Robust genomic copy number predictor of pan cancer metastasis.
Copy number alterations(CNAs) are the most common genetic changes observed in many cancers, reflecting the innate chromosomal instability of this disorder. Yet, how these alterations affect gene function to promote metastases across different tumor types has not been established. In this study, we developed a pan-cancer metastasis potential score (panMPS) based on observed CNAs. panMPS predicts metastasis and metastasis-free survival in cohorts of patients with prostate cancer, triple negative breast cancer and lung adenocarcinoma, and overall survival in the Metabric breast cancer cohort and three cohorts from The Cancer Genome Atlas (TCGA), including prostate, breast and lung adenocarcinoma. These CNAs are present in cell lines of metastatic tumors from eight different origins, reflected by an elevated panMPS for all cell lines. Many copy number alterations involve large chromosomal segments that encompass multiple genes ("clumps"). We show that harnessing this structural information to select only one gene per clump captures the contributions of other genes within the clump, resulting in a robust predictor of metastasis outcome. These sets of selected genes are distinct from cancer drivers that undergo mutation, and in fact, metastasis-related functions have been published for over half of them.


Can we apply lessons learned from Ebola experience in West Africa for COVID-19 in lower income countries?
The COVID-19 affects vulnerable groups disproportionally in a society where inequities are long-standing issue. Weak health system, especially the shortage and maldistribution of capable health workforce will be the main challenge in lower income countries to fight against the COVID-19. Applying the lesson learned and success from the Ebola outbreak in West Africa is important. International collaboration with already well functioned local mechanisms, such as the Network of Managers of Health Workforce in Francophone Africa is the key to provide prompt support. This approach contributes not only to the short-term COVID-19 control but also long-term strengthening of the sustainable and resilient health system in the lower income countries.


DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor.
Nuclease activity present within respiratory tissues contributes to the rapid clearance of injected DNA and therefore may reduce the transfection activity of directly injected transgenes. Most gene transfer technologies transduce or transfect murine tissues more efficiently than corresponding primate tissues. Therefore, it is prudent to assess the utility of novel gene transfer strategies in both rodent and primate models before proceeding with further development. This study analyzed the effects of ATA (a nuclease inhibitor) on the direct transfection of macaque and murine lung tissue; compared the levels of DNase activity in murine, primate, and human lung fluids; and tested the inhibitory activity of ATA on the DNase activity present in these samples. Fluorescent microspheres were used to detect areas of transfection in lung. Intratracheal administration of a nuclease inhibitor (ATA) with naked DNA (0.5 microg ATA/g body weight) enhanced direct transfection efficacy in macaque lung by over 86-fold and by over 54-fold in mouse lung. Hematoxylin and eosin staining showed no apparent tissue toxicity. Moreover, macaque, human, and mouse lung fluids were found to possess similar levels of DNase activity and this activity was inhibited by similar concentrations of ATA. The authors also successfully pioneered the use of carboxylate-modified microsphere tracers to identify areas of transfection and/or treatment. This work provides evidence that using direct nuclease inhibitors will enhance lung transfection and that nuclease activity is present in all lung fluids tested, which can be inhibited by the use of direct DNase inhibitors.


Hypertriglyceridemia: a too long unfairly neglected major cardiovascular risk factor.
The existence of an independent association between elevated triglyceride (TG) levels, cardiovascular (CV) risk and mortality has been largely controversial. The main difficulty in isolating the effect of hypertriglyceridemia on CV risk is the fact that elevated triglyceride levels are commonly associated with concomitant changes in high density lipoprotein (HDL), low density lipoprotein (LDL) and other lipoproteins. As a result of this problem and in disregard of the real biological role of TG, its significance as a plausible therapeutic target was unfoundedly underestimated for many years. However, taking epidemiological data together, both moderate and severe hypertriglyceridaemia are associated with a substantially increased long term total mortality and CV risk. Plasma TG levels partially reflect the concentration of the triglyceride-carrying lipoproteins (TRL): very low density lipoprotein (VLDL), chylomicrons and their remnants. Furthermore, hypertriglyceridemia commonly leads to reduction in HDL and increase in atherogenic small dense LDL levels. TG may also stimulate atherogenesis by mechanisms, such excessive free fatty acids (FFA) release, production of proinflammatory cytokines, fibrinogen, coagulation factors and impairment of fibrinolysis. Genetic studies strongly support hypertriglyceridemia and high concentrations of TRL as causal risk factors for CV disease. The most common forms of hypertriglyceridemia are related to overweight and sedentary life style, which in turn lead to insulin resistance, metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM). Intensive lifestyle therapy is the main initial treatment of hypertriglyceridemia. Statins are a cornerstone of the modern lipids-modifying therapy. If the primary goal is to lower TG levels, fibrates (bezafibrate and fenofibrate for monotherapy, and in combination with statin; gemfibrozil only for monotherapy) could be the preferable drugs. Also ezetimibe has mild positive effects in lowering TG. Initial experience with en ezetimibe/fibrates combination seems promising. The recently released IMPROVE-IT Trial is the first to prove that adding a non-statin drug (ezetimibe) to a statin lowers the risk of future CV events. In conclusion, the classical clinical paradigm of lipids-modifying treatment should be changed and high TG should be recognized as an important target for therapy in their own right. Hypertriglyceridemia should be treated.


Transcriptional regulation by competing transcription factor modules.
Gene regulatory networks lie at the heart of cellular computation. In these networks, intracellular and extracellular signals are integrated by transcription factors, which control the expression of transcription units by binding to cis-regulatory regions on the DNA. The designs of both eukaryotic and prokaryotic cis-regulatory regions are usually highly complex. They frequently consist of both repetitive and overlapping transcription factor binding sites. To unravel the design principles of these promoter architectures, we have designed in silico prokaryotic transcriptional logic gates with predefined input-output relations using an evolutionary algorithm. The resulting cis-regulatory designs are often composed of modules that consist of tandem arrays of binding sites to which the transcription factors bind cooperatively. Moreover, these modules often overlap with each other, leading to competition between them. Our analysis thus identifies a new signal integration motif that is based upon the interplay between intramodular cooperativity and intermodular competition. We show that this signal integration mechanism drastically enhances the capacity of cis-regulatory domains to integrate signals. Our results provide a possible explanation for the complexity of promoter architectures and could be used for the rational design of synthetic gene circuits.


Isolation and characterization of carnocyclin a, a novel circular bacteriocin produced by Carnobacterium maltaromaticum UAL307.
Carnobacterium maltaromaticum UAL307, isolated from fresh pork, exhibits potent activity against a number of gram-positive organisms, including numerous Listeria species. Three bacteriocins were isolated from culture supernatant, and using matrix-assisted laser desorption ionization-time of flight mass spectrometry and Edman sequencing, two of these bacteriocins were identified as piscicolin 126 and carnobacteriocin BM1, both of which have previously been described. The remaining bacteriocin, with a molecular mass of 5,862 Da, could not be sequenced by traditional methods, suggesting that the peptide was either cyclic or N-terminally blocked. This bacteriocin showed remarkable stability over a wide temperature and pH range and was unaffected by a variety of proteases. After digestion with trypsin and alpha-chymotrypsin, the peptide was de novo sequenced by tandem mass spectrometry and a linear sequence deduced, consisting of 60 amino acids. Based on this sequence, the molecular mass was predicted to be 5,880 Da, 18 units higher than the observed molecular mass, which suggested that the peptide has a cyclic structure. Identification of the genetic sequence revealed that this peptide is circular, formed by a covalent linkage between the N and C termini following cleavage of a 4-residue peptide leader sequence. The results of structural studies suggest that the peptide is highly structured in aqueous conditions. This bacteriocin, named carnocyclin A, is the first reported example of a circular bacteriocin produced by Carnobacterium spp.


CSF N-glycan profiles to investigate biomarkers in brain developmental disorders: application to leukodystrophies related to eIF2B mutations.
Primary or secondary abnormalities of glycosylation have been reported in various brain diseases. Decreased asialotransferrin to sialotransferrin ratio in cerebrospinal fluid (CSF) is a diagnostic marker of leukodystrophies related to mutations of genes encoding translation initiation factor, EIF2B. We investigated the CSF glycome of eIF2B-mutated patients and age-matched normal individuals in order to further characterize the glycosylation defect for possible use as a biomarker. We conducted a differential N-glycan analysis using MALDI-TOF/MS of permethylated N-glycans in CSF and plasma of controls and eIF2B-mutated patients. We found in control CSF that tri-antennary/bisecting and high mannose structures were highly represented in samples obtained between 1 to 5 years of age, whereas fucosylated, sialylated structures were predominant at later age. In CSF, but not in plasma, of eIF2B-mutated patient samples, we found increased relative intensity of bi-antennary structures and decreased tri-antennary/bisecting structures in N-glycan profiles. Four of these structures appeared to be biomarker candidates of glycomic profiles of eIF2B-related disorders. Our results suggest a dynamic development of normal CSF N-glycan profiles from high mannose type structures to complex sialylated structures that could be correlated with postnatal brain maturation. CSF N-glycome analysis shows relevant quantitative changes associated with eIF2B related disorders. This approach could be applied to other neurological disorders involving developmental gliogenesis/synaptogenesis abnormalities.


Anaplastic Lymphoma Kinase as a Cancer Target in Pediatric Malignancies.
In this era of more rational therapies, substantial efforts are being made to identify optimal targets. The discovery of translocations involving the anaplastic lymphoma kinase (ALK) receptor tyrosine kinase in a subset of non-small cell lung cancers has become a paradigm for precision medicine. Notably, ALK was initially discovered as the fusion gene in anaplastic large cell non-Hodgkin lymphoma, a disease predominantly of childhood. The discovery of activating kinase domain mutations of the full-length ALK receptor as the major cause of hereditary neuroblastoma, and that somatically acquired mutations and amplification events often drive the malignant process in a subset of sporadic tumors, has established ALK as a tractable molecular target across histologically diverse tumors in which ALK is a critical mediator of oncogenesis. We are now uncovering the reexpression of this developmentally regulated protein in a broader subset of pediatric cancers, providing therapeutic targeting opportunities for diseases with shared molecular etiology. This review focuses on the role of ALK in pediatric malignancies, alongside the prospects and challenges associated with the development of effective ALK-inhibition strategies.


