Carotid stiffness, extra-media thickness and visceral adiposity in young adults.
Carotid extra-media thickness (EMT) encompasses arterial adventitia and perivascular adipose tissue (PAT). Adventitial remodeling and PAT may contribute independently to functional (stiffness) and structural (remodeling) changes in artery wall properties. Visceral adiposity may contribute to PAT, thereby affecting artery stiffness. We investigated the relationships between carotid artery stiffness, EMT, and visceral adiposity in young, healthy individuals. 135 healthy males (20 ± 2 yr, body mass index [BMI] 24.8 ± 3.3 kg/m2) underwent anthropometric and vascular measures on two separate days. Visceral adiposity was assessed using waist circumference and sagittal abdominal diameter (SAD). Brachial and carotid systolic, diastolic, and pulsatile (PP) blood pressures were assessed using an oscillometric cuff and applanation tonometry, respectively. Carotid intima-media thickness (IMT) and EMT were assessed using Doppler ultrasound. Carotid artery stiffness was calculated as β-stiffness and calibrated to carotid pressures. Separate stepwise multiple regression models demonstrated that carotid PP (β = 0.205) and EMT (β = 0.267) accounted for 12.6% of variance in β-stiffness, while carotid PP (β = 0.195) and SAD (β = 0.226) accounted for 10.5% of variance in EMT (p < 0.05). Mediation analyses revealed carotid PP partially mediated the relationship between a) EMT and β-stiffness, and b) SAD and EMT (p < 0.05). Carotid PP and EMT, but not IMT, are related to carotid β-stiffness. Carotid PP and visceral adiposity (SAD) are related to EMT. Carotid PP partially mediates the association between a) EMT and carotid β-stiffness, and b) SAD and EMT. Our findings suggest visceral adiposity may detrimentally affect subclinical markers of cardiovascular disease risk (carotid PP, EMT) and contribute to artery stiffness.


Capsule endoscopy: a cause of late small bowel obstruction and perforation.
Case Report. A 71-year-old man was admitted to the department of gastroenterology with diffuse abdominal pain. Through the previous 12 months, the patient had experienced episodes of vomiting and watery diarrhea of increasing intensity as well as weight loss. The patient was evaluated with ultrasound, MRI, and subsequently a capsule endoscopy. Six months later, the patient presented, and an abdominal CT-scan showed mechanical small bowel obstruction with suspicion of metallic foreign body and perforation. Laparotomy showed perforation, stenosis, and foreign body, approximately 5 cm from the ileocecal valve. A right hemicolectomy and distal ileectomy (60 cm) with an ileostomy were performed. On further inspection of resection, a capsule endoscope was found impacted in a stenosis. The ileostomy was later reversed without complications. Conclusion. It is important to be aware of the possibility of capsule retention, especially in patients with known or suspected Crohn's disease, due to the propensity of Crohn's disease to form stenosis of the bowel. In cases where a stenosis is suspected, it is warranted to perform a patency capsule swallow before subjecting the patient to a capsule endoscopy.


Insertion of dNTPs opposite the 1,N2-propanodeoxyguanosine adduct by Sulfolobus solfataricus P2 DNA polymerase IV.
1, N (2)-Propanodeoxyguanosine (PdG) is a stable structural analogue for the 3-(2'-deoxy-beta- d- erythro-pentofuranosyl)pyrimido[1,2-alpha]purin-10(3 H)-one (M 1dG) adduct derived from exposure of DNA to base propenals and to malondialdehyde. The structures of ternary polymerase-DNA-dNTP complexes for three template-primer DNA sequences were determined, with the Y-family Sulfolobus solfataricus DNA polymerase IV (Dpo4), at resolutions between 2.4 and 2.7 A. Three template 18-mer-primer 13-mer sequences, 5'-d(TCACXAAATCCTTCCCCC)-3'.5'-d(GGGGGAAGGATTT)-3' (template I), 5'-d(TCACXGAATCCTTCCCCC)-3'.5'-d(GGGGGAAGGATTC)-3' (template II), and 5'-d(TCATXGAATCCTTCCCCC)-3'.5'-d(GGGGGAAGGATTC)-3' (template III), where X is PdG, were analyzed. With templates I and II, diffracting ternary complexes including dGTP were obtained. The dGTP did not pair with PdG, but instead with the 5'-neighboring template dC, utilizing Watson-Crick geometry. Replication bypass experiments with the template-primer 5'-TCACXAAATCCTTACGAGCATCGCCCCC-3'.5'-GGGGGCGATGCTCGTAAGGATTT-3', where X is PdG, which includes PdG in the 5'-CXA-3' template sequence as in template I, showed that the Dpo4 polymerase inserted dGTP and dATP when challenged by the PdG adduct. For template III, in which the template sequence was 5'-TXG-3', a diffracting ternary complex including dATP was obtained. The dATP did not pair with PdG, but instead with the 5'-neighboring T, utilizing Watson-Crick geometry. Thus, all three ternary complexes were of the "type II" structure described for ternary complexes with native DNA [Ling, H., Boudsocq, F., Woodgate, R., and Yang, W. (2001) Cell 107, 91-102]. The PdG adduct remained in the anti conformation about the glycosyl bond in each of these threee ternary complexes. These results provide insight into how -1 frameshift mutations might be generated for the PdG adduct, a structural model for the exocylic M 1dG adduct formed by malondialdehyde.


Long-term outcomes after disease activity-guided dose reduction of TNF inhibition in rheumatoid arthritis: 3-year data of the DRESS study - a randomised controlled pragmatic non-inferiority strategy trial.
Tumour necrosis factor inhibitors (TNFi) are effective in rheumatoid arthritis (RA), but disadvantages include adverse events (AEs) and high costs. This can be improved by disease activity-guided dose reduction (DR). We aimed to assess long-term outcomes of TNFi DR in RA by using 3-year data from the DRESS study (Dose REduction Strategy of Subcutaneous TNF inhibitors study). In the intervention phase (month 0-18) of the DRESS study (Dutch trial register, NTR 3216), patients were randomised to DR or usual care (UC). In the extension phase (month 18-36), treatment strategies in both groups converged to continuation of protocolised tight control and allowed dose optimisation. Intention-to-treat analyses were done on flare, disease activity (28 joint count-based disease activity score with C reactive protein (DAS28-CRP)), functioning (health assessment questionnaire-disability index (HAQ-DI)), quality of life (Euroqol 5 dimensions 5 levels questionnaire (EQ5D-5L)), medication use, radiographic progression (Sharp van der Heijde score (SvdH)) and AE. 172/180 patients included in the DRESS study were included in the extension phase. Cumulative incidences of major flare were 10% and 12% (-2%, 95% CI -8 to 15) in DR and UC groups in the extension phase, and 17% and 14% (3%, 95% CI -9 to 13) from 0 to 36 months. Cumulative incidences of short-lived flares were 43% (33 to 52%)%) and 35% (23 to 49%)%) in DR and UC groups in the extension phase, and 83% (75 to 90%)%) and 44% (31 to 58%)%) from 0 to 36 months. Mean DAS28-CRP, HAQ-DI, EQ5D-5L and SvdH remained stable and not significantly different between groups. TNFi use remained low in the DR group and decreased in the UC group. Cumulative incidences of AE were not significantly different between groups. Safety and efficacy of disease activity guided TNFi DR in RA are maintained up to 3 years, with a large reduction in TNFi use, but no other benefits. Implementation of DR would vastly improve the cost-effective use of TNFi.


Chronic administration of membrane sealant prevents severe cardiac injury and ventricular dilatation in dystrophic dogs.
Duchenne muscular dystrophy (DMD) is a fatal disease of striated muscle deterioration caused by lack of the cytoskeletal protein dystrophin. Dystrophin deficiency causes muscle membrane instability, skeletal muscle wasting, cardiomyopathy, and heart failure. Advances in palliative respiratory care have increased the incidence of heart disease in DMD patients, for which there is no cure or effective therapy. Here we have shown that chronic infusion of membrane-sealing poloxamer to severely affected dystrophic dogs reduced myocardial fibrosis, blocked increased serum cardiac troponin I (cTnI) and brain type natriuretic peptide (BNP), and fully prevented left-ventricular remodeling. Mechanistically, we observed a markedly greater primary defect of reduced cell compliance in dystrophic canine myocytes than in the mildly affected mdx mouse myocytes, and this was associated with a lack of utrophin upregulation in the dystrophic canine cardiac myocytes. Interestingly, after chronic poloxamer treatment, the poor compliance of isolated canine myocytes remained evident, but this could be restored to normal upon direct application of poloxamer. Collectively, these findings indicate that dystrophin and utrophin are critical to membrane stability-dependent cardiac myocyte mechanical compliance and that poloxamer confers a highly effective membrane-stabilizing chemical surrogate in dystrophin/utrophin deficiency. We propose that membrane sealant therapy is a potential treatment modality for DMD heart disease and possibly other disorders with membrane defect etiologies.


Switching off the interactions between graphene oxide and doxorubicin using vitamin C: combining simplicity and efficiency in drug delivery.
Well-controlled, low-toxicity and highly efficient delivery systems for anticancer drugs are a key challenge for the development of a new class of nanocarrier systems for cancer chemotherapy. Graphene oxide (GO) has been developed to be a nanocarrier of anti-cancer drugs due to its large surface area and biocompatibility; however, understanding of the interface chemistry is very limited. In this work, we report efficient loading and controlled release of doxorubicin (DOX) using the tunable surface of GO. A deep understanding of the surface chemistry between GO and DOX is achieved using spectroscopies and atomic force microscopy. Hydrogen bonding and π-π stacking are confirmed to be the non-covalent interactions between the drugs and the carriers. As a result, improvement of DOX delivery from the GO surface can be achieved using vitamin C.


