Atogepant for the Preventive Treatment of Migraine.
Atogepant is an oral, small-molecule, calcitonin gene-related peptide receptor antagonist that is being investigated for the preventive treatment of migraine. In a phase 3, double-blind trial, we randomly assigned adults with 4 to 14 migraine days per month in a 1:1:1:1 ratio to receive a once-daily dose of oral atogepant (10 mg, 30 mg, or 60 mg) or placebo for 12 weeks. The primary end point was the change from baseline in the mean number of migraine days per month across the 12 weeks. Secondary end points included headache days per month, a reduction from baseline of at least 50% in the 3-month average of migraine days per month, quality of life, and scores on the Activity Impairment in Migraine-Diary (AIM-D). A total of 2270 participants were screened, 910 were enrolled, and 873 were included in the efficacy analysis; 214 were assigned to the 10-mg atogepant group, 223 to the 30-mg atogepant group, 222 to the 60-mg atogepant group, and 214 to the placebo group. The mean number of migraine days per month at baseline ranged from 7.5 to 7.9 in the four groups. The changes from baseline across 12 weeks were -3.7 days with 10-mg atogepant, -3.9 days with 30-mg atogepant, -4.2 days with 60-mg atogepant, and -2.5 days with placebo. The mean differences from placebo in the change from baseline were -1.2 days with 10-mg atogepant (95% confidence interval [CI], -1.8 to -0.6), -1.4 days with 30-mg atogepant (95% CI, -1.9 to -0.8), and -1.7 days with 60-mg atogepant (95% CI, -2.3 to -1.2) (P<0.001 for all comparisons with placebo). Results for the secondary end points favored atogepant over placebo with the exceptions of the AIM-D Performance of Daily Activities score and the AIM-D Physical Impairment score for the 10-mg dose. The most common adverse events were constipation (6.9 to 7.7% across atogepant doses) and nausea (4.4 to 6.1% across atogepant doses). Serious adverse events included one case each of asthma and optic neuritis in the 10-mg atogepant group. Oral atogepant once daily was effective in reducing the number of migraine days and headache days over a period of 12 weeks. Adverse events included constipation and nausea. Longer and larger trials are needed to determine the effect and safety of atogepant for migraine prevention. (Funded by Allergan; ADVANCE ClinicalTrials.gov number, NCT03777059.).


Probing the structure and function of the protease domain of botulinum neurotoxins using single-domain antibodies.
Botulinum neurotoxins (BoNTs) are among the deadliest of bacterial toxins. BoNT serotype A and B in particular pose the most serious threat to humans because of their high potency and persistence. To date, there is no effective treatment for late post-exposure therapy of botulism patients. Here, we aim to develop single-domain variable heavy-chain (VHH) antibodies targeting the protease domains (also known as the light chain, LC) of BoNT/A and BoNT/B as antidotes for post-intoxication treatments. Using a combination of X-ray crystallography and biochemical assays, we investigated the structures and inhibition mechanisms of a dozen unique VHHs that recognize four and three non-overlapping epitopes on the LC of BoNT/A and BoNT/B, respectively. We show that the VHHs that inhibit the LC activity occupy the extended substrate-recognition exosites or the cleavage pocket of LC/A or LC/B and thus block substrate binding. Notably, we identified several VHHs that recognize highly conserved epitopes across BoNT/A or BoNT/B subtypes, suggesting that these VHHs exhibit broad subtype efficacy. Further, we identify two novel conformations of the full-length LC/A, that could aid future development of inhibitors against BoNT/A. Our studies lay the foundation for structure-based engineering of protein- or peptide-based BoNT inhibitors with enhanced potencies and cross-subtypes properties.


A randomised trial of early palliative care for maternal stress in infants prenatally diagnosed with single-ventricle heart disease.
Children with single-ventricle disease experience high mortality and complex care. In other life-limiting childhood illnesses, paediatric palliative care may mitigate maternal stress. We hypothesised that early palliative care in the single-ventricle population may have the same benefit for mothers. In this pilot randomised trial of early palliative care, mothers of infants with prenatal single-ventricle diagnoses completed surveys measuring depression, anxiety, coping, and quality of life at a prenatal visit and neonatal discharge. Infants were randomised to receive early palliative care - structured evaluation, psychosocial/spiritual, and communication support before surgery - or standard care. Among 56 eligible mothers, 40 enrolled and completed baseline surveys; 38 neonates were randomised, 18 early palliative care and 20 standard care; and 34 postnatal surveys were completed. Baseline Beck Depression Inventory-II and State-Trait Anxiety Index scores exceeded normal pregnant sample scores (mean 13.76±8.46 versus 7.0±5.0 and 46.34±12.59 versus 29.8±6.35, respectively; p=0.0001); there were no significant differences between study groups. The early palliative care group had a decrease in prenatal to postnatal State-Trait Anxiety Index scores (-7.6 versus 0.3 in standard care, p=0.02), higher postnatal Brief Cope Inventory positive reframing scores (p=0.03), and a positive change in PedsQL Family Impact Module communication and family relationships scores (effect size 0.46 and 0.41, respectively). In conclusion, these data show that mothers of infants with single-ventricle disease experience significant depression and anxiety prenatally. Early palliative care resulted in decreased maternal anxiety, improved maternal positive reframing, and improved communication and family relationships.


Statics and dynamics of adhesion between two soap bubbles.
An original set-up is used to study the adhesive properties of two hemispherical soap bubbles put into contact. The contact angle at the line connecting the three films is extracted by image analysis of the bubbles profiles. After the initial contact, the angle rapidly reaches a static value slightly larger than the standard 120 degrees angle expected from Plateau rule. This deviation is consistent with previous experimental and theoretical studies: it can be quantitatively predicted by taking into account the finite size of the Plateau border (the liquid volume trapped at the vertex) in the free energy minimization. The visco-elastic adhesion properties of the bubbles are further explored by measuring the deviation Delta theta (d)(t) of the contact angle from the static value as the distance between the two bubbles supports is sinusoidally modulated. It is found to linearly increase with Delta r(c) / r(c) , where r(c) is the radius of the central film and Delta r(c) the amplitude of modulation of this length induced by the displacement of the supports. The in-phase and out-of-phase components of Delta theta (d)(t) with the imposed modulation frequency are systematically probed, which reveals a transition from a viscous to an elastic response of the system with a crossover pulsation of the order 1rad x s(-1). Independent interfacial rheological measurements, obtained from an oscillating bubble experiment, allow us to develop a model of dynamic adhesion which is confronted to our experimental results. The relevance of such adhesive dynamic properties to the rheology of foams is briefly discussed using a perturbative approach to the Princen 2D model of foams.


Annual summary of vital statistics--2002.
The crude birth rate in 2002 was 13.9 births per 1000 population, the lowest ever reported for the United States. The number of births, the crude birth rate, and the fertility rate (64.8) all declined slightly (by 1% or less) from 2001 to 2002. Fertility rates were highest for Hispanic women (94.0), followed by black (65.4), Asian or Pacific Islander (63.9), Native American (58.0), and non-Hispanic white women (57.5). Fertility rates declined slightly for all race/ethnic groups from 2001 to 2002. The birth rate for teen mothers continued to fall, dropping 5% from 2001 to 2002 to 42.9 births per 1000 women aged 15 to 19 years, another record low. The teen birth rate has fallen 31% since 1991; declines were more rapid for younger teens aged 15 to 17 (40%) than for older teens aged 18 to 19 (23%). The proportion of all births to unmarried women remained approximately the same at one third. Smoking during pregnancy continued to decline; smoking rates were highest among teen mothers. In 2002, 26.1% of births were delivered by cesarean section, up 7% since 2001 and 26% since 1996. The primary cesarean rate has risen 23% since 1996, whereas the rate of vaginal birth after a previous cesarean delivery has fallen 55%. The use of timely prenatal care increased slightly to 83.8% in 2002. From 1990 to 2002, the use of timely prenatal care increased by 6% (to 88.7%) for non-Hispanic white women, by 24% (to 75.2%) for black women, and by 28% (to 76.8%) for Hispanic women, thus narrowing racial disparities. The percentage of preterm births rose to 12.0% in 2002, from 10.6% in 1990 and 9.4% in 1981. Increases were largest for non-Hispanic white women. The percentage of low birth weight (LBW) births also increased to 7.8% in 2002, up from 6.7% in 1984. Twin and triplet/+ birth rates both increased by 3% from 2000 to 2001. Multiple births accounted for 3.2% of all births in 2001. The infant mortality rate (IMR) was 6.9 per 1000 live births (provisional data) in 2002 compared with 6.8 in 2001 (final data). The ratio of the IMR among black infants to that for white infants was 2.5 in 2001, the same as in 2000. Racial differences in infant mortality remain a major public health concern. The role of LBW in infant mortality remains a major issue. New Hampshire, Utah, and Massachusetts had the lowest IMRs. State-by-state differences in IMR reflect racial composition, the percentage of LBW, and birth weight-specific neonatal mortality rates for each state. The United States continues to rank poorly in international comparisons of infant mortality. Expectation of life at birth reached a record high of 77.2 years for all sex and race groups combined in 2001. Death rates in the United States continue to decline. Between 2000 and 2001, death rates declined for the 3 leading causes of death: diseases of the heart, malignant neoplasms, and cerebrovascular diseases. Death rates for children ages 1 to 19 years decreased for unintentional injuries by 3.3% in 2001; the death rate for chronic lower respiratory diseases decreased by 25% in 2001. Cancer and suicide levels did not change for children ages 1 to 19. A large proportion of childhood deaths continue to occur as a result of preventable injuries.