Guidance of B cells by the orphan G protein-coupled receptor EBI2 shapes humoral immune responses.
Humoral immunity depends on both rapid and long-term antibody production against invading pathogens. This is achieved by the generation of spatially distinct extrafollicular plasmablast and follicular germinal center (GC) B cell populations, but the signals that guide responding B cells to these alternative compartments have not been fully elucidated. Here, we show that expression of the orphan G protein-coupled receptor Epstein-Barr virus-induced gene 2 (EBI2, also known as GPR183) by activated B cells was essential for their movement to extrafollicular sites and induction of early plasmablast responses. Conversely, downregulation of EBI2 enabled B cells to access the center of follicles and promoted efficient GC formation. EBI2 therefore provides a previously uncharacterized dimension to B cell migration that is crucial for coordinating rapid versus long-term antibody responses.


Next-Generation Beneficial Microbes: The Case of Akkermansia muciniphila.
Metabolic disorders associated with obesity and cardiometabolic disorders are worldwide epidemic. Among the different environmental factors, the gut microbiota is now considered as a key player interfering with energy metabolism and host susceptibility to several non-communicable diseases. Among the next-generation beneficial microbes that have been identified, Akkermansia muciniphila is a promising candidate. Indeed, A. muciniphila is inversely associated with obesity, diabetes, cardiometabolic diseases and low-grade inflammation. Besides the numerous correlations observed, a large body of evidence has demonstrated the causal beneficial impact of this bacterium in a variety of preclinical models. Translating these exciting observations to human would be the next logic step and it now appears that several obstacles that would prevent the use of A. muciniphila administration in humans have been overcome. Moreover, several lines of evidence indicate that pasteurization of A. muciniphila not only increases its stability but more importantly increases its efficacy. This strongly positions A. muciniphila in the forefront of next-generation candidates for developing novel food or pharma supplements with beneficial effects. Finally, a specific protein present on the outer membrane of A. muciniphila, termed Amuc_1100, could be strong candidate for future drug development. In conclusion, as plants and its related knowledge, known as pharmacognosy, have been the source for designing drugs over the last century, we propose that microbes and microbiomegnosy, or knowledge of our gut microbiome, can become a novel source of future therapies.


Knowledge, Attitude and Practice Concerning Human Papilloma Virus Infection and its Health Effects among Rural Women, Karnataka, South India.
Cervical cancer is one of the commonest cancers among women all over the world. The association of cervical cancer with human papilloma virus (HPV) is well established. Knowledge about the causal relationship between HPV and cervical cancer is important to make appropriate, evidence-based health care choices. In this context we conducted a community based study among women about the knowledge, attitude and practice about HPV infections and their health effects. A cross sectional interview based house to house survey was conducted with a validated data collection tool covering sociodemographic factors, knowledge, attitude and practice about HPV and its health effects, among 1020 women from a rural village, Perdoor, in Udupi district, Karnataka, India in 2013-14. The mean age of participants was 38.9 years (SD=12.6). Study participants showed a high literacy rate (85.7%). Only 2.4% of sexually exposed women had undergone Pap smear testing. Partners of 4.4%women had undergone circumcision and they belonged to the Muslim community. Male condom usage was reported by 26 women (2.6%). However, none of the participants had heard of HPV and its health effects. This community based study found complete ignorance about HPV among rural South Indian women in spite of a high literacy level.


In situ structural analysis of the Yersinia enterocolitica injectisome.
Injectisomes are multi-protein transmembrane machines allowing pathogenic bacteria to inject effector proteins into eukaryotic host cells, a process called type III secretion. Here we present the first three-dimensional structure of Yersinia enterocolitica and Shigella flexneri injectisomes in situ and the first structural analysis of the Yersinia injectisome. Unexpectedly, basal bodies of injectisomes inside the bacterial cells showed length variations of 20%. The in situ structures of the Y. enterocolitica and S. flexneri injectisomes had similar dimensions and were significantly longer than the isolated structures of related injectisomes. The crystal structure of the inner membrane injectisome component YscD appeared elongated compared to a homologous protein, and molecular dynamics simulations documented its elongation elasticity. The ring-shaped secretin YscC at the outer membrane was stretched by 30-40% in situ, compared to its isolated liposome-embedded conformation. We suggest that elasticity is critical for some two-membrane spanning protein complexes to cope with variations in the intermembrane distance. DOI:http://dx.doi.org/10.7554/eLife.00792.001.


First Report of Vibrio tubiashii Associated with a Massive Larval Mortality Event in a Commercial Hatchery of Scallop Argopecten purpuratus in Chile.
The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of this pathology. The pathogenic strain was identified as Vibrio tubiashii based on its phenotypic properties and the sequence analysis of its 16S rRNA and housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA). When triplicate cultures of healthy 10-day-old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.87 ± 3.33%, 34.32 ± 4.94%, and 0% were observed at 12, 24, and 36 h, respectively; whereas uninfected larval cultures showed survival rates of 97.4 ± 1.24% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.66%, significantly (P < 0.001) lower than the control group (97.4 ± 1.20%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.84% after 48 h of exposure. This is the first report of the isolation of V. tubiashii from the diseased larvae of the scallop A. purpuratus, occurring in a commercial culture in Chile, and it was demonstrated that the VPAP30 strain exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds.


Non-functional thyroid cystadenoma in three boxer dogs.
Thyroid neoplasia is a common endocrine neoplasm in dogs. The boxer is one of the reported breeds predisposed to malignant thyroid neoplasia. However, the association between thyroid neoplasia, malignancy and breed should be considered with caution. This article describes the presentation, clinical pathological findings, computed tomographic (CT) imaging findings and histopathological features of benign cystic thyroid tumour (cystadenoma) diagnosed in three boxers. These three dogs were presented for investigation of unilateral (n = 2) or bilateral (n = 1) cervical masses with no associated clinical signs of thyroid dysfunction. In each case, post-contrast CT scan identified a large, lateralised, non-invasive, well-defined homogeneous cystic structure with a hyperattenuating contrast-enhancing capsule of suspected thyroid origin displacing the surrounding cervical tissues. Ultrasound-guided fine needle aspiration of the cysts yielded fluid with a high thyroxine concentration in each case. Histopathology was consistent with thyroid cystadenoma in all cases. One dog was concurrently diagnosed with oral melanoma and euthanased. Two dogs underwent surgical excision with one lost to follow-up after 36 months and the other euthanased after 16 months following diagnosis of mast cell tumour. To the authors' knowledge, this is the first detailed report of non-functional benign thyroid cystadenoma in dogs and provides relevant information about case management for this type of tumour. The presence of a large cystic structure associated with benign non-functional thyroid neoplasia may be a condition to which boxer dogs are predisposed.


Ecological Potential for Rabies Virus Transmission via Scavenging of Dead Bats by Mesocarnivores.
Multiple species of bats are reservoirs of rabies virus in the Americas and are occasionally the source of spillover infections into mesocarnivore species. Although rabies transmission generally is assumed to occur via bite, laboratory studies have demonstrated the potential for rabies transmission via ingestion of rabid animals. We investigated the ecological potential for this mode of transmission by assessing mesocarnivore scavenging behavior of dead bats in suburban habitats of Flagstaff, Arizona, US. In autumn 2013, summer 2014, and autumn 2015, we placed 104 rabies-negative bat carcasses either near buildings, in wildland areas, or in residential yards and then monitored them with trail cameras for 5 d. Overall, 52 (50%) bat carcasses were scavenged, with 39 (75%) of those scavenged by striped skunks ( Mephitis mephitis ). Within our study area, striped skunks had a higher ecological potential to contract rabies via ingestion of bat carcasses compared to other mesocarnivore species, due both to a greater number of encounters and a higher probability of ingestion per encounter (91%), and they were significantly more likely to approach bat carcasses in yards than in wildland areas. Raccoons ( Procyon lotor ) and gray foxes ( Urocyon cinereoargenteus ) had fewer encounters (nine and 13, respectively) and lower probability of ingesting bats (33% and 8%, respectively).


Antrodia cinnamomea sensitizes radio-/chemo-therapy of cancer stem-like cells by modulating microRNA expression.
The discovery of many tissue-specific cancer stem cells (CSCs) continues to attract scientific attention. These CSCs are considered to be associated with chemo- and radio-resistance, and consequently, failure of conventional anticancer therapies. The recent demonstration of several microRNAs as enhancers of tumorigenicity via modulation of epithelial-mesenchymal transition and cancer stemness, makes them putative novel therapeutic target in oncology. Antrodia cinnamomea is a Chinese traditional medicine with several biological functions including anti-inflammation, antioxidant, and cancer prevention. However, the anti-CSC capability of A. Cinnamomea is not clear yet. To investigate the inhibitory effect of A. cinnamomea mycelium and extract on CSCs derived from various human cancer cell lines using our in-house therapeutics and human genome-wide miRNA screening panels. A broad range of human cancer cell lines, including the acute monocytic leukemia (THP-1), glioblastoma multiforme (GBM 8401), lung carcinoma (A549), breast adenocarcinoma (MDA-MB-231), hepatoblastoma (HepG2), colorectal adenocarcinoma (SW620), and foreskin fibroblast (HS68), were exposed to A. cinnamomea in this study. CD133+ CSCs generated from the cell lines were characterized and isolated by flow cytometry, effect of chemo- and radiotherapy was assessed using the MTT assay, while the RT-PCR and human genome wide qRT-PCR determined the differential gene expression patterns. A comparative analysis of the anticancer effect of A. cinnamomea and Cisplatin, Taxol, or irradiation was also performed. Our results indicated that A. cinnamomea mycelium and its ethyl acetate extracts showed anti-proliferation effects against all types of CSCs, especially the lung, breast, and head and neck squamous cell carcinoma CSCs. Furthermore, CSCs treatment with A. cinnamomea combined with irradiation or chemotherapeutics demonstrated significant anti-cancer effect. We also established an association between the CSC-inhibitory effect of A. cinnamomea and significant downregulation of several microRNAs and cancer stemness expression levels in brain and breast CSCs. More importantly, higher CD133 expression is associated with poor prognosis in glioblastoma and breast cancer patients. Herein, we demonstrate the putative role of A. cinnamomea as an effective ethnopharmacologic therapeutic agent for cancer treatment.