Do baby boomers feel healthier than earlier cohorts after retirement age? The Lausanne cohort Lc65+ study.
Despite the popular belief that baby boomers are ageing in better health than previous generations, limited scientific evidence is available since baby boomers have turned retirement age only recently. This study aimed to compare self-reported health status at ages 65-70 years among three cohorts of older people born before, during and at the end (baby boomers) of the Second World War. Repeated cross-sectional population-based study. Community in a region of French-speaking Switzerland. Community-dwelling older adults who enrolled in the Lausanne cohort 65+ study at ages 65-70 years in 2004 (n=1561), 2009 (n=1489) or 2014 (n=1678). Number of self-reported chronic conditions (from a list of 11) and chronic symptoms (from a list of 11); depressive symptoms; self-rated health (very good, good, average, poor or very poor); fear of disease (not afraid at all, barely afraid, a bit afraid, quite afraid or very afraid); self-perception of ageing; disability in basic and instrumental activities of daily living. There was no significant difference between cohorts in the number of self-reported chronic conditions and chronic symptoms as well as the presence of difficulty in basic activities of daily living, depressive symptoms, fear of disease and negative self-perception of ageing. In women only, significant differences between cohorts were observed in self-rated health (p=0.005) and disability in instrumental activities of daily living (p=0.003), but these associations did not remain significant in logistic regression models adjusted for sociodemographic characteristics and unhealthy behaviours. Despite important sociodemographic differences between older baby boomers and earlier cohorts, most health indicators did not suggest any trend towards a compression of morbidity. Future studies comparing these three cohorts at more advanced age are required to further investigate whether differences emerge later in life.


Emissions from a Diesel Engine using Fe-based Fuel Additives and a Sintered Metal Filtration System.
A series of laboratory tests were conducted to assess the effects of Fe-containing fuel additives on aerosols emitted by a diesel engine retrofitted with a sintered metal filter (SMF) system. Emission measurements performed upstream and downstream of the SMF system were compared, for cases when the engine was fueled with neat ultralow sulfur diesel (ULSD) and with ULSD treated with two formulations of additives containing Fe-based catalysts. The effects were assessed for four steady-state engine operating conditions and one transient cycle. The results showed that the SMF system reduced the average total number and surface area concentrations of aerosols by more than 100-fold. The total mass and elemental carbon results confirmed that the SMF system was indeed very effective in the removal of diesel aerosols. When added at the recommended concentrations (30 p.p.m. of iron), the tested additives had minor adverse impacts on the number, surface area, and mass concentrations of filter-out (FOut) aerosols. For one of the test cases, the additives may have contributed to measurable concentrations of engine-out (EOut) nucleation mode aerosols. The additives had only a minor impact on the concentration and size distribution of volatile and semi-volatile FOut aerosols. Metal analysis showed that the introduction of Fe with the additives substantially increased Fe concentration in the EOut, but the SMF system was effective in removal of Fe-containing aerosols. The FOut Fe concentrations for all three tested fuels were found to be much lower than the corresponding EOut Fe concentrations for the case of untreated ULSD fuel. The results support recommendations that these additives should not be used in diesel engines unless they are equipped with exhaust filtration systems. Since the tested SMF system was found to be very efficient in removing Fe introduced by the additives, the use of these additives should not result in a measurable increase in emissions of de novo generated Fe-containing aerosols. The findings from this study should promote a better understanding of the benefits and challenges of using sintered metal systems and fuel additives to control the exposure of underground miners and other workers to diesel aerosols and gases.


Utility of the theory of reasoned action and theory of planned behavior for predicting Chinese adolescent smoking.
One third of smokers worldwide live in China. Identifying predictors of smoking is important for prevention program development. This study explored whether the Theory of Reasoned Action (TRA) and Theory of Planned Behavior (TPB) predict adolescent smoking in China. Data were obtained from 14,434 middle and high school students (48.6% boys, 51.4% girls) in seven geographically varied cities in China. TRA and TPB were tested by multilevel mediation modeling, and compared by multilevel analyses and likelihood ratio tests. Perceived behavioral control was tested as a main effect in TPB and a moderation effect in TRA. The mediation effects of smoking intention were supported in both models (p<0.001). TPB accounted for significantly more variance than TRA (p<0.001). Perceived behavioral control significantly interacted with attitudes and social norms in TRA (p<0.001). Therefore, TRA and TPB are applicable to China to predict adolescent smoking. TPB is superior to TRA for the prediction and TRA can better predict smoking among students with lower than higher perceived behavioral control.


The prognostic role of a gene signature from tumorigenic breast-cancer cells.
Breast cancers contain a minority population of cancer cells characterized by CD44 expression but low or undetectable levels of CD24 (CD44+CD24-/low) that have higher tumorigenic capacity than other subtypes of cancer cells. We compared the gene-expression profile of CD44+CD24-/low tumorigenic breast-cancer cells with that of normal breast epithelium. Differentially expressed genes were used to generate a 186-gene "invasiveness" gene signature (IGS), which was evaluated for its association with overall survival and metastasis-free survival in patients with breast cancer or other types of cancer. There was a significant association between the IGS and both overall and metastasis-free survival (P<0.001, for both) in patients with breast cancer, which was independent of established clinical and pathological variables. When combined with the prognostic criteria of the National Institutes of Health, the IGS was used to stratify patients with high-risk early breast cancer into prognostic categories (good or poor); among patients with a good prognosis, the 10-year rate of metastasis-free survival was 81%, and among those with a poor prognosis, it was 57%. The IGS was also associated with the prognosis in medulloblastoma (P=0.004), lung cancer (P=0.03), and prostate cancer (P=0.01). The prognostic power of the IGS was increased when combined with the wound-response (WR) signature. The IGS is strongly associated with metastasis-free survival and overall survival for four different types of tumors. This genetic signature of tumorigenic breast-cancer cells was even more strongly associated with clinical outcomes when combined with the WR signature in breast cancer.


Modelling experimental uveitis: barrier effects in autoimmune disease.
A mathematical analysis of leukocytes accumulating in experimental autoimmune uveitis (EAU), using ordinary differential equations (ODEs) and incorporating a barrier to cell traffic. Data from an analysis of the kinetics of cell accumulation within the eye during EAU. We applied a well-established mathematical approach that uses ODEs to describe the behaviour of cells on both sides of the blood-retinal barrier and compared data from the mathematical model with experimental data from animals with EAU. The presence of the barrier is critical to the ability of the model to qualitatively reproduce the experimental data. However, barrier breakdown is not sufficient to produce a surge of cells into the eye, which depends also on asymmetry in the rates at which cells can penetrate the barrier. Antigen-presenting cell (APC) generation also plays a critical role and we can derive from the model the ratio for APC production under inflammatory conditions relative to production in the resting state, which has a value that agrees closely with that found by experiment. Asymmetric trafficking and the dynamics of APC production play an important role in the dynamics of cell accumulation in EAU.


Hematologic toxicity during craniospinal irradiation: the impact of prior chemotherapy.
The purpose of this work was to determine the frequency of hematologic toxicity during craniospinal radiation (CSI) and the impact of preirradiation chemotherapy on this frequency. The charts of 37 patients who received CSI were reviewed. Twenty did not have prior chemotherapy (CT-), while 17 did receive 1 to 18 (mean 5) cycles of multi-agent systemic chemotherapy (CT+). Leukopenia/thrombocytopenia necessitating treatment interruptions occurred in 1/20 (5%) in the CT- group, compared to 8/17 (47%) among the CT+ group. This difference was statistically significant, P < 0.0001 (Fisher's exact two-tailed test). The duration of treatment interruption in the CT+ patients was 4-24 days (mean 14). Compared to the CT- group, the CT+ group had a statistically significant greater decline in their white blood cell count (WBC), platelet count, and hematocrit (HCT) during CSI (percent reduction per Gy; (P = 0.018, 0.006, and 0.047, respectively). Although not statistically significant, the CT+ group also experienced lower nadir ratios (nadir count/baseline count) in terms of WBC and platelets (P = 0.07 and 0.22, respectively). While the mean pretreatment baseline blood counts were lower in the CT+ group compared to the CT- group, these differences reached statistical significance for the HCT (P = 0.02), but not the WBC (P = 0.59) or platelets (P = 0.43). Leukopenia and thrombocytopenia are very common in patients who receive craniospinal irradiation following multi-agent systemic chemotherapy. This appears to be due to more rapid and marked reductions in counts during CSI. Since this toxicity may cause treatment interruptions that are potentially therapeutically disadvantageous, aggressive hematologic support with transfusions and growth factors may be necessary. This problem may become more common as combined modality therapy is used more frequently.


The interactomes of POU5F1 and SOX2 enhancers in human embryonic stem cells.
The genes POU5F1 and SOX2 are critical for pluripotency and reprogramming, yet the chromosomal organization around these genes remains poorly understood. We assayed long-range chromosomal interactions on putative enhancers of POU5F1 and SOX2 genes in human embryonic stem cells (hESCs) using 4C-Seq technique. We discovered that their frequent interacting regions mainly overlap with early DNA replication domains. The interactomes are associated with active histone marks and enriched with 5-hydroxymethylcytosine sites. In hESCs, genes within the interactomes have elevated expression. Additionally, some genes associated with the POU5F1 enhancer contribute to pluripotency. Binding sites for multiple DNA binding proteins, including ATF3, CTCF, GABPA, JUND, NANOG, RAD21 and YY1, are enriched in both interactomes. The RARG locus, frequently interacting with the POU5F1 locus, has abundant RAD21 binding sites co-localized with other protein binding sites. Thus the interactomes of these two pluripotency genes could be an important part of the regulatory network in hESCs.


Targeted, High-Resolution RNA Sequencing of Non-coding Genomic Regions Associated With Neuropsychiatric Functions.
The human brain is one of the last frontiers of biomedical research. Genome-wide association studies (GWAS) have succeeded in identifying thousands of haplotype blocks associated with a range of neuropsychiatric traits, including disorders such as schizophrenia, Alzheimer's and Parkinson's disease. However, the majority of single nucleotide polymorphisms (SNPs) that mark these haplotype blocks fall within non-coding regions of the genome, hindering their functional validation. While some of these GWAS loci may contain cis-acting regulatory DNA elements such as enhancers, we hypothesized that many are also transcribed into non-coding RNAs that are missing from publicly available transcriptome annotations. Here, we use targeted RNA capture ('RNA CaptureSeq') in combination with nanopore long-read cDNA sequencing to transcriptionally profile 1,023 haplotype blocks across the genome containing non-coding GWAS SNPs associated with neuropsychiatric traits, using post-mortem human brain tissue from three neurologically healthy donors. We find that the majority (62%) of targeted haplotype blocks, including 13% of intergenic blocks, are transcribed into novel, multi-exonic RNAs, most of which are not yet recorded in GENCODE annotations. We validated our findings with short-read RNA-seq, providing orthogonal confirmation of novel splice junctions and enabling a quantitative assessment of the long-read assemblies. Many novel transcripts are supported by independent evidence of transcription including cap analysis of gene expression (CAGE) data and epigenetic marks, and some show signs of potential functional roles. We present these transcriptomes as a preliminary atlas of non-coding transcription in human brain that can be used to connect neurological phenotypes with gene expression.


