Discrimination of hybrid classes using cross-species amplification of microsatellite loci: methodological challenges and solutions in Daphnia.
Microsatellite markers are important tools in population, conservation and forensic studies and are frequently used for species delineation, the detection of hybridization and introgression. Therefore, marker sets that amplify variable DNA regions in two species are required; however, cross-species amplification is often difficult, as genotyping errors such as null alleles may occur. To estimate the level of potential misidentifications based on genotyping errors, we compared the occurrence of parental alleles in laboratory and natural Daphnia hybrids (Daphnia longispina group). We tested a set of 12 microsatellite loci with regard to their suitability for unambiguous species and hybrid class identification using F(1) hybrids bred in the laboratory. Further, a large set of 44 natural populations of D. cucullata, D. galeata and D. longispina (1715 individuals) as well as their interspecific hybrids were genotyped to validate the discriminatory power of different marker combinations. Species delineation using microsatellite multilocus genotypes produced reliable results for all three studied species using assignment tests. Daphnia galeata × cucullata hybrid detection was limited due to three loci exhibiting D. cucullata-specific null alleles, which most likely are caused by differences in primer-binding sites of parental species. Overall, discriminatory power in hybrid detection was improved when a subset of markers was identified that amplifies equally well in both species.


Pathogenesis of polymyalgia rheumatica.
Polymyalgia rheumatica (PMR) is a chronic, inflammatory disorder of unknown cause, almost exclusively occurring in people aged over 50 and often associated with giant cell arteritis. The evidence that PMR occurs almost exclusively in individuals aged over 50 may indicate that age-related immune alterations in genetically predisposed subjects contribute to development of the disease. Several infectious agents have been investigated as possible triggers of PMR even though the results are inconclusive. Activation of the innate and adaptive immune systems has been proved in PMR patients as demonstrated by the activation of dendritic cells and monocytes/macrophages and the altered balance between Th17 and Treg cells. Disturbed B cell distribution and function have been also demonstrated in PMR patients suggesting a pathogenesis more complex than previously imagined. In this review we will discuss the recent findings regarding the pathogenesis of PMR.


[An analysis of the degree of unexplained small-area variation in costs of mental health care: towards a model of evidence-based commissioning of care?]
Part of the quality of a mental health service is the degree in which the service can demonstrate that its output is aligned with indicators of population need.
 AIM: To investigate to what degree small areas in the Netherlands (PC3-level, 798 areas) differ in the level of mental health care consumption; to what degree this is reducible to demographic and socioeconomic indicators representing known predictors of population mental health needs; and to what degree associations between indicators of need and mental health consumption vary across regions.
 METHOD: Analysis of public mental health cost data in the Netherlands over de period 2011-2017 for the age group 18-65 years.
 RESULTS: Mental health care costs displayed a pattern of small area variation that for 77% was reducible to socioeconomic and demographic factors at the regional level. Mental health care costs were associated independently with younger age, female sex and a range of socioeconomic factors including urbanicity, ethnic density and a socioeconomic index variable. However, the degree to which these indicators of mental health needs impacted mental health costs varied from region to region.
 CONCLUSION: Regional mental health care output in the Netherlands is partially aligned with population needs These analyses validate the notion that mental health care commissioning may be guided by benchmarked evidence of service output being proportional to the needs predicted by population socioeconomic indices.


Desquamative interstitial pneumonia associated with chrysotile asbestos fibres.
The drywall construction trade has in the past been associated with exposure to airborne asbestos fibres. This paper reports a drywall construction worker with 32 years of dust exposure who developed dyspnoea and diminished diffusing capacity, and showed diffuse irregular opacities on chest radiography. He did not respond to treatment with corticosteroids. Open lung biopsy examination showed desquamative interstitial pneumonia. Only a single ferruginous body was seen on frozen section, but tissue examination by electron microscopy showed an extraordinary pulmonary burden of mineral dust with especially high concentrations of chrysotile asbestos fibres. This report emphasises the need to consider asbestos fibre as an agent in the aetiology of desquamative interstitial pneumonia. The coexistent slight interstitial fibrosis present in this case is also considered to have resulted from exposure to mineral dust, particularly ultramicroscopic asbestos fibres.


Clinical significance of the low normal sperm morphology value as proposed in the fifth edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen.
The very low cut-off value for sperm morphology of 4% morphologically normal spermatozoa, as proposed in the new edition of the World Health Organization (WHO) manual on semen analysis, is in agreement with recently published values and reflects the trend of a decline in reported mean values for normal sperm morphology. The reduced value for morphologically normal spermatozoa over the years may be due to several factors. The first is the introduction of strict criteria for the evaluation of sperm morphology. Other reasons may include the introduction of additional criteria for sperm morphology abnormalities and the suggested decrease in semen parameters because of increasing negative environmental influences. Although on its own the newly proposed very low normal value may not provide the strong predictive value for a males' fertility potential, as originally reported for sperm morphology evaluated according to strict criteria, a good predictive value can still be obtained if the holistic, strict approach for sperm morphology evaluation is followed together with additional sperm morphology parameters now available, because certain morphology patterns and sperm abnormalities are now known to be of strong prognostic value. In addition, better international standardization of the technical methodology, consensus on the interpretation of sperm morphology evaluation criteria and standardized international external quality control (EQC) schemes, are of utmost importance to maintain the good predictive value of sperm morphology.


Socioeconomic Status And Quality Of Care In Rural India: New Evidence From Provider And Household Surveys.
To assess socioeconomic differences in access to high-quality health care services, we collected novel data on illnesses that required primary care from 23,275 households in 100 villages in Madhya Pradesh, India. We matched the primary care visits for those illnesses to characteristics of the health care providers that members of the households visited. People in the average village in our sample could access eleven providers, of whom 71 percent were in the private sector and 49 percent had no formal medical training. The private sector accounted for 89 percent of the primary care visits in our sample, with 77 percent of the visits made to providers with no formal training. Both access to and use of more knowledgeable providers increased with socioeconomic status, mostly as a result of differences across districts and villages. Strikingly, people in high- and low-socioeconomic-status households in the same village visited equally knowledgeable providers. It was the poor people who lived in poor communities who received especially low-quality care.


Peptides modeled after the alpha-domain of metallothionein induce neurite outgrowth and promote survival of cerebellar granule neurons.
Metallothionein (MT) is a metal-binding protein capable of preventing oxidative stress and apoptotic cell death in the central nervous system of mammals, and hence is of putative therapeutic value in the treatment of neurodegenerative disorders. Recently, we demonstrated that a peptide modeled after the beta-domain of MT, EmtinB, induced neurite outgrowth and increased neuronal survival through binding to receptors of the low-density lipoprotein receptor family (LDLR). The present study identified two MT alpha-domain-derived peptide sequences termed EmtinAn and EmtinAc, each consisting of 14 amino acids, as potent stimulators of neuronal differentiation and survival of primary neurons. In addition, we show that a peptide derived from the N-terminus of the MT beta-domain, EmtinBn, promotes neuronal survival. The neuritogenic and survival promoting effects of EmtinAc, similar to MT and EmtinB but not EmtinAn, were dependent on the functional integrity of LDLR. Moreover, EmtinAn and EmtinAc induced activation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB/Akt). We suggest that multiple functional sites of MT serve to cross-link MT receptor(s), thereby transducing signals leading to an increase in neurite outgrowth and survival.


The Influence of Label Design on Search Performance and Noticeability in Wide Field of View Augmented Reality Displays.
In Augmented Reality (AR), search performance for outdoor tasks is an important metric for evaluating the success of a large number of AR applications. Users must be able to find content quickly, labels and indicators must not be invasive but still clearly noticeable, and the user interface should maximize search performance in a variety of conditions. To address these issues, we have set up a series of experiments to test the influence of virtual characteristics such as color, size, and leader lines on the performance of search tasks and noticeability in both real and simulated environments. We evaluate two primary areas, including 1) the effects of peripheral field of view (FOV) limitations and labeling techniques on target acquisition during outdoor mobile search, and 2) the influence of local characteristics such as color, size, and motion on text labels over dynamic backgrounds. The first experiment showed that limited FOV will severely limit search performance, but that appropriate placement of labels and leaders within the periphery can alleviate this problem without interfering with walking or decreasing user comfort. In the second experiment, we found that different types of motion are more noticeable in optical versus video see-through displays, but that blue coloration is most noticeable in both. Results can aid in designing more effective view management techniques, especially for wider field of view displays.


Nursing highlights from the 2015 European Cancer Congress (ECCO18-ESMO40), 25-29 September 2015, Vienna: reinforcing multidisciplinarity.
The 2015 European Cancer Congress (ECC2015) was the widest European platform for every professional involved in the struggle against cancer (17,656 participants, 2482 abstracts submitted). In this context, the nursing contribution was very relevant, involving high quality research and experience. The major nursing issues were: online information and support; innovations in cancer nursing roles; patient safety and the nursing workforce; managing older people with cancer and other co-morbidities; living with and beyond cancer; nutrition and body image changes; the changing face of cancer care for oncology nurses. Indeed, an important amount of research was presented during proffered papers and poster presentations by nurses from all over the world, concerning challenging issues, such as advanced nursing roles, end of life care, impact of cancer on patients and families, new developments, supportive and palliative care, survivorship and rehabilitation, symptom management and transitions in care. Nurses' presentations were mainly focused on understanding patients' needs and on sharing the best evidence-based approach to meet those needs. This is particularly significant in a field where innovation develops rapidly in every area of clinical practice, such as cancer care, bridging the weaknesses between different approaches and profiles, within the paradigm of multidisciplinarity.


Hemoglobin subunit beta interacts with the capsid protein and antagonizes the growth of classical swine fever virus.
The capsid (C) protein of the Flaviviridae family members is involved in nucleocapsid formation and virion assembly. However, the influence of C protein-interacting partners on the outcome of pestivirus infections is poorly defined. In this study, hemoglobin subunit beta (HB) was identified as a C protein-binding protein by glutathione S-transferase pulldown and subsequent mass spectrometry analysis of PK-15 cells, which are permissive cells for classical swine fever virus (CSFV). Coimmunoprecipitation and confocal microscopy confirmed that HB interacts and colocalizes with the C protein in the cytoplasm. Silencing of HB with small interfering RNAs promoted CSFV growth and replication, whereas overexpression of HB suppressed CSFV replication and growth. Interestingly, HB was found to interact with retinoic acid-inducible gene I and increase its expression, resulting in increased production of type I interferon (IFN). However, HB was unable to suppress CSFV growth when the RIG-I pathway was blocked. Overall, our results suggest that cellular HB antagonizes CSFV growth and replication by triggering IFN signaling, and might represent a novel antiviral restriction factor. This study reports for the first time the novel role of HB in innate immunity.