Predictors of Outcome in Ulcerative Colitis.
Approximately 80% of patients with ulcerative colitis (UC) have intermittently active disease and up to 20% will require a colectomy, but little data available on predictors of poor disease course. The aim of this study was to identify clinical and genetic markers that can predict prognosis. Medical records of patients with UC with ≥5 years of follow-up and available DNA and serum were retrospectively assessed. Immunochip was used to genotype loci associated with immune mediated inflammatory disorders (IMIDs), inflammatory bowel diseases, and other single nucleotide polypmorphisms previously associated with disease severity. Serum levels of pANCA, ASCA, CBir1, and OmpC were also evaluated. Requirement for colectomy, medication, and hospitalization were used to group patients into 3 prognostic groups. Six hundred one patients with UC were classified as mild (n = 78), moderate (n = 273), or severe disease (n = 250). Proximal disease location frequencies at diagnosis were 13%, 21%, and 30% for mild, moderate, and severe UC, respectively (P = 0.001). Disease severity was associated with greater proximal extension rates on follow-up (P < 0.0001) and with shorter time to extension (P = 0.03) and to prednisone initiation (P = 0.0004). When comparing severe UC with mild and moderate UC together, diagnosis age >40 and proximal disease location were associated with severe UC (odds ratios = 1.94 and 2.12, respectively). None of the single nucleotide polypmorphisms or serum markers tested was associated with severe UC, proximal disease extension or colectomy. Older age and proximal disease location at diagnosis, but not genetic and serum markers, were associated with a more severe course. Further work is required to identify biomarkers that will predict outcomes in UC.


[The affective disorder-related burden imposed on the family environment--an overview].
The family burden (FB) has been defined as a multidimensional impact imposed by an illness on caregivers. FB can be divided into objective (i.e. related to measurable problems) and subjective one (i.e. related to caregivers' emotions arising in response to objective difficulties). FB is known to be related to disturbances in the functioning of the family system, higher level of stress, and the presence of financial problems. Some gender-dependent differences in the characteristics of FB have been found. Since family member's illness can be not only a ballast, but also a potential source of satisfaction, it has been found that the level of caregiving-related satisfaction is a significant predictor of FB severity. FB dynamics does not seem to be parallel to the course of illness. Problem-focused and task-focused coping strategies are known to be related to lower values of FB. There is evidence suggesting that in families of patients with BD depressive episodes trigger substantially higher severity of FB, as compared to manic episodes. Data on FB related to major depressive disorder (MDD) are scarce. Assertive community treatment strategies are the main option of reducing FB in the context of affective disorders, yet data on their effectiveness are inconclusive.


Mothers' parenting and child sex differences in behavior problems among African American preschoolers.
Sex differences in rates of behavior problems, including internalizing and externalizing problems, begin to emerge during early childhood. These sex differences may occur because mothers parent their sons and daughters differently, or because the impact of parenting on behavior problems is different for boys and girls. In this study, we examined whether associations between observations of mothers' positive and negative parenting and children's externalizing and internalizing behaviors vary as a function of child sex. The sample consisted of 137 African American low-income families with one sibling approximately 2 years old and the closest-aged older sibling who was approximately 4 years old. Results from fixed-effects within-family models indicate clear sex differences regardless of child age. Mothers were observed to use less positive parenting with sons than with daughters. Higher levels of observed negative parenting were linked to more externalizing behaviors for boys, whereas lower levels of positive parenting were linked to more externalizing behaviors for girls. No child sex differences emerged regarding associations between observed positive and negative parenting and internalizing behaviors.


Lithium use and discontinuation in a health maintenance organization.
For decades, lithium has been prescribed to treat bipolar disorder. Its efficacy has been documented by clinical trials and by data from specialized clinics. An issue of considerable interest is the use of lithium in managed care systems. This project examined the use of lithium and its continuity of use by enrollees of a health maintenance organization (HMO). In a 6-year longitudinal cohort study, 1,594 lithium users aged 15 years and older who were enrolled in an HMO were followed from 1986-1991. Data were obtained from automated databases and from medical record review of a random 5% subsample of the group (N = 74). Bipolar disorder was the most frequent morbidity treated. Eighty-four percent (N = 62 or 74) received services from a psychiatrist. Annual prevalence of lithium use increased over the 6-year period from 1.4 to 2.3 persons per thousand HMO members. Duration of use varied widely with some long-term but many short-term users. Discontinuation of treatment was associated with evaluated rates of psychiatric hospitalization and use of psychiatric emergency services. Lithium users were much more likely to receive a variety of other psychotropic drugs than nonusers of the same sex and age. Lithium use in this population was more often sporadic than continuous. Discontinuation of lithium was associated with psychiatric hospitalization. If lithium is to be a cost-effective treatment for bipolar disorder, managed care systems will need to improve adherence to drug treatment regimens.


Distribution of plantar pressure on the second metatarsal head during walking.
Although high pressure occurs on the second metatarsal head during waking, the distribution of this plantar pressure has not been reported. The purpose of the study was to analyze the distribution of plantar pressure on the second to the fourth metatarsal heads during walking in healthy adults. Plantar pressure was measured at the second and adjacent metatarsal heads by 2.54 mm x 2.54 mm transducers during walking in 72 healthy adults (32 women and 40 men). Data at the instant of peak pressure were analyzed using spatial statistics. The length and width of 90% predicted peak pressure (PPP), and anterior and posterior extents to 80, 70, 60, and 50% PPP were regressed by peak pressure, body height, body weight, gender, and foot length of the subjects. The distribution parameters of plantar pressure decreased proportionally from the second to the fourth metatarsal heads, and were different from the first metatarsal head. The distribution parameters were inversely correlated to peak pressure. Besides peak pressure, the length and width of 90% PPP, and anterior extent to 80% PPP could be predicted by body weight. The anterior extent to 70% PPP could be predicted by foot length. The posterior extent to 80, 70, 60, and 50% PPP could be predicted by body height of the subjects. Besides peak pressure, the distribution of plantar pressure was correlated with body weight, body height, and foot length at different areas on the second metatarsal head in normal adults.


Detection of Vero Cells Infected with Herpes Simplex Types 1 and 2 and Varicella Zoster Viruses Using Raman Spectroscopy and Advanced Statistical Methods.
Of the eight members of the herpes family of viruses, HSV1, HSV2, and varicella zoster are the most common and are mainly involved in cutaneous disorders. These viruses usually are not life-threatening, but in some cases they might cause serious infections to the eyes and the brain that can lead to blindness and possibly death. An effective drug (acyclovir and its derivatives) is available against these viruses. Therefore, early detection and identification of these viral infections is highly important for an effective treatment. Raman spectroscopy, which has been widely used in the past years in medicine and biology, was used as a powerful spectroscopic tool for the detection and identification of these viral infections in cell culture, due to its sensitivity, rapidity and reliability. Our results showed that it was possible to differentiate, with a 97% identification success rate, the uninfected Vero cells that served as a control, from the Vero cells that were infected with HSV-1, HSV-2, and VZV. For that, linear discriminant analysis (LDA) was performed on the Raman spectra after principal component analysis (PCA) with a leave one out (LOO) approach. Raman spectroscopy in tandem with PCA and LDA enable to differentiate among the different herpes viral infections of Vero cells in time span of few minutes with high accuracy rate. Understanding cell molecular changes due to herpes viral infections using Raman spectroscopy may help in early detection and effective treatment.


The dentist's role in bereavement support.
An aging population that is retaining more dentition and an increasing number of persons affected by terminal illnesses have resulted in dentists encountering dying patients and patient deaths with increasing frequency. These events call upon dentists and their staffs to provide support for the terminally ill and for survivors of the deceased. Dentists are effective in providing bereavement support and survivors benefit greatly from it. However, this type of support is stressful for dentists and many report the need for more formal education in this area. The ethical basis for dealing effectively with a dying patient's need for open and honest discussion derives from the obligations of the doctor-patient relationship. These ethical requirements carry over to providing some level of bereavement support for the surviving loved ones of the deceased and address the professional duties of compassion and care. Sending a sympathy card with a personal note, attending the patient's funeral, or making a follow-up telephone call to the patient's survivors are all appropriate, beneficial, and appreciated.


No effect of short-term exposure to GSM-modulated low-power microwaves on benzo(a)pyrene-induced tumours in rat.
In view of current interest in the biological effects of amplitude-modulated microwaves arising from the rapid development of mobile communications, the effects of low-level microwaves on cancer development were investigated using a rat sarcoma model. Two-month-old female Sprague-Dawley rats were treated by injection of benzo(a)pyrene and irradiated with GSM (Global System for Mobile)-modulated 900-MHz microwaves in an anechoic chamber at 55 or 200 microW cm(-2) (75 and 270 mW kg(-1) average whole-body SAR, 2h daily for 2 weeks). Rats were exposed from day 20, 40 or 75 after carcinogen injection. Additional groups of rats were sham-exposed in a second anechoic chamber. Anti-phosphatidylinositol autoantibody levels were evaluated in sera to monitor malignant transformation. Microwave exposure had no effect on the development of tumours. No acceleration or delays in tumour onset were observed. Animal survival was not modified and serum autoantibody levels were similar in exposed and sham-exposed groups. Low-level GSM microwave exposure of rat bearing benzo(a)pyrene-induced tumours had no effect on auto-antibody levels, tumour appearance and survival. The low exposure levels used here correspond to exposure limits for whole-body exposure of humans.