Selective alterations in gene expression for NMDA receptor subunits in prefrontal cortex of schizophrenics.
NMDA receptor antagonists can induce a schizophrenia-like psychosis, but the role of NMDA receptors in the pathophysiology of schizophrenia remains unclear. Expression patterns of mRNAs for five NMDA receptor subunits (NR1/NR2A-D) were determined by in situ hybridization in prefrontal, parieto-temporal, and cerebellar cortex of brains from schizophrenics and from neuroleptic-treated and nonmedicated controls. In the cerebral cortex of both schizophrenics and controls, mRNAs for NR1, NR2A, NR2B, and NR2D subunits were preferentially expressed in layers II/III, Va, and VIa, with much higher levels in the prefrontal than in the parieto-temporal cortex. Levels of mRNA for the NR2C subunit were very low overall. By contrast, the cerebellar cortex of both schizophrenics and controls contained very high levels of NR2C subunit mRNA, whereas levels for the other subunit mRNAs were very low, except NR1, for which levels were moderate. Significant alterations in the schizophrenic cohort were confined to the prefrontal cortex. Here there was a shift in the relative proportions of mRNAs for the NR2 subunit family, with a 53% relative increase in expression of the NR2D subunit mRNA. No comparable changes were found in neuroleptic-treated or untreated controls. These findings indicate regional heterogeneity of NMDA receptor subunit expression in human cerebral and cerebellar cortex. In schizophrenics, the alterations in expression of NR2 subunit mRNA in prefrontal cortex are potential indicators of deficits in NMDA receptor-mediated neurotransmission accompanying functional hypoactivity of the frontal lobes.


Very early passive cycling exercise in mechanically ventilated critically ill patients: physiological and safety aspects--a case series.
Early mobilization can be performed in critically ill patients and improves outcomes. A daily cycling exercise started from day 5 after ICU admission is feasible and can enhance functional capacity after hospital discharge. In the present study we verified the physiological changes and safety of an earlier cycling intervention (< 72 hrs of mechanical ventilation) in critical ill patients. Nineteen hemodynamically stable and deeply sedated patients within the first 72 hrs of mechanical ventilation were enrolled in a single 20 minute passive leg cycling exercise using an electric cycle ergometer. A minute-by-minute evaluation of hemodynamic, respiratory and metabolic variables was undertaken before, during and after the exercise. Analyzed variables included the following: cardiac output, systemic vascular resistance, central venous blood oxygen saturation, respiratory rate and tidal volume, oxygen consumption, carbon dioxide production and blood lactate levels. We enrolled 19 patients (42% male, age 55 ± 17 years, SOFA = 6 ± 3, SAPS3 score = 58 ± 13, PaO2/FIO2 = 223 ± 75). The median time of mechanical ventilation was 1 day (02), and 68% (n=13) of our patients required norepinephrine (maximum concentration = 0.47 µg.kg(-1).min(-1)). There were no clinically relevant changes in any of the analyzed variables during the exercise, and two minor adverse events unrelated to hemodynamic instability were observed. In our study, this very early passive cycling exercise in sedated, critically ill, mechanically ventilated patients was considered safe and was not associated with significant alterations in hemodynamic, respiratory or metabolic variables even in those requiring vasoactive agents.


Influences of carboxyl functionalization of intercalators on exfoliation of graphite oxide: a molecular dynamics simulation.
In this study, the influences of the carboxyl functionalization of intercalators on exfoliation of graphite oxide were analyzed using molecular dynamics (MD) simulations. Molecular models of four-layered graphene oxide (GO) sheets, four different solvents (ethanol, dimethylformamide, tetrahydrofuran, and N-methyl-2-pyrrolidone), and four different intercalators (anthracene, 2-anthracenecarboxylic acid, 2,3-anthracenedicarboxylic acid, and 2,6-anthracenedicarboxylic acid) were used in the MD simulations. A separation simulation of GO sheets was performed to determine the point at which the GO sheets begin to exfoliate. An insertion simulation was used to obtain the minimum kinetic energy required for exfoliation and to calculate GO-solvent and GO-intercalator interaction energies. As the simulation result, GO-solvent and GO-intercalator interactions affected the minimum kinetic energy required for exfoliation. Having more carboxyl functional groups on the anthracene improved both the GO-intercalator interaction and the efficiency of the intercalators during exfoliation. These results reveal that increasing the interaction energy between the GO sheets and the insertion molecules is an efficient way to improve the performance of the solvents and the intercalators for the exfoliation of GO sheets.


Long-term culture of human liver tissue with advanced hepatic functions.
A major challenge for studying authentic liver cell function and cell replacement therapies is that primary human hepatocytes rapidly lose their advanced function in conventional, 2-dimensional culture platforms. Here, we describe the fabrication of 3-dimensional hexagonally arrayed lobular human liver tissues inspired by the liver's natural architecture. The engineered liver tissues exhibit key features of advanced differentiation, such as human-specific cytochrome P450-mediated drug metabolism and the ability to support efficient infection with patient-derived inoculums of hepatitis C virus. The tissues permit the assessment of antiviral agents and maintain their advanced functions for over 5 months in culture. This extended functionality enabled the prediction of a fatal human-specific hepatotoxicity caused by fialuridine (FIAU), which had escaped detection by preclinical models and short-term clinical studies. The results obtained with the engineered human liver tissue in this study provide proof-of-concept determination of human-specific drug metabolism, demonstrate the ability to support infection with human hepatitis virus derived from an infected patient and subsequent antiviral drug testing against said infection, and facilitate detection of human-specific drug hepatotoxicity associated with late-onset liver failure. Looking forward, the scalability and biocompatibility of the scaffold are also ideal for future cell replacement therapeutic strategies.


Two chloroplast DNA inversions originated simultaneously during the early evolution of the sunflower family (Asteraceae).
The chloroplast DNA (cpDNA) inversion in the Asteraceae has been cited as a classic example of using genomic rearrangements for defining major lineages of plants. We further characterize cpDNA inversions in the Asteraceae using extensive sequence comparisons among 56 species, including representatives of all major clades of the family and related families. We determine the boundaries of the 22-kb (now known as 22.8 kb) inversion that defines a major split within the Asteraceae, and in the process, we characterize the second and a new, smaller 3.3-kb inversion that occurs at one end of the larger inversion. One end point of the smaller inversion is upstream of the trnE-UUC gene, and the other end point is located between the trnC-GCA and rpoB genes. Although a diverse sampling of Asteraceae experienced substantial length variation and base substitution during the long evolutionary history subsequent to the inversion events, the precise locations of the inversion end points are identified using comparative sequence alignments in the inversion regions. The phylogenetic distribution of two inversions is identical among the members of Asteraceae, suggesting that the inversion events likely occurred simultaneously or within a short time period shortly after the origin of the family. Estimates of divergence times based on ndhF and rbcL sequences suggest that two inversions originated during the late Eocene (38-42 MYA). The divergence time estimates also suggest that the Asteraceae originated in the mid Eocene (42-47 MYA).