Analytical, numerical, and experimental studies of viscoelastic effects on the performance of soft piezoelectric nanocomposites.
Piezoelectric composite (p-NC) made of a polymeric matrix and piezoelectric nanoparticles with conductive additives is an attractive material for many applications. As the matrix of p-NC is made of viscoelastic materials, both elastic and viscous characteristics of the matrix are expected to contribute to the piezoelectric response of p-NC. However, there is limited understanding of how viscoelasticity influences the piezoelectric performance of p-NC. Here we combined analytical and numerical analyses with experimental studies to investigate effects of viscoelasticity on piezoelectric performance of p-NC. The viscoelastic properties of synthesized p-NCs were controlled by changing the ratio between monomer and cross-linker of the polymer matrix. We found good agreement between our analytical models and experimental results for both quasi-static and dynamic loadings. It is found that, under quasi-static loading conditions, the piezoelectric coefficients (d33) of the specimen with the lowest Young's modulus (∼0.45 MPa at 5% strain) were ∼120 pC N-1, while the one with the highest Young's modulus (∼1.3 MPa at 5% strain) were ∼62 pC N-1. The results suggest that softer matrices enhance the energy harvesting performance because they can result in larger deformation for a given load. Moreover, from our theoretical analysis and experiments under dynamic loading conditions, we found the viscous modulus of a matrix is also important for piezoelectric performance. For instance, at 40 Hz and 50 Hz the storage moduli of the softest specimen were ∼0.625 MPa and ∼0.485 MPa, while the loss moduli were ∼0.108 MPa and ∼0.151 MPa, respectively. As piezocomposites with less viscous loss can transfer mechanical energy to piezoelectric particles more efficiently, the dynamic piezoelectric coefficient (d'33) measured at 40 Hz (∼53 pC N-1) was larger than that at 50 Hz (∼47 pC N-1) though it has a larger storage modulus. As an application of our findings, we fabricated 3D piezo-shells with different viscoelastic properties and compared the charging time. The results showed a good agreement with the predicted trend that the composition with the smallest elastic and viscous moduli showed the fastest charging rate. Our findings can open new opportunities for optimizing the performance of polymer-based multifunctional materials by harnessing viscoelasticity.


GVHD: biology matters.
Acute graft-versus-host disease (GVHD) targets the crypts in the gastrointestinal (GI) tract that are responsible for the self-renewal of the intestinal mucosa. Recent advances in the identification and culture of intestinal stem cells have improved our understanding of the interactions between the microbiome and the immune system (both innate and adaptive) that are key to the pathophysiology of GVHD. The identification of serum biomarkers that best predict long-term GVHD outcomes derive from the GI tract and have focused attention on cellular elements that act as shields against GVHD as well as its targets. These biomarkers have illuminated new mechanisms of crypt biology and provided insights that should prove useful both in the design of clinical trials and as guides to GVHD prevention and treatment.


Hedgehog signalling in the mouse requires intraflagellar transport proteins.
Intraflagellar transport (IFT) proteins were first identified as essential factors for the growth and maintenance of flagella in the single-celled alga Chlamydomonas reinhardtii. In a screen for embryonic patterning mutations induced by ethylnitrosourea, here we identify two mouse mutants, wimple (wim) and flexo (fxo), that lack ventral neural cell types and show other phenotypes characteristic of defects in Sonic hedgehog signalling. Both mutations disrupt IFT proteins: the wim mutation is an allele of the previously uncharacterized mouse homologue of IFT172; and fxo is a new hypomorphic allele of polaris, the mouse homologue of IFT88. Genetic analysis shows that Wim, Polaris and the IFT motor protein Kif3a are required for Hedgehog signalling at a step downstream of Patched1 (the Hedgehog receptor) and upstream of direct targets of Hedgehog signalling. Our data show that IFT machinery has an essential and vertebrate-specific role in Hedgehog signal transduction.


Injectable microgel-hydrogel composites for prolonged small-molecule drug delivery.
The design and application of soft nanocomposite injectable hydrogels containing entrapped microgels for small-molecule drug delivery is demonstrated. Copolymer microgels based on N-isopropylacrylamide and acrylic acid were synthesized that exhibited both ionic and hydrophobic affinity for binding to bupivacaine, a cationic local anesthetic used as a model drug. Microgels were subsequently immobilized within an in situ-gelling hydrogel network cross-linked via hydrazide-aldehyde chemistry to generate hydrogel-microgel soft nanocomposites. Drug release could be sustained for up to 60 days from these nanocomposite hydrogels, significantly longer than that achievable using the constituent hydrogel or microgels alone (<1 week). Drug release kinetics could be readily tuned by varying the affinity of the microgel and hydrogel phases for drug-polymer interactions and the network density of the hydrogel phase.


C5a alters blood-brain barrier integrity in experimental lupus.
The blood-brain barrier (BBB) is a crucial anatomic location in the brain. Its dysfunction complicates many neurodegenerative diseases, from acute conditions, such as sepsis, to chronic diseases, such as systemic lupus erythematosus (SLE). Several studies suggest an altered BBB in lupus, but the underlying mechanism remains unknown. In the current study, we observed a definite loss of BBB integrity in MRL/MpJ-Tnfrsf6(lpr) (MRL/lpr) lupus mice by IgG infiltration into brain parenchyma. In line with this result, we examined the role of complement activation, a key event in this setting, in maintenance of BBB integrity. Complement activation generates C5a, a molecule with multiple functions. Because the expression of the C5a receptor (C5aR) is significantly increased in brain endothelial cells treated with lupus serum, the study focused on the role of C5a signaling through its G-protein-coupled receptor C5aR in brain endothelial cells, in a lupus setting. Reactive oxygen species production increased significantly in endothelial cells, in both primary cells and the bEnd3 cell line treated with lupus serum from MRL/lpr mice, compared with those treated with control serum from MRL(+/+) mice. In addition, increased permeability monitored by changes in transendothelial electrical resistance, cytoskeletal remodeling caused by actin fiber rearrangement, and increased iNOS mRNA expression were observed in bEnd3 cells. These disruptive effects were alleviated by pretreating cells with a C5a receptor antagonist (C5aRant) or a C5a antibody. Furthermore, the structural integrity of the vasculature in MRL/lpr brain was maintained by C5aR inhibition. These results demonstrate the regulation of BBB integrity by the complement system in a neuroinflammatory setting. For the first time, a novel role of C5a in the maintenance of BBB integrity is identified and the potential of C5a/C5aR blockade highlighted as a promising therapeutic strategy in SLE and other neurodegenerative diseases.


Demonstration of an SOA-assisted open metro-access infrastructure for heterogeneous services.
An open converged metro-access network approach allows for sharing optical layer resources like fibers and optical spectrum among different services and operators. We demonstrated experimentally the feasibility of such a concept by the simultaneous operation of multiple services showing different modulation formats and multiplexing techniques. Flexible access nodes are implemented including semiconductor optical amplifiers to create a transparent and reconfigurable optical ring network. The impact of cascaded optical amplifiers on the signal quality is studied along the ring. In addition, the influence of high power rival signals in the same waveband and in the same fiber is analyzed.


Intention-to-treat analysis of pulmonary artery banding in conditions with a morphological right ventricle in the systemic circulation with a view to anatomic biventricular repair.
Some patients with a morphological right ventricle (mRV) in the systemic circulation require early intervention because of progressive systemic ventricular dysfunction or atrioventricular valve regurgitation. They may be eligible for anatomic repair (correction of atrioventricular and ventriculoarterial discordance) but require prior training of the morphological left ventricle (mLV). Forty-one patients with congenitally corrected transposition of the great arteries or a previous atrial switch procedure embarked on a protocol of pulmonary artery (PA) banding with a view to anatomic repair. All had an mRV in the systemic circulation and a subpulmonary mLV that was not conditioned by either volume or pressure load. Two patients were not banded, and 39 were followed up for a median of 4.3 years (range, 25 days to 12.6 years). Sixteen patients achieved anatomic repair, with 3 in the early stages of the training protocol. After 2 years, 12 patients were not suitable for anatomic repair and persisted with palliative banding; 8 were functionally improved; and 4 died, underwent transplantation, or required debanding. PA banding improved functional class but did not improve tricuspid regurgitation in the long term for patients not achieving anatomic repair. mLV function was a critical determinant of survival with a PA band as well as survival after anatomic repair. Patients >16 years were unlikely to achieve anatomic repair. PA banding is a safe and effective method of training the mLV before anatomic repair. It is also an effective palliative procedure for those who do not attain this goal.


Ecdysone induced gene expression is associated with acetylation of histone H3 lysine 23 in Drosophila melanogaster.
Posttranslational modification of histones regulates transcription but the exact role that acetylation of specific lysine residues plays in biological processes in vivo is still not clearly understood. To assess the contribution of different histone modifications to transcriptional activation in vivo, we determined the acetylation patterns on the ecdysone induced Eip74EF and Eip75B genes in Drosophila melanogaster larvae by chromatin immunoprecipitation. We found that acetylation of histone H3 lysine 23 is localized to promoters and correlates with endogenous ecdysone induced gene activation. In contrast, acetylation of lysines 8, 12 and 16 of histone H4 and lysine 9 of histone H3 showed minor differences in their distribution on the regulatory and transcribed regions tested, and had limited or no correlation with ecdysone induced transcriptional activity. We found that dCBP, which is encoded by the nejire gene, acetylates H3 lysine 23 in vivo, and silencing of nejire leads to reduced expression of the Eip74EF and Eip75B genes. Our results suggest that acetylation of specific lysine residues of histones contribute specifically to the dynamic regulation of transcription. Furthermore, along with previous studies identify CBP dependent H3 lysine 23 acetylation as an evolutionarily conserved chromatin modification involved in steroid induced gene activation.