Management of Degenerative Lumbar Spinal Stenosis in the Elderly.
Lumbar spinal stenosis can cause symptomatic neurogenic claudication alongside radicular pain and weakness. In appropriately selected patients, surgical intervention has been demonstrated to provide for improvement in pain, disability, and quality of life. This systematic review sought to define the utility and safety of such decompression with or without arthrodesis in the management of symptomatic lumbar spinal stenosis for elderly patients older than 65 years of age. A systematic review was conducted using MEDLINE for literature published through December 2014. The first question focused on the effectiveness of lumbar spinal surgery for symptomatic lumbar spinal stenosis in elderly patients (over age 65 y). The second question focused on safety of surgical intervention on this elderly population with emphasis on perioperative complication rates. Review of 11 studies reveals that the majority of elderly patients exhibit significant symptomatic improvement, with overall benefits observed for pain (change visual analog scale 4.4 points) and disability (change Oswestry Disability Index 23 points). Review of 11 studies reveals that perioperative complications were infrequent and acceptable with pooled estimates of mortality (0.5%), inadvertent durotomy (5%), and wound infection (2%). Outcomes seem less favorable with greater complication rates among patients with diabetes or obesity. Based on largely low-quality, retrospective evidence, we recommend that elderly patients should not be excluded from surgical intervention for symptomatic lumbar spinal stenosis.


Social-ecological correlates of excessive TV viewing: difference by race and sex.
The primary and secondary purposes were to examine social-ecological correlates of excessive TV viewing (>2hr/day) in American children 1) between race/ethnic groups and 2) between boys and girls. Children (n = 48,505) aged 6 to 18 years from the 2007 National Survey of Children's Health were included. Social-ecological correlates included individual-, family-, and community-level variables. Logistic regression analyses were used for race/ethnicity [Non-Hispanic White (NHW), Non-Hispanic Black (NHB), Hispanic, other] and sex groups (boys, girls), to determine associated correlates. By race/ethnicity, 16.6%, 37.8%, and 24.5% of NHW, NHB, and Hispanic exceeded recommendations. For boys and girls, 23.7% and 18.2% viewed excessive TV. Having a TV in the bedroom and higher poverty level were associated with excessive TV in all groups. Differences by race/ethnicity were age, sex, TV in the bedroom, extracurricular activities, physical activity, adequate sleep, family structure, family meals, knowing child's friends, parent/child communication, and neighborhood characteristics. Differences by sex were family structure, parent/child communication, family meals, and neighborhood elements. Social-ecological correlates and associated prevalence of excessive TV viewing differed across subgroups. These specific correlates can be targeted in tailored interventions.


Primary, Secondary, and Tertiary Effects of Carbohydrate Ingestion During Exercise.
The purpose of this current opinion paper is to describe the journey of ingested carbohydrate from 'mouth to mitochondria' culminating in energy production in skeletal muscles during exercise. This journey is conveniently described as primary, secondary, and tertiary events. The primary stage is detection of ingested carbohydrate by receptors in the oral cavity and on the tongue that activate reward and other centers in the brain leading to insulin secretion. After digestion, the secondary stage is the transport of monosaccharides from the small intestine into the systemic circulation. The passage of these monosaccharides is facilitated by the presence of various transport proteins. The intestinal mucosa has carbohydrate sensors that stimulate the release of two 'incretin' hormones (GIP and GLP-1) whose actions range from the secretion of insulin to appetite regulation. Most of the ingested carbohydrate is taken up by the liver resulting in a transient inhibition of hepatic glucose release in a dose-dependent manner. Nonetheless, the subsequent increased hepatic glucose (and lactate) output can increase exogenous carbohydrate oxidation rates by 40-50%. The recognition and successful distribution of carbohydrate to the brain and skeletal muscles to maintain carbohydrate oxidation as well as prevent hypoglycaemia underpins the mechanisms to improve exercise performance.


Mechanical properties of mesh materials used for hernia repair and soft tissue augmentation.
Hernia repair is the most common surgical procedure in the world. Augmentation with synthetic meshes has gained importance in recent decades. Most of the published work about hernia meshes focuses on the surgical technique, outcome in terms of mortality and morbidity and the recurrence rate. Appropriate biomechanical and engineering terminology is frequently absent. Meshes are under continuous development but there is little knowledge in the public domain about their mechanical properties. In the presented experimental study we investigated the mechanical properties of several widely available meshes according to German Industrial Standards (DIN ISO). Six different meshes were assessed considering longitudinal and transverse direction in a uni-axial tensile test. Based on the force/displacement curve, the maximum force, breaking strain, and stiffness were computed. According to the maximum force the values were assigned to the groups weak and strong to determine a base for comparison. We discovered differences in the maximum force (11.1±6.4 to 100.9±9.4 N/cm), stiffness (0.3±0.1 to 4.6±0.5 N/mm), and breaking strain (150±6% to 340±20%) considering the direction of tension. The measured stiffness and breaking strength vary widely among available mesh materials for hernia repair, and most of the materials show significant anisotropy in their mechanical behavior. Considering the forces present in the abdominal wall, our results suggest that some meshes should be implanted in an appropriate orientation, and that information regarding the directionality of their mechanical properties should be provided by the manufacturers.


Type I interferon plays opposing roles in cytotoxicity and interferon-γ production by natural killer and CD8 T cells after influenza A virus infection in mice.
Type I interferons (IFNs) promote natural killer (NK) and CD8(+) T-cell responses, which play a role not only in the resolution of infection but also in the induction of acute lung injury following influenza A virus infection. We show here that IFN-α receptor knock-out (Ifnar1(-/-)) mice exhibited impaired cytotoxic activity as well as an increased ability of NK and CD8(+) T cells to produce IFN-γ after infection with influenza virus A/FM/1/47 (H1N1, a mouse-adapted strain). A deficiency in IFNAR signaling significantly impaired IL-10 production in influenza virus-infected lungs and enhanced IFN-γ production by NK cells, which were suppressed by exogenous IL-10. Depletion of NK cells but not CD8(+) T cells in Ifnar1(-/-) mice improved the survival rate after A/FM/1/47 infection, indicating that NK cells are responsible for acute lung injury in Ifnar1(-/-) mice following influenza A virus infection, although the depletion of IFN-γ did not improve the outcome. Thus, type I IFN signaling plays a role not only in the upregulation of cytotoxicity but also in the downregulation of some effector mechanisms including IFN-γ production by NK and CD8(+) T cells via IL-10 production.


The Spineless Origins of Prefrontal Cortex Dysfunction and Psychiatric Disorders.
Fundamental research into early circuits of the neocortex provides insight into the etiology of mental illness. In this issue of Neuron, Chini et al. (2020) probe the consequences of combined genetic and environmental perturbation on emergent network activity in the prefrontal cortex, identifying a window for possible intervention.


Intake and sources of added sugars among Australian children and adolescents.
To examine the intake and sources of added sugars (AS) of Australian children and adolescents, and compare their intake of free sugars (FS) to the recommended limit set by the World Health Organization (<10 % energy from FS). Data of 4140 children and adolescents aged 2-16 years with plausible intakes based on 2 × 24 h recalls from the 2007 Australian National Children Nutrition and Physical Activity Survey were used. AS content of foods was estimated based on a published method. Intakes of AS and FS, as well as food sources of AS, were calculated. One-way ANOVA was used for comparisons between age groups and gender. The mean (SD) AS intake was 58.9 (35.1) g/day, representing 11.9 (5.6) % of daily energy intake and 46.9 (17.5) % of daily total sugars intake. More than 80 % of the subjects had % energy from FS > 10 %. Significant increasing trends for AS intake, % energy from AS, % energy from FS across age groups were observed. Sugar-sweetened beverages (19.6 %), cakes, biscuits, pastries and batter-based products (14.3 %), and sugar and sweet spreads (10.5 %) were the top three contributors of AS intake in the whole sample. Higher contribution of AS from sugar-sweetened beverages was observed in adolescents (p trend < 0.001). A large proportion of Australian youths are consuming excessive amounts of energy from AS. Since the main sources of AS were energy-dense, nutrient-poor foods, interventions which target the reduction in these foods would reduce energy and AS intake with minimal impact to core nutrient intake.


The relationship between active travel to school and health-related fitness in children and adolescents: a systematic review.
Active travel to school (ATS) has been identified as an important source of physical activity for youth. However, the relationship between ATS and health-related fitness (HRF) among youth remains unclear. A systematic search of seven electronic databases (EMBASE, OVID MEDLINE, PsycINFO, PubMed, Scopus, SPORTDiscus and TRIS on line) was conducted in December 2009 and studies published since 1980 were considered for inclusion. Twenty seven articles were identified that explored the relationship between ATS and the following aspects of HRF: weight status/body composition, cardiorespiratory fitness, muscular fitness and flexibility. Forty-eight percent of the studies that examined the relationship between ATS and weight status/body composition reported significant associations, this increased to 55% once poor quality studies were removed. Furthermore, the findings from five studies, including one longitudinal study, indicate that ATS is positively associated with cardiorespiratory fitness in youth. However, the evidence for the relationships between ATS and muscular fitness or flexibility is equivocal and limited by low study numbers. There is some evidence to suggest that ATS is associated with a healthier body composition and level of cardiorespiratory fitness among youth. Strategies to increase ATS are warranted and should be included in whole-of-school approaches to the promotion of physical activity.


Nuclear transfer in the rhesus monkey: practical and basic implications.
In early 1997, the birth of a lamb after transfer of the nucleus from an adult mammary gland cell into an enucleated oocyte, along with the production of rhesus monkeys by nuclear transfer of embryonic cells, marked a reemergence of the field of mammalian cloning. Clonally derived rhesus monkeys would be invaluable in biomedical research, and the commercial interests in transgenic sheep and cattle propagated by cloning are substantial. Nuclear transfer technology is under consideration in human in vitro fertilization clinics to overcome infertility secondary to advanced maternal age or mitochondrial-based genetic disease. Nuclear transfer involves preparing a cytoplast as a recipient cell, in most cases a mature metaphase II oocyte from which the chromosomes have been removed. A donor nucleus cell is then placed between the zona and the cytoplast, and fusion, as well as cytoplast activation, is initiated by electrical stimulation. Successful reprogramming of the donor cell nucleus by the cytoplast is critical--a step that may be influenced by cell cycle stage. Embryos produced by nuclear transfer are cultured in vitro for several cell divisions before cryopreservation or transfer to the oviduct or uterus of a host mother. The efficiency of producing live young by nuclear transfer in domestic species is low, with a high frequency of developmental abnormalities in both preterm and term animals. However, a number of pregnancies have now been established using fetal cells as the source of donor nuclei. The use of cell lines not only allows large clone sizes but also supports the ability to genetically manipulate cells in vitro before nuclear transfer. Ongoing research focused on the production of clonally derived rhesus monkeys using fetal fibroblasts and embryonic stem cells as the source of donor nuclei will be reviewed.


