underlying such interactions in nociceptive neurons. We first excluded physical interactions of both channels using radioligand binding studies. By microfluorimetry, electrophysiological experiments, cAMP measurements, and site-directed mutagenesis we found a sensitization of TRPV1 after TRPA1 stimulation with mustard oil in a calcium and cAMP/protein kinase A (PKA)-dependent manner. TRPA1 stimulation enhanced TRPV1 phosphorylation via the putative PKA phosphorylation site serine 116. We also detected calcium-sensitive increased TRPV1 activity after TRPA1 activation in dorsal root ganglion neurons. The inhibition of TRPA1 by HC-030031 (1,2,3,6-tetrahydro-1,3-dimethyl-N-[4-(1-methylethyl)phenyl]-2,6-dioxo-7H-purine-7-acetamide, 2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-7H-purin-7-yl)-N-(4-isopropylphenyl)acetamide) after its initial stimulation (and the calcium-insensitive TRPA1 mutant D477A) still showed increased capsaicin-induced TRPV1 activity. This excludes a calcium-induced additive TRPA1 current after TRPV1 stimulation. Our study shows sensitization of TRPV1 via activation of TRPA1, which involves adenylyl cyclase, increased cAMP, subsequent translocation and activation of PKA, and phosphorylation of TRPV1 at PKA phosphorylation residues. This suggests that cross-sensitization of TRP channels contributes to enhanced pain sensitivity in inflamed tissues.


Heparan sulfate side chains have a critical role in the inhibitory effects of perlecan on vascular smooth muscle cell response to arterial injury.
Perlecan is a proteoglycan composed of a 470-kDa core protein linked to three heparan sulfate (HS) glycosaminoglycan chains. The intact proteoglycan inhibits the smooth muscle cell (SMC) response to vascular injury. Hspg2(Δ3/Δ3) (MΔ3/Δ3) mice produce a mutant perlecan lacking the HS side chains. The objective of this study was to determine differences between these two types of perlecan in modifying SMC activities to the arterial injury response, in order to define the specific role of the HS side chains. In vitro proliferative and migratory activities were compared in SMC isolated from MΔ3/Δ3 and wild-type mice. Proliferation of MΔ3/Δ3 SMC was 1.5× greater than in wild type (P < 0.001), increased by addition of growth factors, and showed a 42% greater migratory response than wild-type cells to PDGF-BB (P < 0.001). In MΔ3/Δ3 SMC adhesion to fibronectin, and collagen types I and IV was significantly greater than wild type. Addition of DRL-12582, an inducer of perlecan expression, decreased proliferation and migratory response to PDGF-BB stimulation in wild-type SMC compared with MΔ3/Δ3. In an in vivo carotid artery wire injury model, the medial thickness, medial area/lumen ratio, and macrophage infiltration were significantly increased in the MΔ3/Δ3 mice, indicating a prominent role of the HS side chain in limiting vascular injury response. Mutant perlecan that lacks HS side chains had a marked reduction in the inhibition of in vitro SMC function and the in vivo arterial response to injury, indicating the critical role of HS side chains in perlecan function in the vessel wall.


Conjugate chemistry, iodination and cellular binding of mEGF-dextran-tyrosine: preclinical tests in preparation for clinical trials.
A conjugate with specific binding to the epidermal growth factor receptor, EGFR, and of interest for clinical tests was prepared using mouse epidermal growth factor, mEGF, and dextran. The mEGF was first coupled to dextran by reductive amination in which the free amino group on the N-terminal of mEGF was reacted with the aldehyde group on the reductive end of the dextran chain. The end-end coupled intermediate was further activated by the cyanopyridinium agent CDAP and tyrosines introduced to the dextran part of the conjugate. The mEGF-dextran-tyrosine conjugate was, with high efficiency, iodinated with the chloramine-T method. Approximately 25-35% of the radioactivity could be removed from the conjugate after exposure to protease K while 65-75% of the radioactivity could be removed after exposure to dextranase. Thus, the largest amount of the iodine was on the dextran part of the conjugate. The iodinated mEGF-dextran-tyrosine had EGFR specific binding since the binding to an EGFR rich human glioma cell line could be displaced by an excess of non-radioactive mEGF. The conjugate was to a large extent internalized in these cells and the administrated radioactivity was thereby retained inside the cells for at least up to 50 h.


[Clear cell carcinoma of the thyroid; diagnostic difficulties].
Thyroid carcinomas with clear cell change are rare neoplasms, and always bring up the possibility of a metastasis from another primary clear cell carcinoma. We report the case of a 33-year-old woman operated on for a solitary cold thyroid nodule. Histologic and ultrastructural studies showed a clear cell carcinoma with a trabecular and follicular pattern. Neoplastic cells included lipid droplets, glycogen granules and intranuclear cytoplasmic inclusions, and were stained with one of the two antithyroglobulin polyclonal antibodies used. Radiologic investigations then showed a tumor mass of the right kidney. Surgical management proved it to be a benign cyst, and assessed the absence of kidney neoplasm. Thus, this case exemplifies the pitfalls in the diagnosis of clear cell thyroid neoplasms, and points out the fact that immunohistochemical staining for thyroglobulin should be systematically performed.