Rapid creation of a temporary isolation ward for patients with severe acute respiratory syndrome in Taiwan.
To rapidly establish a temporary isolation ward to handle an unexpected sudden outbreak of severe acute respiratory syndrome (SARS) and to evaluate the implementation of exposure control measures by healthcare workers (HCWs) for SARS patients. Rapid creation of 60 relatively negative pressure isolation rooms for 196 suspected SARS patients transferred from 19 hospitals and daily temperature recordings of 180 volunteer HCWs from 6 medical centers. A military hospital. Of the 196 patients, 34 (17.3%) met the World Health Organization criteria for probable SARS with positive results of serologic testing for SARS-associated coronavirus (SARS-CoV), reverse transcriptase polymerase chain reaction (RT-PCR) from nasopharyngeal or throat swabs for SARS-CoV, or both. Seventy-four patients had suspected SARS based on unprotected exposure to SARS patients; three of them had positive results on RT-PCR but negative serologic results. The remaining 88 patients did not meet the criteria for a probable or suspected SARS diagnosis. Of the 34 patients with probable SARS, 13 were transferred to medical centers to receive mechanical ventilation due to rapid deterioration of chest x-ray results, and three patients died of SARS despite intensive therapy in medical centers. During the study period, one nurse developed probable SARS due to violation of infection control measures, but there was no evidence of cross-transmission to other HCWs. Despite the use of full personal protection equipment, the facility failed to totally prevent exposures of HCWs to SARS but minimized the risk of nosocomial transmission. Better training and improvements in infection control infrastructure may limit the impact of SARS.


Fast-RCM: Fast Tree-Based Unsupervised Rare-Class Mining.
Rare classes are usually hidden in an imbalanced dataset with the majority of the data examples from major classes. Rare-class mining (RCM) aims at extracting all the data examples belonging to rare classes. Most of the existing approaches for RCM require a certain amount of labeled data examples as input. However, they are ineffective in practice since requesting label information from domain experts is time consuming and human-labor extensive. Thus, we investigate the unsupervised RCM problem, which to the best of our knowledge is the first such attempt. To this end, we propose an efficient algorithm called Fast-RCM for unsupervised RCM, which has an approximately linear time complexity with respect to data size and data dimensionality. Given an unlabeled dataset, Fast-RCM mines out the rare class by first building a rare tree for the input dataset and then extracting data examples of the rare classes based on this rare tree. Compared with the existing approaches which have quadric or even cubic time complexity, Fast-RCM is much faster and can be extended to large-scale datasets. The experimental evaluation on both synthetic and real-world datasets demonstrate that our algorithm can effectively and efficiently extract the rare classes from an unlabeled dataset under the unsupervised settings, and is approximately five times faster than that of the state-of-the-art methods.


Antiviral activities of hemp cannabinoids.
Hemp is an understudied source of pharmacologically active compounds and many unique plant secondary metabolites including more than 100 cannabinoids. After years of legal restriction, research on hemp has recently demonstrated antiviral activities in silico, in vitro, and in vivo for cannabidiol (CBD), Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), and several other cannabinoids against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), human immunodeficiency virus (HIV), and γ-herpes viruses. Mechanisms of action include inhibition of viral cell entry, inhibition of viral proteases, and stimulation of cellular innate immune responses. The anti-inflammatory properties of cannabinoids are also under investigation for mitigating the cytokine storm of COVID-19 and controlling chronic inflammation in people living with HIV. Retrospective clinical studies support antiviral activities of CBD, Δ9-THC, and cannabinoid mixtures as do some prospective clinical trials, but appropriately designed clinical trials of safety and efficacy of antiviral cannabinoids are urgently needed.


Strategies and synthetic methods directed toward the preparation of libraries of substituted isoquinolines.
Strategies for the production of substituted isoquinoline libraries were developed and explored. Routes involving microwave-assisted variants of the Bischler-Napieralski or Pictet-Spengler reaction allowed for cyclization of substituted beta-arylethylamine derivatives. The dihydroisoquinolines and tetrahydroisoquinolines thus generated could then be oxidized to their corresponding isoquinoline analogues. An alternate strategy, however, involving the preparation and activation of isoquinolin-1(2H)-ones is demonstrated to be a more practical, rapid, and efficient route to C1- and C4-substituted isoquinoline libraries.


New strategy for the synthesis of chemically modified RNA constructs exemplified by hairpin and hammerhead ribozymes.
The CuAAC reaction (click chemistry) has been used in conjunction with solid-phase synthesis to produce catalytically active hairpin ribozymes around 100 nucleotides in length. Cross-strand ligation through neighboring nucleobases was successful in covalently linking presynthesized RNA strands with high efficiency (trans-ligation). In an alternative strategy, intrastrand click ligation was employed to produce a functional hammerhead ribozyme containing a novel nucleic acid backbone mimic at the catalytic site (cis-ligation). The ability to synthesize long RNA strands by a combination of solid-phase synthesis and click ligation is an important addition to RNA chemistry. It is compatible with a plethora of site-specific modifications and is applicable to the synthesis of many biologically important RNA molecules.


[Assessment of attention deficit hyperactivity: SNAP-IV scale adapted to Argentina].
Assess the usefulness of the SNAP-IV scale as an instrument for detecting attention deficit hyperactivity disorder (ADHD) in Argentine children aged 4 to 14 years. The SNAP-IV scale was adapted and administered to a group of 1 230 schoolchildren in the province of Buenos Aires, Argentina. The diagnosis was determined with the clinical control, based on the criteria of the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. The sensitivity and specificity, as well as the cut-off scores for the SNAP-IV scale in the population studied, were determined. The score on the SNAP-IV scale with the best correlation between sensitivity and specificity was established in order to determine the true positive cases that in fact had a clinical diagnosis. The cut-off scores obtained were: 1.66 (15/27 points) for the attention deficit subscale and 1.77 (16/27 points) for hyperactivity/impulsivity in the population studied. The SNAP-IV scale for detection of ADHD is considered to be valid in the population studied as long as the cut-off scores are modified to obtain the best sensitivity/specificity ratio based on the cultural and socioeconomic features of the population.


Associations of steps per day and step intensity with the risk of diabetes: the Hispanic Community Health Study / Study of Latinos (HCHS/SOL).
Higher levels of moderate-to-vigorous physical activity have been associated with a lower risk of diabetes, but less is known about how daily step counts (steps/day) are associated with diabetes risk. Therefore, we examined the association of steps/day and step intensity with incident diabetes. We included 6634 adults from the population-based prospective cohort Hispanic Community Health Study/Study of Latinos (HCHS/SOL) (2008-2017). Cox proportional hazard models that accounted for complex survey design and sampling weights were used to estimate the association of baseline accelerometer-assessed steps/day and step intensity with 6-year risk of incident diabetes as hazard ratios (HR) and 95% confidence intervals (CI). We further examined whether the percent of intense steps at a given accumulation of steps/day was associated with diabetes risk, and if associations were modified by specific cohort characteristics. The average age of cohort members was 39 years and 52% were female. Adults had an average of 8164 steps/day and spent 12 min/day in brisk ambulation (> 100 steps/min). Over 6 years of follow-up, there were 1115 cases of diabetes. There was a suggestive lower risk of diabetes with more steps/day- adults had a 2% lower risk per 1000 steps/day (HR = 0.98 (95% CI 0.95, 1.00)). Inverse associations between average steps/day and diabetes incidence were observed across many cohort characteristics, but most importantly among adults at high risk for diabetes - those who were older, or had obesity or prediabetes. Adults who accumulated 17 min/day in brisk ambulation compared to < 2 min/day had a 31% lower risk of diabetes (HR = 0.69 (95% CI 0.53, 0.89)). A greater percent of intense steps for a given accumulation of steps/day was associated with further risk reduction. Adults who accumulate more daily steps may have a lower risk of diabetes. Accumulating more steps/day and greater step intensity appear to be important targets for preventing diabetes.


Development and Feasibility Testing of Internet-Delivered Acceptance and Commitment Therapy for Severe Health Anxiety: Pilot Study.
Severe health anxiety (hypochondriasis), or illness anxiety disorder according to the Diagnostic and Statistical Manual of Mental Disorders, 5th edition, is characterized by preoccupation with fear of suffering from a serious illness in spite of medical reassurance. It is a debilitating, prevalent disorder associated with increased health care utilization. Still, there is a lack of easily accessible specialized treatment for severe health anxiety. The aims of this paper were to (1) describe the development and setup of a new internet-delivered acceptance and commitment therapy (iACT) program for patients with severe health anxiety using self-referral and a video-based assessment; and (2) examine the feasibility and potential clinical efficacy of iACT for severe health anxiety. Self-referred patients (N=15) with severe health anxiety were diagnostically assessed by a video-based interview. They received 7 sessions of clinician-supported iACT comprising self-help texts, video clips, audio files, and worksheets over 12 weeks. Self-report questionnaires were obtained at baseline, post-treatment, and at 3-month follow-up. The primary outcome was Whiteley-7 Index (WI-7) measuring health anxiety severity. Depressive symptoms, health-related quality of life (HRQoL), life satisfaction, and psychological flexibility were also assessed. A within-group design was employed. Means, standard deviations, and effect sizes using the standardized response mean (SRM) were estimated. Post-treatment interviews were conducted to evaluate the patient experience of the usability and acceptability of the treatment setup and program. The self-referral and video-based assessments were well received. Most patients (12/15, 80%) completed the treatment, and only 1 (1/15, 7%) dropped out. Post-treatment (14/15, 93%) and 3-month follow-up (12/15, 80%) data were available for almost all patients. Paired t tests showed significant improvements on all outcome measures both at post-treatment and 3-month follow-up, except on one physical component subscale of HRQoL. Health anxiety symptoms decreased with 33.9 points at 3-month follow-up (95% CI 13.6-54.3, t11= 3.66, P=.004) with a large within-group effect size of 1.06 as measured by the SRM. Treatment adherence and potential efficacy suggest that iACT may be a feasible treatment for health anxiety. The uncontrolled design and small sample size of the study limited the robustness of the findings. Therefore, the findings should be replicated in a randomized controlled trial. Potentially, iACT may increase availability and accessibility of specialized treatment for health anxiety. Danish Data Protection Agency, Central Denmark Region: 1-16-02-427-14; https://www.rm.dk/sundhed/faginfo/forskning/datatilsynet/ (Archived by Webcite at http://www.webcitation.org/6yDA7WovM).