A tricky water molecule coordinated to a verdazyl radical-iron(II) complex: a multitechnique approach.
The first iron complexes of high-spin iron(II) species directly coordinated to verdazyl radicals, [Fe(II)(vdCOO)2(H2O)2]·2H2O (1; vdCOO(-) = 1,5-dimethyl-6-oxo-verdazyl-3-carboxylate) and [Fe(II)(vdCOO)2(D2O)2]·2D2O (2), were synthesized. The crystal structure of 1 was investigated by single-crystal X-ray diffraction at room temperature and at 90 K. The compound crystallizes in the P1 space group with no phase transition between 300 and 90 K. The crystals are composed of discrete [Fe(II)(vdCOO)2(H2O)2] complexes and crystallization water molecules. In the complex, two vdCOO(-) ligands coordinate to the iron(II) ion in a head-to-tail arrangement and two water molecules complete the coordination sphere. The Fe-X (X = O, N) distances vary in the 2.069-2.213 Å range at 300 K and in the 2.0679-2.2111 Å range at 90 K, indicating that the iron(II) ion is in its high-spin (HS) state at both temperatures. At 300 K, one of the coordinated water molecules is H-bonded to two crystallization water molecules whereas the second one appears as loosely H-bonded to the two oxygen atoms of the carboxylate group of two neighboring complexes. At 90 K, the former H-bonds remain essentially the same whereas the second coordinated water molecule reveals a complicated behavior appearing simultaneously as tightly H-bonded to two oxygen atoms and non-H-bonded. The (57)Fe Mössbauer spectra, recorded between 300 K and 10 K, give a clue to this situation. They show two sets of doublets typical of HS iron(II) species whose intensity ratio varies smoothly with temperature. It demonstrates the existence of an equilibrium between the high temperature and low temperature forms of the compounds. The solid-state magic angle spinning (2)H NMR spectra of 2 were recorded between 310 K and 193 K. The spectra suggest the existence of a strongly temperature-dependent motion of one of the coordinated water molecules in the whole temperature range. Variable-temperature magnetic susceptibility measurements indicate an antiferromagnetic interaction (J(Fe-vd) = -27.1 cm(-1); H = -J(ij)S(i)S(j)) of the HS iron(II) ion and the radical spins with high g(Fe) and D(Fe) values (g(Fe) = 2.25, D(Fe) = +3.37 cm(-1)) for the HS iron(II) ion. Moreover, the radicals are strongly antiferromagnetically coupled through the iron(II) center (J(vd-vd) = -42.8 cm(-1)). These last results are analysed based on the framework of the magnetic orbitals formalism.


MultiFacTV: module detection from higher-order time series biological data.
Identifying modules from time series biological data helps us understand biological functionalities of a group of proteins/genes interacting together and how responses of these proteins/genes dynamically change with respect to time. With rapid acquisition of time series biological data from different laboratories or databases, new challenges are posed for the identification task and powerful methods which are able to detect modules with integrative analysis are urgently called for. To accomplish such integrative analysis, we assemble multiple time series biological data into a higher-order form, e.g., a gene × condition × time tensor. It is interesting and useful to develop methods to identify modules from this tensor. In this paper, we present MultiFacTV, a new method to find modules from higher-order time series biological data. This method employs a tensor factorization objective function where a time-related total variation regularization term is incorporated. According to factorization results, MultiFacTV extracts modules that are composed of some genes, conditions and time-points. We have performed MultiFacTV on synthetic datasets and the results have shown that MultiFacTV outperforms existing methods EDISA and Metafac. Moreover, we have applied MultiFacTV to Arabidopsis thaliana root(shoot) tissue dataset represented as a gene × condition × time tensor of size 2395 × 9 × 6(3454 × 8 × 6), to Yeast dataset and Homo sapiens dataset represented as tensors of sizes 4425 × 6 × 6 and 2920 × 14 × 9 respectively. The results have shown that MultiFacTV indeed identifies some interesting modules in these datasets, which have been validated and explained by Gene Ontology analysis with DAVID or other analysis. Experimental results on both synthetic datasets and real datasets show that the proposed MultiFacTV is effective in identifying modules for higher-order time series biological data. It provides, compared to traditional non-integrative analysis methods, a more comprehensive and better view on biological process since modules composed of more than two types of biological variables could be identified and analyzed.


Pharmacological properties of the receptor(s) involved in the 5-hydroxytryptamine-induced contraction of the feline middle cerebral artery.
The characterization of the receptor(s) involved in the 5-hydroxytryptamine (5-HT)-induced contraction was studied in the cat middle cerebral artery. 5-HT agonists and antagonists were tested on isolated arterial segments and their vascular potency correlated with their affinity value for 5-HT receptor subtypes as defined in the literature. 5-Carboxamidotryptamine (5-CT) and RU 24969 (5-HT1A-1B agonists) were more potent and as potent as 5-HT, respectively. Agonists at the 5-HT1C-2 (alpha-methyl-5-HT) and 5-HT3 (2-methyl-5-HT) sites were significantly less potent than 5-HT. All these agents induced similar maximal effects. The selective 5-HT1A agonist, 8-hydroxy-2(di-n-propylamino)tetralin, was the least potent but elicited a more intense vasoconstriction. The 5-HT-induced contraction was potently inhibited by compounds with high affinity at the 5-HT2 site in an apparently noncompetitive manner. The order of potency was: pizotifen greater than ritanserin greater than or equal to dihydroergotamine greater than or equal to ketanserin greater than or equal to methysergide. The nonselective agent methiothepin inhibited strongly the vasoconstriction induced by both 5-HT and 5-CT although a decrease in the maximal responses was observed together with a shift of the dose-response curves. In contrast to 5-HT, the contraction induced by 5-CT was only slightly affected by micromolar concentrations of ketanserin. 5-HT1A-1B (propranolol and cyanopindolol), 5-HT1C (mesulergine) and 5-HT3 (MDL 7222 and GR 38032F) selective drugs were almost completely devoid of significant antagonistic activity. Thus, the pharmacology of the feline cerebrovascular receptor(s) shares agonist similarities with the 5-HT1B/5-HT1D receptor subtypes whereas its antagonist profile relates more to that of the 5-HT2 site described in mammalian brain. Although these results may suggest the presence of two receptors in this cerebrovascular smooth muscle, they do not exclude the possibility that a single 5-HT receptor, not yet described by radioligand binding techniques, mediates the contractions induced by 5-HT. A definitive classification of this site, however, appears premature and would require novel and more selective agents.


Open-source, community-driven microfluidics with Metafluidics.
Microfluidic devices have the potential to automate and miniaturize biological experiments, but open-source sharing of device designs has lagged behind sharing of other resources such as software. Synthetic biologists have used microfluidics for DNA assembly, cell-free expression, and cell culture, but a combination of expense, device complexity, and reliance on custom set-ups hampers their widespread adoption. We present Metafluidics, an open-source, community-driven repository that hosts digital design files, assembly specifications, and open-source software to enable users to build, configure, and operate a microfluidic device. We use Metafluidics to share designs and fabrication instructions for both a microfluidic ring-mixer device and a 32-channel tabletop microfluidic controller. This device and controller are applied to build genetic circuits using standard DNA assembly methods including ligation, Gateway, Gibson, and Golden Gate. Metafluidics is intended to enable a broad community of engineers, DIY enthusiasts, and other nontraditional participants with limited fabrication skills to contribute to microfluidic research.


Establishing a protocol for building a pan-Canadian population-based monitoring system for early childhood development for children with health disorders: Canadian Children's Health in Context Study (CCHICS).
Health disorders early in life have tremendous impact on children's developmental trajectories. Almost 80% of children with health disorders lack the developmental skills to take full advantage of school-based education relative to 27% of children without a health disorder. In Canada, there is currently a dearth of nationally representative data on the social determinants of early childhood development for children with health disorders. Evidence from Canada and other countries indicate that poorer developmental outcomes in typically developing children are associated with lower socioeconomic status (SES). However, to date, it is not known whether this relationship is stronger among children with health disorders. The study's objectives are to estimate the prevalence and to investigate social determinants of developmental outcomes for young children with health disorders, using the Early Development Instrument (EDI). Study objectives will be achieved through three steps. First, using existing EDI data for 10 provinces and 2 territories collected from 2004 to 2015, we will investigate differences in developmental health outcomes among children with identified health disorders. Second, population-level EDI data will be linked with neighbourhood sociodemographic census data to explore associations between socioeconomic characteristics and rates of specific diagnoses among children aged 5-6 years, including trends over time. Third, for 3 of these 12 regions, additional health and/or education databases will be linked at an individual level. These data will be used to establish differences in EDI outcomes in relation to the age-of-onset of diagnosis, and presence of intervention or treatment. Study methodologies have been approved by the Hamilton Integrated Research Ethics Board. The results of the analyses of developmental health outcomes for children with health disorders combined with SES will have implications for both health service delivery and school-based intervention strategies. Results will contribute to a framework for public policy.


Enterocytozoon bieneusi infection in acquired immunodeficiency syndrome-related sclerosing cholangitis.
Acalculous cholecystitis and sclerosing cholangitis due to Cryptosporidium sp, and cytomegalovirus have been described in patients with the acquired immunodeficiency syndrome (AIDS). However, in about 40% of cases of AIDS-related biliary disease, no opportunistic pathogen is identified. The current case report describes the first case, to the best of the authors' knowledge, of AIDS-related sclerosing cholangitis associated with microsporidiosis. Enterocytozoon bieneusi was detected in the duodenum and bile by means of light microscopy and confirmed by electron microscopy. Microsporidian infection should be suspected in patients with AIDS-related sclerosing cholangitis as well as in cases of diarrhea in which none of the usual pathogens are found.