HTR2C Gene Variant and Salivary Cortisol Levels after Endurance Physical Activity: A Pilot Study.
The 5-hydroxytryptamine 2C receptor (HTR2C) rs6318 polymorphism has been associated with increased sensitivity to stress. This study investigated whether the rs6318 genotype modified the cortisol response to endurance physical activity. The HTR2C SNP was genotyped in a population of agonistic cyclists, and salivary cortisol levels were measured before and after an endurance competition. Salivary cortisol levels increased after the competition (from 20.72 ± 12.36 ng/mL to 33.80 ± 21.53 ng/mL; p = 3.189 × 10-5). rs6318 C carriers displayed higher baseline cortisol levels compared to G carriers (26.60 ± 9.35 ng/mL vs. 19.50 ± 12.63 ng/mL; p = 0.04). Baseline cortisol levels were able to predict the cortisol response to exercise (β = -0.846; p = 1.2 × 10-5). Although regression analysis did not identify an association between HTR2C genotype and change in cortisol levels, a secondary analysis in which the population was classified by median cortisol changes suggested that they might be weakly associated, thus warranting further investigation.


Surface Plasmon Resonance for Biomarker Detection: Advances in Non-invasive Cancer Diagnosis.
Biomarker-based cancer analysis has great potential to lead to a better understanding of disease at the molecular level and to improve early diagnosis and monitoring. Unlike conventional tissue biopsy, liquid biopsy allows the detection of a large variety of circulating biomarkers, such as microRNA (miRNA), exosomes, circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and proteins, in an easily accessible and minimally invasive way. In this review, we describe and evaluate the relevance and applicability of surface plasmon resonance (SPR) and localized SPR (LSPR)-based platforms for the detection of different classes of cancer biomarkers in liquid biopsy samples. Firstly, we critically discuss unsolved problems and issues in capturing and analyzing biomarkers. Secondly, we highlight current challenges which need to be resolved in applying SPR biosensors into clinical practice. Then, we mainly focus on applications of SPR-based platforms that process a patient sample aiming to detect and quantify biomarkers as a minimally invasive liquid biopsy tool for cancer patients appearing over the last 5 years. Finally, we describe the analytical performances of selected SPR biosensor assays and their significant advantages in terms of high sensitivity and specificity as well as accuracy and workflow simplicity.


High-Resolution Ion Mobility Spectrometry for Rapid Cannabis Potency Testing.
We report initial results from an ion mobility spectrometry (IMS)-based analysis of natural cannabis samples and explore the possibility of using this technique to distinguish medical marijuana from illegal forms of the drug, as defined by Swiss legislation. We analyzed cannabis extracts by electrospray ionization IMS-MS and found that high-resolution drift-tube IMS ( R > 150) can effectively isolate and quantify the controlled substance, Δ9-tetrahydrocannabinol (THC), even in the presence of other noncontrolled cannabinoid isomers including cannabidiol (CBD). We used this information to determine whether the THC content of a given sample surpassed the legal limit, which is 1% by weight in Switzerland. Our IMS-MS methodology produced equivalent quantification results to standard HPLC-based methods and offers the additional advantage of significantly shorter time requirements for the analysis. In addition, IMS-based analysis offers flexibility over HPLC in that it can be performed on portable devices. As such, these findings may have implications for cannabis testing in police laboratories.


Lack of association between hypothyroxinemia of prematurity and transient thyroid abnormalities with adverse long term neurodevelopmental outcome in very low birth weight infants.
The association between hypothyroxinemia of prematurity with neurodevelopment was controversial. To compare 5 year neurodevelopmental outcomes of very low birth weight (VLBW) infants with hypothyroxinemia of prematurity against those without. Retrospective cohort study in a single tertiary neonatal centre of VLBW infants born between the year 2008 to 2011. Comparisons were made between all abnormal and normal thyroid function controls using cord thyroid function tests, thyroid function tests during admission and pre-discharge thyroid function test done at term equivalent age. At 2 years corrected age, Bayley scales of infant and toddler development-third edition and Vineland II adaptive behaviour scales (VABS) were collected. At 5 years, Wechsler Preschool and Primary Scale of Intelligence (WPPSI-III), Bracken School Readiness Assessment, VABS and Beery Test of Visual-Motor Integration were collected. 110 subjects were studied at 2 years corrected age and 80 subjects at 5 years old. 29 infants had abnormal thyroid function test (10 infants with hypothyroxinemia of prematurity and 19 infants with transient thyroid abnormalities). There were no significant difference in the 2 years and 5 years developmental outcome between infants with and without hypothyroxinemia of prematurity (p-value>0.05); and between infants with and without transient thyroid abnormalities (p-value>0.05). There were no significant difference in neurological, visual and hearing impairment between infants with or without hypothyroxinemia of prematurity (p-value>0.05). Hypothyroxinemia of prematurity or transient thyroid abnormalities in VLBW infants were not associated with poorer neurodevelopment and did not support the need for levothyroxine supplementation.


Automated data-intensive forecasting of plant phenology throughout the United States.
Phenology, the timing of cyclical and seasonal natural phenomena such as flowering and leaf out, is an integral part of ecological systems with impacts on human activities like environmental management, tourism, and agriculture. As a result, there are numerous potential applications for actionable predictions of when phenological events will occur. However, despite the availability of phenological data with large spatial, temporal, and taxonomic extents, and numerous phenology models, there have been no automated species-level forecasts of plant phenology. This is due in part to the challenges of building a system that integrates large volumes of climate observations and forecasts, uses that data to fit models and make predictions for large numbers of species, and consistently disseminates the results of these forecasts in interpretable ways. Here, we describe a new near-term phenology-forecasting system that makes predictions for the timing of budburst, flowers, ripe fruit, and fall colors for 78 species across the United States up to 6 months in advance and is updated every four days. We use the lessons learned in developing this system to provide guidance developing large-scale near-term ecological forecast systems more generally, to help advance the use of automated forecasting in ecology.