Synonymous variants in holoprosencephaly alter codon usage and impact the Sonic Hedgehog protein.
Synonymous single nucleotide variants (sSNVs) have been implicated in various genetic disorders through alterations of pre-mRNA splicing, mRNA structure and miRNA regulation. However, their impact on synonymous codon usage and protein translation remains to be elucidated in clinical context. Here, we explore the functional impact of sSNVs in the Sonic Hedgehog (SHH) gene, identified in patients affected by holoprosencephaly, a congenital brain defect resulting from incomplete forebrain cleavage. We identified eight sSNVs in SHH, selectively enriched in holoprosencephaly patients as compared to healthy individuals, and systematically assessed their effect at both transcriptional and translational levels using a series of in silico and in vitro approaches. Although no evidence of impact of these sSNVs on splicing, mRNA structure or miRNA regulation was found, five sSNVs introduced significant changes in codon usage and were predicted to impact protein translation. Cell assays demonstrated that these five sSNVs are associated with a significantly reduced amount of the resulting protein, ranging from 5% to 23%. Inhibition of the proteasome rescued the protein levels for four out of five sSNVs, confirming their impact on protein stability and folding. Remarkably, we found a significant correlation between experimental values of protein reduction and computational measures of codon usage, indicating the relevance of in silico models in predicting the impact of sSNVs on translation. Considering the critical role of SHH in brain development, our findings highlight the clinical relevance of sSNVs in holoprosencephaly and underline the importance of investigating their impact on translation in human pathologies.


Sympathectomy via a posterior approach after a failed trans-thoracic approach: a case of its use for arrhythmia.
Congenital long QT syndrome (LQTS) provides an opportunity for neurosurgical intervention. Medication and implantable cardiac defibrillator (ICD)-refractory patients often require left cardiac sympathetic denervation (LCSD) via anterior video-assisted thoracoscopic surgery (VATS). However, this approach has major pulmonary contraindications and risks, with a common concern in children being their inability to tolerate single-lung ventilation. At Oregon Health & Science University, the authors have developed a posterior approach-extrapleural, minimally invasive, T1-5 LCSD-that minimizes this risk. A 9-year-old girl with LQTS type III presented to the emergency department while experiencing ventricular tachycardia (VT) and ventricular fibrillation (VF) with multiple ICD firings. Medical management failed to resolve the VF/VT. VATS was attempted but could not be safely performed due to respiratory insufficiency. The patient was reintubated for dual-lung ventilation and repositioned prone. Her respiratory insufficiency resolved. Using METRx serial dilating tubes under the microscope, the left T1-5 sympathetic ganglia were sectioned and removed. Postoperatively, the patient had no episodes of VF/VT, pneumothorax, hemothorax, or Horner syndrome. With mexiletine and propranolol, she has remained largely VF/VT free, with only one VT episode during the 2-year follow-up period. Minimally invasive, posterior, extrapleural, T1-5 LCSD is safe and effective for treating congenital LQTS in children, while minimizing the risks associated with VATS.


Genes of domestic mammals augmented by backcrossing with wild ancestors.
Both archaeological data and the presence of few mitochondrial DNA lineages suggest that most widespread domestic mammals (cattle, sheep, goats, pigs and dogs) derive from only a handful of domestication events. However, each of these species shows a high level of diversity at the nuclear genes of the major histocompatibility complex (MHC). Through simulations incorporating various degrees of population subdivision, growth rate and selection, we demonstrate that the numerous MHC DRB alleles that are present in modern domestic mammals implies that substantial backcrossing with wild ancestors, either accidental or intentional, has been important in shaping the genetic diversity of our domesticates. These results support the view that, contrary to common assumption, domestic and wild lineages might not have been clearly separated throughout their history.


Yeast Rev1 protein promotes complex formation of DNA polymerase zeta with Pol32 subunit of DNA polymerase delta.
Yeast DNA polymerase (Pol) delta, essential for DNA replication, is comprised of 3 subunits, Pol3, Pol31, and Pol32. Of these, the catalytic subunit Pol3 and the second subunit Pol31 are essential, whereas the Pol32 subunit is not essential for DNA replication. Although Pol32 is an integral component of Pol delta, it is also required for translesion synthesis (TLS) by Pol zeta. To begin to decipher the bases of Pol32 involvement in Pol zeta-mediated TLS, here we examine whether Pol32 physically interacts with Pol zeta or its associated proteins and provide evidence for the physical interaction of Pol32 with Rev1. Rev1 plays an indispensable structural role in Pol zeta-mediated TLS and it binds the Rev3 catalytic subunit of Pol zeta. Here, we show that although Pol32 does not directly bind Pol zeta, Pol32 can bind the Rev1-Pol zeta complex through its interaction with Rev1. We find that Pol32 binding has no stimulatory effect on DNA synthesis either by Rev1 in the Rev1-Pol32 complex or by Pol zeta in the Pol zeta-Rev1-Pol32 complex, irrespective of whether proliferating cell nuclear antigen has been loaded onto DNA or not. We discuss evidence for the biological significance of Rev1 binding to Pol32 for Pol zeta function in TLS and suggest a structural role for Rev1 in modulating the binding of Pol zeta with Pol32 in Pol delta stalled at a lesion site.


Overcoming Translational Barriers in Acute Kidney Injury: A Report from an NIDDK Workshop.
AKI is a complex clinical condition associated with high mortality, morbidity, and health care costs. Despite improvements in methodology and design of clinical trials, and advances in understanding the underlying pathophysiology of rodent AKI, no pharmacologic agent exists for the prevention or treatment of AKI in humans. To address the barriers that affect successful clinical translation of drug targets identified and validated in preclinical animal models of AKI in this patient population, the National Institute of Diabetes and Digestive and Kidney Diseases convened the "AKI Outcomes: Overcoming Barriers in AKI" workshop on February 10-12, 2015. The workshop used a reverse translational medicine approach to identify steps necessary to achieve clinical success. During the workshop, breakout groups were charged first to design feasible, phase 2, proof-of-concept clinical trials for delayed transplant graft function, prevention of AKI (primary prevention), and treatment of AKI (secondary prevention and recovery). Breakout groups then were responsible for identification of preclinical animal models that would replicate the pathophysiology of the phase 2 proof-of-concept patient population, including primary and secondary end points. Breakout groups identified considerable gaps in knowledge regarding human AKI, our understanding of the pathophysiology of AKI in preclinical animal models, and the fidelity of cellular and molecular targets that have been evaluated preclinically to provide information regarding human AKI of various etiologies. The workshop concluded with attendees defining a new path forward to a better understanding of the etiology, pathology, and pathophysiology of human AKI.


Non-invasive monitoring of intra-tumor drug concentration and therapeutic response using optical spectroscopy.
Optical spectroscopy was used to monitor changes in tumor physiology with therapy, and its influence on drug delivery and treatment efficacy for hyperthermia treatment combined with free doxorubicin or a low-temperature sensitive liposomal formulation. Monte Carlo-based modeling techniques were used to characterize the intrinsic absorption, scattering, and fluorescence properties of tissue. Fluorescence assessment of drug concentration was validated against HPLC and found to be significantly linearly correlated (r=0.88). Cluster analysis on the physiologic data obtained by optical spectroscopy revealed two physiologic phenotypes prior to treatment. One of these was relatively hypoxic, with relatively low total hemoglobin content. This hypoxic group was found to have a significantly shorter time to reach 3 times pre-treatment volume, indicating a more treatment resistant phenotype (p=0.003). Influence of tumor physiology was assessed in more detail for the liposomal doxorubicin+hyperthermia group, which demonstrated a highly significant correlation between pre-treatment hemoglobin saturation and tumor growth delay, and also between post-hyperthermia total hemoglobin content and tumor drug delivery. Finally, it was found that the doxorubicin concentration, measured in vivo using fluorescence techniques significantly predicted for chemoresponse (hazard ratio: 0.34, p=0.0007). The ability to characterize drug delivery and tumor physiology in vivo makes this a potentially useful tool for evaluating the efficacy of targeted delivery systems in preclinical studies, and may be translatable for monitoring and predicting individual treatment responses in the clinic.