Tumor capacitance: electrical measurements of renal neoplasia.
Studies have demonstrated that biological tissues possess unique electrical properties. We evaluate the electrical properties of renal tumors using a specialized probe with the capability of measuring intra-tissue capacitance in an ex vivo model of fresh surgically excised tissue. An electrical monitoring device was used to measure tissue capacitance at a frequency of 1 MHz on 34 ex vivo kidney specimens freshly obtained after surgical excision. Tissue capacitance was promptly measured in the excised tumor as well as surrounding normal parenchyma and fat. Dielectric permittivity in each tissue was calculated using the measured capacitance data. These data were compared and correlated to pathological findings. The final pathology on the 34 specimens revealed 28 renal cell carcinomas (RCC), 3 oncocytomas and 3 angiomyolipomas. In patients with RCC dielectric permittivity of tumor tissue was 1.43 +/- 0.39 times greater than that of surrounding normal parenchyma (p < 0.001). The average tumor-to-normal tissue dielectric permittivity ratio for RCC was significantly greater than that for angiomyolipoma (1.43 +/- 0.39 vs 0.73 +/- 0.77, p < 0.05) but similar to that for oncocytoma (1.43 +/- 0.39 vs 1.63 +/- 0.77, p = 0.39). Tissue capacitance measurements may be used to differentiate renal tumor from surrounding normal tissue. In vivo studies will ultimately determine the clinical use of this technology in localizing renal neoplasms and differentiating between malignant and benign tissues.


Social networking profile correlates of schizotypy.
Social networking sites, such as Facebook, are extremely popular and have become a primary method for socialization and communication. Despite a report of increased use among those on the schizophrenia-spectrum, few details are known about their actual practices. In the current research, undergraduate participants completed measures of schizotypy and personality, and provided access to their Facebook profiles. Information from the profiles were then systematically coded and compared to the questionnaire data. As predicted, social anhedonia (SocAnh) was associated with a decrease in social participation variables, including a decrease in number of friends and number of photos, and an increase in length of time since communication with a friend, but SocAnh was also associated with an increase in profile length. Also, SocAnh was highly correlated with extraversion. Relatedly, extraversion uniquely predicted the number of friends and photos and length of time since communication with a friend. In addition, perceptual aberration/magical ideation (PerMag) was associated with an increased number of "black outs" on Facebook profile print-outs, a measure of paranoia. Overall, results from this naturalistic-like study show that SocAnh and extraversion are associated with decreased social participation and PerMag with increased paranoia related to information on social networking sites.


Characterization of Sulfur and Nanostructured Sulfur Battery Cathodes in Electron Microscopy Without Sublimation Artifacts.
Lithium sulfur (Li-S) batteries have the potential to provide higher energy storage density at lower cost than conventional lithium ion batteries. A key challenge for Li-S batteries is the loss of sulfur to the electrolyte during cycling. This loss can be mitigated by sequestering the sulfur in nanostructured carbon-sulfur composites. The nanoscale characterization of the sulfur distribution within these complex nanostructured electrodes is normally performed by electron microscopy, but sulfur sublimates and redistributes in the high-vacuum conditions of conventional electron microscopes. The resulting sublimation artifacts render characterization of sulfur in conventional electron microscopes problematic and unreliable. Here, we demonstrate two techniques, cryogenic transmission electron microscopy (cryo-TEM) and scanning electron microscopy in air (airSEM), that enable the reliable characterization of sulfur across multiple length scales by suppressing sulfur sublimation. We use cryo-TEM and airSEM to examine carbon-sulfur composites synthesized for use as Li-S battery cathodes, noting several cases where the commonly employed sulfur melt infusion method is highly inefficient at infiltrating sulfur into porous carbon hosts.


Removal by sorption and in situ biodegradation of oil spills limits damage to marine biota: a laboratory simulation.
This study examined the efficiency of cotton grass fibers in removing diesel oil from the surface of water in conditions prevailing in the Baltic Sea. The effect of low temperature, salinity, and bacterial amendments were tested in laboratory-scale set-ups, whereas 600-L mesocosms filled with Baltic Sea water were used for testing the effects of diesel oil and rapid removal of the oil on microorganisms, phytoplankton, and mussels. Cotton grass proved to be an excellent sorbent for diesel oil from the water surface at a low temperature. Inoculation with diesel-enriched microorganisms enhanced degradation of oil significantly in laboratory-scale experiments. In mesocosm experiments, the addition of diesel oil (0.66 mg L(-1), 0.533 L m(-2)) to the basins resulted in higher microbial density than in all other basins, including inoculated ones, suggesting that the Baltic Sea contains indigenous hydrocarbon degraders. The removal of oil with cotton grass significantly improved the survival of mussels in the mesocosm tests: 100% mortality in diesel basins versus 0% mortality in basins with cotton grass, respectively. However, the surviving mussels suffered from histopathological changes such as inflammatory responses, degenerations, and cell death. The observed rescuing effect was observable even when the cotton grass-bound oil was left in the water. The results underline the importance of rapid action in limiting damage caused by oil spills.


Adsorption of trimethyl phosphate and triethyl phosphate on dry and water pre-covered hematite, maghemite, and goethite nanoparticles.
Adsorption of trimethyl phosphate (TMP) and triethyl phosphate (TEP) on well-characterized nanoparticles of hematite (α-Fe(2)O(3)), maghemite (γ-Fe(2)O(3)), and goethite (α-FeOOH) has been studied by in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), 2D correlation DRIFTS analysis, and X-ray photoelectron spectroscopy (XPS) on dry and water pre-covered surfaces. It is shown that, at room temperature and low coverage, both TMP and TEP coordinate to Lewis acid Fe sites through the O phosphoryl atom on hematite and maghemite, while hydrogen bonding to Brønstedt acid surface OH groups dominates on goethite. At room temperature, slow dissociation of TMP occurs on the iron (hydr)oxide nanoparticles, whereby a methoxy group is displaced to form surface methoxy, leaving adsorbed dimethyl phosphate (DMP). Methoxy is further decomposed to formate, suggesting an oxidative degradation pathway in synthetic air on the oxide particles. Relatively, larger amounts of DMP and surface methoxy form on maghemite, while more formate is produced on hematite. Upon TMP adsorption on dry goethite nanoparticles, no oxidation surface products were detected. Instead, a slow TMP hydrolysis pathway is observed, yielding orthophosphate. It is found that pre-adsorbed water stimulates the hydrolysis of TMP. In contrast to TMP, TEP adsorbs molecularly on all iron hydr(oxide) nanoparticles. This is attributed to the longer aliphatic chain, which stabilizes the loss of charge on the methoxy CO bonds by charge redistribution upon phosphoryl O coordination to Fe surface atoms. The presented results implicate different reactivity depending on specific molecular structure of the organophosphorus compound (larger functional groups can compensate loss of charge due to surface coordination) and iron (hydr)oxide surface structure (exposing Lewis acid or Brønstedt acid sites).


Comparison of purple carrot juice and β-carotene in a high-carbohydrate, high-fat diet-fed rat model of the metabolic syndrome.
Anthocyanins, phenolic acids and carotenoids are the predominant phytochemicals present in purple carrots. These phytochemicals could be useful in treatment of the metabolic syndrome since anthocyanins improve dyslipidaemia, glucose tolerance, hypertension and insulin resistance; the phenolic acids may also protect against CVD and β-carotene may protect against oxidative processes. In the present study, we have compared the ability of purple carrot juice and β-carotene to reverse the structural and functional changes in rats fed a high-carbohydrate, high-fat diet as a model of the metabolic syndrome induced by diet. Cardiac structure and function were defined by histology, echocardiography and in isolated hearts and blood vessels; liver structure and function, oxidative stress and inflammation were defined by histology and plasma markers. High-carbohydrate, high-fat diet-fed rats developed hypertension, cardiac fibrosis, increased cardiac stiffness, endothelial dysfunction, impaired glucose tolerance, increased abdominal fat deposition, altered plasma lipid profile, liver fibrosis and increased plasma liver enzymes together with increased plasma markers of oxidative stress and inflammation as well as increased inflammatory cell infiltration. Purple carrot juice attenuated or reversed all changes while β-carotene did not reduce oxidative stress, cardiac stiffness or hepatic fat deposition. As the juice itself contained low concentrations of carotenoids, it is likely that the anthocyanins are responsible for the antioxidant and anti-inflammatory properties of purple carrot juice to improve glucose tolerance as well as cardiovascular and hepatic structure and function.


Evolution of the gene network underlying wing polyphenism in ants.
Wing polyphenism in ants evolved once, 125 million years ago, and has been a key to their amazing evolutionary success. We characterized the expression of several genes within the network underlying the wing primordia of reproductive (winged) and sterile (wingless) ant castes. We show that the expression of several genes within the network is conserved in the winged castes of four ant species, whereas points of interruption within the network in the wingless castes are evolutionarily labile. The simultaneous evolutionary lability and conservation of the network underlying wing development in ants may have played an important role in the morphological diversification of this group and may be a general feature of polyphenic development and evolution in plants and animals.


Molecular Epigenetics: Chemical Biology Tools Come of Age.
The field of epigenetics has exploded over the last two decades, revealing an astonishing level of complexity in the way genetic information is stored and accessed in eukaryotes. This expansion of knowledge, which is very much ongoing, has been made possible by the availability of evermore sensitive and precise molecular tools. This review focuses on the increasingly important role that chemistry plays in this burgeoning field. In an effort to make these contributions more accessible to the nonspecialist, we group available chemical approaches into those that allow the covalent structure of the protein and DNA components of chromatin to be manipulated, those that allow the activity of myriad factors that act on chromatin to be controlled, and those that allow the covalent structure and folding of chromatin to be characterized. The application of these tools is illustrated through a series of case studies that highlight how the molecular precision afforded by chemistry is being used to establish causal biochemical relationships at the heart of epigenetic regulation.