Intracellular amyloid and the neuronal origin of Alzheimer neuritic plaques.
Genetic analysis of familial forms of Alzheimer's disease (AD) causally links the proteolytic processing of the amyloid precursor protein (APP) and AD. However, the specific type of amyloid and mechanisms of amyloid pathogenesis remain unclear. We conducted a detailed analysis of intracellular amyloid with an aggregation specific conformation dependent monoclonal antibody, M78, raised against fibrillar Aß42. M78 immunoreactivity colocalizes with Aß and the carboxyl terminus of APP (APP-CTF) immunoreactivities in perinuclear compartments at intermediate times in 10month 3XTg-AD mice, indicating that this represents misfolded and aggregated protein rather than normally folded APP. At 12months, M78 immunoreactivity also accumulates in the nucleus. Neuritic plaques at 12months display the same spatial organization of centrally colocalized M78, diffuse chromatin and neuronal nuclear NeuN staining surrounded by peripheral M78 and APP-CTF immunoreactivity as observed in neurons, indicating that neuritic plaques arise from degenerating neurons with intracellular amyloid immunoreactivity. The same staining pattern was observed in neuritic plaques in human AD brains, showing elevated intracellular M78 immunoreactivity at intermediate stages of amyloid pathology (Braak A and B) compared to no amyloid pathology and late stage amyloid pathology (Braak 0 and C, respectively). These results indicate that intraneuronal protein aggregation and amyloid accumulation is an early event in AD and that neuritic plaques are initiated by the degeneration and death of neurons by a mechanism that may be related to the formation of extracellular traps by neutrophils.


Comparison of different approaches for assessment of HER2 expression on protein and mRNA level: prediction of chemotherapy response in the neoadjuvant GeparTrio trial (NCT00544765).
Human epidermal growth factor receptor 2 (HER2) testing is an essential part of pathological assessment in breast cancer patients, as HER2 provides not only prognostic but also predictive information on response to targeted therapy. So far, HER2 test accuracy of immunohistochemistry/in situ-hybridization techniques is still under debate, and more reliable and robust technologies are needed. To address this issue and to evaluate the predictive value of HER2 on chemotherapy, we investigated a cohort of 278 patients from the GeparTrio trial, a prospective neoadjuvant anthracycline/taxane-based multicenter study. In the GeparTrio trial, patients were not treated with any anti-HER2 therapy, as this was not standard therapy at this time. The HER2 status was analyzed by three different approaches: local and central evaluation using immunohistochemistry combined with in situ-hybridization as well as evaluation of HER2 mRNA expression using kinetic RT-PCR from formalin-fixed, paraffin-embedded (FFPE) tissue samples using a predefined cutoff. HER2 overexpression/amplification was observed in 37.3% (91/244) and 17.9% (41/229) of the informative samples in the local and central evaluations, respectively. Positive HER2 mRNA levels were found in 19.8% (55/278). We observed a highly significant correlation between central HER2 expression and HER2 status measured by kinetic RT-PCR (r = 0.856, P < 0.0001) and an overall agreement of 95.6% (κ statistic, 0.862, CI 0.77-0.94). Further, central HER2 as well as HER2 mRNA expression were predictors for a pathological complete response after neoadjuvant anthracycline/taxane-based primary chemotherapy in a univariate binary logistic regression analysis (OR 3.29, P = 0.002; OR 2.65, P = 0.004). The predictive value could be confirmed for the central HER2 status by multivariate analysis (OR 3.04, P = 0.027). The locally assessed HER2 status was not predictive of response to chemotherapy. Our results suggest that standardized methods are preferable for evaluation of HER2 status. The kinetic RT-PCR from FFPE tissue might be an additional approach for assessment of this important prognostic and predictive parameter but has to be confirmed by other studies.


Maximizing the Benefit of Life-Saving Treatments for Pompe Disease, Spinal Muscular Atrophy, and Duchenne Muscular Dystrophy Through Newborn Screening: Essential Steps.
Newborn screening (NBS) identifies infants with specific congenital disorders for which earlier intervention cannot only prevent a lifetime of chronic disability but also, most importantly, save lives. In this article, we discuss complexities associated with NBS processes in the United States, with a focus on challenges in neuromuscular disorders. As new interventions for neuromuscular disorders become available, the clinical community must prepare to overcome the challenges of adding new diseases to screening panels and understand the rigorous evidence review at the federal level and the complex process of state-level implementation. In this regard, NBS programs for Pompe disease and spinal muscular atrophy can guide the path of Duchenne muscular dystrophy and other neuromuscular disorders as future candidates for NBS. The availability of advanced screening methods, the emergence of effective treatment, and the support of professional organizations may facilitate the expansion of NBS, such that an increasing number of infants can be identified in the newborn period who will benefit from life-saving interventions.


Quantum dots: an insight and perspective of their biological interaction and how this relates to their relevance for clinical use.
Due to their novel physico-chemical characteristics, semi-conductor nanocrystal quantum dots (QDs) provide an advantageous perspective towards numerous different consumer and medical applications. The most notable potential application of QDs is their use as therapeutic and diagnostic tools in nanomedicine. Despite the many benefits posed by QDs, the proposed, intentional exposure to humans has raised concerns towards their potential impact upon human health. These concerns are predominantly based upon the heterogeneous composition of QDs, which most commonly comprises of a cadmium-based core and zinc sulphide shell. Whilst other nanoparticle (NP) types possess a similar structure to QDs (i.e. core-shell technology (e.g. Fe(2)O(3), Au and superparamagnetic iron oxide NPs)), the importance of the concerns surrounding human exposure to QDs is amplified further since, due to the sophisticated chemical and light-emitting properties of QDs, the use of these NPs within any (nano)medical setting/application could be suggested as realistic, rather than simply an advantageous possibility. It is therefore imperative that a thorough understanding of how QDs interact with various biological systems, predominantly those relative to humans and what the consequences of such interactions are is gained with extreme alacrity. It is the aim of this review to highlight the current knowledge base of QD-biological system interactions, where the knowledge gaps (still) remain and how the understanding of this interaction relates to the most notable of applications for QDs; their clinical relevance.


Biomarkers of Alzheimer's disease risk in peripheral tissues; focus on buccal cells.
Alzheimer's disease (AD) is a progressive degenerative disorder of the brain and is the most common form of dementia. To-date no simple, inexpensive and minimally invasive procedure is available to confirm with certainty the early diagnosis of AD prior to the manifestations of symptoms characteristic of the disease. Therefore, if population screening of individuals is to be performed, more suitable, easily accessible tissues would need to be used for a diagnostic test that would identify those who exhibit cellular pathology indicative of mild cognitive impairment (MCI) and AD risk so that they can be prioritized for primary prevention. This need for minimally invasive tests could be achieved by targeting surrogate tissues, since it is now well recognized that AD is not only a disorder restricted to pathology and biomarkers within the brain. Human buccal cells for instance are accessible in a minimally invasive manner, and exhibit cytological and nuclear morphologies that may be indicative of accelerated ageing or neurodegenerative disorders such as AD. However, to our knowledge there is no review available in the literature covering the biology of buccal cells and their applications in AD biomarker research. Therefore, the aim of this review is to summarize some of the main findings of biomarkers reported for AD in peripheral tissues, with a further focus on the rationale for the use of the buccal mucosa (BM) for biomarkers of AD and the evidence to date of changes exhibited in buccal cells with AD.


The Social Determinants of Trauma: A Trauma Disparities Scoping Review and Framework.
The drivers of trauma disparities are multiple and complex; yet, understanding the causes will direct needed interventions. The aims of this article are to (1) explore how the injured patient, his or her social environment, and the health care system interact to contribute to trauma disparities and examine the evidence in support of interventions and (2) develop a conceptual framework that captures the socioecological context of trauma disparities. Using a scoping review methodology, articles were identified through PubMed and CINAHL between 2000 and 2015. Data were extracted on the patient population, social determinants of health, and interventions targeting trauma disparities and violence. Based on the scoping review of 663 relevant articles, we inductively developed a conceptual model, The Social Determinants of Trauma: A Trauma Disparities Framework, based on the categorization of articles by: institutional power (n = 9), social context-place (n = 117), discrimination experiences (n = 59), behaviors and comorbidities (n = 57), disparities research (n = 18), and trauma outcomes (n = 85). Intervention groupings included social services investment (n = 54), patient factors (n = 88), hospital factors (n = 27), workforce factors (n = 31), and performance improvement (n = 118). This scoping review produced a needed taxonomy scheme of the drivers of trauma disparities and known interventions that in turn informed the development of The Social Determinants of Trauma: A Trauma Disparities Framework. This study adds to the trauma disparities literature by establishing social context as a key contributor to disparities in trauma outcomes and provides a road map for future trauma disparities research.


Evaluating the impacts of new walking and cycling infrastructure on carbon dioxide emissions from motorized travel: a controlled longitudinal study.
Walking and cycling is widely assumed to substitute for at least some motorized travel and thereby reduce energy use and carbon dioxide (CO2) emissions. While the evidence suggests that a supportive built environment may be needed to promote walking and cycling, it is unclear whether and how interventions in the built environment that attract walkers and cyclists may reduce transport CO2 emissions. Our aim was therefore to evaluate the effects of providing new infrastructure for walking and cycling on CO2 emissions from motorised travel. A cohort of 1849 adults completed questionnaires at baseline (2010) and one-year follow-up (2011), before and after the construction of new high-quality routes provided as part of the Sustrans Connect2 programme in three UK municipalities. A second cohort of 1510 adults completed questionnaires at baseline and two-year follow-up (2012). The participants reported their past-week travel behaviour and car characteristics from which CO2 emissions by mode and purpose were derived using methods described previously. A set of exposure measures of proximity to and use of the new routes were derived. Overall transport CO2 emissions decreased slightly over the study period, consistent with a secular trend in the case study regions. As found previously the new infrastructure was well used at one- and two-year follow-up, and was associated with population-level increases in walking, cycling and physical activity at two-year follow-up. However, these effects did not translate into sizeable CO2 effects as neither living near the infrastructure nor using it predicted changes in CO2 emissions from motorised travel, either overall or disaggregated by journey purpose. This lack of a discernible effect on travel CO2 emissions are consistent with an interpretation that some of those living nearer the infrastructure may simply have changed where they walked or cycled, while others may have walked or cycled more but few, if any, may have substituted active for motorised modes of travel as a result of the interventions. While the findings to date cannot exclude the possibility of small effects of the new routes on CO2 emissions, a more comprehensive approach of a higher 'dosage' of active travel promotion linked with policies targeted at mode shift away from private motorized transport (such as urban car restraint and parking pricing, car sharing/pooling for travel to work, integrating bike sharing into public transport system) may be needed to achieve the substantial CO2 savings needed to meet climate change mitigation and energy security goals.