Treatment results of radical radiotherapy of carcinoma uterine cervix using external beam radiotherapy and high dose rate intracavitary radiotherapy.
To report the outcome of carcinoma cervix patients treated radically by external beam radiotherapy and high dose rate intracavitary radiotherapy. From January 2005 to December 2006, a total of 709 newly diagnosed cases of carcinoma cervix were reported in our department. All cases were staged according to the International Federation of Gynecologist and Oncologist staging system. Out of 709 cases, 342 completed radical radiotherapy and were retrospectively analyzed for the presence of local residual disease, local recurrence, distant metastases, radiation reaction, and disease free survival. There were 11(3.22%), 82(23.98%), 232(67.83%), and 17(4.97%) patients in stages I, II, III, and IV, respectively. The median follow up time for all patients was 36 months (range 3 -54 months). The overall treatment time (OTT) ranged from 52 to 69 days (median 58 days). The 3 year disease free survival rate was 81.8%, 70.7%, 40.08%, and 11.76% for stages I, II, III, and IV, respectively. There were 91 (26.6%) cases with local residual diseases, 27(7.9%) developed distant metastasis, and 18(5.26%) pts had local recurrence. The results of this study suggest that radical radiotherapy with HDR brachytherapy was appropriate for the treatment of early staged cancer of uterine cervix. For locally advanced cancer of cervix addition of concurrent chemotherapy, higher radiation doses, reduction of overall treatment time to less than 8 weeks, and use of latest radiotherapy techniques such as IMRT is recommended to improve the results.


Cross-cultural adaptation to Brazilian Portuguese of the Waterloo Footedness Questionnaire-Revised: WFQ-R-Brazil.
Post-stroke hemiparesis causes compensated postures, which can modify the footedness established before the impairment. Recently, a paresis severity-modulated dominance hypothesis stated that measures to detect footedness become crucial to float new ideas for neurorehabilitation strategies. The Waterloo Footedness Questionnaire-Revised (WFQ-R) represents the most acceptable measure but it had not yet been cross-culturally adapted for Portuguese spoken in Brazil. Our aim was to cross-culturally adapt the WFQ-R to Brazilian Portuguese, verifying its reliability. We completed the essential steps to cross-culturally adapt one version, tested in 12 patients with post-stroke hemiparesis and 12 able-bodied individuals, sampled by convenience, to verify reliability. Measurements were taken by two independent raters during the test and by one of them at the one-week retest. No great semantic, linguistic or cultural differences were found, and acceptable reliability was recorded. The WFQ-R-Brazil is reliable and ready for use in the Brazilian able-bodied and post-stroke hemiparesis population.


Linkages between public and non-government sectors in healthcare: A case study from Uttar Pradesh, India.
Effective utilisation of collaborative non-governmental organisation (NGO)-public health system linkages in pluralistic health systems of developing countries can substantially improve equity and quality of services. This study explores level and types of linkages between public health sector and NGOs in Uttar Pradesh (UP), an underprivileged state of India, using a social science model for the first time. It also identifies gaps and challenges for effective linkage. Two NGOs were selected as case studies. Data collection included semi-structured in-depth interviews with senior staff and review of records and reporting formats. Formal linkages of NGOs with the public health system related to registration, participation in district level meetings, workforce linkages and sharing information on government-supported programmes. Challenges included limited data sharing, participation in planning and limited monitoring of regulatory compliances. Linkage between public health system and NGOs in UP was moderate, marked by frequent interaction and some reciprocity in information and resource flows, but weak participation in policy and planning. The type of linkage could be described as 'complementarity', entailing information and resource sharing but not joint action. Stronger linkage is required for sustained and systematic collaboration, with joint planning, implementation and evaluation.


Simultaneous Testing for 6 Lysosomal Storage Disorders and X-Adrenoleukodystrophy in Dried Blood Spots by Tandem Mass Spectrometry.
Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and cost-effective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for β-glucosidase, acid α-glucosidase, α-galactosidase A, galactocerebrosidase, and α-L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n = 33), X-adrenoleukodystrophy (n = 9), or a peroxisomal biogenesis disorder (n = 5), as well as carriers for Fabry disease (n = 17) and X-adrenoleukodystrophy (n = 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS.


"Get back to where you once belonged": monitoring the AIDS pandemic in the 21st century.
This paper reviews the concepts and methodological strategies that have shaped the monitoring of the AIDS pandemic, today in its third decade. A deliberate effort was made to highlight aspects usually forgotten by the canon. The paper aims to track the footsteps of the evolving strategies in the field of surveillance & monitoring, with the help of disciplines such as epidemiology, molecular biology, social, and behavioral sciences. The deep divide that opposes societies severely affected by the epidemic and affluent societies much less affected by the epidemic is contrasted with the scarce human and financial resources of the societies facing harshest epidemic vis-à-vis the comprehensiveness of the response to the epidemic in their affluent counterparts in terms of the scope and high standards of their initiatives on monitoring, prevention, management & care. The pressing need to implement feasible alternatives to the current sophisticated and expensive ones is briefly discussed. Beyond the renewed challenge posed to the creativity of scientists and health professionals, the AIDS pandemic is described as a major public health crisis, compromising the social fabric in some contexts, and as a never fulfilled calling for an ethics of solidarity between different societies and different social strata of each given society.


Two-character motion analysis and synthesis.
In this paper, we deal with the problem of synthesizing novel motions of standing-up martial arts such as Kickboxing, Karate, and Taekwondo performed by a pair of human-like characters while reflecting their interactions. Adopting an example-based paradigm, we address three non-trivial issues embedded in this problem: motion modeling, interaction modeling, and motion synthesis. For the first issue, we present a semi-automatic motion labeling scheme based on force-based motion segmentation and learning-based action classification. We also construct a pair of motion transition graphs each of which represents an individual motion stream. For the second issue, we propose a scheme for capturing the interactions between two players. A dynamic Bayesian network is adopted to build a motion transition model on top of the coupled motion transition graph that is constructed from an example motion stream. For the last issue, we provide a scheme for synthesizing a novel sequence of coupled motions, guided by the motion transition model. Although the focus of the present work is on martial arts, we believe that the framework of the proposed approach can be conveyed to other two-player motions as well.


A Genomics-Based Approach Identifies a Thioviridamide-Like Compound with Selective Anticancer Activity.
Thioviridamide is a structurally novel ribosomally synthesized and post-translational modified peptide (RiPP) produced by Streptomyces olivoviridis NA005001. It is characterized by a structure that features a series of thioamide groups and possesses potent antiproliferative activity in cancer cell lines. Its unusual structure allied to its promise as an anticancer compound led us to investigate the diversity of thioviridamide-like pathways across sequenced bacterial genomes. We have isolated and characterized three diverse members of this family of natural products. This characterization is supported by transformation-associated recombination cloning and heterologous expression of one of these compounds, thiostreptamide S4. Our work provides an insight into the diversity of this rare class of compound and indicates that the unusual N-terminus of thioviridamide is not introduced biosynthetically but is instead introduced during acetone extraction. A detailed analysis of the biological activity of one of the newly discovered compounds, thioalbamide, indicates that it is highly cytotoxic to cancer cells, while exhibiting significantly less activity toward a noncancerous epithelial cell line.


Single vs. dual chamber implantable cardioverter-defibrillators or programming of implantable cardioverter-defibrillators in patients without a bradycardia pacing indication: systematic review and meta-analysis.
Implantable cardioverter-defibrillators (ICDs) are key in the prevention of sudden cardiac death, but outcomes may vary by type of device or programming [single chamber (SC) vs. dual chamber (DC)] in patients without a bradycardia pacing indication. We sought to meta-analyse patient outcomes of randomized trials of SC vs. DC devices or programming. We searched PubMed, Embase, Scopus, Web of Science, and Cochrane trials databases for relevant studies excluding those published before 2000, involving children, or not available in English. Endpoints included mortality, inappropriate ICD therapies, and implant complications. Endpoints with at least three reporting studies were meta-analysed. We identified eight studies meeting inclusion criteria representing 2087 patients with 16.1 months mean follow-up. Mean age was 62.7 years (SD 1.92); in six studies reporting sex, most patients were male (85%). Comparing patients with a SC or DC ICD or programming, we found similar rates of mortality [odds ratio (OR) 0.95, 95% confidence interval (CI) 0.54-1.68; P = 0.86] and inappropriate therapies (OR 1.46, 95% CI 0.97-2.19; P = 0.07) in five and six studies, respectively. In three studies of SC vs. DC ICDs (but not programming) rates of pneumothorax and lead dislodgement were not different (OR 2.12, 95% CI 0.18-24.72; P = 0.55 and OR 0.87, 95% CI 0.32-2.47; P = 0.83, respectively). In this meta-analysis of randomized controlled trials comparing SC vs. DC ICD device or programming, there was no significant difference in inappropriate therapies, mortality, pneumothorax, or lead dislodgement. Future studies should compare these devices over longer follow-up and in specific patient populations.