Paleo-demography of the Drosophila melanogaster subgroup: application of the maximum likelihood method.
The species divergence times and demographic histories of Drosophila melanogaster and its three sibling species, D. mauritiana, D. simulans, and D. yakuba, were investigated using a maximum likelihood (ML) method. Thirty-nine orthologous loci for these four species were retrieved from DDBJ/EMBL/GenBank database. Both autosomal and X-linked loci were used in this study. A significant degree of rate heterogeneity across loci was observed for each pair of species. Most loci have the GC content greater than 50% at the third codon position. The codon usage bias in Drosophila loci is considered to result in the high GC content and the heterogenous rates across loci. The chi-square, G, and Fisher's exact tests indicated that data sets with 11, 23, and 9 pairs of DNA sequences for the comparison of D. melanogaster with D. mauritiana, D. simulans, and D. yakuba, respectively, retain homogeneous rates across loci. We applied the ML method to these data sets to estimate the DNA sequence divergences before and after speciation of each species pair along with their standard deviations. Using 1.6 x 10(-8) as the rate of nucleotide substitutions per silent site per year, our results indicate that the D. melanogaster lineage split from D. yakuba approximately 5.1 +/- 0.8 million years ago (mya), D. mauritiana 2.7 +/- 0.4 mya, and D. simulans 2.3 +/- 0.3 mya. It implies that D. melanogaster became distinct from D. mauritiana and D. simulans at approximately the same time and from D. yakuba no earlier than 10 mya. The effective ancestral population size of D. melanogaster appears to be stable over evolutionary time. Assuming 10 generations per year for Drosophila, the effective population size in the ancestral lineage immediately prior to the time of species divergence is approximately 3 x 10(6), which is close to that estimated for the extant D. melanogaster population. The D. melanogaster did not encounter any obvious bottleneck during the past 10 million years.


Self-calibration for transmitted wavefront measurements.
Micro-optic components and subsystems are becoming increasingly important in optical sensors, communications, data storage, and many other diverse applications. To adequately predict the performance of the final system, it is important to understand the element's effect on the wavefront as it propagates through the system. The wavefront can be measured using interferometric means, however, random and systematic errors contribute to the measurement. Self-calibration techniques exploit symmetries of the measurement or averaging techniques to separate the systematic errors of the instrument from the errors in the test lens. If the transmitted wavefront of a ball lens is measured in a number of random orientations and the measurements are averaged, the only remaining deviations from a perfect wavefront will be spherical aberration from the ball lens and the systematic errors of the interferometer. If the radius, aperture, and focal length of the ball lens are known, the spherical aberration can be calculated and subtracted, leaving only the systematic errors of the interferometer. We develop the theory behind the technique and illustrate the approach with a description of the calibration of a microinterferometer used to measure the transmitted wavefront error of micro-optics.


DropletMicroarray: facile formation of arrays of microdroplets and hydrogel micropads for cell screening applications.
We describe a one-step method for creating thousands of isolated pico- to microliter-sized droplets with defined geometry and volume. Arrays of droplets are instantly formed as liquid moves along a superhydrophilic-superhydrophobic patterned surface. Bioactive molecules, nonadherent cells, or microorganisms can be trapped in the fully isolated microdroplets for high-throughput screening, or in hydrogel micropads for screening in 3D microenvironments.


Pancreatic islet transplantation using vascularised chambers containing nerve growth factor ameliorates hyperglycaemia in diabetic mice.
Intraportal islet transplantation has shown initial promise for the treatment of type 1 diabetes. However, the portal vein site is associated with complications such as thrombosis and hepatic steatosis, leading to transplant failure. The aims of this study were to (1) test the feasibility of an alternative islet transplantation method that utilises a FDA-approved gelatin sponge as a novel islet carrier and (2) assess if exogenous addition of nerve growth factor (NGF) has any additional beneficial effects on graft performance in diabetic mice. Mice were rendered diabetic by a single intraperitoneal injection of streptozotocin. Five hundred syngeneic islets were seeded onto a Gelitaspon((R)) disc in the presence or absence of NGF, and placed into a silicone chamber surrounding the femoral neurovascular pedicle. Islet function was assessed by weekly monitoring of blood glucose levels and an intraperitoneal glucose tolerance test performed at the end of the study. Chambers were harvested for further histological analysis. Four of five mice transplanted with islets seeded onto Gelitaspon with NGF showed a significant reduction in blood glucose levels by 4 weeks after transplantation, and demonstrated a response similar to non-diabetic mice when tested with an intraperitoneal glucose tolerance test. Chamber tissue from this group contained islets with insulin-producing beta cells adjacent to the vascular pedicle. Islets seeded onto Gelitaspon with NGF and sited on femoral vessels using a tissue-engineering chamber offer an alternative method for islet transplantation in diabetic mice. This may have potential as a method for clinical islet transplantation.


hedgehog and engrailed: pattern formation and polarity in the Drosophila abdomen.
Like the Drosophila embryo, the abdomen of the adult consists of alternating anterior (A) and posterior (P) compartments. However the wing is made by only part of one A and part of one P compartment. The abdomen therefore offers an opportunity to compare two compartment borders (A/P is within the segment and P/A intervenes between two segments), and ask if they act differently in pattern formation. In the embryo, abdomen and wing P compartment cells express the selector gene engrailed and secrete Hedgehog protein whilst A compartment cells need the patched and smoothened genes in order to respond to Hedgehog. We made clones of cells with altered activities of the engrailed, patched and smoothened genes. Our results confirm (1) that the state of engrailed, whether 'off' or 'on', determines whether a cell is of A or P type and (2) that Hedgehog signalling, coming from the adjacent P compartments across both A/P and P/A boundaries, organises the pattern of all the A cells. We have uncovered four new aspects of compartments and engrailed in the abdomen. First, we show that engrailed acts in the A compartment: Hedgehog leaves the P cells and crosses the A/P boundary where it induces engrailed in a narrow band of A cells. engrailed causes these cells to form a special type of cuticle. No similar effect occurs when Hedgehog crosses the P/A border. Second, we look at the polarity changes induced by the clones, and build a working hypothesis that polarity is organised, in both compartments, by molecule(s) emanating from the A/P but not the P/A boundaries. Third, we show that both the A and P compartments are each divided into anterior and posterior subdomains. This additional stratification makes the A/P and the P/A boundaries fundamentally distinct from each other. Finally, we find that when engrailed is removed from P cells (of, say, segment A5) they transform not into A cells of the same segment, but into A cells of the same parasegment (segment A6).


CCR2 acts as scavenger for CCL2 during monocyte chemotaxis.
Leukocyte migration is essential for effective host defense against invading pathogens and during immune homeostasis. A hallmark of the regulation of this process is the presentation of chemokines in gradients stimulating leukocyte chemotaxis via cognate chemokine receptors. For efficient migration, receptor responsiveness must be maintained whilst the cells crawl on cell surfaces or on matrices along the attracting gradient towards increasing concentrations of agonist. On the other hand agonist-induced desensitization and internalization is a general paradigm for chemokine receptors which is inconsistent with the prolonged migratory capacity. Chemotaxis of monocytes was monitored in response to fluorescent CCL2-mCherry by time-lapse video microscopy. Uptake of the fluorescent agonist was used as indirect measure to follow the endogenous receptor CCR2 expressed on primary human monocytes. During chemotaxis CCL2-mCherry becomes endocytosed as cargo of CCR2, however, the internalization of CCR2 is not accompanied by reduced responsiveness of the cells due to desensitization. During chemotaxis CCR2 expressed on monocytes internalizes with the bound chemoattractant, but cycles rapidly back to the plasma membrane to maintain high responsiveness. Moreover, following relocation of the source of attractant, monocytes can rapidly reverse their polarization axis organizing a new leading edge along the newly formed gradient, suggesting a uniform distribution of highly receptive CCR2 on the plasma membrane. The present observations further indicate that during chemotaxis CCR2 acts as scavenger consuming the chemokine forming the attracting cue.


IQGAP1 regulates NR2A signaling, spine density, and cognitive processes.
General or brain-region-specific decreases in spine number or morphology accompany major neuropsychiatric disorders. It is unclear, however, whether changes in spine density are specific for an individual mental process or disorder and, if so, which molecules confer such specificity. Here we identify the scaffolding protein IQGAP1 as a key regulator of dendritic spine number with a specific role in cognitive but not emotional or motivational processes. We show that IQGAP1 is an important component of NMDAR multiprotein complexes and functionally interacts with the NR2A subunits and the extracellular signal-regulated kinase 1 (ERK1) and ERK2 signaling pathway. Mice lacking the IQGAP1 gene exhibited significantly lower levels of surface NR2A and impaired ERK activity compared to their wild-type littermates. Accordingly, primary hippocampal cultures of IQGAP1(-/-) neurons exhibited reduced surface expression of NR2A and disrupted ERK signaling in response to NR2A-dependent NMDAR stimulation. These molecular changes were accompanied by region-specific reductions of dendritic spine density in key brain areas involved in cognition, emotion, and motivation. IQGAP1 knock-outs exhibited marked long-term memory deficits accompanied by impaired hippocampal long-term potentiation (LTP) in a weak cellular learning model; in contrast, LTP was unaffected when induced with stronger stimulation paradigms. Anxiety- and depression-like behavior remained intact. On the basis of these findings, we propose that a dysfunctional IQGAP1 gene contributes to the cognitive deficits in brain disorders characterized by fewer dendritic spines.


Medical costs of secondhand-smoke exposure in North Carolina.
The health hazards of exposure to secondhand smoke (SHS) are well-defined. Less is known about the economic costs. We performed an analysis of the medical costs of SHS in North Carolina that was based on a similar study conducted in Minnesota. We used 2006 Blue Cross and Blue Shield of North Carolina claims data and national and state surveillance data to calculate the treated prevalence of medical conditions that have been found to be related to exposure to SHS, as established by a 2006 report from the US surgeon general. We used the population attributable risk for these conditions to calculate the number of individuals whose episodes of illness could be attributed to exposure to SHS. We adjusted these treatment costs for other types of insurance provided in the state, using Medical Expenditure Panel Survey data. The total annual cost of treatment for conditions related to SHS exposure in North Carolina was estimated to be $293,304,430, in 2009 inflation-adjusted dollars. Sensitivity analysis showed a range of $208.2 million to $386.3 million. The majority of individuals affected were children, but the greatest costs were for cardiovascular conditions. These cost data provide additional rationale for regulating smoking in all work sites and public places.