The amphioxus FoxQ1 gene is expressed in the developing endostyle.
The FoxQ1 genes form a distinct group within the Fox (also known as forkhead) gene family. We have isolated a gene from the amphioxus Branchiostoma floridae that encodes a forkhead domain with high identity to FoxQ1 genes in other chordates. Molecular phylogenetic analysis places AmphiFoxQ1 in a robust grouping with vertebrate FoxQ1 genes and with Ciona intestinalis Ci-FoxQ1. This group is separate from that containing AmphiFoxQ2, which instead groups with other invertebrate Fox genes. The expression of AmphiFoxQ1 was analysed by whole mount in situ hybridisation. The results show that AmphiFoxQ1 expression is confined to the developing endoderm, and specifically marks the endostyle and associated peripharyngeal bands of amphioxus larvae. Ci-FoxQ1 is also expressed in the endostyle, highlighting this as a conserved site of FoxQ1 gene expression in basal chordates.


Points for Consideration for dengue vaccine introduction - recommendations by the Dengue Vaccine Initiative.
Dengue is a public health problem in the tropics and subtropics. There are several vaccine candidates in clinical development. However, there may be gaps in the new vaccine introduction after vaccine licensure before it becomes available in developing countries. In anticipation of the first dengue vaccine candidate to be licensed, Dengue Vaccine Initiative (DVI) and, its predecessor, Pediatric Dengue Vaccine Initiative (PDVI) have been working on points for consideration to accelerate evidence-based dengue vaccine introduction, once a vaccine becomes available. In this paper, we review the history of PDVI and its successor, the DVI, and elaborate on the points of consideration for dengue vaccine introduction.


Genetic and environmental predictors of latent trajectories of alcohol use from adolescence to adulthood: a male twin study.
Adolescence is characterized by higher levels of novelty-seeking and risk-taking behavior, including initiation of alcohol use. Also, there is considerable heterogeneity in the change and continuity of alcohol use over time, which emphasizes the need to examine factors predicting alcohol use and the patterns of use over time. Retrospective data on average monthly alcohol use and risk and protective factors were obtained through interviews and questionnaires in 1,560 adult male twins. Latent class growth analysis in Mplus was performed on data of alcohol use over ages 15 to 36. Second, logistic regression analyses were used to associate risk and protective characteristics with membership in distinct latent trajectories of alcohol use. Six trajectories of alcohol use were identified, varying in the level of alcohol use, the rate of change in use in early adolescence and the persistence of use into adulthood. Genetic risk of externalizing disorder and peer deviance showed the greatest risks for unfavorable alcohol trajectories with higher levels of use and higher rates of early increase in use. Parental monitoring and involvement in social activities showed protective effects. Involvement in religious activities was strongly associated with reduced persistence of high-level drinking in univariate but not multivariate regression analyses. Risk and protective factors impacted differentially on level of alcohol use, rate of increase in use during adolescence, and persistence of heavy alcohol use over time. Insight into the different ways in which predictors impact on alcohol use is relevant for the development of new intervention strategies. For this purpose, causality of the associations should be further examined.


Possible genetic influence on the strength of human muscle nerve sympathetic activity at rest.
Large reproducible interindividual differences in the strength of human muscle nerve sympathetic activity have been demonstrated previously without satisfactory explanation. We undertook the present study to investigate whether a genetic influence may be a factor of importance. Microneurographic recordings of sympathetic impulse traffic were made in the peroneal nerve in nine pairs of monozygotic male twins and eight pairs of age-matched male subjects without family relationship. The strength of the sympathetic activity was quantitated as number of sympathetic bursts per 100 heart beats and bursts per minute. Group mean values of muscle sympathetic activity, heart rate, and blood pressure were similar in the two groups. Intrapair differences (mean +/- SEM) of sympathetic activity were 5.4 +/- 1.7 bursts per 100 heart beats (1.7 +/- 0.5 bursts per minute) for the twins and 19.4 +/- 3.2 bursts per 100 heart beats (11.8 +/- 2.5 bursts per minute) for the control subjects (P < .01 for both). The degree of reproducibility between twins is similar to that reported previously between repeated recordings in the same subject. The finding may indicate that the strength of sympathetic outflow to muscle is controlled genetically. If so, we speculate that this may contribute to the heritability of blood pressure in both normotensive and hypertensive subjects.


Comparison of full-sky polarization and radiance observations to radiative transfer simulations which employ AERONET products.
Visible-band and near infrared polarization and radiance images measured with a ground-based full-sky polarimeter are compared against a successive orders of scattering (SOS) radiative transfer model for 2009 summer cloud-free days in Bozeman, Montana, USA. The polarimeter measures radiance and polarization in 10-nm bands centered at 450 nm, 490 nm, 530 nm, 630 nm, and 700 nm. AERONET products are used to represent aerosols in the SOS model, while MISR satellite BRF products are used for the surface reflectance. While model results generally agree well with observation, the simulated degree of polarization is typically higher than observed data. Potential sources of this difference may include cloud contamination and/or underestimation of the AERONET-retrieved aerosol real refractive index. Problems with the retrieved parameters are not unexpected given the low aerosol optical depth range (0.025 to 0.17 at 500 nm) during the study and the corresponding difficulties that these conditions pose to the AERONET inversion algorithm.


Parthenolide induces significant apoptosis and production of reactive oxygen species in high-risk pre-B leukemia cells.
We investigated whether parthenolide, the principal bioactive component of the herb feverfew (Tanacetum parthenium) induced apoptosis in pre-B acute lymphoblastic leukemia (ALL) lines, including cells carrying the t(4;11)(q21;q23) chromosomal translocation. Parthenolide induced rapid apoptotic cell death distinguished by loss of nuclear DNA, externalization of cell membrane phosphatidylserine, and depolarization of mitochondrial membranes at concentrations ranging from 5 to 100 microM. Using reactive oxygen species (ROS)-specific dyes, an increase in nitric oxide and superoxide anion was detected in the cells by 4 h after exposure to parthenolide. Parthenolide-induced elevation of hypochlorite anion was observed only in the two t(4;11) lines. These data suggest parthenolide may have potential as a potent and novel therapeutic agent against pre-B ALLs.


MMP13 mediates cell cycle progression in melanocytes and melanoma cells: in vitro studies of migration and proliferation.
Melanoma cells are usually characterized by a strong proliferative potential and efficient invasive migration. Among the multiple molecular changes that are recorded during progression of this disease, aberrant activation of receptor tyrosine kinases (RTK) is often observed. Activation of matrix metalloproteases goes along with RTK activation and usually enhances RTK-driven migration. The purpose of this study was to examine RTK-driven three-dimensional migration of melanocytes and the pro-tumorigenic role of matrix metalloproteases for melanocytes and melanoma cells. Using experimental melanocyte dedifferentiation as a model for early melanomagenesis we show that an activated EGF receptor variant potentiates migration through three-dimensional fibrillar collagen. EGFR stimulation also resulted in a strong induction of matrix metalloproteases in a MAPK-dependent manner. However, neither MAPK nor MMP activity were required for migration, as the cells migrated in an entirely amoeboid mode. Instead, MMPs fulfilled a function in cell cycle regulation, as their inhibition resulted in strong growth inhibition of melanocytes. The same effect was observed in the human melanoma cell line A375 after stimulation with FCS. Using sh- and siRNA techniques, we could show that MMP13 is the protease responsible for this effect. Along with decreased proliferation, knockdown of MMP13 strongly enhanced pigmentation of melanocytes. Our data show for the first time that growth stimuli are mediated via MMP13 in melanocytes and melanoma, suggesting an autocrine MMP13-driven loop. Given that MMP13-specific inhibitors are already developed, these results support the evaluation of these inhibitors in the treatment of melanoma.


Using self-report assessment methods to explore facets of mindfulness.
The authors examine the facet structure of mindfulness using five recently developed mindfulness questionnaires. Two large samples of undergraduate students completed mindfulness questionnaires and measures of other constructs. Psychometric properties of the mindfulness questionnaires were examined, including internal consistency and convergent and discriminant relationships with other variables. Factor analyses of the combined pool of items from the mindfulness questionnaires suggested that collectively they contain five clear, interpretable facets of mindfulness. Hierarchical confirmatory factor analyses suggested that at least four of the identified factors are components of an overall mindfulness construct and that the factor structure of mindfulness may vary with meditation experience. Mindfulness facets were shown to be differentially correlated in expected ways with several other constructs and to have incremental validity in the prediction of psychological symptoms. Findings suggest that conceptualizing mindfulness as a multifaceted construct is helpful in understanding its components and its relationships with other variables.


GEOSAT Altimeter Observations of Kelvin Waves and the 1986-87 El Nino.
Two years of GEOSAT altimeter observations are used to investigate the response of sea level to anomalous westerly wind bursts in the tropical Pacific Ocean before and during the 1986-87 El Niño. Sea level time series along the equator show examples of both positive and negative anomalies of 10-centimeter amplitude and 2- to 4-week time scale propagating across the Pacific with phase speeds of 2.4 to 2.8 meters per second, suggesting downwelling and upwelling Kelvin waves, respectively. A comparison of island wind observations with sea level indicates one instance (May 1986) in which a positive sea level anomaly can be related to westerly winds caused by a cross-equatorial cyclone pair in the western Pacific. This episode was followed by additional wind bursts later in the year, and finally by sustained westerlies in the western Pacific during November-December 1986, at the height of El Niño. The GEOSAT observations reveal the sea level response to these meteorological events and provide a synoptic description of the El Niño oceanographic phenomenon.