Developmental demands contribute to early neuromuscular degeneration in CMT2D mice.
Dominantly inherited, missense mutations in the widely expressed housekeeping gene, GARS1, cause Charcot-Marie-Tooth type 2D (CMT2D), a peripheral neuropathy characterised by muscle weakness and wasting in limb extremities. Mice modelling CMT2D display early and selective neuromuscular junction (NMJ) pathology, epitomised by disturbed maturation and neurotransmission, leading to denervation. Indeed, the NMJ disruption has been reported in several different muscles; however, a systematic comparison of neuromuscular synapses from distinct body locations has yet to be performed. We therefore analysed NMJ development and degeneration across five different wholemount muscles to identify key synaptic features contributing to the distinct pattern of neurodegeneration in CMT2D mice. Denervation was found to occur along a distal-to-proximal gradient, providing a cellular explanation for the greater weakness observed in mutant Gars hindlimbs compared with forelimbs. Nonetheless, muscles from similar locations and innervated by axons of equivalent length showed significant differences in neuropathology, suggestive of additional factors impacting on site-specific neuromuscular degeneration. Defective NMJ development preceded and associated with degeneration, but was not linked to a delay of wild-type NMJ maturation processes. Correlation analyses indicate that muscle fibre type nor synaptic architecture explain the differential denervation of CMT2D NMJs, rather it is the extent of post-natal synaptic growth that predisposes to neurodegeneration. Together, this work improves our understanding of the mechanisms driving synaptic vulnerability in CMT2D and hints at pertinent pathogenic pathways.


Effect of supraglottic and super-supraglottic swallows on tongue pressure production against hard palate.
The objectives of this study were to evaluate the state of tongue pressure production during supraglottic swallow (SS) and super-supraglottic swallow (SSS) performed by healthy adults, and to investigate the effects of these swallowing maneuvers on the oral stage of swallowing. The participants were 19 healthy individuals. Tongue pressure against the hard palate during swallowing was measured using a tongue pressure sensor sheet system with five pressure-sensitive points. The tasks comprised swallowing 5 mL of water by normal wet swallow, SS, and SSS, and the parameters for analysis were the duration, the maximal magnitude, and the integrated value of tongue pressure during swallowing. The duration of tongue pressure was significantly longer at the anterior-median part of the hard palate during both SS and SSS than with normal wet swallow. The maximal magnitude increased significantly only at the posterior part of the hard palate during SS, but at all points during SSS. The integrated value increased significantly only at the posterior-median part of the hard palate during SS, but at all points except the mid-median part of the hard palate during SSS. The maximal magnitude and integrated value were also significantly higher at the anterior-median and posterior circumferential parts during SSS than during SS. These results show that these two swallowing maneuvers, which are known primarily as techniques to protect the airway, also function to strengthen the tongue pressure produced by the contact between the tongue and the hard palate during swallowing and this effect is more pronounced during SSS.


Prognostic implications of the systolic to diastolic duration ratio in children with idiopathic or familial dilated cardiomyopathy.
Childhood dilated cardiomyopathy (DCM) carries high morbidity and mortality. The echocardiographic systolic to diastolic (S:D) duration ratio, an indicator of global cardiac performance, is elevated in DCM; however, its prognostic implications have not been investigated in this population. We investigated systolic and diastolic durations and the resultant S:D ratio using pulsed tissue Doppler imaging in children with idiopathic or familial DCM. We studied serial echocardiograms from presentation until the last follow-up echo. Results were compared with heart rate-matched controls and between DCM subgroups based on an acute or insidious presentation. The association between S:D ratio and death or need for transplant was analyzed. All analyses were adjusted for repeated measures per patient. We studied 200 serial echocardiograms of 48 children with DCM (7.0±6.0 years) and 25 controls. Adjusted for repeated measures through a compound symmetry covariance structure, the S:D ratio was higher in DCM patients (-0.425 [0.072]; P<0.001) because of shortened diastole. A S:D ratio >1.2 at presentation and on serial evaluation was associated with a hazard ratio of 10.5 (95% confidence interval, 3.9-27.8; P<0.001) for death or transplant. In combined multivariable analysis, a S:D ratio >1.2 remained significantly associated with hazard of death/transplant (hazard ratio, 9.1; P=0.04) after adjustment for ejection fraction (hazard ratio: 2.2 per -10%; P<0.001). A high S:D ratio is associated with increased risk for death or need for transplant in children with DCM across the spectrum of heart rates and may be a useful prognostic index for serial evaluation of children with DCM.


Enhancing sulforaphane absorption and excretion in healthy men through the combined consumption of fresh broccoli sprouts and a glucoraphanin-rich powder.
Sulforaphane (SF) is a chemopreventive isothiocyanate (ITC) derived from glucoraphanin (GRP) hydrolysis by myrosinase, a thioglucoside present in broccoli. The ability of broccoli powders sold as supplements to provide dietary SF is often of concern as many supplements contain GRP, but lack myrosinase. In a previous study, biomarkers of SF bioavailability from a powder rich in GRP, but lacking myrosinase, were enhanced by co-consumption of a myrosinase-containing air-dried broccoli sprout powder. Here, we studied the absorption of SF from the GRP-rich powder used in the previous study, but in combination with fresh broccoli sprouts, which are commercially available and more applicable to the human diet than air-dried sprout powder. A total of four participants each consumed four meals (separated by 1 week) consisting of dry cereal and yogurt with sprouts equivalent to 70 μmol SF, GRP powder equivalent to 120 μmol SF, both or neither. Metabolites of SF were analysed in blood and urine. The 24 h urinary SF-N-acetylcysteine recovery was 65, 60 and 24 % of the dose ingested from combination, broccoli sprout and GRP powder meals, respectively. In urine and plasma, ITC appearance was delayed following the GRP powder meal compared with the sprout and combination meals. Compared with the GRP powder or sprouts alone, combining broccoli sprouts with the GRP powder synergistically enhanced the early appearance of SF, offering insight into the combination of foods for improved health benefits of foods that reduce the risk for cancer.


Measurement of phasic velocities in vessels of intact freely beating hearts.
The study of flow dynamics on the coronary microcirculation of the intact beating heart has been greatly hampered by the thickness of the tissue and its continual movement. Using fluorescent microspheres and video-image processing, we have devised a means to measure an index of blood velocities. Measurements were made on open-chest anesthetized cats without any artificial restraints to the heart motion. However, advantages exist for minimizing mechanical motion, therefore, a heart holder based on a vacuum concept, along with modified versions of previously demonstrated methods, were tested. Results showed that the vacuum holder provided more restraint to epicardial motion than the pericardium alone, and despite large differences in epicardial motion between the two, overall flow measurements of the microspheres were comparable. Other methods of immobilizing the heart usually did not yield normal flow velocities. This video analysis method allows for the reproducible measurement of particle velocities at time points throughout the cardiac cycle. In keeping the normal heart dynamics intact, more realistic results from the study of coronary blood flows can be obtained.


First Report of Canker Disease Caused by Botryosphaeria parva on Cork Oak Trees in Italy.
A survey was carried out in the spring of 2003 to study the fungi associated with declining trees in a cork oak (Quercus suber L.) forest located in Sassari Province, Sardinia, Italy (40°52'N, 9°01'E) at an altitude of 150 m (above sea level). Several isolates obtained from live twigs and branches showing sunken necrotic bark lesions were identified as Fusicoccum parvum Pennycook & Samuels (teleomorph Botryosphaeria parva Pennycook & Samuels). Neither pycnidia nor ascomata were observed on the symptomatic samples collected. On potato dextrose agar (PDA) at 25°C, the isolates developed an aerial and compact mycelium, initially white but becoming gray after 4 to 6 days, and produced pycnidia after 1 month on sterile cork oak twigs placed on the surface of PDA. Conidia from culture were hyaline, ellipsoidal to fusiform, externally smooth, thin walled, nonseptate, 12 to 19 (15.5) × 5.5 to 8.5 (6.5) μm, with length/width ratio of 2.4 ± 0.1 (mean ± standard error). Identity was confirmed by analysis of the nucleotide sequences of the internal transcribed spacer (ITS) from the rRNA repeat and the translation elongation factor 1-alpha (EF1-α), as done elsewhere (1,4). BLAST searches at GenBank showed a high identity with reference sequences (ITS: >99%; EF1-α: 100%). Representative sequences of both regions were deposited at GenBank (ITS: Accession No. DQ487157; EF1-α: Accession No. DQ487158). Pathogenicity tests were carried out on seven 2-year-old cork oak seedlings maintained in a greenhouse at 14 to 26°C with the B. parva strain CBS 119937 obtained in this study. A mycelial plug (3 to 4 mm2) taken from the margin of an actively growing colony on PDA was put in a shallow wound made by a scalpel on the basal part of the stem of each seedling. Sterile PDA plugs were placed into similar wounds on three control seedlings. The inoculation points were wrapped in Parafilm to retain moisture for 1 week. After 4 weeks, all seedlings inoculated with B. parva died and showed a collapse of the stem cortical tissues associated with dark brown discolorations and vascular necrosis measuring 10.9 ± 0.4 cm. No symptoms were visible in the control seedlings. The pathogen was reisolated from all the inoculated seedlings, thus fulfilling Koch's postulates. The results confirm the virulence of this fungus and point to its possible involvement in the aetiology of cork oak decline. B. parva is a cosmopolitan, plurivorous pathogen causing disease in several hosts of economic importance, such as grapevine (3), kiwi (2), and Eucalyptus spp. trees (1). To our knowledge, this is the first report of B. parva causing canker disease on cork oak trees. References: (1) A. Gezahgne et al. S. Afr. J. Bot. 70:241, 2004. (2) S. R. Pennycook and G. J. Samuels. Mycotaxon 24:445, 1985. (3) A. J. L. Phillips. Phytopathol. Mediterr. 41:3, 2002. (4) B. Slippers et al. Mycologia 96:83, 2004.


Genetic predisposition for tympanosclerotic degeneration.
Atherosclerosis and tympanosclerosis are disorders that affect different organs in the body. When tissue samples taken from atherosclerotic and tympanosclerotic lesions are compared under light and electron microscopes, common pathological characteristics become apparent in both tissues. These similarities indicate that genetic predisposition to atherosclerosis is also seen in the middle ear. For the purpose of evaluating this genetic predisposition, the incidence of tympanosclerosis in atherosclerotic patients was examined and compared with the normal population. Otoscopic examination of 1,024 atherosclerotic patients was performed, and myringosclerosis was revealed in 66.6%. Myringosclerosis was found in 36 out of 300 non-atherosclerotic patients (12%). In the group of atherosclerotic patients, 174 individuals had past histories of previous ear infections. Myringosclerosis was detected in 147 of these patients (84.5%) during ear examinations. In 637 non-atherosclerotic patients with histories of ear infections, this figure was 114 (17.7%). The high rate of tympanosclerosis observed in atherosclerotic patients in comparison to the normal population was found to be statistically significant. These findings indicate that there must be a genetic predisposition for sclerotic degeneration in some individuals.