Cloning and characterization of short-chain N-acyl homoserine lactone-producing Enterobacter asburiae strain L1 from lettuce leaves.
In gram-negative bacteria, bacterial communication or quorum sensing (QS) is achieved using common signaling molecules known as N-acyl homoserine lactones (AHL). We have previously reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. In silico analysis of the strain L1 genome revealed the presence of a pair of luxI/R genes responsible for AHL-type QS, designated as easIR. In this work, the 639 bp luxI homolog, encoding 212 amino acids, have been cloned and overexpressed in Escherichia coli BL21 (DE3)pLysS. The purified protein (~25 kDa) shares high similarity to several members of the LuxI family among different E asburiae strains. Our findings showed that the heterologously expressed EasI protein has activated violacein production by AHL biosensor Chromobacterium violaceum CV026 as the wild-type E. asburiae. The mass spectrometry analysis showed the production of N-butanoyl homoserine lactone and N-hexanoyl homoserine lactone from induced E. coli harboring the recombinant EasI, suggesting that EasI is a functional AHL synthase. E. asburiae strain L1 was also shown to possess biofilm-forming characteristic activity using crystal violet binding assay. This is the first report on cloning and characterization of the luxI homolog from E. asburiae.


Long-acting inhaled therapy (beta-agonists, anticholinergics and steroids) for COPD: a network meta-analysis.
Pharmacological therapy for chronic obstructive pulmonary disease (COPD) is aimed at relieving symptoms, improving quality of life and preventing or treating exacerbations.Treatment tends to begin with one inhaler, and additional therapies are introduced as necessary. For persistent or worsening symptoms, long-acting inhaled therapies taken once or twice daily are preferred over short-acting inhalers. Several Cochrane reviews have looked at the risks and benefits of specific long-acting inhaled therapies compared with placebo or other treatments. However for patients and clinicians, it is important to understand the merits of these treatments relative to each other, and whether a particular class of inhaled therapies is more beneficial than the others. To assess the efficacy of treatment options for patients whose chronic obstructive pulmonary disease cannot be controlled by short-acting therapies alone. The review will not look at combination therapies usually considered later in the course of the disease.As part of this network meta-analysis, we will address the following issues.1. How does long-term efficacy compare between different pharmacological treatments for COPD?2. Are there limitations in the current evidence base that may compromise the conclusions drawn by this network meta-analysis? If so, what are the implications for future research? We identified randomised controlled trials (RCTs) in existing Cochrane reviews by searching the Cochrane Database of Systematic Reviews (CDSR). In addition, we ran a comprehensive citation search on the Cochrane Airways Group Register of trials (CAGR) and checked manufacturer websites and reference lists of other reviews. The most recent searches were conducted in September 2013. We included parallel-group RCTs of at least 6 months' duration recruiting people with COPD. Studies were included if they compared any of the following treatments versus any other: long-acting beta2-agonists (LABAs; formoterol, indacaterol, salmeterol); long-acting muscarinic antagonists (LAMAs; aclidinium, glycopyrronium, tiotropium); inhaled corticosteroids (ICSs; budesonide, fluticasone, mometasone); combination long-acting beta2-agonist (LABA) and inhaled corticosteroid (LABA/ICS) (formoterol/budesonide, formoterol/mometasone, salmeterol/fluticasone); and placebo. We conducted a network meta-analysis using Markov chain Monte Carlo methods for two efficacy outcomes: St George's Respiratory Questionnaire (SGRQ) total score and trough forced expiratory volume in one second (FEV1). We modelled the relative effectiveness of any two treatments as a function of each treatment relative to the reference treatment (placebo). We assumed that treatment effects were similar within treatment classes (LAMA, LABA, ICS, LABA/ICS). We present estimates of class effects, variability between treatments within each class and individual treatment effects compared with every other.To justify the analyses, we assessed the trials for clinical and methodological transitivity across comparisons. We tested the robustness of our analyses by performing sensitivity analyses for lack of blinding and by considering six- and 12-month data separately. We identified 71 RCTs randomly assigning 73,062 people with COPD to 184 treatment arms of interest. Trials were similar with regards to methodology, inclusion and exclusion criteria and key baseline characteristics. Participants were more often male, aged in their mid sixties, with FEV1 predicted normal between 40% and 50% and with substantial smoking histories (40+ pack-years). The risk of bias was generally low, although missing information made it hard to judge risk of selection bias and selective outcome reporting. Fixed effects were used for SGRQ analyses, and random effects for Trough FEV1 analyses, based on model fit statistics and deviance information criteria (DIC). SGRQ SGRQ data were available in 42 studies (n = 54,613). At six months, 39 pairwise comparisons were made between 18 treatments in 25 studies (n = 27,024). Combination LABA/ICS was the highest ranked intervention, with a mean improvement over placebo of -3.89 units at six months (95% credible interval (CrI) -4.70 to -2.97) and -3.60 at 12 months (95% CrI -4.63 to -2.34). LAMAs and LABAs were ranked second and third at six months, with mean differences of -2.63 (95% CrI -3.53 to -1.97) and -2.29 (95% CrI -3.18 to -1.53), respectively. Inhaled corticosteroids were ranked fourth (MD -2.00, 95% CrI -3.06 to -0.87). Class differences between LABA, LAMA and ICS were less prominent at 12 months. Indacaterol and aclidinium were ranked somewhat higher than other members of their classes, and formoterol 12 mcg, budesonide 400 mcg and formoterol/mometasone combination were ranked lower within their classes. There was considerable overlap in credible intervals and rankings for both classes and individual treatments. Trough FEV1 Trough FEV1 data were available in 46 studies (n = 47,409). At six months, 41 pairwise comparisons were made between 20 treatments in 31 studies (n = 29,271). As for SGRQ, combination LABA/ICS was the highest ranked class, with a mean improvement over placebo of 133.3 mL at six months (95% CrI 100.6 to 164.0) and slightly less at 12 months (mean difference (MD) 100, 95% CrI 55.5 to 140.1). LAMAs (MD 103.5, 95% CrI 81.8 to 124.9) and LABAs (MD 99.4, 95% CrI 72.0 to 127.8) showed roughly equivalent results at six months, and ICSs were the fourth ranked class (MD 65.4, 95% CrI 33.1 to 96.9). As with SGRQ, initial differences between classes were not so prominent at 12 months. Indacaterol and salmeterol/fluticasone were ranked slightly better than others in their class, and formoterol 12, aclidinium, budesonide and formoterol/budesonide combination were ranked lower within their classes. All credible intervals for individual rankings were wide. This network meta-analysis compares four different classes of long-acting inhalers for people with COPD who need more than short-acting bronchodilators. Quality of life and lung function were improved most on combination inhalers (LABA and ICS) and least on ICS alone at 6 and at 12 months. Overall LAMA and LABA inhalers had similar effects, particularly at 12 months. The network has demonstrated the benefit of ICS when added to LABA for these outcomes in participants who largely had an FEV1 that was less than 50% predicted, but the additional expense of combination inhalers and any potential for increased adverse events (which has been established by other reviews) require consideration. Our findings are in keeping with current National Institute for Health and Care Excellence (NICE) guidelines.


Genetic heterogeneity of the dengue vector Aedes aegypti in Martinique.
In Martinique, Aedes aegypti, the vector of dengue viruses has been the target of insecticide control for more than 35 years. Despite significant control efforts, dengue has become a major disease of public health concern. We conducted a population genetic analysis based on isoenzyme variations combined with an estimation of infection rate to a dengue virus among 26 Ae. aegypti samples. Aedes aegypti samples could be differentiated for their susceptibility to dengue infection (infection rates ranging from 42.8% to 98.6%) and showed important genetic variation (significant F(ST) values).


Nuclear translocation of FGFR1 and FGF2 in pancreatic stellate cells facilitates pancreatic cancer cell invasion.
Pancreatic cancer is characterised by desmoplasia, driven by activated pancreatic stellate cells (PSCs). Over-expression of FGFs and their receptors is a feature of pancreatic cancer and correlates with poor prognosis, but whether their expression impacts on PSCs is unclear. At the invasive front of human pancreatic cancer, FGF2 and FGFR1 localise to the nucleus in activated PSCs but not cancer cells. In vitro, inhibiting FGFR1 and FGF2 in PSCs, using RNAi or chemical inhibition, resulted in significantly reduced cell proliferation, which was not seen in cancer cells. In physiomimetic organotypic co-cultures, FGFR inhibition prevented PSC as well as cancer cell invasion. FGFR inhibition resulted in cytoplasmic localisation of FGFR1 and FGF2, in contrast to vehicle-treated conditions where PSCs with nuclear FGFR1 and FGF2 led cancer cells to invade the underlying extra-cellular matrix. Strikingly, abrogation of nuclear FGFR1 and FGF2 in PSCs abolished cancer cell invasion. These findings suggest a novel therapeutic approach, where preventing nuclear FGF/FGFR mediated proliferation and invasion in PSCs leads to disruption of the tumour microenvironment, preventing pancreatic cancer cell invasion.


Standardized human pedigree nomenclature: update and assessment of the recommendations of the National Society of Genetic Counselors.
In 1995, the Pedigree Standardization Task Force (PSTF) of the National Society of Genetic Counselors (NSGC) proposed a system of pedigree nomenclature. Recently, the PSTF (now called the Pedigree Standardization Work Group or PSWG) sought evidence that the published symbols met the needs of health professionals, were incorporated into health professional training and were utilized in publications. We searched PubMed and reference lists of select publications, reviewed the Instructions for Authors of several journals, searched the websites of professional societies, sought comment from the membership of the NSGC, and looked at recommendations and training practices of various health professional organizations. Many journals still do not cite specific standards for pedigrees, but those found cited the PSTF nomenclature. We did not find significant objections or alternatives to the 1995 nomenclature. Based on our review, we propose only a few minor stylistic changes to the pedigree symbols. The pedigree nomenclature of the NSGC is the only consistently acknowledged standard for drawing a family health history. We recommend regular and continued review of these pedigree standards to determine if additional symbols are needed to accommodate changes in clinical practice to ensure that the symbols continue to meet the needs of health professionals and researchers as well as adhere to evolving ethical and privacy standards. All health professionals, trainees, and researchers should be made aware of the utility of using a common pedigree nomenclature in clinical practice and publication. This will become particularly important as electronic medical records become more widely utilized.