Management of anaplastic thyroid cancer.
Anaplastic thyroid cancer (ATC) is a deadly disease with a dismal prognosis. Molecular analyses of ATC tumors have yielded interesting results, which could help in understanding the underlying mechanisms of this aggressive disease process. Managing ATC can be challenging and includes rapid diagnosis, adequate staging, and interdisciplinary, multimodal treatments to optimize patient outcome. Treatments include surgical resection to gross negative margins when possible, as well as neo- or adjuvant treatment with chemotherapy or external beam radiation (XRT) for locoregional disease. New treatment strategies include evaluating the benefits of vascular disrupting agents and tyrosine kinase inhibitors for advanced ATC with driver mutations, which can be targeted. This review summarizes key concepts in managing ATC.


Intratracheal administration of liposomal clodronate accelerates alveolar macrophage reconstitution following fetal liver transplantation.
To facilitate study of alveolar macrophages in vivo, we developed a method to rapidly and efficiently replace resident alveolar macrophages with macrophages of a different (donor) genotype. Chimeric mice were generated by lethal irradiation followed by fetal liver transplantation (FLT) using green fluorescent protein (GFP) transgenic reporter mice as donors. Kinetics of peripheral blood monocyte (PBM) and alveolar macrophage reconstitution was determined 4 and 10 weeks post-FLT by quantifying the percentage of GFP+ cells. To enhance the recruitment of donor monocytes into the lung after FLT, mice were treated with intratracheal administration of liposomal clodronate to deplete host alveolar macrophages at 6 weeks post-FLT. PBM reconstitution occurred by 4 weeks after FLT (85.7+/-1.6% of CD11b+/Gr-1+ monocytes were GFP+), and minimal alveolar macrophage repopulation was observed (9.5% GFP+). By 10 weeks following FLT, 48% of alveolar macrophages were GFP+ by immunostaining of macrophages on lung tissue sections, and 55.1 +/- 1.6% of lung lavage macrophages were GFP+ by fluorescein-activated cell sorter analysis. Clodronate treatment resulted in a significant increase in GFP+ alveolar macrophages 10 weeks after FLT. By immunostaining, 90% of macrophages were GFP+ on lung tissue sections and 87.5 +/- 1.1% GFP+ in lung lavage (compared with GFP-transgenic controls). The ability of newly recruited alveolar macrophages to clear Pseudomonas aeruginosa and activate nuclear factor-kappaB in response to Eschericia coli lipopolysaccharide demonstrated normal macrophage function. Optimizing this methodology provides an important tool for the study of specific genes and their contribution to alveolar macrophage function in vivo.


The temporal dynamics and infectiousness of subpatent Plasmodium falciparum infections in relation to parasite density.
Malaria infections occurring below the limit of detection of standard diagnostics are common in all endemic settings. However, key questions remain surrounding their contribution to sustaining transmission and whether they need to be detected and targeted to achieve malaria elimination. In this study we analyse a range of malaria datasets to quantify the density, detectability, course of infection and infectiousness of subpatent infections. Asymptomatically infected individuals have lower parasite densities on average in low transmission settings compared to individuals in higher transmission settings. In cohort studies, subpatent infections are found to be predictive of future periods of patent infection and in membrane feeding studies, individuals infected with subpatent asexual parasite densities are found to be approximately a third as infectious to mosquitoes as individuals with patent (asexual parasite) infection. These results indicate that subpatent infections contribute to the infectious reservoir, may be long lasting, and require more sensitive diagnostics to detect them in lower transmission settings.


Intradural, extramedullary spinal cord granular cell tumor: a case report and clinicopathologic review of the literature.
Granular cell tumor (GCT) of the spine is uncommon, with intradural extramedullary location being exceptionally rare. The non-specific clinical presentation and variable histologic patterns can make recognition of this tumor challenging. Two previous reports of GCT of the spine were reviewed (Medline 1960-2009) and analyzed with respect to this case report. The patients included two women and one man (mean age, 28.7 years). Patients presented with 3 to 4 months of lower back pain and/or lower extremity radiculopathy. The lesions appeared radiographically to be intradural and extramedullary or intramedullary. The tumors were found at T10 or L1-L2 space. Radiographically, all tumors enhanced homogenously on T1 post-gadolinium imaging with a mean tumor size of approximately 1.6 cm. Histologically, the tumors were composed of large, polygonal granular cells. The abundant cytoplasm was fine or coarsely granular, surrounding small, pale-staining nuclei, which were eccentrically located in the cell. The tumor cells were periodic acid Schiff positive, diastase resistant, and were positive with S-100 protein, CD68, inhibin, and neuron-specific enolase immunohistochemistry. The clinical and histologic differential diagnosis includes schwannoma, neurofibroma, meningioma, astrocytoma, melanocytoma, and metastatic tumors. Patients were managed with excision. One patient had symptomatic and radiographic local recurrence that was subsequently treated with radiation, resulting in stabilization of disease and symptoms. Intradural GCTs of the spine are rare and radiographically indistinguishable from tumors that more commonly arise in this location. Histologic recognition of this rare tumor is important because the subsequent clinical course of the disease differs from other similar lesions.


Identification of multi-target effects of Huaier aqueous extract via microarray profiling in triple-negative breast cancer cells.
Breast cancer is one of the most common malignant tumors in the world. Long-term maintenance treatment is important for breast cancer. However, effective maintenance treatment is lacking for triple-negative breast cancer (TNBC). Traditional Chinese medicine (TCM) has shown its potential anticancer roles as an effective maintenance treatment for TNBC. However its mechanisms remained unclear. In this study, we detected the differentially expressed genes (DEGs) after treatment with Huaier aqueous extract by using microarray profiling in MDA-MB-231 cells. Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and gene-gene interaction network were conducted to confirm the altered biological functions induced by Huaier extract. Screening of DEGs gave 387 genes (226 upregulated and 161 downregulated) in MDA-MB-231 cells which were regulated significantly by Huaier extract. GO and KEGG pathway analysis suggested that a number of functions were affected by Huaier, including proliferation, apoptosis, migration, and angiogenesis. Gene-gene interaction network showed the detailed molecular signal-net. Based on microarray data, we studied several functions of Huaier extract and in return verified the results of microarray profiling. This study had important guidance roles and indicated new research directions.


Quantitative trait locus for reading disability: correction.
In the news article "Can risky mergers save hospital-based research?" by Wade Roush (19 May, p. 968), the statement that University Hospitals of Cleveland rose from 20th in the rankings of teaching hospitals funded by the National Institutes of Health (NIH) in 1991 to 12th at present was incorrect. In fact, it was Case Western Reserve University (CWRU), with which University Hospitals of Cleveland is affiliated, that received $69 million in NIH grants in 1993, making it the 20th largest recipient of such grants among medical centers; the university then received $97 million in 1994, raising its rank to 12th. About $15 million of the increase, or 53%, was attributable to CWRU's 1992 affiliation with Henry Ford Hospital in Detroit. Other hospitals affiliated with Case Western include MetroHealth Medical Center, Mount Sinai Medical Center, St. Luke's Medical Center, and Cleveland Veterans' Affairs Medical Center.


Jak3 negatively regulates dendritic-cell cytokine production and survival.
Cytokines are critical in regulating the development and function of diverse cells. Janus kinase 3 (Jak3) is a tyrosine kinase expressed in hematopoietic cells that associates with the common gamma chain (gammac) and is required for signaling for a family of cytokines including interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15, and IL-21; deficiency of either Jak3 or gammac results in severe combined immunodeficiency (SCID). While Jak3 is essential for lymphoid-cell development, the potential roles for Jak3 in regulating dendritic cells (DCs) were unclear. Herein, we show that although CD8+CD11c+ splenic DCs are absent in Jak3-/- mice, bone marrow-derived DCs developed normally in vitro from Jak3-/- precursor cells. In fact, the survival of Jak3-/- DCs was enhanced, and they expressed lower levels of proapoptotic proteins. Jak3-/- DCs exhibited normal antigen uptake and up-regulation of costimulatory molecules. However, Jak3-/- DCs produced more IL-12 and IL-10 in response to Toll-like receptor ligands, which correlated with enhanced T helper 1 (Th1) differentiation in vivo. In summary, Jak3 is not essential for DC development but unexpectedly appears to be an important negative regulator. These results may be relevant clinically for patients with SCID who have undergone hematopoietic stem cell transplantation and for patients who might be treated with a Jak3 inhibitor.


Collagen-binding VEGF targeting the cardiac extracellular matrix promotes recovery in porcine chronic myocardial infarction.
An effective therapy for chronic myocardial infarction (MI) has yet to be developed. Vascular endothelial growth factor (VEGF) promotes angiogenesis and improves cardiac function after MI. However, non-targeted delivery of VEGF decreases its therapeutic efficacy. In this study, for targeting the cardiac extracellular matrix, a collagen-binding domain (CBD) VEGF was used to bind specifically to the collagen-rich cardiac extracellular matrix. When intramyocardially injected into the peri-infarct region of a chronically infarcted porcine heart, CBD-VEGF attenuated the remodeling of the left ventricle with a decreased infarct size and promoted cardiomyocyte survival and angiogenesis 3 months after injection. In the 12-month trial, mature vessel networks and myocardium-like tissues were observed in the infarct region after CBD-VEGF injection. Also these beneficial effects might derive from CBD-VEGF significantly protecting cardiomyocytes from apoptosis and recruiting cardiac progenitor cells to the infarcted region. These results demonstrated that CBD-VEGF could be a promising therapeutic strategy for chronic MI.


Recent advances in controlled synthesis of two-dimensional transition metal dichalcogenides via vapour deposition techniques.
In recent years there have been many breakthroughs in two-dimensional (2D) nanomaterials, among which the transition metal dichalcogenides (TMDs) attract significant attention owing to their unusual properties associated with their strictly defined dimensionalities. TMD materials with a generalized formula of MX2, where M is a transition metal and X is a chalcogen, represent a diverse and largely untapped source of 2D systems. Semiconducting TMD monolayers such as MoS2, MoSe2, WSe2 and WS2 have been demonstrated to be feasible for future electronics and optoelectronics. The exotic electronic properties and high specific surface areas of 2D TMDs offer unlimited potential in various fields including sensing, catalysis, and energy storage applications. Very recently, the chemical vapour deposition technique (CVD) has shown great promise to generate high-quality TMD layers with a scalable size, controllable thickness and excellent electronic properties. Wafer-scale deposition of mono to few layer TMD films has been obtained. Despite the initial success in the CVD synthesis of TMDs, substantial research studies on extending the methodology open up a new way for substitution doping, formation of monolayer alloys and producing TMD stacking structures or superlattices. In this tutorial review, we will introduce the latest development of the synthesis of monolayer TMDs by CVD approaches.