Physical examination tests for screening and diagnosis of cervicogenic headache: A systematic review.
It has been suggested that differential diagnosis of headaches should consist of a robust subjective examination and a detailed physical examination of the cervical spine. Cervicogenic headache (CGH) is a form of headache that involves referred pain from the neck. To our knowledge, no studies have summarized the reliability and diagnostic accuracy of physical examination tests for CGH. The aim of this study was to summarize the reliability and diagnostic accuracy of physical examination tests used to diagnose CGH. A systematic review following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines was performed in four electronic databases (MEDLINE, Web of Science, Embase and Scopus). Full text reports concerning physical tests for the diagnosis of CGH which reported the clinometric properties for assessment of CGH, were included and screened for methodological quality. Quality Appraisal for Reliability Studies (QAREL) and Quality Assessment of Studies of Diagnostic Accuracy (QUADAS-2) scores were completed to assess article quality. Eight articles were retrieved for quality assessment and data extraction. Studies investigating diagnostic reliability of physical examination tests for CGH scored poorer on methodological quality (higher risk of bias) than those of diagnostic accuracy. There is sufficient evidence showing high levels of reliability and diagnostic accuracy of the selected physical examination tests for the diagnosis of CGH. The cervical flexion-rotation test (CFRT) exhibited both the highest reliability and the strongest diagnostic accuracy for the diagnosis of CGH.


Ethical practice in my work: community health workers' perspectives using photovoice in Wakiso district, Uganda.
Health service delivery should ensure ethical principles are observed at all levels of healthcare. Working towards this goal requires understanding the ethics-related priorities and concerns in the day-to-day activities among different health practitioners. These practitioners include community health workers (CHWs) who are involved in healthcare delivery in communities in many low-and middle-income countries such as Uganda. In this study, we used photovoice, an innovative community based participatory research method that uses photography, to examine CHWs' perspectives on ethical concerns in their work. We explored perspectives of 10 CHWs (5 females and 5 males) on ethical dimensions of their work for 5 months using photovoice in a rural community in Wakiso district, Uganda. As part of the study, we: 1. Oriented CHWs on photovoice research and ethics; 2. Asked CHWs to take photographs of key ethical dimensions of their work; 3. Held monthly meetings to discuss and reflect on the photos; and 4. Disseminated the findings. The discussions from the monthly meetings were audio recorded, transcribed, and emerging data analysed using conventional content analysis with the help of Atlas ti version 6.0.15. CHWs were aware of and highly concerned about the need to observe ethical principles while carrying out their roles. The ethical principles CHWs were aware of and endeavoured to observe during their work were: maintaining professional integrity and abiding by ethical principles of practice; ethical responsibility in patient care; maintaining confidentiality while handling clients; respect for persons and communities; and enhancing their knowledge and skills for better practice. However, CHWs also identified challenges concerning their observance of ethical principles including: low commitment to their work due to other obligations; availability of some reference materials and guidelines in English yet majority could only read in the local language; and minimal avenues for knowledge enhancement such as trainings. CHWs were aware of and keen to discuss ethical issues in their work. However, there is need to address the challenges they face so as to facilitate observing ethical principles during the course of their work in communities.


A novel cysteine-linked antibacterial surface coating significantly inhibits bacterial colonization of nasal silicone prongs in a phase one pre-clinical trial.
Ventilator associated pneumonia and sepsis are frequent complications in neonatal care. Bacterial colonization of medical devices and interfaces used for respiratory support may contribute by functioning as a bacterial reservoir seeding bacteria into airways. We have developed an antibacterial surface coating based on a cysteine ligand covalently coupled via a spacer to a carboxylic backbone layer on an acrylic acid grafted silicone surface. This coating was applied on a commercially available nasal prong and the antibacterial effect was evaluated both in vitro and in vivo in a first-in-human phase 1 trial. The coated nasal prongs had strong antibacterial activity against both Gram-negative and Gram-positive bacteria in vitro. In a randomized pre-clinical trial study of 24 + 24 healthy adult volunteers who carried coated or non-coated nasal prongs for 18 h, a 10log difference in mean bacterial colonization of 5.82 (p < 0.0001) was observed. These results show that this coating technique can prevent colonization by the normal skin and mucosal flora, and thus represent a promising novel technology for reduction of medical device-associated hospital acquired infections.


Characterisation and performance of a Terfenol-D coated femtosecond laser inscribed optical fibre Bragg sensor with a laser ablated microslot for the detection of static magnetic fields.
We present a novel device for the characterisation of static magnetic fields through monitoring wavelength shifts of femtosecond inscribed fibre Bragg grating and micromachined slot, coated with Terfenol-D. The device was sensitive to static magnetic fields and can be used as a vectoral sensor for the detection of magnetic fields as low as 0.046 mT with a resolution of ± 0.3mT in transmission and ± 0.7mT in reflection. The use of a femtosecond laser to both inscribe the FBGs and micromachine the slot in a single stage prior to coating the device significantly simplifies the fabrication.


The Urokinase Plasminogen Activation System in Rheumatoid Arthritis: Pathophysiological Roles and Prospective Therapeutic Targets.
Rheumatoid arthritis (RA) is a chronic and progressive inflammatory disease characterized in its early stages by synovial hyperplasia and inflammatory cell infiltration and later by irreversible joint tissue destruction. The plasminogen activation system (PAS) is associated with a wide range of physiological and pathophysiological states involving fibrinolysis, inflammation and tissue remodeling. Various components of the PAS are implicated in the pathophysiology of RA. Urokinase plasminogen activator (uPA) in particular is a pro-inflammatory mediator that appears to play an important role in the bone and cartilage destruction associated with RA. Clinical studies have shown that uPA and its receptor uPAR are overexpressed in synovia of patients with rheumatoid arthritis. Further, genetic knockdown and antibody-mediated neutralization of uPA have been shown to be protective against induction or progression of arthritis in animal models. The pro-arthritic role of uPA is differentiated from its haemodynamic counterpart, tissue plasminogen activator (tPA), which appears to play a protective role in RA animal models. This review summarises available evidence supporting the PAS as a critical determinant of RA pathogenesis and highlights opportunities for the development of novel uPAS-targeting therapeutics.


Indirect effects of elevated body mass index on memory performance through altered cerebral metabolite concentrations.
Elevated body mass index (BMI) at midlife is associated with increased risk of cognitive decline in later life. The goal of the current study was to assess mechanisms of early brain vulnerability by examining if higher BMI at midlife affects current cognitive performance through alterations in cerebral neurochemistry. Fifty-five participants, aged 40 to 60 years, underwent neuropsychological testing, health screen, and proton magnetic resonance spectroscopy examining N-acetylaspartate, creatine (Cr), myo-inositol (mI), choline, and glutamate concentrations in occipitoparietal gray matter. Concentrations of N-acetylaspartate, choline, mI, and glutamate were calculated as a ratio over Cr and examined in relation to BMI using multivariate regression analyses. Structural equation modeling was used to determine if BMI had an indirect effect on cognition through cerebral metabolite levels. Higher BMI was associated with elevations in mI/Cr (F(5,45) = 3.843, p = .006, β = 0.444, p = .002), independent of age, sex, fasting glucose levels, and systolic blood pressure. Moreover, a χ(2) difference test of the direct and indirect structural equation models revealed that BMI had an indirect effect on global cognitive performance (Δχ(2) = 19.939, df = 2, p < .001). Subsequent follow-up analyses revealed that this effect was specific to memory (Δχ(2) = 22.027, df = 2, p < .001). Higher BMI was associated with elevations in mI/Cr concentrations in the occipitoparietal gray matter and indirectly related to poorer memory performance through mI/Cr levels, potentially implicating plasma hypertonicity and neuroinflammation as mechanisms underlying obesity-related brain vulnerability.


Commonly studied single-nucleotide polymorphisms and breast cancer: results from the Breast Cancer Association Consortium.
The Breast Cancer Association Consortium (BCAC) is an international collaboration that was established to provide large sample sizes for examining genetic associations. We conducted combined analyses on all single-nucleotide polymorphisms (SNPs) whose associations with breast cancer have been investigated by at least three participating groups. Data from up to 12 studies were pooled for each SNP (ADH1C I350V, AURKA F31I, BRCA2 N372H, CASP8 D302H, ERCC2 D312N, IGFBP3 -202 c>a, LIG4 D501D, PGR V660L, SOD2 V16A, TGFB1 L10P, TP53 R72P, XRCC1 R399Q, XRCC2 R188H, XRCC3 T241M, XRCC3 5' UTR, and XRCC3 IVS7-14). Genotype frequencies in case and control subjects were compared, and genotype-specific odds ratios for the risk of breast cancer in heterozygotes and homozygotes for the rare allele compared with homozygotes for the common allele were estimated with logistic regression. Statistical tests were two-sided. The total number of subjects for analysis of each SNP ranged from 12,013 to 31,595. For five SNPs--CASP8 D302H, IGFBP3 -202 c>a, PGR V660L, SOD2 V16A, and TGFB1 L10P--the associations with breast cancer were of borderline statistical significance (P = .016, .060, .047, .056, and .0088 respectively). The remaining 11 SNPs were not associated with breast cancer risk; genotype-specific odds ratios were close to unity. There was some evidence for between-study heterogeneity (P<.05) for four of the 11 SNPs (ADH1C I350V, ERCC2 D312N, XRCC1 R399Q, and XRCC3 IVS5-14). Pooling data within a large consortium has helped to clarify associations of SNPs with breast cancer. In the future, consortia such as the BCAC will be important in the analysis of rare polymorphisms and gene x gene or gene x environment interactions, for which individual studies have low power to identify associations, and in the validation of associations identified from genome-wide association studies.