Pulmonary artery thrombosis in experimental Angiostrongylus vasorum infection does not result in pulmonary hypertension and echocardiographic right ventricular changes.
Dogs experimentally inoculated with Angiostrongylus vasorum develop severe pulmonary parenchymal lesions and arterial thrombosis at the time of patency. A. vasorum-induced thrombosis results in arterial hypoxemia, pulmonary hypertension (PH), and altered cardiac morphology and function. Six healthy Beagles experimentally inoculated with A. vasorum. Thoracic radiographs and arterial blood gas analyses were performed 8 and 13 weeks postinoculation (wpi) and 9 weeks posttherapy (wpt). Echocardiography was done before and 2, 5, 8, 13 wpi and 9 wpt. Invasive pulmonary artery pressure (PAP) measurements were obtained 8 wpi. Two untreated dogs were necropsied 13 wpi and 4 treated dogs 9 wpt. All dogs had patent infections at 7 wpi and clinical respiratory signs at 8 wpi. Moderate hypoxemia (median PaO2 of 73 and 74 mmHg) present at 8 and 13 wpi had resolved by 9 wpt. Echocardiographically, no evidence of PH and no abnormalities in cardiac size and function were discernible at any time point. PAP invasively measured at 8 wpi was not different from that of control dogs. Severe radiographic pulmonary parenchymal and suspected thrombotic lesions at 13 wpi were corroborated by necropsy. Most histopathologic changes had resolved at 9 wpt, but focal inflammatory, thrombotic, and fibrotic changes still were present in all dogs. In experimentally infected Beagles, pulmonary and vascular changes induced by A. vasorum are reflected by marked radiographic changes and arterial hypoxemia. These did not result in PH and echocardiographic changes in cardiac size and function.


Measures of Aging Biology in Saliva and Blood as Novel Biomarkers for Stroke and Heart Disease in Older Adults.
The role of aging biology as a novel risk factor and biomarker for vascular outcomes in different accessible body tissues such as saliva and blood remain unclear. We aimed to (1) assess the role of aging biology as a risk factor of stroke and heart disease among individuals of same chronologic age and sex and (2) compare aging biology biomarkers measured in different accessible body tissues as novel biomarkers for stroke and heart disease in older adults. This study included individuals who consented for blood and saliva draw in the Venous Blood Substudy and Telomere Length Study of the Health and Retirement Study (HRS). The HRS is a population-based, nationally representative longitudinal survey of individuals aged 50 years and older in the United States. Saliva-based measures included telomere length. Blood-based measures included DNA methylation and physiology biomarkers. Propensity scores-matched analyses and Cox regression models were conducted. This study included individuals aged 50 years and older, who consented for blood (N = 9,934) and saliva (N = 5,808) draw in the HRS. Blood-based biomarkers of aging biology showed strong associations with incident stroke as follows: compared with the lowest tertile of blood-based biomarkers of aging, biologically older individuals had significantly higher risk of stroke based on DNA methylation Grim Age clock (adjusted hazard ratio [aHR] = 2.64, 95% CI 1.90-3.66, p < 0.001) and Physiology-based Phenotypic Age clock (aHR = 1.75, 95% CI 1.27-2.42, p < 0.001). In secondary analysis, biologically older individuals had increased risk of heart disease as follows: DNA methylation Grim Age clock (aHR = 1.77, 95% CI 1.49-2.11, p < 0.001) and Physiology-based Phenotypic Age clock (aHR = 1.61, 95% CI 1.36-1.90, p < 0.001). Compared with saliva-based telomere length, blood-based aging physiology and some DNA methylation biomarkers are strongly associated with vascular disorders including stroke and are more precise and sensitive biomarkers of aging. Saliva-based telomere length and blood-based DNA methylation and physiology biomarkers likely represent different aspects of biological aging and accordingly vary in their precision as novel biomarkers for optimal vascular health.


Decorin accumulation contributes to the stromal opacities found in congenital stromal corneal dystrophy.
Congenital stromal corneal dystrophy (CSCD) is characterized by stromal opacities that morphologically are seen as interlamellar layers of amorphous substance with small filaments, the nature of which has hitherto been unknown. CSCD is associated with truncating mutations in the decorin gene (DCN). To understand the molecular basis for the corneal opacities we analyzed the expression of decorin in this disease, both at the morphologic and the molecular level. Corneal specimens were examined after contrast enhancement with cuprolinic blue and by immunoelectron microscopy. Decorin protein from corneal tissue and keratocyte culture was studied by immunoblot analysis before and after O- and N-deglycosylation. The relative level of DCN mRNA expression was examined using Q-RT-PCR, and cDNA was sequenced. Recombinant wild-type and truncated decorin transiently expressed in HEK293 cells were analyzed by gel filtration and immunoblotting. The areas of interlamellar filaments were stained by cuprolinic blue. Immunoelectron microscopy using decorin antibodies revealed intense labeling of these areas. Both wild-type and truncated decorin protein was expressed in corneal tissue and keratocytes of affected persons. When decorin expressed in HEK293 cells was examined by gel filtration, the truncated decorin eluted as high molecular weight aggregates. Accumulation of decorin was found in the interlamellar areas of amorphous substance. The truncated decorin is present in CSCD corneas, and there is evidence it may aggregate in vitro. Thus, decorin accumulation appears to contribute to the stromal opacities that are characteristic of CSCD.