Carcino-embryonic antigen may function as a chemo-attractant in colorectal-carcinoma cell lines.
Locomotion of colorectal-carcinoma cells was tested in order to establish whether it might be affected by carcino-embryonic antigen (CEA). CEA production, cell growth and DNA ploidy were measured in 22 colorectal-carcinoma cell lines. A cell-invasion assay was adapted using a transfilter chamber, the lower surface of which was coated with various substrates in the amount of 5 microgram/filter (CEA, type-IV collagen, laminin). Cells infiltrated into the lower surface of the filter were counted over 9-microscope fields (x400). All cell lines produced CEA, 9 producing more than 100 ng/ml medium. Of the total, 8 cell lines were diploid and 14 were aneuploid. Invasiveness, measured by the number of infiltrated cells, was highest in CEA-coated filters, and next highest in type-IV-collagen- and laminin-coated filters, in descending order (p < 0.001-0.05). Invasiveness of each cell line was closely correlated with 2 substrates. Poorly differentiated or advanced-stage tumors were more invasive than well-differentiated or early-stage tumors (p < 0.001-0. 05). However, invasiveness was not associated with DNA ploidy or CEA production. CEA may function as a chemo-attractant as well as an adhesion molecule in colorectal-carcinoma cell lines. In addition, adhesion to CEA appears to be related to type-IV collagen and laminin.


Cerebral Mitochondrial Microangiopathy Leads to Leukoencephalopathy in Mitochondrial Neurogastrointestinal Encephalopathy.
Mitochondrial neurogastrointestinal encephalopathy is a rare disorder due to recessive mutations in the thymidine phosphorylase gene, encoding thymidine phosphorylase protein required for mitochondrial DNA replication. Clinical manifestations include gastrointestinal dysmotility and diffuse asymptomatic leukoencephalopathy. This study aimed to elucidate the mechanisms underlying brain leukoencephalopathy in patients with mitochondrial neurogastrointestinal encephalopathy by correlating multimodal neuroradiologic features to postmortem pathology. Seven patients underwent brain MR imaging, including single-voxel proton MR spectroscopy and diffusion imaging. Absolute concentrations of metabolites calculated by acquiring unsuppressed water spectra at multiple TEs, along with diffusion metrics based on the tensor model, were compared with those of healthy controls using unpaired t tests in multiple white matters regions. Brain postmortem histologic, immunohistochemical, and molecular analyses were performed in 1 patient. All patients showed bilateral and nearly symmetric cerebral white matter hyperintensities on T2-weighted images, extending to the cerebellar white matter and brain stem in 4. White matter, N-acetylaspartate, creatine, and choline concentrations were significantly reduced compared with those in controls, with a prominent increase in the radial water diffusivity component. At postmortem examination, severe fibrosis of brain vessel smooth muscle was evident, along with mitochondrial DNA replication depletion in brain and vascular smooth-muscle and endothelial cells, without neuronal loss, myelin damage, or gliosis. Prominent periependymal cytochrome C oxidase deficiency was also observed. Vascular functional and histologic alterations account for leukoencephalopathy in mitochondrial neurogastrointestinal encephalopathy. Thymidine toxicity and mitochondrial DNA replication depletion may induce microangiopathy and blood-brain-barrier dysfunction, leading to increased water content in the white matter. Periependymal cytochrome C oxidase deficiency could explain prominent periventricular impairment.


Effect of dextromethorphan, diphenhydramine, and placebo on nocturnal cough and sleep quality for coughing children and their parents.
To determine whether the commonly used over-the-counter medications dextromethorphan and diphenhydramine are superior to placebo for the treatment of nocturnal cough and sleep difficulty associated with upper respiratory infections and to determine whether parents have improved sleep quality when their children receive the medications when compared with placebo. Parents of 100 children with upper respiratory infections were questioned to assess the frequency, severity, and bothersome nature of the nocturnal cough. Their answers were recorded on 2 consecutive days, initially on the day of presentation, when no medication had been given the previous evening, and then again on the subsequent day, when either medication or placebo was given before bedtime. Sleep quality for both the child and the parent were also assessed for both nights. For the entire cohort, all outcomes were significantly improved on the second night of the study when either medication or placebo was given. However, neither diphenhydramine nor dextromethorphan produced a superior benefit when compared with placebo for any of the outcomes studied. Insomnia was reported more frequently in those who were given dextromethorphan, and drowsiness was reported more commonly in those who were given diphenhydramine. Diphenhydramine and dextromethorphan are not superior to placebo in providing nocturnal symptom relief for children with cough and sleep difficulty as a result of an upper respiratory infection. Furthermore, the medications given to children do not result in improved quality of sleep for their parents when compared with placebo. Each clinician should consider these findings, the potential for adverse effects, and the individual and cumulative costs of the drugs before recommending them to families.


Patients' opinions about referral from a tertiary specialist psychiatric hospital to primary healthcare.
Referral of patients from tertiary specialist psychiatric hospitals to primary healthcare settings is a worldwide goal. This is of particular importance in South Africa with its considerable burden of mental disorders and limited resources. However, patients are often reluctant to be referred and studies have shown that patients may prefer a dedicated psychiatric service over an integrated primary healthcare service. This study explored the opinions of patients receiving care at a tertiary psychiatric hospital's outpatient department (OPD) about referral to a primary healthcare clinic (PHCC). The study was conducted at Weskoppies Psychiatric Hospital OPD. This was a qualitative study based on grounded theory. Participants were recruited through purposive-theoretical sampling. Data were collected by means of individual interviews and mini-essays. From the 80 participants, 18 had individual interviews and 62 wrote mini-essays. Thirty-nine participants had previously attended a PHCC, while 41 had not. Perceived advantages of referral to PHCCs included: close proximity to participants' homes, resulting in saving on travelling time and transport costs, as well as the convenience of receiving psychiatric and other medical treatment at the same healthcare facility. Perceived disadvantages of PHCCs included: unavailability of treatment; lack of doctor-based care; lack of specialised care; loss of established relationships with hospital healthcare workers; mistreatment by PHCC nursing staff; longer waiting times; more stigmatisation. The perceived disadvantages of referral from a tertiary psychiatric hospital to a PHCC outweighed the perceived advantages. Nonetheless, participants expressed willingness for such a referral if their concerns were addressed.


Soft self-assembled sub-5 nm scale chessboard and snub-square tilings with oligo(para-phenyleneethynylene) rods.
X-Shaped bolapolyphiles, comprising a linear polyaromatic core with glycerol groups at each end and two chemically different and incompatible chains, fixed to it at opposite sides, were synthesized and found to self-assemble into honeycomb-type liquid crystalline phases with square symmetry. The polyaromatic π-conjugated rods form the cell walls and the resulting prismatic cells of sub-5 nm size are alternatively filled with perfluorocarbon (RF) and the carbosilane chains (RSi). The resulting structures can be represented as either a two-colour snub-square tiling with triangular and square cells or as a chessboard tiling of squares.


Tobacco use among middle and high school students--United States, 2013.
Tobacco use is the leading preventable cause of disease and death in the United States, and nearly all tobacco use begins during youth and young adulthood. Among U.S. youths, cigarette smoking has declined in recent years; however, the use of some other tobacco products has increased, and nearly half of tobacco users use two or more tobacco products. CDC analyzed data from the 2013 National Youth Tobacco Survey to determine the prevalence of ever (at least once) and current (at least 1 day in the past 30 days) use of one or more of 10 tobacco products (cigarettes, cigars, hookahs, smokeless tobacco, electronic cigarettes [e-cigarettes], pipes, snus, bidis, kreteks, and dissolvable tobacco) among U.S. middle school (grades 6-8) and high school (grades 9-12) students. In 2013, 22.9% of high school students reported current use of any tobacco product, and 12.6% reported current use of two or more tobacco products; current use of combustible products (i.e., cigarettes, cigars, pipes, bidis, kreteks, and/or hookahs) was substantially greater (20.7%) than use of other types of tobacco. Also, 46.0% of high school students reported having ever tried a tobacco product, and 31.4% reported ever trying two or more tobacco products. Among middle school students, 3.1% reported current use of cigars, and 2.9% reported current use of cigarettes, with non-Hispanic black students more than twice as likely to report current use of cigars than cigarettes. Monitoring the prevalence of the use of all available tobacco products, including new and emerging products, is critical to support effective population-based interventions to prevent and reduce tobacco use among youths as part of comprehensive tobacco prevention and control programs.


INC280, an orally available small molecule inhibitor of c-MET, reduces migration and adhesion in ovarian cancer cell models.
5-year survival rates for ovarian cancer are approximately 40%, and for women diagnosed at late stage (the majority), just 27%. This indicates a dire need for new treatments to improve survival rates. Recent molecular characterization has greatly improved our understanding of the disease and allowed the identification of potential new targets. One such pathway of interest is the HGF/c-MET axis. Activation of the HGF/c-MET axis has been demonstrated in certain ovarian tumours, and been found to be associated with decreased overall survival, suggesting its potential as a therapeutic target. The objective of this study was to determine the efficacy of a novel, highly potent, orally-bioavailable c-MET inhibitor, INC280, in blocking cell phenotypes important in ovarian cancer metastasis. Using in vitro and ex vivo models, we demonstrate that INC280 inhibits HGF-induced c-MET, and reduces downstream signalling. HGF-stimulated chemotactic and random migration are decreased by INC280 treatment, to levels seen in non-stimulated cells. Additionally, HGF-induced adhesion of cancer cells to peritoneal tissue is significantly decreased by INC280 treatment. Overall, these data indicate that INC280 inhibits many cell behaviours that promote ovarian cancer metastasis, and merits further investigation as a therapeutic candidate in the treatment of patients with ovarian cancer.