No effect of natural transformation on the evolution of resistance to bacteriophages in the Acinetobacter baylyi model system.
The adaptive benefits of natural transformation, the active uptake of free DNA molecules from the environment followed by incorporation of this DNA into the genome, may be the improved response to selection resulting from increased genetic variation. Drawing analogies with sexual reproduction, transformation may be particularly beneficial when selection rapidly fluctuates during coevolution with virulent parasites ('the Red Queen Hypothesis'). Here we test this hypothesis by experimentally evolving the naturally transformable and recombinogenic species Acinetobacter baylyi with a cocktail of lytic phages. No increased levels of resistance to phage were found in the wild type compared to a recombination deficient ΔdprA strain after five days of evolution. When exposed to A. baylyi DNA and phage, naturally transformable cells show greater levels of phage resistance. However, increased resistance arose regardless of whether they were exposed to DNA from phage-sensitive or -resistant A. baylyi, suggesting resistance was not the result of transformation, but was related to other benefits of competence. Subsequent evolution in the absence of phages did not show that recombination could alleviate the cost of resistance. Within this study system we found no support for transformation-mediated recombination being an advantage to bacteria exposed to parasitic phages.


Nano-fabrication of molecular electronic junctions by targeted modification of metal-molecule bonds.
Reproducibility, stability and the coupling between electrical and molecular properties are central challenges in the field of molecular electronics. The field not only needs devices that fulfill these criteria but they also need to be up-scalable to application size. In this work, few-molecule based electronics devices with reproducible electrical characteristics are demonstrated. Our previously reported 5 nm gold nanoparticles (AuNP) coated with ω-triphenylmethyl (trityl) protected 1,8-octanedithiol molecules are trapped in between sub-20 nm gap spacing gold nanoelectrodes forming AuNP-molecule network. When the trityl groups are removed, reproducible devices and stable Au-thiol junctions are established on both ends of the alkane segment. The resistance of more than 50 devices is reduced by orders of magnitude as well as a reduction of the spread in the resistance histogram is observed. By density functional theory calculations the orders of magnitude decrease in resistance can be explained and supported by TEM observations thus indicating that the resistance changes and strongly improved resistance spread are related to the establishment of reproducible and stable metal-molecule bonds. The same experimental sequence is carried out using 1,6-hexanedithiol functionalized AuNPs. The average resistances as a function of molecular length, demonstrated herein, are comparable to the one found in single molecule devices.


Retinal pigment epithelial cells display specific transcriptional responses upon TNF-α stimulation.
Tumour necrosis factor-α (TNF-α) is a key mediator of ocular inflammation and its interaction with the retinal pigment epithelium (RPE) may be a driving force in vitreoretinal disorders such as age-related macular degeneration, proliferative vitreoretinopathy (PVR) and diabetic retinopathy. Under inflammatory conditions, the ability of RPE cells to maintain the blood-retinal barrier and immune privilege may be lost and proliferation of RPE cells is facilitated. To gain insight into the effects of TNF-α on RPE cells, a gene expression study was performed. ARPE-19 and HT-29 cells were stimulated with 50 ng/mL TNF-α for 6 h. Gene expression patterns were compared between stimulated and control cells using whole genome gene expression arrays. Data were analysed using Partek and OmniViz and validated using quantitative RT-PCR. Functional annotation analysis was performed using Ingenuity and DAVID. A total of 97 genes were uniquely modulated by TNF-α in ARPE-19 cells compared with HT-29 cells (86 upregulated and 11 downregulated). Most commonly affected biological processes were apoptosis, cell motility and cell signalling. The highest upregulated gene was EFNA1. Among the downregulated genes were transcription factors implicated in ocular development (SIX3, PAX6) and modulation of p53-mediated apoptosis (CITED2). This study provides insight into the unique responses of RPE cells to TNF-α stimulation and suggests a role for genes involved in apoptosis and retinal epithelial development. These findings contribute to our understanding of the behaviour of RPE cells under inflammatory conditions and the crucial role of RPE cells in vitreoretinal diseases.


Vitamin C deficiency in early postnatal life impairs spatial memory and reduces the number of hippocampal neurons in guinea pigs.
The neonatal brain is particularly vulnerable to imbalances in redox homeostasis because of rapid growth and immature antioxidant systems. Vitamin C has been shown to have a key function in the brain, and during states of deficiency it is able to retain higher concentrations of vitamin C than other organs. However, because neurons maintain one of the highest intracellular concentrations of vitamin C in the organism, the brain may still be more sensitive to deficiency despite these preventive measures. The objective was to study the potential link between chronic vitamin C deficiency and neuronal damage in newborn guinea pigs. Thirty 6- to 7-d-old guinea pigs were randomly assigned to 2 groups to receive either a vitamin C-sufficient diet or the same diet containing a low concentration of vitamin C (but adequate to prevent scurvy) for 2 mo. Spatial memory was assessed by the Morris Water Maze, and hippocampal neuron numbers were quantified by stereologic techniques. The results showed a reduction in spatial memory (P < 0.05) and an increased time to first platform hit (P < 0.05) in deficient animals compared with controls. The deficient animals had a lower total number of neurons in hippocampal subdivisions (dentate gyrus, cornu ammonis 1, and cornu ammonis 2-3) than did the normal controls (P < 0.05). Our data show that vitamin C deficiency in early postnatal life results in impaired neuronal development and a functional decrease in spatial memory in guinea pigs. We speculate that this unrecognized effect of vitamin C deficiency may have clinical implications for high-risk individuals, such as in children born from vitamin C-deficient mothers.


Sugar-decorated mesoporous silica nanoparticles as delivery vehicles for the poorly soluble drug celastrol enables targeted induction of apoptosis in cancer cells.
Cancerous cells have a rapid metabolism by which they take up sugars, such as glucose, at significantly higher rates than normal cells. Celastrol is a traditional herbal medicine known for its anti-inflammatory and anti-cancer activities. The poor aqueous solubility and lack of target selectivity of celastrol result in low therapeutic concentration of the drug reaching subcellular compartments of the target tissue, making it an interesting candidate for nanoparticulate delivery. The goal of this study was to utilize glucose as an affinity ligand decorated on mesoporous silica nanoparticles (MSNs), with the aim of delivering these celastrol-loaded MSNs with high specificity to cancer cells and inducing minimal off-target effects in healthy cells. MSNs were thus functionalized with sugar moieties by two different routes, either by conjugation directly to the MSN surface or mediated by a hyperbranched poly(ethylene imine), PEI layer; the latter to increase the cellular uptake by providing an overall positive surface charge as well as to increase the reaction sites for sugar conjugation. The effect of surface functionalization on the target-specific efficacy of the particles was assessed by analyzing the uptake in HeLa and A549 cells as cancer cell models, as compared to mouse embryonic fibroblasts (MEF) as a representative for normal cells. To this end a comprehensive analysis strategy was employed, including flow cytometry, confocal microscopy, and spectrophotometry. When the apoptotic effect of celastrol was evaluated, the anti-cancer activity of celastrol was shown to be significantly enhanced when it was loaded into the specifically designed MSNs. The particles themselves did not induce any toxicity, and normal cells displayed minimal off-target effects. In summary, we show that glucose-functionalized MSNs can be used as efficient carriers for targeted celastrol delivery to achieve specific induction of apoptosis in cancer cells.


Decreasing dietary lauric and myristic acids improves plasma lipids more favorably than decreasing dietary palmitic acid in rhesus monkeys fed AHA step 1 type diets.
Current dietary recommendations advocate reductions in saturated fatty acids (SFA) and cholesterol (C) as a primary intervention for achieving a more desirable plasma lipid profile. To ascertain whether it is more efficacious to decrease dietary lauric and myristic acids (12:0 + 14:0) or dietary palmitic acid (16:0) in conjunction with a reduction in dietary C, 11 rhesus monkeys (8 males, 3 females) were initially fed a control diet rich in SFA + C for 14 wk [dietary fat approximately 38% of energy (%en), SFA 16%en and C at 180 mg/1000 kcal]. Plasma lipids were measured between the 9th and 13th wk, and LDL metabolism was assessed after 13 wk. Monkeys were then split into two groups and fed one of two American Heart Association (AHA) Step 1 diets (approximately 30%en fat, 10%en SFA, 75 mg cholesterol/1000 kcal) for an additional 14 wk, and plasma lipids and LDL metabolism were re-evaluated. Group 1 received a 16:0-rich diet in which most 12:0 + 14:0 were deleted (approximately 8.6%en from 16:0 and approximately 0.3%en from 12:0 + 14:0), whereas Group 2 received a diet rich in 12:0 + 14:0 from which 16:0 was selectively removed (2.6%en from 16:0 and approximately 6.3%en 12:0 + 14:0). In all three diets, oleic and linoleic acid were held relatively constant so that only SFA, the level of total fat and cholesterol were manipulated. Only the Step 1 diet that selectively removed 12:0 + 14:0 (the 16:0-rich diet) significantly reduced all lipid fractions, including total cholesterol (TC), HDL-C, LDL-C, apolipoprotein B (apoB) and the LDL pool size. Plasma triglyceride (TG) and the ratio of TC/HDL-C were not altered by either Step 1 diet. The smaller LDL pool size following the 16:0-rich diet in Group 1 was attributable to a significantly higher fractional catabolic rate (FCR) of LDL because the transport rate of LDL apoB was unaffected. Although the FCR was increased with the 12:0 + 14:0-rich diet, the LDL apoB pool was not affected because the transport rate of LDL tended to increase as well. The data suggest that a Step 1 diet that reduces total fat by decreasing 12:0 + 14:0 in conjunction with dietary C, improves plasma lipids more favorably than a similar diet that selectively removes 16:0 and C. Previous data would imply that the benefit resulted from removal of 12:0 + 14:0 per se, but the possibility is not eliminated that removal of C (independent of 12:0 + 14:0) muted the potential interaction between C and palmitic acid that tends to raise TC.


Possible brain injury events and symptoms in elementary school children.
The purpose of this study was to determine the prevalence of potential mild brain injury incidents accompanied by behavioural changes in a non-clinical population of elementary school-aged children. The researchers also examined data regarding the occurrence of possible brain injuries among regular vs special education students. Survey study. The researchers distributed questionnaires through which parents or guardians of 692 first-to-fifth grade students provided information about possible brain injury incidents and associated behavioural changes. Findings suggest that as many as one-third of children experience one or more incidents potentially resulting in brain injury before completing elementary school. Despite this, only 7.66% of the sample received positive screen results because of behavioural changes associated with a potential incident. None of the 53 children with positive screens were receiving special education services through the traumatic brain injury category of IDEA at the time of the screening. Positive screens were more common among students verified for special education than those enrolled in regular education. Specifically, 25.40% of positive screen results were for children identified with a disability warranting special education services; in contrast, only 5.90% of regular education students received positive screen results.