Host immune responses to a viral immune modulating protein: immunogenicity of viral interleukin-10 in rhesus cytomegalovirus-infected rhesus macaques.
Considerable evidence has accumulated that multiple viruses, bacteria, and protozoa manipulate interleukin-10 (IL-10)-mediated signaling through the IL-10 receptor (IL-10R) in ways that could enable establishment of a persistent microbial infection. This suggests that inhibition of pathogen targeting of IL-10/IL-10R signaling could prevent microbial persistence. Human cytomegalovirus (HCMV) and rhesus cytomegalovirus (RhCMV) express a viral interleukin-10 (cmvIL-10 and rhcmvIL-10, respectively) with comparable immune modulating properties in vitro to that of their host's cellular IL-10 (cIL-10). A prior study noted that rhcmvIL-10 alters innate and adaptive immunity to RhCMV in vivo, consistent with a central role for rhcmvIL-10 during acute virus-host interactions. Since cmvIL-10 and rhcmvIL-10 are extremely divergent from the cIL-10 of their respective hosts, vaccine-mediated neutralization of their function could inhibit establishment of viral persistence without inhibition of cIL-10. As a prelude to evaluating cmvIL-10-based vaccines in humans, the rhesus macaque model of HCMV was used to interrogate peripheral and mucosal immune responses to rhcmvIL-10 in RhCMV-infected animals. ELISA were used to detect rhcmvIL-10-binding antibodies in plasma and saliva, and an IL-12-based bioassay was used to quantify plasma antibodies that neutralized rhcmvIL-10 function. rhcmvIL-10 is highly immunogenic during RhCMV infection, stimulating high avidity rhcmvIL-10-binding antibodies in the plasma of all infected animals. Most infected animals also exhibited plasma antibodies that partially neutralized rhcmvIL-10 function but did not cross-neutralize the function of rhesus cIL-10. Notably, minimally detectable rhcmvIL-10-binding antibodies were detected in saliva. This study demonstrates that rhcmvIL-10, as a surrogate for cmvIL-10, is a viable vaccine candidate because (1) it is highly immunogenic during natural RhCMV infection, and (2) neutralizing antibodies to rhcmvIL-10 do not cross-react with rhesus cIL-10. Exceedingly low rhcmvIL-10 antibodies in saliva further suggest that the oral mucosa, which is critical in RhCMV natural history, is associated with suboptimal anti-rhcmvIL-10 antibody responses.


Flavonoids, cognition, and dementia: actions, mechanisms, and potential therapeutic utility for Alzheimer disease.
There is increasing evidence that the consumption of flavonoid-rich foods can beneficially influence normal cognitive function. In addition, a growing number of flavonoids have been shown to inhibit the development of Alzheimer disease (AD)-like pathology and to reverse deficits in cognition in rodent models, suggestive of potential therapeutic utility in dementia. The actions of flavonoid-rich foods (e.g., green tea, blueberry, and cocoa) seem to be mediated by the direct interactions of absorbed flavonoids and their metabolites with a number of cellular and molecular targets. For example, their specific interactions within the ERK and PI3-kinase/Akt signaling pathways, at the level of receptors or kinases, have been shown to increase the expression of neuroprotective and neuromodulatory proteins and increase the number of, and strength of, connections between neurons. Concurrently, their effects on the vascular system may also lead to enhancements in cognitive performance through increased brain blood flow and an ability to initiate neurogenesis in the hippocampus. Additional mechanisms have been suggested for the ability of flavonoids to delay the initiation of and/or slow the progression of AD-like pathology and related neurodegenerative disorders, including a potential to inhibit neuronal apoptosis triggered by neurotoxic species (e.g., oxidative stress and neuroinflammation) or disrupt amyloid β aggregation and effects on amyloid precursor protein processing through the inhibition of β-secretase (BACE-1) and/or activation of α-secretase (ADAM10). Together, these processes act to maintain the number and quality of synaptic connections in key brain regions and thus flavonoids have the potential to prevent the progression of neurodegenerative pathologies and to promote cognitive performance.


A diffusion theory model of spatially resolved fluorescence from depth-dependent fluorophore concentrations.
A photon diffusion model has been developed to calculate the steady-state spatially resolved fluorescence from pencil beam excitation in layered tissue. The model allows the calculation of both the excitation reflectance and the fluorescence escape for an arbitrary continuous depth distribution of tissue optical properties and fluorophore concentration. The validity of this model was verified by comparison with Monte Carlo simulations and experimental measurements using phantoms with tissue-like optical properties. The potential usefulness of the spatially resolved fluorescence was explored using the model and simulations of realistic drug distributions. It was shown that using this technique it may be possible to quantify the diffusion of a topically administered drug into the skin, or the photobleaching of a sensitizer during photodynamic therapy.


The sources of atmospheric black carbon at a European gateway to the Arctic.
Black carbon (BC) aerosols from incomplete combustion of biomass and fossil fuel contribute to Arctic climate warming. Models-seeking to advise mitigation policy-are challenged in reproducing observations of seasonally varying BC concentrations in the Arctic air. Here we compare year-round observations of BC and its δ(13)C/Δ(14)C-diagnosed sources in Arctic Scandinavia, with tailored simulations from an atmospheric transport model. The model predictions for this European gateway to the Arctic are greatly improved when the emission inventory of anthropogenic sources is amended by satellite-derived estimates of BC emissions from fires. Both BC concentrations (R(2)=0.89, P<0.05) and source contributions (R(2)=0.77, P<0.05) are accurately mimicked and linked to predominantly European emissions. This improved model skill allows for more accurate assessment of sources and effects of BC in the Arctic, and a more credible scientific underpinning of policy efforts aimed at efficiently reducing BC emissions reaching the European Arctic.


Combining Extracellular miRNA Determination with Microfluidic 3D Cell Cultures for the Assessment of Nephrotoxicity: a Proof of Concept Study.
Drug-induced kidney injury is often observed in the clinics and can lead to long-term organ failure. In this work, we evaluated a novel in vitro system that aims at detecting whether compounds can cause renal proximal tubule damage in man. For this, we implemented organotypic cultures of human conditionally immortalized proximal tubule epithelial cells overexpressing the organic anion transporter 1 (ciPTEC-OAT1) in a three-channel OrganoPlate under microfluidic conditions. Cells were exposed to four known nephrotoxicants (cisplatin, tenofovir, cyclosporine A, and tobramycin). The effect on cell viability and NAG release into the medium was determined. A novel panel of four miRNAs (mir-21, mir-29a, mir-34a, and mir-192) was selected as potential biomarkers of proximal tubule damage. After nephrotoxicant treatment, miRNA levels in culture medium were earlier indicators than cell viability (WST-8 assay) and outperformed NAG for proximal tubule damage. In particular, mir-29a, mir-34a, and mir-192 were highly reproducible between experiments and across compounds, whereas mir-21 showed more variability. Moreover, similar data were obtained in two different laboratories, underlining the reproducibility and technical transferability of the results, a key requirement for the implementation of novel biomarkers. In conclusion, the selected miRNAs behaved like sensitive biomarkers of damage to tubular epithelial cells caused by several nephrotoxicity mechanisms. This biomarker panel, in combination with the 3D cultures of ciPTEC-OAT1 in the OrganoPlate, represents a novel tool for in vitro nephrotoxicity detection. These results pave the way for the application of miRNAs in longitudinal, time-course in vitro toxicity studies.


Virulence, cultivating conditions, and phylogenetic analyses of oomycete parasites in Daphnia.
We describe the infectivity, virulence, cultivating conditions, and phylogenetic positions of naturally occurring oomycete parasites of Daphnia, invertebrates which play a major role in aquatic food webs. Daphnia pulex individuals were found dead and covered by oomycete mycelia when exposed to pond sediments. We were able to extract 4 oomycete isolates from dead Daphnia and successfully cultivate them. Using the ITS and LSU rDNA sequences, we further showed these isolates to be distinct species. The isolates were experimentally demonstrated to be parasitic and not saprobic. After exposure to the parasites, Daphnia mortality was much higher than that reported for Daphnia infected with other known parasite species. Therefore, it is likely that oomycete parasites are important selective pressures in natural Daphnia populations. Moreover, their close phylogenetic relationship to parasites of fish and algae suggests that the stability of aquatic food webs (i.e. fish-Daphnia-algae) might be influenced by the shared parasite communities.


Multimode interference demultiplexers and splitters in metal-insulator-metal waveguides.
Multimode interference (MMI) effect in metal-insulator-metal (MIM) waveguides is studied in this paper. Theoretical calculation based on the self-imaging principle and the finite element method (FEM) simulation are used to analyze the effect and further guide the design of plasmonic MMI devices. We show that wavelength-selective routing and splitting with high extinction ratios can be realized in the multimode waveguides, and active control by refractive index modulation is also demonstrated.