Disappearance and formation rates of microaneurysms in early diabetic retinopathy.
To analyse the formation and disappearance rates of individual microaneurysms in mild background retinopathy. Three fluorescein angiograms were taken at 1 year intervals during a 2 year follow up from 24 type 1 diabetics with mild background retinopathy. Microaneurysms were identified and localised twice from each angiogram using a computerised system for retrieval of the coordinates for each microaneurysm. Microaneurysms identified similarly in both sessions were then processed further to obtain rates of microaneurysm formation and disappearance, and microaneurysm count changes. In the whole material the total number of microaneurysms increased from 298 to 436 from baseline to 2 years. During the 2 year follow up 395 new microaneurysms formed and 258 microaneurysms disappeared. Of the microaneurysms present at baseline 174 (58%) were still present at the 1 year and 142 (48%) at the 2 year follow up. In patients with good glucose control (HbA1c < or = 7.5 mmol) microaneurysm formation showed a trend of being decreased whereas microaneurysm disappearance did not correlate with glucose control. Background diabetic retinopathy is a dynamic process. A significant proportion of microaneurysms present disappear within 2 years. This is compensated for by formation of new microaneurysms, the resultant net changes in microaneurysm counts being small. Microaneurysm formation and disappearance rates are new variables of diabetic retinopathy and may prove to be more sensitive indicators of the progression patterns of background diabetic retinopathy than microaneurysm count changes.


Why inner city mothers take their children for routine medical and dental examinations.
The preventive behaviour relating to visits for dental and medical examination of 100 mothers of pre-school children was investigated. The theory of reasoned action of Fishbein and Ajzen was used to predict the intention of 100 mothers to visit doctors and dentists. The study was conducted in one of the most socially deprived wards of the London Borough of Bloomsbury. The relationship between beliefs and attitudes held by mothers on preventive dental and repeated medical visits was investigated. In younger mothers aged 16-24 years, individual attitudes and subjective norms were better predictors of intention to visit the dentist every 6 months than were the total attitude and total subjective norm scores. The prediction of intention to visit the doctor and the dentist showed a similar pattern, but the variance accounted for by intention to visit the doctor was lower. Younger mothers showed stronger attitudes associated with dental treatment outcome, whereas older mothers showed a more positive preventive dental orientation. This finding is supported by principal components analysis of both the medical and dental data. The theory of reasoned action was successfully applied, but the external variable age was a very important prediction factor. This modifying factor, in relation to Ajzen and Fishbein's theory, gives a better insight into preventive dental and medical health behaviours.


The proofreading exonuclease subunit epsilon of Escherichia coli DNA polymerase III is tethered to the polymerase subunit alpha via a flexible linker.
Escherichia coli DNA polymerase III holoenzyme is composed of 10 different subunits linked by noncovalent interactions. The polymerase activity resides in the alpha-subunit. The epsilon-subunit, which contains the proofreading exonuclease site within its N-terminal 185 residues, binds to alpha via a segment of 57 additional C-terminal residues, and also to theta, whose function is less well defined. The present study shows that theta greatly enhances the solubility of epsilon during cell-free synthesis. In addition, synthesis of epsilon in the presence of theta and alpha resulted in a soluble ternary complex that could readily be purified and analyzed by NMR spectroscopy. Cell-free synthesis of epsilon from PCR-amplified DNA coupled with site-directed mutagenesis and selective 15N-labeling provided site-specific assignments of NMR resonances of epsilon that were confirmed by lanthanide-induced pseudocontact shifts. The data show that the proofreading domain of epsilon is connected to alpha via a flexible linker peptide comprising over 20 residues. This distinguishes the alpha : epsilon complex from other proofreading polymerases, which have a more rigid multidomain structure.


Direct Observation of α-Synuclein Amyloid Aggregates in Endocytic Vesicles of Neuroblastoma Cells.
Aggregation of α-synuclein has been linked to both familial and sporadic Parkinson's disease. Recent studies suggest that α-synuclein aggregates may spread from cell to cell and raise questions about the propagation of neurodegeneration. While continuous progress has been made characterizing α-synuclein aggregates in vitro, there is a lack of information regarding the structure of these species inside the cells. Here, we use confocal fluorescence microscopy in combination with direct stochastic optical reconstruction microscopy, dSTORM, to investigate α-synuclein uptake when added exogenously to SH-SY5Y neuroblastoma cells, and to probe in situ morphological features of α-synuclein aggregates with near nanometer resolution. We demonstrate that using dSTORM, it is possible to follow noninvasively the uptake of extracellularly added α-synuclein aggregates by the cells. Once the aggregates are internalized, they move through the endosomal pathway and accumulate in lysosomes to be degraded. Our dSTORM data show that α-synuclein aggregates remain assembled after internalization and they are shortened as they move through the endosomal pathway. No further aggregation was observed inside the lysosomes as speculated in the literature, nor in the cytoplasm of the cells. Our study thus highlights the super-resolution capability of dSTORM to follow directly the endocytotic uptake of extracellularly added amyloid aggregates and to probe the morphology of in situ protein aggregates even when they accumulate in small vesicular compartments.


Algorithm for backrub motions in protein design.
The Backrub is a small but kinematically efficient side-chain-coupled local backbone motion frequently observed in atomic-resolution crystal structures of proteins. A backrub shifts the C(alpha)-C(beta) orientation of a given side-chain by rigid-body dipeptide rotation plus smaller individual rotations of the two peptides, with virtually no change in the rest of the protein. Backrubs can therefore provide a biophysically realistic model of local backbone flexibility for structure-based protein design. Previously, however, backrub motions were applied via manual interactive model-building, so their incorporation into a protein design algorithm (a simultaneous search over mutation and backbone/side-chain conformation space) was infeasible. We present a combinatorial search algorithm for protein design that incorporates an automated procedure for local backbone flexibility via backrub motions. We further derive a dead-end elimination (DEE)-based criterion for pruning candidate rotamers that, in contrast to previous DEE algorithms, is provably accurate with backrub motions. Our backrub-based algorithm successfully predicts alternate side-chain conformations from < or = 0.9 A resolution structures, confirming the suitability of the automated backrub procedure. Finally, the application of our algorithm to redesign two different proteins is shown to identify a large number of lower-energy conformations and mutation sequences that would have been ignored by a rigid-backbone model. Contact authors for source code.


Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa, using CRISPR/Cas9 gene editing.
The CRISPR/Cas9 nuclease system is a powerful and flexible tool for genome editing, and novel applications of this system are being developed rapidly. Here, we used CRISPR/Cas9 to target the FAD2 gene in Arabidopsis thaliana and in the closely related emerging oil seed plant, Camelina sativa, with the goal of improving seed oil composition. We successfully obtained Camelina seeds in which oleic acid content was increased from 16% to over 50% of the fatty acid composition. These increases were associated with significant decreases in the less desirable polyunsaturated fatty acids, linoleic acid (i.e. a decrease from ~16% to <4%) and linolenic acid (a decrease from ~35% to <10%). These changes result in oils that are superior on multiple levels: they are healthier, more oxidatively stable and better suited for production of certain commercial chemicals, including biofuels. As expected, A. thaliana T2 and T3 generation seeds exhibiting these types of altered fatty acid profiles were homozygous for disrupted FAD2 alleles. In the allohexaploid, Camelina, guide RNAs were designed that simultaneously targeted all three homoeologous FAD2 genes. This strategy that significantly enhanced oil composition in T3 and T4 generation Camelina seeds was associated with a combination of germ-line mutations and somatic cell mutations in FAD2 genes in each of the three Camelina subgenomes.


An aptamer that neutralizes R5 strains of HIV-1 binds to core residues of gp120 in the CCR5 binding site.
We have previously isolated nucleic acid ligands (aptamers) that bind the surface envelope glycoprotein, gp120, of HIV-1, and neutralize infection of diverse sub-types of virus. Our earlier studies have identified the overall structure of one of these aptamers, B40, and have indicated that it binds to gp120 in a manner that competes with that of the HIV-1 coreceptor, CCR5, and select "CD4i" antibodies with epitopes overlapping this region. Here, we sought to map the B40 binding site on gp120 more precisely by analysing its interaction with a panel of alanine substitution mutants of gp120. Furthermore, we tested our hypothesis concerning the structure of the 40 nucleotide functional core of the aptamer by the solid-phase synthesis of truncated and chemically modified derivatives. The results confirm our structural predictions and demonstrate that aptamer B40 neutralizes a diverse range of HIV-1 isolates as a result of binding to relatively conserved residues on gp120 at the heart of the CCR5-binding site. These structural insights may provide the basis for the development of potential anti-viral agents with high specificity and robustness.


Interpreting two-photon imaging data of lymphocyte motility.
Recently, using two-photon imaging it has been found that the movement of B and T cells in lymph nodes can be described by a random walk with persistence of orientation in the range of 2 minutes. We interpret this new class of lymphocyte motility data within a theoretical model. The model considers cell movement to be composed of the movement of subunits of the cell membrane. In this way movement and deformation of the cell are correlated to each other. We find that, indeed, the lymphocyte movement in lymph nodes can best be described as a random walk with persistence of orientation. The assumption of motility induced cell elongation is consistent with the data. Within the framework of our model the two-photon data suggest that T and B cells are in a single velocity state with large stochastic width. The alternative of three different velocity states with frequent changes of their state and small stochastic width is less likely. Two velocity states can be excluded.


Towards a chemo-enzymatic method for the asymmetric synthesis of beta-amino tertiary alcohols.
The synthesis of a number of beta-amino tertiary alcohols has been achieved via ring-opening of an unsymmetrical epoxide with primary and secondary amines. The results revealed that primary amines give symmetrical triol products following an undesired acyl migration reaction, whereas secondary amines give the desired chiral (racemic) products. Furthermore, we demonstrate that asymmetric products can be formed using enantiomerically pure epoxide and aromatic amines without any loss of enantiomeric excess.


Surgical management of coccidioidomycosis of the spine: clinical article.
Coccidioides immitis is a dimorphous fungus endemic in the southwestern US and northern Mexico. While its primary presentation is pulmonary, it can have devastating neurological sequelae. The authors provide a retrospective review with long-term follow-up between 1986 and 2008 at a single institution. The authors identified 27 patients between 13 and 81 years old (mean 41.4 years) with spinal coccidioides who were treated surgically at the Barrow Neurological Institute between 1986 and 2008. There were 24 males (89%) and 3 females (11%). Eleven patients (41%) had cervical spine involvement, 15 (56%) had thoracic involvement, 7 (26%) had lumbar involvement, and 2 (7%) had sacral involvement. All 27 patients presented with localized or radiating pain. Nine patients (33%) had myelopathic symptoms at presentation, 5 (19%) had radiculopathy, 4 (15%) had fever, and 12 (44%) had progressive kyphosis. The disease was most frequently seen among African American patients (14 patients [52%]), followed by Caucasians (5 patients [19%]), Asians (3 patients [11%]), and Hispanics (3 patients [11%]). Ten patients (37%) required multiple operations at the same level. Follow-up was available in 19 patients (70%) (mean 9.8 months, range 1-39 months). Sixteen (84%) of these 19 patients improved from their preoperative baseline states, 1 (5%) was stable on examination, 1 patient's condition (5%) deteriorated compared with the preoperative examination, and 1 patient (5%) died in the postoperative period. Although spinal involvement of coccidioidomycosis is relatively uncommon, a high index of suspicion and aggressive therapy are warranted to prevent devastating neurological injury, and lifelong antifungal therapy is often warranted.