Osteopathic interventions via telehealth in a pediatric population: a retrospective case series.
Healthcare delivery was dramatically affected during the coronavirus disease 2019 (COVID-19) pandemic. Many outpatient visits were cancelled or forgone for fear of exposure to the virus, allowing telemedicine to take on a much larger role in healthcare. The delivery of manual therapies, such as osteopathic manipulative treatment (OMT), via telehealth posed a unique challenge as these are typically provided in-person by a trained osteopathic physician. This study provides a description of one osteopathic pediatrician's experience in delivering osteopathic interventions to pediatric patients via telehealth. To our knowledge, these techniques have not previously been described in the literature. To detail the experience of one osteopathic pediatrician's experience in delivering osteopathic interventions via telehealth. Patients were offered the option of converting their existing OMT appointment to a telehealth visit. Prior to the appointment, instructions were emailed to the patient's parent or guardian along with a voluntary survey to provide feedback. Thirty-minute telehealth visits were conducted during which the provider gave verbal and visual instructions to a parent or guardian over a video platform to guide them in providing treatment to the patient based on osteopathic principles. Patients aged 3 and older rated their pain before and after the appointment using the Wong-Baker FACES scale. Deidentified patient demographics, chief complaints, treatments, anatomic locations, and pain scores were recorded in a REDcap database. Descriptive statistics were analyzed and paired samples t-tests were used with a p-value of <0.05 used to determine significance. Eighteen patients ranging from 6 months to 19 years of age were treated utilizing osteopathic interventions via telehealth during 54 distinct visits. The most common chief complaints treated were back (n=31; 26.3%) and neck (n=28; 23.7%) pain. The most common osteopathic techniques upon which instruction was based were inhibition (n=131; 29.7%) soft tissue (n=127; 28.8%) and counterstrain (n=78; 17.7%). The average post-treatment pain score (2.57) was significantly lower than the average pre-treatment pain score (6.77) p<0.01. No serious complications were observed. In our small retrospective case series, osteopathic interventions via telehealth resulted in decreased average pain scores following treatment while minimizing risk of viral exposure and transmission. Further study is needed to determine if such treatment methods could be effective on a larger scale when distance or illness preclude an in-person OMT visit.


Effects of α-blocker therapy on active duty military and military retirees for benign prostatic hypertrophy on diabetic complications.
Determine if men with type 2 diabetes who take α-blockers (ABs) for benign prostatic hypertrophy gain additional benefit with reduced diabetic complications. Chart review of 1,100 men with type 2 diabetes and benign prostatic hypertrophy taking either an angiotensin-converting enzyme inhibitor or angiotensin-receptor blocker. Of the 1,100 men, 330 took ABs and 770 did not take ABs. Despite no difference in blood pressure between men taking or not taking ABs, those taking them had more evidence of renal and cardiovascular disease. The prevalence of complications varied among the AB types with tamsulosin users having more coronary artery disease diagnoses and doxazosin users having more renal disease diagnoses. ABs when prescribed for benign prostatic hypertrophy not only failed to give additional protection against developing diabetic complications but were associated with more cardiovascular and renal disease diagnoses. Prospective randomized controlled trials are necessary to determine if there is a causal relationship between ABs and adverse outcomes in patients with type 2 diabetes and benign prostatic hypertrophy already on an angiotensin converting enzyme inhibitor or angiotensin-receptor blocker.


Do urban open spaces provide refugia for frogs in urban environments?
Urbanization is among the largest threats to wildlife populations through factors such as fragmentation, isolation, and habitat destruction. Urban open spaces, such as parks and golf courses, have the potential to provide wildlife with suitable habitat within an urbanized matrix. These refugia may be particularly important for amphibians, which represent one of the most endangered and least vagile vertebrate groups on earth. During the spring and summer of 2018, we conducted surveys to determine the presence of anurans at 51 wetland sites within the Piedmont ecoregion of South Carolina. Nearly one-third of these wetlands were located within urban open spaces, one-third in low development areas, and one-third in highly developed areas. Impervious surface and total road length surrounding the wetlands were measured at two scales, a core habitat scale (300 m) and average maximum migration scale (750 m), and we measured several within-wetland habitat variables. Urban Open Space wetlands had levels of surrounding impervious surface similar to High Urbanization wetlands at the larger scale and were intermediate between Low and High Urbanization wetlands at the smaller scale. The total length of road segments occurring within buffers (at both scales) surrounding our study wetlands was higher for Urban Open Space compared to Low and High Urbanization sites. Among the within-wetland variables measured, Low Urbanization sites had higher canopy cover and were more likely to have a terrestrial buffer zone relative to the other categories. Species richness decreased significantly as total road length increased among all wetlands. Wetland category was not a significant driver explaining species richness, but β-diversity was more variable among Urban Open Space wetlands than either Low or High Urbanization wetlands. Urban Open Space wetlands did not appear to increase suitability for anurans relative to High Urbanization wetlands. Urban Open Space wetlands had higher variability in species composition, which was perhaps attributable to the diversity among sites represented in the Urban Open Space category.