Paraplegic standing supported by FES-controlled ankle stiffness.
The objective of this paper was to investigate whether a paraplegic subject is able to maintain balance during standing by means of voluntary and reflex activity of the upper body while being supported by closed loop controlled ankle stiffness using FES. The knees and hips of the subject were held in extended positions by a mechanical apparatus, which restricted movement to the sagittal plane. The subject underwent several training sessions where the appropriate level of stiffness around the ankles was maintained by the mechanical apparatus. This enabled the subject to learn how to use the upper body for balancing. After the subject gained adequate skills closed-loop FES was employed to regulate ankle stiffness, replacing the stiffness provided by the apparatus. A method to control antagonist muscle moment was implemented. In subsequent standing sessions, the subject had no difficulties in maintaining balance. When the FES support was withheld, the ability to balance was lost.


Orphans and schooling in Africa: a longitudinal analysis.
AIDS deaths could have a major impact on economic development by affecting the human capital accumulation of the next generation. We estimate the impact of parent death on primary school participation using an unusual five-year panel data set of over 20,000 Kenyan children. There is a substantial decrease in school participation following a parent death and a smaller drop before the death (presumably due to pre-death morbidity). Estimated impacts are smaller in specifications without individual fixed effects, suggesting that estimates based on cross-sectional data are biased toward zero. Effects are largest for children whose mothers died and, in a novel finding, for those with low baseline academic performance.


The cannabinergic system as a target for anti-inflammatory therapies.
Habitual cannabis use has been shown to affect the human immune system, and recent advances in endocannabinoid research provide a basis for understanding these immunomodulatory effects. Cell-based experiments or in vivo animal testing suggest that regulation of the endocannabinoid circuitry can impact almost every major function associated with the immune system. These studies were assisted by the development of numerous novel molecules that exert their biological effects through the endocannabinoid system. Several of these compounds were tested for their effects on immune function, and the results suggest therapeutic opportunities for a variety of inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, atherosclerosis, allergic asthma, and autoimmune diabetes through modulation of the endocannabinoid system.


Primary Percutaneous Coronary Intervention Angioplasty of Occluded Twin Circumflex Coronary Artery in a Patient of Acute Inferior Wall Myocardial Infarction: A Rare Anomaly.
Anomalies of the coronary arteries are reported in 1-2% of patients among diagnostic angiogram. Dual origin of a circumflex from both sinuses is extremely rare among them. We report a case of a patient who underwent primary percutaneous coronary intervention for acute inferior wall myocardial infarction where left coronary injection demonstrated normal obtuse marginal and right coronary injection demonstrated normal right coronary artery (RCA). On further probing, an anomalous left circumflex (LCx) artery was seen arising from RCA ostium which was subsequently cannulated and revascularized by deployment of 2.75 × 26 mm Xience Prime drug-eluting stent (Abott Vascular, USA). Herein, we report for the first time primary percutaneous coronary intervention of twin circumflex and also illustrate that anomalous circumflex can be missed if it arises from RCA ostium and if not probed carefully.


The Y-chromosome point mutation rate in humans.
Mutations are the fundamental source of biological variation, and their rate is a crucial parameter for evolutionary and medical studies. Here we used whole-genome sequence data from 753 Icelandic males, grouped into 274 patrilines, to estimate the point mutation rate for 21.3 Mb of male-specific Y chromosome (MSY) sequence, on the basis of 1,365 meioses (47,123 years). The combined mutation rate for 15.2 Mb of X-degenerate (XDG), X-transposed (XTR) and ampliconic excluding palindromes (rAMP) sequence was 8.71 × 10(-10) mutations per position per year (PPPY). We observed a lower rate (P = 0.04) of 7.37 × 10(-10) PPPY for 6.1 Mb of sequence from palindromes (PAL), which was not statistically different from the rate of 7.2 × 10(-10) PPPY for paternally transmitted autosomes. We postulate that the difference between PAL and the other MSY regions may provide an indication of the rate at which nascent autosomal and PAL de novo mutations are repaired as a result of gene conversion.


Cytokine production by bone marrow mononuclear cells in inherited bone marrow failure syndromes.
Fanconi anaemia (FA), dyskeratosis congenita (DC), Diamond-Blackfan anaemia (DBA), and Shwachman-Diamond syndrome (SDS) are characterized by the progressive development of bone marrow failure. Overproduction of tumour necrosis factor-α (TNF-α) from activated bone marrow T-cells has been proposed as a mechanism of FA-related aplasia. Whether such overproduction occurs in the other syndromes is unknown. We conducted a comparative study on bone marrow mononuclear cells to examine the cellular subset composition and cytokine production. We found lower proportions of haematopoietic stem cells in FA, DC, and SDS, and a lower proportion of monocytes in FA, DC, and DBA compared with controls. The T- and B-lymphocyte proportions were similar to controls, except for low B-cells in DC. We did not observe overproduction of TNF-α or IFN-γ by T-cells in any patients. Induction levels of TNF-α, interleukin (IL)-6, IL-1β, IL-10, granulocyte colony-stimulating factor, and granulocyte-macrophage colony-stimulating factor in monocytes stimulated with high-dose lipopolysaccharide (LPS) were similar at 4 h but lower at 24 h when compared to controls. Unexpectedly, patient samples showed a trend toward higher cytokine level in response to low-dose (0·001 μg/ml) LPS. Increased sensitivity to LPS may have clinical implications and could contribute to the development of pancytopenia by creating a chronic subclinical inflammatory micro-environment in the bone marrow.


Uterine rupture by intended mode of delivery in the UK: a national case-control study.
Recent reports of the risk of morbidity due to uterine rupture are thought to have contributed in some countries to a decrease in the number of women attempting a vaginal birth after caesarean section. The aims of this study were to estimate the incidence of true uterine rupture in the UK and to investigate and quantify the associated risk factors and outcomes, on the basis of intended mode of delivery. A UK national case-control study was undertaken between April 2009 and April 2010. The participants comprised 159 women with uterine rupture and 448 control women with a previous caesarean delivery. The estimated incidence of uterine rupture was 0.2 per 1,000 maternities overall; 2.1 and 0.3 per 1,000 maternities in women with a previous caesarean delivery planning vaginal or elective caesarean delivery, respectively. Amongst women with a previous caesarean delivery, odds of rupture were also increased in women who had ≥ two previous caesarean deliveries (adjusted odds ratio [aOR] 3.02, 95% CI 1.16-7.85) and <12 months since their last caesarean delivery (aOR 3.12, 95% CI 1.62-6.02). A higher risk of rupture with labour induction and oxytocin use was apparent (aOR 3.92, 95% CI 1.00-15.33). Two women with uterine rupture died (case fatality 1.3%, 95% CI 0.2-4.5%). There were 18 perinatal deaths associated with uterine rupture among 145 infants (perinatal mortality 124 per 1,000 total births, 95% CI 75-189). Although uterine rupture is associated with significant mortality and morbidity, even amongst women with a previous caesarean section planning a vaginal delivery, it is a rare occurrence. For women with a previous caesarean section, risk of uterine rupture increases with number of previous caesarean deliveries, a short interval since the last caesarean section, and labour induction and/or augmentation. These factors should be considered when counselling and managing the labour of women with a previous caesarean section.


Changing emotional engagement with running through communal self-tracking: the implications of 'teleoaffective shaping' for public health.
Emerging research explores the role of self-tracking in supporting healthy behaviour. Self-tracking comprises a number of interrelated practices; some individual some communal. In this article, we focus on practices that enable interaction between self-trackers through data sharing and communication around personal data. For public health, communal self-tracking has been explored for the additional benefits it provides in addition to self-knowledge. However, under-explored is the emotional entanglement of self-tracking and tracked activities, or the role of practitioners in the dynamic evolution of tracked practices. Qualitative, mixed methods data were collected from leisure-time runners in the SW England who self-track using social fitness app 'Strava', and were interpreted through the lens of practice theory. We find that communal self-tracking affords the active shaping of the emotion and purpose of running. This 'teleoaffective shaping' allows practitioners to negotiate and reconstitute appealing meanings associated with running to protect their practice loyalty. We identify three mechanisms for teleoaffective shaping afforded by Strava: labelling, reward and materialising effort. Findings advance our understanding of how social fitness apps work to retain practitioners of physically active leisure practices. Future research should further explore the multiple ways that associations with tracked physical activity evolve through entanglement with self-tracking practices.


Measures of condom and safer sex social norms and stigma towards HIV/AIDS among Beijing MSM.
Social norms around condom use and safe sex as well as HIV/AIDS stigma are used to identify persons at higher risk for HIV. These measures have been developed and tested in a variety of settings and populations. While efforts have been undertaken to develop context specific measures of these domains among Chinese MSM, the feasibility of using existing measures is unknown. A survey of MSM, based on respondent-driven sampling, was conducted in Beijing. Existing measures of condom social norms, attitudes towards safer sex and HIV/AIDS stigma were piloted. Internal consistency of all measures was high. As expected higher levels of condom social norms and positive attitudes towards safer sex were associated with condom use. HIV/AIDS stigma and discrimination had a significant relationship with never having an HIV test and lack of discussion of HIV/AIDS with male partners. Correlates of low condom social norms were age, education, employment and resident status. Existing measures of condom social norms, attitudes towards safer sex and HIV/AIDS stigma appear to be appropriate for use among Chinese MSM. Using existing measures as opposed to developing new measures has the potential to expedite investigations into psychosocial correlates of HIV risk behavior.


Assessing the Russian Internet Research Agency's impact on the political attitudes and behaviors of American Twitter users in late 2017.
There is widespread concern that Russia and other countries have launched social-media campaigns designed to increase political divisions in the United States. Though a growing number of studies analyze the strategy of such campaigns, it is not yet known how these efforts shaped the political attitudes and behaviors of Americans. We study this question using longitudinal data that describe the attitudes and online behaviors of 1,239 Republican and Democratic Twitter users from late 2017 merged with nonpublic data about the Russian Internet Research Agency (IRA) from Twitter. Using Bayesian regression tree models, we find no evidence that interaction with IRA accounts substantially impacted 6 distinctive measures of political attitudes and behaviors over a 1-mo period. We also find that interaction with IRA accounts were most common among respondents with strong ideological homophily within their Twitter network, high interest in politics, and high frequency of Twitter usage. Together, these findings suggest that Russian trolls might have failed to sow discord because they mostly interacted with those who were already highly polarized. We conclude by discussing several important limitations of our study-especially our inability to determine whether IRA accounts influenced the 2016 presidential election-as well as its implications for future research on social media influence campaigns, political polarization, and computational social science.