Effect of subspecialty training and volume on outcome after pediatric inguinal hernia repair.
Inguinal hernia repair is the most common operation performed in children. The aim of this study was to determine if there are any differences in outcome when this procedure is performed by subspecialist pediatric surgeons when compared with general surgeons. All pediatric inguinal hernias repaired in the province of Ontario between 1993 and 2000 were reviewed using a population-based database. Children with complex medical conditions or prematurity were excluded. Cases done by general surgeons were compared with those done by pediatric surgeons. The chi2 test was used for nominal data and the Student's t test was used for continuous variables. Probabilities were calculated based on a logistic regression model. Of 20,545 eligible hernia repairs, 50.3% were performed by pediatric surgeons and 49.7% were performed by general surgeons. Pediatric surgeons operated on 62.4% of children younger than 2 years, 51.8% of children aged 26 years, and 37% of children older than 7 years. Duration of operation, length of hospital stay, and incidence of early postoperative complications were similar among pediatric and general surgeons. The rate of recurrent inguinal hernia was higher in the general surgeon group compared with pediatric surgeons (1.10% vs 0.45%, P < .001). Among pediatric surgeons, the estimated risk of hernia recurrence was independent of surgical volume. There was a significant inverse correlation between surgeon volume and recurrence risk among general surgeons, with the highest volume general surgeons achieving recurrence rates similar to pediatric surgeons. Pediatric surgeons have a lower rate of recurrence after inguinal hernia repair in children. General surgeons with high volumes have similar outcomes to pediatric surgeons.


Promotion of osteogenic differentiation by amnion/chorion membrane extracts.
The amniotic membrane is a favorable biomaterial to apply in the field of tissue engineering because of its unique biological properties. Human amniotic membranes consist of 2-layered sheets containing numerous growth factors, cytokines and other bioactive substances. In this study, we explored the potential of amnion membrane extracts (AME) and amnion/chorion membrane extracts (A/CME) to promote osteogenic differentiation of osteoblast-like (MG-63) cells. MG-63 cells were cultured in osteogenic induction medium (OIM) with or without 100 µg/mL of AME or A/CME. To determine the early and late differentiation of osteogenesis, alkaline phosphatase (ALP) activity and calcium deposition were measured at 3, 7, 10 and 24 days. Expression of specific genes associated with osteogenic differentiation, including osteocalcin (OCN), osteopontin (OPN), runt domain-containing transcription factor (Runx2) and osterix (OSX) was also determined. In vitro experiments demonstrated that A/CME increased ALP activity, osteogenic gene expression and mineralization under osteogenic-inducing conditions. Notably, we found that A/CME contained growth factors related to osteogenesis, including fibroblast growth factors and transforming growth factors, which potentially promoted osteogenic differentiation of MG-63 cells to a greater extent than AME. These results indicate that A/CME is capable of providing growth factors and other substrates for osteogenic differentiation, which significantly increased the efficacy of osteogenesis in MG-63 cells. Taken together, the results of this study suggest that human A/CME is a promising biomaterial with therapeutic potential in bone regeneration applications.


Accelerometer Quantification of Physical Activity and Activity Patterns in Patients with Ankylosing Spondylitis and Population Controls.
To compare the total amount of physical activity (TPA) and time spent in various activity intensities of patients with ankylosing spondylitis (AS) and population controls, and to explore factors related to physical activity (PA). Subjects were asked to wear a triaxial accelerometer for 7 days and to complete a series of questionnaires. Multivariable regressions were used to assess generic determinants of TPA in patients and controls, and in patients to explore demographic and disease-specific determinants of various PA intensities. One hundred and thirty-five patients [51 ± 13 yrs, 60% men, body mass index (BMI) 26.0 ± 4.3 kg/m(2)] and 99 controls (45 ± 12 yrs, 67% men, BMI 25.1 ± 4.3 kg/m(2)) were included. Patients did not differ from controls regarding TPA (589 vs 608 vector count/min, p = 0.98), minutes/day spent in sedentary (524 vs 541, p = 0.17), and light PA (290 vs 290 p = 0.95), but spent fewer minutes/day in moderate to vigorous PA (MVPA; 23 vs 30 min/day, p = 0.006). Perceived functional ability (physical component summary of the Medical Outcomes Study Short Form-36) and BMI were associated with TPA independent of having AS (p interaction = 0.21 and 0.94, respectively). Additional analyses in patients showed that time spent in MVPA was negatively influenced by BMI, physical function (Bath AS Functional Index), and disease duration. In patients ≥ 52 years old, a higher Bath AS Disease Activity Index was associated with less time spent in sedentary and more time spent in light activities. Compared with controls, patients with AS had similar TPA, but may avoid engagement in higher intensities of PA. Lower levels of functional ability and higher BMI were associated with lower TPA in both patients and controls.


Diacetatodi-tert-butyltin(IV).
The title compound, [Sn(C4H9)2(CH3COO)2], was synthesized in order to study the influence of large organic groups on the mol-ecular structure of diorganotin di-acetates. The title compound exhibits the same structure type as other diorganotin(IV) di-acetates characterized by an unsymmetrical bidentate bonding mode of the two acetate groups to tin. The influence of the t-butyl groups on this mol-ecular structure is expressed in two significant differences: tin-carbon bond lengths are much more longer than in the other di-acetates, as are the additional inter-actions of the acetate groups with the tin atom. Inter-molecular inter-actions are restricted to C-H⋯O ones similar to those in the other di-acetates, giving rise to a chain-like arrangement of the molecules with the tin atoms and acetate groups in the propagation plane.


Health consequences of shift work and insufficient sleep.
This review summarises the literature on shift work and its relation to insufficient sleep, chronic diseases, and accidents. It is based on 38 meta-analyses and 24 systematic reviews, with additional narrative reviews and articles used for outlining possible mechanisms by which shift work may cause accidents and adverse health. Evidence shows that the effect of shift work on sleep mainly concerns acute sleep loss in connection with night shifts and early morning shifts. A link also exists between shift work and accidents, type 2 diabetes (relative risk range 1.09-1.40), weight gain, coronary heart disease (relative risk 1.23), stroke (relative risk 1.05), and cancer (relative risk range 1.01-1.32), although the original studies showed mixed results. The relations of shift work to cardiometabolic diseases and accidents mimic those with insufficient sleep. Laboratory studies indicate that cardiometabolic stress and cognitive impairments are increased by shift work, as well as by sleep loss. Given that the health and safety consequences of shift work and insufficient sleep are very similar, they are likely to share common mechanisms. However, additional research is needed to determine whether insufficient sleep is a causal pathway for the adverse health effects associated with shift work.


Injectable shear-thinning hydrogels for delivering osteogenic and angiogenic cells and growth factors.
Bone nonunion may occur when the fracture is unstable, or blood supply is impeded. To provide an effective treatment for the healing of nonunion defects, we introduce an injectable osteogenic hydrogel that can deliver cells and vasculogenic growth factors. We used a silicate-based shear-thinning hydrogel (STH) to engineer an injectable scaffold and incorporated polycaprolactone (PCL) nanoparticles that entrap and release vasculogenic growth factors in a controlled manner. By adjusting the solid composition of gelatin and silicate nanoplatelets in the STH, we defined optimal conditions that enable injection of STHs, which can deliver cells and growth factors. Different types of STHs could be simultaneously injected into 3D constructs through a single extrusion head composed of multiple syringes and needles, while maintaining their engineered structure in a continuous manner. The injected STHs were also capable of filling any irregularly shaped defects in bone. Osteogenic cells and endothelial cells were encapsulated in STHs with and without vasculogenic growth factors, respectively, and when co-cultured, their growth and differentiation were significantly enhanced compared to cells grown in monoculture. This study introduces an initial step of developing a new platform of shape-tunable materials with controlled release of angiogenic growth factors by utilizing PCL nanoparticles.