A Novel Mitochondrial Inhibitor Blocks MAPK Pathway and Overcomes MAPK Inhibitor Resistance in Melanoma.
The purpose of this study is to determine if inhibition of mitochondrial oxidative phosphorylation (OxPhos) is an effective strategy against MAPK pathway inhibitor (MAPKi)-resistant BRAF-mutant melanomas.Experimental Design: The antimelanoma activity of IACS-010759 (OPi), a novel OxPhos complex I inhibitor, was evaluated in vitro and in vivo. Mechanistic studies and predictors of response were evaluated using molecularly and metabolically stratified melanoma cell lines. 13C-labeling and targeted metabolomics were used to evaluate the effect of OPi on cellular energy utilization. OxPhos inhibition in vivo was evaluated noninvasively by [18F]-fluoroazomycin arabinoside (FAZA) PET imaging. OPi potently inhibited OxPhos and the in vivo growth of multiple MAPKi-resistant BRAF-mutant melanoma models with high OxPhos at well-tolerated doses. In vivo tumor regression with single-agent OPi treatment correlated with inhibition of both MAPK and mTOR complex I activity. Unexpectedly, antitumor activity was not improved by combined treatment with MAPKi in vitro or in vivo. Signaling and growth-inhibitory effects were mediated by LKB1-AMPK axis, and proportional to AMPK activation. OPi increased glucose incorporation into glycolysis, inhibited glucose and glutamine incorporation into the mitochondrial tricarboxylic acid cycle, and decreased cellular nucleotide and amino acid pools. Early changes in [18F]-FAZA PET uptake in vivo, and the degree of mTORC1 pathway inhibition in vitro, correlated with efficacy. Targeting OxPhos with OPi has significant antitumor activity in MAPKi-resistant, BRAF-mutant melanomas, and merits further clinical investigation as a potential new strategy to overcome intrinsic and acquired resistance to MAPKi in patients.


Student, teacher, and classroom predictors of between-teacher variance of students' teacher-rated behavior.
The current study examined between-teacher variance in teacher ratings of student behavioral and emotional risk to identify student, teacher and classroom characteristics that predict such differences and can be considered in future research and practice. Data were taken from seven elementary schools in one school district implementing universal screening, including 1,241 students rated by 68 teachers. Students were mostly African America (68.5%) with equal gender (female 50.1%) and grade-level distributions. Teachers, mostly White (76.5%) and female (89.7%), completed both a background survey regarding their professional experiences and demographic characteristics and the Behavior Assessment System for Children (Second Edition) Behavioral and Emotional Screening System-Teacher Form for all students in their class, rating an average of 17.69 students each. Extant student data were provided by the district. Analyses followed multilevel linear model stepwise model-building procedures. We detected a significant amount of variance in teachers' ratings of students' behavioral and emotional risk at both student and teacher/classroom levels with student predictors explaining about 39% of student-level variance and teacher/classroom predictors explaining about 20% of between-teacher differences. The final model fit the data (Akaike information criterion = 8,687.709; pseudo-R2 = 0.544) significantly better than the null model (Akaike information criterion = 9,457.160). Significant predictors included student gender, race ethnicity, academic performance and disciplinary incidents, teacher gender, student-teacher gender interaction, teacher professional development in behavior screening, and classroom academic performance. Future research and practice should interpret teacher-rated universal screening of students' behavioral and emotional risk with consideration of the between-teacher variance unrelated to student behavior detected. (PsycINFO Database Record


Prevalence and impact of cardiovascular metabolic diseases on COVID-19 in China.
Studies have reminded that cardiovascular metabolic comorbidities made patients more susceptible to suffer 2019 novel corona virus (2019-nCoV) disease (COVID-19), and exacerbated the infection. The aim of this analysis is to determine the association of cardiovascular metabolic diseases with the development of COVID-19. A meta-analysis of eligible studies that summarized the prevalence of cardiovascular metabolic diseases in COVID-19 and compared the incidences of the comorbidities in ICU/severe and non-ICU/severe patients was performed. Embase and PubMed were searched for relevant studies. A total of six studies with 1527 patients were included in this analysis. The proportions of hypertension, cardia-cerebrovascular disease and diabetes in patients with COVID-19 were 17.1%, 16.4% and 9.7%, respectively. The incidences of hypertension, cardia-cerebrovascular diseases and diabetes were about twofolds, threefolds and twofolds, respectively, higher in ICU/severe cases than in their non-ICU/severe counterparts. At least 8.0% patients with COVID-19 suffered the acute cardiac injury. The incidence of acute cardiac injury was about 13 folds higher in ICU/severe patients compared with the non-ICU/severe patients. Patients with previous cardiovascular metabolic diseases may face a greater risk of developing into the severe condition and the comorbidities can also greatly affect the prognosis of the COVID-19. On the other hand, COVID-19 can, in turn, aggravate the damage to the heart.


Kinetics of functional beta cell mass decay in a diphtheria toxin receptor mouse model of diabetes.
Functional beta cell mass is an essential biomarker for the diagnosis and staging of diabetes. It has however proven technically challenging to study this parameter during diabetes progression. Here we have detailed the kinetics of the rapid decline in functional beta cell mass in the RIP-DTR mouse, a model of hyperglycemia resulting from diphtheria toxin induced beta cell ablation. A novel combination of imaging modalities was employed to study the pattern of beta cell destruction. Optical projection tomography of the pancreas and longitudinal in vivo confocal microscopy of islets transplanted into the anterior chamber of the eye allowed to investigate kinetics and tomographic location of beta cell mass decay in individual islets as well as at the entire islet population level. The correlation between beta cell mass and function was determined by complementary in vivo and ex vivo characterizations, demonstrating that beta cell function and glucose tolerance were impaired within the first two days following treatment when more than 50% of beta cell mass was remaining. Our results illustrate the importance of acquiring quantitative functional and morphological parameters to assess the functional status of the endocrine pancreas.