Doubly robust testing and estimation of model-adjusted effect-measure modification with complex survey data.
Model-based standardization enables adjustment for confounding of a population-averaged exposure effect on an outcome. It requires either a model for the probability of the exposure conditional on the confounders (an exposure model) or a model for the expectation of the outcome conditional on the exposure and the confounders (an outcome model). The methodology can also be applied to estimate averaged exposure effects within categories of an effect modifier and to test whether these effects differ or not. Recently, we extended that methodology for use with complex survey data, to estimate the effects of disability status on cost barriers to health care within three age categories and to test for differences. We applied the methodology to data from the 2007 Florida Behavioral Risk Factor Surveillance System Survey (BRFSS). The exposure modeling and outcome modeling approaches yielded two contrasting sets of results. In the present paper, we develop and apply to the BRFSS example two doubly robust approaches to testing and estimating effect modification with complex survey data; these approaches require that only one of these two models be correctly specified. Furthermore, assuming that at least one of the models is correctly specified, we can use the doubly robust approaches to develop and apply goodness-of-fit tests for the exposure and outcome models. We compare the exposure modeling, outcome modeling, and doubly robust approaches in terms of a simulation study and the BRFSS example.


Cholesterol removal from various samples by cholesterol-imprinted monosize microsphere-embedded cryogels.
Cholesterol-imprinted monosize poly(glycidyl methacrylate-N-methacryloyl-(L)-tyrosine methylester) microspheres were embedded into the poly(hydroxyethyl methacrylate) (PHEMA) cryogels and the resulting composite cryogel was used for the selective removal of cholesterol. Composite cryogels were characterized by swelling tests, multipoint BET apparatus, SEM, FTIR and elemental analysis studies. Specific surface area of the PHEMA cryogel was increased from 13 to 72.7 m(2)/g by embedding of microspheres. Composite cryogels removed 80% of cholesterol from homogenized milk. The maximum adsorption capacity was found as 42.7 mg/g for intestinal mimicking solution. After 20 adsorption-desorption cycles, there was no remarkable decrease in the adsorption capacity.


First report of ranavirus mortality in a common snapping turtle Chelydra serpentina.
An adult male snapping turtle with marked palpebral edema and multifocal skin ulceration was found alive in a marsh in southern Ontario in summer 2017. The turtle was transported to a rehabilitation facility and died 4 d after arrival. The carcass was submitted to the Canadian Wildlife Health Cooperative for post-mortem examination. Gross lesions included ulcerative conjunctivitis, necrotizing stomatitis, and splenomegaly. Microscopically, this corresponded to multisystemic fibrinonecrotizing vasculitis and severe fibrinous splenic necrosis. Liver tissue tested positive for frog virus 3-like ranavirus and negative for herpesvirus via polymerase chain reaction. The gross and microscopic lesions were consistent with previous reports of ranavirus infection in turtles and were severe enough to have been the cause of death in this case. This is the first report of morbidity and mortality in a common snapping turtle with a ranavirus infection, and the first reported case of ranavirus infection in a reptile in Canada. Ranaviruses are considered to be an emerging infectious disease in chelonians as they are increasing in distribution, prevalence, and host range.


Feasibility and perceived benefits of a framework for physician-parent follow-up meetings after a child's death in the PICU.
To evaluate the feasibility and perceived benefits of conducting physician-parent follow-up meetings after a child's death in the PICU according to a framework developed by the Collaborative Pediatric Critical Care Research Network. Prospective observational study. Seven Collaborative Pediatric Critical Care Research Network-affiliated children's hospitals. Critical care attending physicians, bereaved parents, and meeting guests (i.e., parent support persons, other health professionals). Physician-parent follow-up meetings using the Collaborative Pediatric Critical Care Research Network framework. Forty-six critical care physicians were trained to conduct follow-up meetings using the framework. All meetings were video recorded. Videos were evaluated for the presence or absence of physician behaviors consistent with the framework. Present behaviors were evaluated for performance quality using a 5-point scale (1 = low, 5 = high). Participants completed meeting evaluation surveys. Parents of 194 deceased children were mailed an invitation to a follow-up meeting. Of these, one or both parents from 39 families (20%) agreed to participate, 80 (41%) refused, and 75 (39%) could not be contacted. Of 39 who initially agreed, three meetings were canceled due to conflicting schedules. Thirty-six meetings were conducted including 54 bereaved parents, 17 parent support persons, 23 critical care physicians, and 47 other health professionals. Physician adherence to the framework was high; 79% of behaviors consistent with the framework were rated as present with a quality score of 4.3 ± 0.2. Of 50 evaluation surveys completed by parents, 46 (92%) agreed or strongly agreed the meeting was helpful to them and 40 (89%) to others they brought with them. Of 36 evaluation surveys completed by critical care physicians (i.e., one per meeting), 33 (92%) agreed or strongly agreed the meeting was beneficial to parents and 31 (89%) to them. Follow-up meetings using the Collaborative Pediatric Critical Care Research Network framework are feasible and viewed as beneficial by meeting participants. Future research should evaluate the effects of follow-up meetings on bereaved parents' health outcomes.


Molecular mapping of poly(methyl methacrylate) super-helix stereocomplexes.
In this study, by using X-ray powder diffraction profiles as blueprints, we successfully mapped the most probable molecular-level structural arrangement of the PMMA super-helix stereocomplexes through molecular dynamic simulations. Molecular-level resolution of the PMMA triple-helix supramolecule was not previously achievable by experimental methods. After constructing molecular models of stereo-regular complexes composed of linear it- and st-PMMAs, our all-atom molecular dynamics simulations identified the stereocomplex structure that best reproduces experimental diffraction profiles and thermodynamic properties as a double helix of isotactic (it-)PMMA with a helical pitch of 1.8 nm and 9 units per turn surrounded by a single helix of syndiotactic (st-)PMMA with an average helical pitch of 0.9 nm and 20 units per turn. The it-/st- complexing stoichiometry in the PMMA triple-helix is therefore 9 : 20. This presents the first all-atom model of the it-/st-PMMA triple-helix stereocomplex that accurately fits experimental X-ray diffraction profiles. In addition, the simulation results revealed the outer st-PMMA helix of the PMMA stereocomplex has a fiber diameter of at least 1.85 nm and adopts a non-ideal helical geometry. Furthermore, through dynamic simulations, surprising new sights into the effect of the structural configuration of the PMMA stereocomplex (i.e., helical pitch and direction, and tilt angle) on the physical properties of their crystal structures were obtained. Those crystal properties include X-ray diffraction profile, packing density, chain-chain spacing, chain width and cohesive energy density.


Guided internet treatment for anxiety disorders. As effective as face-to-face therapies?
Guided Internet-delivered treatments were developed in the late 1990s and have since been tested in numerous controlled trials. While promising, there are yet few direct comparisons between Internet treatments and traditional face-to-face treatments. The aim of the present study is to present an overview of the evidence in the field of anxiety disorders. Studies were located, including unpublished trials from our research group in Sweden. Results of direct comparative trials on panic disorder (n=3) and social anxiety disorder (n=3) show equivalent outcomes. One study on specific phobia did not show equivalent outcomes with an advantage for face-to-face treatment. However, a systematic review by Cuijpers et al. (2010) found equivalent outcomes across several self-help formats, suggesting that guided self-help overall can be as affective as face-to-face treatments. Overall, there are still few large-scale trials and statistical power is often limited. A preliminary conclusion is that guided Internet treatment can be as effective as face-to-face treatments, but there is a need to investigate moderators and mediators of the outcome.


Characterisation and optimisation of individual wastewater treatment systems.
Onsite individual wastewater treatment systems can provide a financially attractive alternative to a sewer connection in locations far from the existing sewer network. These systems are, however, relatively new, and practical experiences, especially long-term field studies, are lacking. Therefore, a thorough study of two compact biofilm-based, aerobic onsite systems, both of five population equivalents, was started in 2001. The assessment of the treatment performance of these systems, as well as the maintenance requirements and the characterisation of the feed are of great importance for the better understanding of the systems in order to optimise their design and performance. This paper presents an evaluation and discussion of the start-up and a starvation period of the two studied systems, followed by a characterisation of the incoming wastewater using activated sludge respirometry experiments in the context of the assessment and improvement of the denitrification process. Individual wastewater treatment systems are characterised by a rather long start-up period of 70-120 days. An important characteristic during the start-up is the nitrite peak, which indicates the initiation of the nitrification process. The respirometric experiments reveal that the failing denitrification is probably caused by an insufficient amount of readily biodegradable COD in the influent.


Dentin sialophosphoprotein knockout mouse teeth display widened predentin zone and develop defective dentin mineralization similar to human dentinogenesis imperfecta type III.
Dentin sialophosphoprotein (Dspp) is mainly expressed in teeth by the odontoblasts and preameloblasts. The Dspp mRNA is translated into a single protein, Dspp, and cleaved into two peptides, dentin sialoprotein and dentin phosphoprotein, that are localized within the dentin matrix. Recently, mutations in this gene were identified in human dentinogenesis imperfecta II (Online Mendelian Inheritance in Man (OMIM) accession number 125490) and in dentin dysplasia II (OMIM accession number 125420) syndromes. Herein, we report the generation of Dspp-null mice that develop tooth defects similar to human dentinogenesis imperfecta III with enlarged pulp chambers, increased width of predentin zone, hypomineralization, and pulp exposure. Electron microscopy revealed an irregular mineralization front and a lack of calcospherites coalescence in the dentin. Interestingly, the levels of biglycan and decorin, small leucine-rich proteoglycans, were increased in the widened predentin zone and in void spaces among the calcospherites in the dentin of null teeth. These enhanced levels correlate well with the defective regions in mineralization and further indicate that these molecules may adversely affect the dentin mineralization process by interfering with coalescence of calcospherites. Overall, our results identify a crucial role for Dspp in orchestrating the events essential during dentin mineralization, including potential regulation of proteoglycan levels.