Resting heart rate and all-cause and cardiovascular mortality in the general population: a meta-analysis.
Data on resting heart rate and risk of all-cause and cardiovascular mortality are inconsistent; the magnitude of associations between resting heart rate and risk of all-cause and cardiovascular mortality varies across studies. We performed a meta-analysis of prospective cohort studies to quantitatively evaluate the associations in the general population. We searched PubMed, Embase and MEDLINE from inception to Jan. 1, 2015. We used a random-effects model to combine study-specific relative risks and 95% confidence intervals (CIs). We used restricted cubic spline functions to assess the dose-response relation. A total of 46 studies were included in the meta-analysis, involving 1 246 203 patients and 78 349 deaths for all-cause mortality, and 848 320 patients and 25 800 deaths for cardiovascular mortality. The relative risk with 10 beats/min increment of resting heart rate was 1.09 (95% CI 1.07-1.12) for all-cause mortality and 1.08 (95% CI 1.06-1.10) for cardiovascular mortality. Compared with the lowest category, patients with a resting heart rate of 60-80 beats/min had a relative risk of 1.12 (95% CI 1.07-1.17) for all-cause mortality and 1.08 (95% CI 0.99-1.17) for cardiovascular mortality, and those with a resting heart rate of greater than 80 beats/min had a relative risk of 1.45 (95% CI 1.34-1.57) for all-cause mortality and 1.33 (95% CI 1.19-1.47) for cardiovascular mortality. Overall, the results did not differ after adjustment for traditional risk factors for cardiovascular disease. Compared with 45 beats/min, the risk of all-cause mortality increased significantly with increasing resting heart rate in a linear relation, but a significantly increased risk of cardiovascular mortality was observed at 90 beats/min. Substantial heterogeneity and publication bias were detected. Higher resting heart rate was independently associated with increased risks of all-cause and cardiovascular mortality. This indicates that resting heart rate is a predictor of all-cause and cardiovascular mortality in the general population.


Hierarchical Bayesian modelling of gene expression time series across irregularly sampled replicates and clusters.
Time course data from microarrays and high-throughput sequencing experiments require simple, computationally efficient and powerful statistical models to extract meaningful biological signal, and for tasks such as data fusion and clustering. Existing methodologies fail to capture either the temporal or replicated nature of the experiments, and often impose constraints on the data collection process, such as regularly spaced samples, or similar sampling schema across replications. We propose hierarchical Gaussian processes as a general model of gene expression time-series, with application to a variety of problems. In particular, we illustrate the method's capacity for missing data imputation, data fusion and clustering.The method can impute data which is missing both systematically and at random: in a hold-out test on real data, performance is significantly better than commonly used imputation methods. The method's ability to model inter- and intra-cluster variance leads to more biologically meaningful clusters. The approach removes the necessity for evenly spaced samples, an advantage illustrated on a developmental Drosophila dataset with irregular replications. The hierarchical Gaussian process model provides an excellent statistical basis for several gene-expression time-series tasks. It has only a few additional parameters over a regular GP, has negligible additional complexity, is easily implemented and can be integrated into several existing algorithms. Our experiments were implemented in python, and are available from the authors' website: http://staffwww.dcs.shef.ac.uk/people/J.Hensman/.


Quantitative clinical pharmacology practice for optimal use of antibiotics during the neonatal period.
For safe and effective neonatal antibiotic therapy, knowledge of the pharmacokinetic parameters of antibacterial agents in neonates is a prerequisite. Fast maturational changes during the neonatal period influence pharmacokinetic and pharmacodynamic parameters and their variability. Consequently, the need for applying quantitative clinical pharmacology and determining optimal drug dosing regimens in neonates has become increasingly recognized. Modern quantitative approaches, such as pharmacometrics, are increasingly utilized to characterize, understand and predict the pharmacokinetics of a drug and its effect, and to quantify the variability in the neonatal population. Individual factors, called covariates in modeling, are integrated in such approaches to explain inter-individual pharmacokinetic variability. Pharmacometrics has been shown to be a relevant tool to evaluate, optimize and individualize drug dosing regimens. Challenges for optimal use of antibiotics in neonates can largely be overcome with quantitative clinical pharmacology practice. Clinicians should be aware that there is a next step to support the clinical decision-making based on clinical characteristics and therapeutic drug monitoring, through Bayesian-based modeling and simulation methods. Pharmacometric modeling and simulation approaches permit us to characterize population average, inter-subject and intra-subject variability of pharmacokinetic parameters such as clearance and volume of distribution, and to identify and quantify key factors that influence the pharmacokinetic behavior of antibiotics during the neonatal period.


Completion of Kunjin virus RNA sequence and recovery of an infectious RNA transcribed from stably cloned full-length cDNA.
Completion of the Kunjin virus (KUN) RNA sequence showed that it is the longest flavivirus sequence reported (11,022 bases), commencing with a 5' noncoding region of 96 bases. The 3' noncoding sequence of 624 nucleotides included a unique insertion sequence of 46 bases adjacent to the stop codon, but otherwise it had properties similar to those of RNAs of closely related flaviviruses. A full-length KUN cDNA clone which could be stably propagated in Escherichia coli DH5 alpha was constructed; SP6 polymerase RNA transcripts from amplified cDNA were infectious when transfected into BHK-21 cells. A mutational change abolishing the BamHI restriction site at position 4049, leading to a conservative amino acid change of Arg-175 to Lys in the NS2A protein, was introduced into the cDNA during construction and was retained in the recovered virus. Extra terminal nucleotides introduced during cloning of the cDNA were shown to be present in the in vitro RNA transcripts but absent in the RNA of recovered virus. Although recovered virus differed from the parental KUN by a smaller plaque phenotype and delayed growth rate in BHK-21 cells and mice, it was very similar as assessed by several other criteria, such as peak titer during growth in cells, infectivity titer in cells and in mice, rate of adsorption and penetration in cells, replication at 39 degrees C, and neurovirulence after intraperitoneal injection in mice. The KUN stably cloned cDNA will provide a useful basis for future studies in defining and characterizing functional roles of all the gene products.


Outcome following thiopurine use in children with ulcerative colitis: a prospective multicenter registry study.
Despite little supporting data, thiopurine use is common in pediatric ulcerative colitis (UC). Our aim was to determine outcome following thiopurine use in a multicenter inception cohort of children diagnosed with UC. Data were obtained from a prospective observational study of newly diagnosed children <16 years of age. Data are recorded at diagnosis, 30 days, and quarterly. Patients are managed by physician dictates not protocol. Disease activity is classified by physician global assessment. The primary outcome was corticosteroid (CS)-free inactive UC at 1 year following thiopurine initiation without the need for rescue therapy (infliximab, calcineurin inhibitors, or colectomy). Of 1,490 patients in our registry, 394 have UC (mean age at diagnosis 11.3±3.7 years); 197 (50%) received thiopurine (49% ≤3 months from diagnosis). Also, 84% were receiving CSs and 60% 5-aminosalicylates at thiopurine start. Of the 197 patients, there was insufficient follow-up (41), previous or concomitant use of infliximab (16), or calcineurin inhibitor (7), leaving 133 patients evaluable at 1 year. Of these, 65 (49%) had CS-free inactive UC without rescue therapy. CS-free inactive disease at 1 year after initiating thiopurine was not affected by starting thiopurine ≤3 months vs. >3 months from diagnosis, gender, age, or concomitant treatment with 5-aminosalicylates. Kaplan-Meier analysis showed that the likelihood of remaining free of rescue therapy in the thiopurine-treated patients was 73% at 1 year. Approximately 50% of children with UC starting thiopurine without previous or concomitant biologic or calcineurin inhibitor therapy have CS-free inactive disease 1 year later without the need for rescue therapy.


Medial habenula cholinergic signaling regulates cocaine-associated relapse-like behavior.
Propensity to relapse, even following long periods of abstinence, is a key feature in substance use disorders. Relapse and relapse-like behaviors are known to be induced, in part, by re-exposure to drug-associated cues. Yet, while many critical nodes in the neural circuitry contributing to relapse have been identified and studied, a full description of the networks driving reinstatement of drug-seeking behaviors is lacking. One area that may provide further insight to the mechanisms of relapse is the habenula complex, an epithalamic region composed of lateral and medial (MHb) substructures, each with unique cell and target populations. Although well conserved across vertebrate species, the functions of the MHb are not well understood. Recent research has demonstrated that the MHb regulates nicotine aversion and withdrawal. However, it remains undetermined whether MHb function is limited to nicotine and aversive stimuli or if MHb circuit regulates responses to other drugs of abuse. Advances in circuit-level manipulations now allow for cell-type and temporally specific manipulations during behavior, specifically in spatially restrictive brain regions, such as the MHb. In this study, we focus on the response of the MHb to reinstatement of cocaine-associated behavior, demonstrating that cocaine-primed reinstatement of conditioned place preference engages habenula circuitry. Using chemogenetics, we demonstrate that MHb activity is sufficient to induce reinstatement behavior. Together, these data identify the MHb as a key hub in the circuitry underlying reinstatement and may serve as a target for regulating relapse-like behaviors.