Heterochronic misexpression of Ascl1 in the Atoh7 retinal cell lineage blocks cell cycle exit.
Retinal neurons and glia arise from a common progenitor pool in a temporal order, with retinal ganglion cells (RGCs) appearing first, and Müller glia last. The transcription factors Atoh7/Math5 and Ascl1/Mash1 represent divergent bHLH clades, and exhibit distinct spatial and temporal retinal expression patterns, with little overlap during early development. Here, we tested the ability of Ascl1 to change the fate of cells in the Atoh7 lineage when misexpressed from the Atoh7 locus, using an Ascl1-IRES-DsRed2 knock-in allele. In Atoh7(Ascl1KI/+) and Atoh7(Ascl1KI/Ascl1KI) embryos, ectopic Ascl1 delayed cell cycle exit and differentiation, even in cells coexpressing Atoh7. The heterozygous retinas recovered, and eventually produced a normal complement of RGCs, while homozygous substitution of Ascl1 for Atoh7 did not promote postnatal retinal fates precociously, nor rescue Atoh7 mutant phenotypes. However, our analyses revealed two unexpected findings. First, ectopic Ascl1 disrupted cell cycle progression within the marked Atoh7 lineage, but also nonautonomously in other retinal cells. Second, the size of the Atoh7 retinal lineage was unaffected, supporting the idea of a compensatory shift of the non-proliferative cohort to maintain lineage size. Overall, we conclude that Ascl1 acts dominantly to block cell cycle exit, but is incapable of redirecting the fates of early RPCs.


A framework of joint graph embedding and sparse regression for dimensionality reduction.
Over the past few decades, a large number of algorithms have been developed for dimensionality reduction. Despite the different motivations of these algorithms, they can be interpreted by a common framework known as graph embedding. In order to explore the significant features of data, some sparse regression algorithms have been proposed based on graph embedding. However, the problem is that these algorithms include two separate steps: (1) embedding learning and (2) sparse regression. Thus their performance is largely determined by the effectiveness of the constructed graph. In this paper, we present a framework by combining the objective functions of graph embedding and sparse regression so that embedding learning and sparse regression can be jointly implemented and optimized, instead of simply using the graph spectral for sparse regression. By the proposed framework, supervised, semisupervised, and unsupervised learning algorithms could be unified. Furthermore, we analyze two situations of the optimization problem for the proposed framework. By adopting an ℓ2,1-norm regularization for the proposed framework, it can perform feature selection and subspace learning simultaneously. Experiments on seven standard databases demonstrate that joint graph embedding and sparse regression method can significantly improve the recognition performance and consistently outperform the sparse regression method.


On/off-switchable LSPR nano-immunoassay for troponin-T.
Regeneration of immunosensors is a longstanding challenge. We have developed a re-usable troponin-T (TnT) immunoassay based on localised surface plasmon resonance (LSPR) at gold nanorods (GNR). Thermosensitive poly(N-isopropylacrylamide) (PNIPAAM) was functionalised with anti-TnT to control the affinity interaction with TnT. The LSPR was extremely sensitive to the dielectric constant of the surrounding medium as modulated by antigen binding after 20 min incubation at 37 °C. Computational modelling incorporating molecular docking, molecular dynamics and free energy calculations was used to elucidate the interactions between the various subsystems namely, IgG-antibody (c.f., anti-TnT), PNIPAAM and/or TnT. This study demonstrates a remarkable temperature dependent immuno-interaction due to changes in the PNIPAAM secondary structures, i.e., globular and coil, at above or below the lower critical solution temperature (LCST). A series of concentrations of TnT were measured by correlating the λLSPR shift with relative changes in extinction intensity at the distinct plasmonic maximum (i.e., 832 nm). The magnitude of the red shift in λLSPR was nearly linear with increasing concentration of TnT, over the range 7.6 × 10-15 to 9.1 × 10-4 g/mL. The LSPR based nano-immunoassay could be simply regenerated by switching the polymer conformation and creating a gradient of microenvironments between the two states with a modest change in temperature.


Incubation with macroalgae induces large shifts in water column microbiota, but minor changes to the epibiota of co-occurring macroalgae.
Macroalgae variably promote and deter microbial growth through release of organic carbon and antimicrobial compounds into the water column. Consequently, macroalgae influence the microbial composition of the surrounding water column and biofilms on nearby surfaces. Here, we use manipulative experiments to test the hypotheses that (i) Nereocystis luetkeana and Mastocarpus sp. macroalgae alter the water column microbiota in species-specific manner, that (ii) neighbouring macroalgae alter the bacterial communities on the surface (epibiota) of actively growing Nereocystis luetkeana meristem fragments (NMFs), and that (iii) neighbours alter NMF growth rate. We also assess the impact of laboratory incubation on macroalgal epibiota by comparing each species to wild counterparts. We find strong differences between the Nereocystis and Mastocarpus epibiota that are maintained in the laboratory. Nereocystis and Mastocarpus alter water column bacterial community composition and richness in a species specific manner, but cause only small compositional shifts on NMF surfaces that do not differ by species, and do not change richness. Co-incubation with macroalgae results in significant change in abundance of fivefold more genera in the water column compared to NMF surfaces, although the direction (i.e., enrichment or reduction) of shift is generally consistent between the water and NMF surfaces. Finally, NMFs grew during the experiment, but growth did not depend on the presence or identity of neighbouring macroalgae. Thus, macroalgae exhibit a strong and species-specific influence on the water column microbiota, but a much weaker influence on the epibiota of neighbouring macroalgae. Overall, these results support the idea that macroalgae surfaces are highly selective and demonstrate that modulations of macroalgal microbiota operate within an overarching paradigm of host species specificity.