Transmission of drug-resistant tuberculosis in HIV-endemic settings.
The emergence and expansion of the multidrug-resistant tuberculosis epidemic is a threat to the global control of tuberculosis. Multidrug-resistant tuberculosis is the result of the selection of resistance-conferring mutations during inadequate antituberculosis treatment. However, HIV has a profound effect on the natural history of tuberculosis, manifesting in an increased rate of disease progression, leading to increased transmission and amplification of multidrug-resistant tuberculosis. Interventions specific to HIV-endemic areas are urgently needed to block tuberculosis transmission. These interventions should include a combination of rapid molecular diagnostics and improved chemotherapy to shorten the duration of infectiousness, implementation of infection control measures, and active screening of multidrug-resistant tuberculosis contacts, with prophylactic regimens for individuals without evidence of disease. Development and improvement of the efficacy of interventions will require a greater understanding of the factors affecting the transmission of multidrug-resistant tuberculosis in HIV-endemic settings, including population-based molecular epidemiology studies. In this Series article, we review what we know about the transmission of multidrug-resistant tuberculosis in settings with high burdens of HIV and define the research priorities required to develop more effective interventions, to diminish ongoing transmission and the amplification of drug resistance.


Sexually transmitted infections and prostate cancer among men in the U.S. military.
Studies of self-reported sexually transmitted infections (STI) suggesting an association with prostate cancer may reflect underreporting of such infections among nondiseased subjects. To reduce such bias, we studied archived sera in a cohort of U.S. military personnel known to have high rates of both STIs and prostate cancer. Using a nested case-control design, serum samples from 534 men who served on active duty between September 1, 1993 and September 1, 2003 were examined. Controls were individually matched to cases based on date of serum collection, date of birth, branch of service, military rank, marital status, and race. Each of the 267 case-control pairs had two serum samples: a recent serum sample, taken approximately 1 year before the case's prostate cancer diagnosis, and an earlier serum sample, taken approximately 8 years before diagnosis. Each serum specimen was studied for antibodies against human papillomavirus, herpes simplex virus-2 (HSV-2), and Chlamydia trachomatis. Logistic regression accounted for matching and potential confounding factors. Study data indicated no association between prostate cancer and serologic evidence of infections just before the reference date. However, a statistically significant association between prostate cancer and serologic evidence of HSV-2 infection was detected in the earlier sample (odds ratio, 1.60; 95% confidence interval, 1.05-2.44). The strength of this association increased when analyses were restricted to sera collected at least 60 months before diagnosis (odds ratio, 2.04; 95% confidence interval, 1.26-3.29; 204 pairs). If this association is causal, then our findings would suggest a long latency period for prostate cancer development after HSV-2 infection.


Contribution of HLA alleles in the regulation of antibody production in Lyme disease.
A small subset of patients infected with Borrelia burgdorferi (Bb) does not produce Bb specific antibody. Our research provides additional evidence of a genetic predisposition for seronegativity in some individuals with Lyme disease. Because human leukocyte antigen (HLA) class II, a heterodimeric glycoprotein, plays an essential role in the regulation of antibody production, we investigated the difference in HLA genes between seropositive and seronegative patients with Lyme disease (LD). Our results show that HLA-DR7 was associated with anti-Bb antibody production. Nine out of the 22 seropositive LD patients (40.9%) had HLA-DRB1*0701, *0703, *0704 (HLA-DR7); only 1 out of the 18 seronegative LD patients (5.6%) had HLA-DR7 (odds ratio (OR)=11.8, P=0.0126). HLA-DRB1*01021 and HLA-DRB1*0101, *0104, *0105 (HLA-DR1) contributed negatively to anti-Bb antibody production. Seven of 18 seronegative LD patients had HLA-DR1, only 1 of 22 seropositive LD patients had HLA-DR1 (38.9% vs. 4.5%, OR=13.4, P=0.0138). These results suggest that the presence and or lack of production of specific antibody to Bb infection may be associated with particular HLA specificities of the Class II.


Identifying a Transcription Factor's Regulatory Targets from its Binding Targets.
ChIP-chip data, which shows binding of transcription factors (TFs) to promoter regions in vivo, are widely used by biologists to identify the regulatory targets of TFs. However, the binding of a TF to a gene does not necessarily imply regulation. Thus, it is important to develop computational methods which can extract a TF's regulatory targets from its binding targets. We developed a method, called REgulatory Targets Extraction Algorithm (RETEA), which uses partial correlation analysis on gene expression data to extract a TF's regulatory targets from its binding targets inferred from ChIP-chip data. We applied RETEA to yeast cell cycle microarray data and identified the plausible regulatory targets of eleven known cell cycle TFs. We validated our predictions by checking the enrichments for cell cycle-regulated genes, common cellular processes and common molecular functions. Finally, we showed that RETEA performs better than three published methods (MA-Network, TRIA and Garten et al's method).


The Multi-kinase Inhibitor Debio 0617B Reduces Maintenance and Self-renewal of Primary Human AML CD34+ Stem/Progenitor Cells.
Acute myelogenous leukemia (AML) is initiated and maintained by leukemia stem cells (LSC). LSCs are therapy-resistant, cause relapse, and represent a major obstacle for the cure of AML. Resistance to therapy is often mediated by aberrant tyrosine kinase (TK) activation. These TKs primarily activate downstream signaling via STAT3/STAT5. In this study, we analyzed the potential to therapeutically target aberrant TK signaling and to eliminate LSCs via the multi-TK inhibitor Debio 0617B. Debio 0617B has a unique profile targeting key kinases upstream of STAT3/STAT5 signaling such as JAK, SRC, ABL, and class III/V receptor TKs. We demonstrate that expression of phospho-STAT3 (pSTAT3) in AML blasts is an independent prognostic factor for overall survival. Furthermore, phospho-STAT5 (pSTAT5) signaling is increased in primary CD34+ AML stem/progenitors. STAT3/STAT5 activation depends on tyrosine phosphorylation, mediated by several upstream TKs. Inhibition of single upstream TKs did not eliminate LSCs. In contrast, the multi-TK inhibitor Debio 0617B reduced maintenance and self-renewal of primary human AML CD34+ stem/progenitor cells in vitro and in xenotransplantation experiments resulting in long-term elimination of human LSCs and leukemia. Therefore, inhibition of multiple TKs upstream of STAT3/5 may result in sustained therapeutic efficacy of targeted therapy in AML and prevent relapses. Mol Cancer Ther; 16(8); 1497-510. ©2017 AACR.


Aggressive mammary carcinoma progression in Nrf2 knockout mice treated with 7,12-dimethylbenz[a]anthracene.
Activation of nuclear factor erythroid 2-related factor (Nrf2), which belongs to the basic leucine zipper transcription factor family, is a strategy for cancer chemopreventive phytochemicals. It is an important regulator of genes induced by oxidative stress, such as glutathione S-transferases, heme oxygenase-1 and peroxiredoxin 1, by activating the antioxidant response element (ARE). We hypothesized that (1) the citrus coumarin auraptene may suppress premalignant mammary lesions via activation of Nrf2/ARE, and (2) that Nrf2 knockout (KO) mice would be more susceptible to mammary carcinogenesis. Premalignant lesions and mammary carcinomas were induced by medroxyprogesterone acetate and 7,12-dimethylbenz[a]anthracene treatment. The 10-week pre-malignant study was performed in which 8 groups of 10 each female wild-type (WT) and KO mice were fed either control diet or diets containing auraptene (500 ppm). A carcinogenesis study was also conducted in KO vs. WT mice (n = 30-34). Comparisons between groups were evaluated using ANOVA and Kaplan-Meier Survival statistics, and the Mann-Whitney U-test. All mice treated with carcinogen exhibited premalignant lesions but there were no differences by genotype or diet. In the KO mice, there was a dramatic increase in mammary carcinoma growth rate, size, and weight. Although there was no difference in overall survival, the KO mice had significantly lower mammary tumor-free survival. Also, in the KO mammary carcinomas, the active forms of NF-κB and β-catenin were increased ~2-fold whereas no differences in oxidized proteins were observed. Many other tumors were observed, including lymphomas. Interestingly, the incidences of lung adenomas in the KO mice were significantly higher than in the WT mice. We report, for the first time, that there was no apparent difference in the formation of premalignant lesions, but rather, the KO mice exhibited rapid, aggressive mammary carcinoma progression.


Tetrabenazine as anti-chorea therapy in Huntington disease: an open-label continuation study. Huntington Study Group/TETRA-HD Investigators.
Tetrabenazine (TBZ) selectively depletes central monoamines by reversibly binding to the type-2 vesicular monoamine transporter. A previous double blind study in Huntington disease (HD) demonstrated that TBZ effectively suppressed chorea, with a favorable short-term safety profile (Neurology 2006;66:366-372). The objective of this study was to assess the long-term safety and effectiveness of TBZ for chorea in HD. Subjects who completed the 13-week, double blind protocol were invited to participate in this open label extension study for up to 80 weeks. Subjects were titrated to the best individual dose or a maximum of 200 mg/day. Chorea was assessed using the Total Maximal Chorea (TMC) score from the Unified Huntington Disease Rating Scale. Of the 75 participants, 45 subjects completed 80 weeks. Three participants terminated due to adverse events (AEs) including depression, delusions with associated previous suicidal behavior, and vocal tics. One subject died due to breast cancer. The other 26 subjects chose not to continue on with each ensuing extension for various reasons. When mild and unrelated AEs were excluded, the most commonly reported AEs (number of subjects) were sedation/somnolence (18), depressed mood (17), anxiety (13), insomnia (10), and akathisia (9). Parkinsonism and dysarthria [corrected] scores were significantly increased at week 80 compared to baseline. At week 80, chorea had significantly improved from baseline with a mean reduction in the TMC score of 4.6 (SD 5.5) units. The mean dosage at week 80 was 63.4 mg (range 12.5-175 mg). TBZ effectively suppresses HD-related chorea for up to 80 weeks. Patients treated chronically with TBZ should be monitored for parkinsonism, dysphagia and other side effects including sleep disturbance, depression, anxiety, and akathisia. Clinicaltrials.gov registration number (initial study): NCT00219804.