Apraxia, autism, attention-deficit hyperactivity disorder: do we have a new spectrum?
Gio is a bilingual 6-year 10-month-old boy new to your practice who presents for an unscheduled visit with concerns for speech and language delay. He was born in Portugal, and his native language is Portuguese. When he was 21 months old, his family moved to Italy and then moved to the United States 3 years later. He had very little contact with other children while living in Italy, but his parents report that he has made friends quickly in the United States. His family speaks Portuguese at home, although his father is fluent in English. He started school 3 months after moving to the United States and is currently repeating kindergarten. He is in a sheltered English classroom with several other students who speak Portuguese. He is able to understand and follow directions in English. A recent school evaluation revealed solidly average nonverbal reasoning skills and relative weaknesses in verbal reasoning and working memory. His speech is described as unintelligible in conversation, both in English and Portuguese. Results of a special education evaluation qualified him for services with a bilingual therapist. His teachers are very concerned that he may have autism spectrum disorder (ASD) and attention-deficit hyperactivity disorder (ADHD). They describe him as having limited interest in other children, poor eye contact, and hypersensitivities. He wanders at recess. He is very skilled at art and seems to prefer to draw rather than interact with others. He needs constant support and redirection throughout the school day. He has difficulty putting on his coat, using playground equipment, and following daily classroom routines. On the Vanderbilt Rating Scale, his teacher endorses 17 of 18 ADHD symptoms as present often or very often and significant impairment in his performance. Gio presents to your clinic as a relatable young boy with childhood apraxia of speech. Only his productions of single words and short routine phrases are intelligible. He attempts to engage in conversation but averts his gaze and becomes frustrated when asked to repeat things. Scores on the Parent Conners Rating Scale and Social Responsiveness Scale are not elevated. When you bring up school's concerns, his father describes feeling somewhat badgered by his teachers about possibility of ASD. School is considering placement in an inclusion classroom for children with ASD. What do you recommend? How would you advise his parents?


Validity of caregivers' reports on prior use of antibacterials in children under five years presenting to health facilities in Gulu, northern Uganda.
Given the frequent initiation of antibacterial treatment at home by caregivers of children under five years in low-income countries, there is a need to find out whether caregivers' reports of prior antibacterial intake by their children before being brought to the healthcare facility are accurate. The aim of this study was to describe and validate caregivers' reported use of antibacterials by their children prior to seeking care at the healthcare facility. A cross sectional study was conducted among children under five years seeking care at healthcare facilities in Gulu district, northern Uganda. Using a researcher administered questionnaire, data were obtained from caregivers regarding reported prior antibacterial intake in their children. These reports were validated by comparing them to common antibacterial agents detected in blood and urine samples from the children using liquid chromatography with tandem mass spectrometry (LC-MS/MS) methods. A total of 355 study participants had a complete set of data on prior antibacterial use collected using both self-report and LC-MS/MS. Of the caregivers, 14.4% (51/355, CI: 10.9-18.5%) reported giving children antibacterials prior to visiting the healthcare facility. However, LC-MS/MS detected antibacterials in blood and urine samples in 63.7% (226/355, CI: 58.4-68.7%) of the children. The most common antibacterials detected from the laboratory analysis were cotrimoxazole (29%, 103/355), ciprofloxacin (13%, 46/355), and metronidazole (9.9%, 35/355). The sensitivity, specificity, positive predictive value (PPV), negative predictive value and agreement of self-reported antibacterial intake prior to healthcare facility visit were 17.3% (12.6-22.8), 90.7% (84.3-95.1), 76.5% (62.5-87.2), 38.5% (33.0-44.2) and 43.9% (k 0.06) respectively. There is low validity of caregivers' reports on prior intake of antibacterials by these children. There is need for further research to understand the factors associated with under reporting of prior antibacterial use.


Biological characteristics of type C viruses isolated from different Friend erythroleukaemic cells.
We have examined the expression of type C RNA viruses in different Friend erythroleukaemic cell types, distinguished on the basis of increasingly malignant characteristics, which arise during the ageing of mice inoculated with the polycythaemic Friend virus complex. Most early appearing Friend cells (type I) expressed ecotropic virus, but cells of later malignant types showed decreased and variable expression. In general, the more malignant cells released less ecotropic virus. Xenotropic virus was detected in low numbers from type I, II, and IV cells. Two viruses were cloned from type II tumour cells: a xenotropic virus (II clone 1) and an N-tropic ecotropic virus (II clone 2). No pathogenic activity was found when II clone 1 was inoculated into newborn and adult DBA/2J and NIH/Swiss mice observed for up to 20 months, whereas II clone 2 caused a rapid anaemic erythroleukaemia in both N- and B-type newborn mouse strains. It caused a similar form of leukaemia in susceptible N-type adult mice, but at a lower frequency and with a longer latency (usually greater than 5 months). This finding demonstrated a lack of NB restriction in newborn mice. The virus was much less active in DBA/2J mice from which it had been originally cloned; it also appeared to cause lymphoma or to shorten the latency of spontaneous lymphoma in DBA/2J mice.


The influence of English proficiency on access to care.
The number of individuals with limited English proficiency in the USA is large and rapidly growing. Consequently, addressing language barriers in access to medical care is becoming increasingly important. Previous studies have reported that individuals with limited English proficiency have more difficulty gaining access to care, compared to English-proficient individuals. We assessed the impact of English language proficiency on access to medical care, accounting for health and socioeconomic status, using nationally representative data. Cross-sectional data from the 2006 National Health Interview Survey (n=29,868). The main outcome measures of interest were self-reported delayed medical care, forgone needed care, and visits to a health care professional. In unadjusted analyses, individuals with limited English proficiency were more likely to forgo needed medical care and less likely to have a health care visit, compared to individuals who were proficient in English. There was no significant association between language proficiency and reports of delayed care. After accounting for individuals' health and socioeconomic status, only the relationship between limited English proficiency and health care visits remained statistically significant. Most associations between language proficiency and access to care did not differ across various racial/ethnic groups. Results indicate that the choice of access measure may influence conclusions about language barriers in health care. Given the growing proportion of US residents with limited English proficiency, health care settings need to better address potential language barriers.


The Influence of Hepatitis C Virus Genetic Region on Phylogenetic Clustering Analysis.
Sequencing is important for understanding the molecular epidemiology and viral evolution of hepatitis C virus (HCV) infection. To date, there is little standardisation among sequencing protocols, in-part due to the high genetic diversity that is observed within HCV. This study aimed to develop a novel, practical sequencing protocol that covered both conserved and variable regions of the viral genome and assess the influence of each subregion, sequence concatenation and unrelated reference sequences on phylogenetic clustering analysis. The Core to the hypervariable region 1 (HVR1) of envelope-2 (E2) and non-structural-5B (NS5B) regions of the HCV genome were amplified and sequenced from participants from the Australian Trial in Acute Hepatitis C (ATAHC), a prospective study of the natural history and treatment of recent HCV infection. Phylogenetic trees were constructed using a general time-reversible substitution model and sensitivity analyses were completed for every subregion. Pairwise distance, genetic distance and bootstrap support were computed to assess the impact of HCV region on clustering results as measured by the identification and percentage of participants falling within all clusters, cluster size, average patristic distance, and bootstrap value. The Robinson-Foulds metrics was also used to compare phylogenetic trees among the different HCV regions. Our results demonstrated that the genomic region of HCV analysed influenced phylogenetic tree topology and clustering results. The HCV Core region alone was not suitable for clustering analysis; NS5B concatenation, the inclusion of reference sequences and removal of HVR1 all influenced clustering outcome. The Core-E2 region, which represented the highest genetic diversity and longest sequence length in this study, provides an ideal method for clustering analysis to address a range of molecular epidemiological questions.


The Mozart study: a relation between dynamic hyperinflation and physical activity in patients with chronic obstructive pulmonary disease?
Many patients with chronic obstructive pulmonary disease (COPD) experience dyspnoea during exercise, resulting in a reduction of physical activity (PA). Dynamic hyperinflation (DH) is seen as a major cause of dyspnoea in COPD. The objective of the current study was to investigate the relationship between DH, in terms of the amount of DH and the development and recovery rate of DH in patients with COPD, and PA. Thirty-five patients with stable COPD were included from an outpatient clinic (14 GOLD II and 21 GOLD III, median age 65). PA was assessed using an accelerometer. Subjects underwent metronome-paced tachypnoea (MPT) to induce DH. To quantify the amount of DH during MPT, a decrease in inspiratory capacity (IC) or a change in IC as percentage of total lung capacity was used. No significant correlations were found between the parameters describing DH and PA. Secondary correlation analyses showed a negative correlation between static hyperinflation (SH) and PA (r = -0·39; P = 0·02). The pattern of breathing during MPT and the test itself showed high interpatient variability. The absence of a significant correlation between DH and PA is contrary to previous studies. SH did show a correlation with PA. The variety in results and the technical difficulties in execution of the measurements ask for a new, more reliable, method to detect DH and investigate its relation with PA in patients with COPD.


Transparency around abortion training on obstetrics and gynecology residency program websites.
The availability of family planning and abortion training in residency is a concern for applicants, particularly following the overturning of Roe v Wade. We aimed to characterize public information on abortion training in obstetrics and gynecology residency programs in states with and without abortion bans. We abstracted residency program data using a publicly available database of obstetrics and gynecology residency programs. We performed a systematic internet search of each program's website to identify information on abortion training. We defined states with abortion bans as those with total or near-total bans as of November 2022. Using Stata SE 16, we used Student t tests and Fisher exact tests to characterize differences between programs with public abortion training information and those without. Of 293 obstetrics and gynecology residency programs, 197 (67.2%) included online information regarding abortion training. Of 64 programs in states with abortion bans, 34 (53%) had public information regarding abortion training, compared to 163 (71%) of the 229 programs in states with legal abortion. Programs with publicly available abortion training information were more likely to be academic (49.8% vs 26%, p < 0.001) and have a higher percentage of female residents (86.6% vs 82.9%, p = 0.003). Programs in states with legal abortion were more likely to use words like "abortion" (36.2% vs 17.7%, p = 0.05). Only two-thirds of obstetrics and gynecology residency programs publicize abortion training information. Almost half of all programs in states with abortion bans do not share this information, revealing a need for improved transparency to better inform residency applicant decision-making. During the 2022-2023 academic cycle, one-third of obstetrics and gynecology residency programs did not include information about family planning or abortion training online. In states with abortion bans, this number increased to almost one-half. These findings serve as a call to action for programs to clarify their commitment to abortion training.