Inferred global connectivity of whale shark Rhincodon typus populations.
Ten years have passed since the last synopsis of whale shark Rhincodon typus biogeography. While a recent review of the species' biology and ecology summarized the vast data collected since then, it is clear that information on population geographic connectivity, migration and demography of R. typus is still limited and scattered. Understanding R. typus migratory behaviour is central to its conservation management considering the genetic evidence suggesting local aggregations are connected at the generational scale over entire ocean basins. By collating available data on sightings, tracked movements and distribution information, this review provides evidence for the hypothesis of broad-scale connectivity among populations, and generates a model describing how the world's R. typus are part of a single, global meta-population. Rhincodon typus occurrence timings and distribution patterns make possible a connection between several aggregation sites in the Indian Ocean. The present conceptual model and validating data lend support to the hypothesis that R. typus are able to move among the three largest ocean basins with a minimum total travelling time of around 2-4 years. The model provides a worldwide perspective of possible R. typus migration routes, and suggests a modified focus for additional research to test its predictions. The framework can be used to trim the hypotheses for R. typus movements and aggregation timings, thereby isolating possible mating and breeding areas that are currently unknown. This will assist endeavours to predict the longer-term response of the species to ocean warming and changing patterns of human-induced mortality.


Vitamin A and Bone Fractures: Systematic Review and Meta-Analysis.
Vitamin A is a generic term for compounds that have biological activity similar to that of retinol and includes carotenoids like β-carotene and α-carotene. Some studies suggest high dietary intake of vitamin A can increase bone fracture risk. This investigation involved a systematic review and meta-analysis examining the association between vitamin A and fracture risk. Published literature was searched to find studies that (1) involved human participants, (2) had prospective cohort or case-control study designs, (3) contained original quantitative data on associations between dietary intake of vitamin A and fractures, and (4) provided either risk ratios (RRs), odds ratios (ORs), or hazard ratios (HRs) with 95% confidence intervals (95% CIs) comparing various levels of vitamin A consumption to fracture risk. Thirteen studies met the review criteria. Meta-analyses indicated that risk of hip fracture was increased by high dietary intake of total vitamin A (RR = 1.29; 95% CI = 1.07-1.57) or retinol (RR = 1.23; 95% CI = 1.02-1.48). Hip fracture risk was reduced by high dietary intake of total carotene (RR = 0.62; 95% CI = 0.42-0.93), β-carotene (RR = 0.72; 95% CI = 0.58-0.89), or α-carotene (RR = 0.81; 95% CI = 0.67-0.97). Total fracture risk was not associated with any vitamin A compound. High intake of total vitamin A or retinol increased hip fracture risk, while high intake of some carotenoids reduced hip fracture risk.


Relevance of individual characteristics for human heat stress response is dependent on exercise intensity and climate type.
Multiple heterogeneous groups of subjects (both sexes and a wide range of maximal oxygen uptake VO2max, body mass, body surface area (AD),% body fat, and AD/mass coefficient) exercised on a cycle ergometer at a relative (%VO2max, REL) or an absolute (60 W) exercise intensity in a cool (CO 21 degrees C, 50% relative humidity), warm humid (WH 35 degrees C, 80%) and a hot dry (HD 45 degrees C, 20%) environment. Rectal temperature (Tre) responses were analysed for the influence of the individual's characteristics, environment and exercise intensity. Exposures consisted of 30-min rest, followed by 60-min exercise. The Tre was negatively correlated with mass in all conditions. Body mass acted as a passive heat sink in all the conditions tested. While negatively correlated with VO2max and VO2max per kilogram body mass in most climates, Tre was positively correlated with VO2max and VO2max per kilogram body mass in the WH/REL condition. Thus, when evaporative heat loss was limited as in WH, the higher heat production of the fitter subjects in the REL trials determined Tre and not the greater efficiency for heat loss associated with high VO2max. Body fatness significantly affected Tre only in the CO condition, where, with low skin blood flows (measured as increases in forearm blood flow), the insulative effect of fat was pronounced. In the warmer environments, high skin blood flows offset the resistance offered by peripheral adipose tissue. Contrary to other studies, Tre was positively correlated with AD/mass coefficient for all conditions tested. For both exercise types used, being big (a high heat loss area and heat capacity) was apparently more beneficial from a heat strain standpoint than having a favourable AD/mass coefficient (high in small subjects). The total amount of variance in Tre responses which could be attributed to individual characteristics was dependent on the climate and the type of exercise. Though substantial for absolute exercise intensities (52%-58%) the variance explained in Tre differed markedly for relative intensities: 72% for the WH climate with its limited evaporative capacity, and only 10%-26% for the HD and CO climates. The results showed that individual characteristics play a significant role in determining the responses of body core temperature in all conditions tested, but their contribution was low for relative exercise intensities when evaporative heat loss was not restricted. This study demonstrated that effects of individual characteristics on human responses to heat stress cannot be interpreted without taking into consideration both the heat transfer properties of the environment and the metabolic heat production resulting from the exercise type and intensity chosen. Their impact varies substantially among conditions.


CETP genotype and changes in lipid levels in response to weight-loss diet intervention in the POUNDS LOST and DIRECT randomized trials.
Little is known about whether cholesteryl ester transfer protein (CETP) genetic variation may modify the effect of weight-loss diets varying in fat content on changes in lipid levels. We analyzed the interaction between the CETP variant rs3764261 and dietary interventions on changes in lipid levels among 732 overweight/obese adults from a 2 year randomized weight-loss trial [Preventing Overweight Using Novel Dietary Strategies (POUNDS LOST)], and replicated the findings in 171 overweight/obese adults from an independent 2 year weight-loss trial [Dietary Intervention Randomized Controlled Trial (DIRECT)]. In the POUNDS LOST, participants with the CETP rs3764261 CC genotype on the high-fat diet had larger increases in HDL cholesterol (P = 0.001) and decreases in triglycerides (P = 0.007) than those on the low-fat diet at 6 months, while no significant difference between these two diets was observed among participants carrying other genotypes. The gene-diet interactions on changes in HDL-cholesterol and tri-glyc-erides were replicated in the DIRECT (pooled P for interaction ≤ 0.01). Similar results on trajectory of changes in HDL cholesterol and triglycerides over the 2 year intervention were observed in both trials. Our study provides replicable evidence that individuals with the CETP rs3764261 CC genotype might derive greater effects on raising HDL cholesterol and lowering triglycerides by choosing a low-carbohydrate/high-fat weight-loss diet instead of a low-fat diet.


Usefulness of Cytological Scoring Method by Breast Fine Needle Aspiration Cytology on Breast Duct Dilatation and Cystic Lesions.
The objective of this study was to apply a scoring method to fine needle aspiration cytology on breast duct dilatation and cystic lesions, to set an optimum cut-off value to differentiate between benign and malignant cases, and to identify features useful for cell judgment. Samples were 23 preparations of specimens (12 benign and 11 malignant cases) suspected with intraductal lesions or cystic change by ultrasonography or mammography and cytology. The scoring system comprised the following 10 items, and each item was scored 1-3, with a total score of 10-30. Three items were concerning structural atypia: 1, scattered epithelial cells; 2, uneven irregular cluster edge; and 3, overlapping nuclei of epithelial cells, and seven items were concerning cellular atypia: 4, irregular nuclear size; 5, irregular nuclear morphology; 6, deep dyeing chromatin; 7, chromatin granularity; 8, chromatin distribution; 9, nucleolus; and 10, absence of myoepithelial cells. (1) Scoring cut-off value: malignancy is to be suspected when the score is 20.75 or higher (diagnostic accuracy: 95.7%). (2) Findings useful for cancer judgment: the sensitivity of the following four findings was high: uneven irregular cluster edge, irregular nuclear overlapping, chromatin granularity, and absence of myoepithelial cells. (3) Correlation among the findings: the findings correlated with malignancy were as follows: scattered epithelial cells versus uneven irregular cluster edge (rs = 0.8). Cytological evaluation by scoring lesions accompanied by intraductal dilatation and cystic change was a useful method capable of differentiating between benign and malignant cases at a high accuracy.


Newborn Screening for Congenital Adrenal Hyperplasia: Review of Factors Affecting Screening Accuracy.
Newborn screening for 21-hydroxylase deficiency (21OHD), the most common form of congenital adrenal hyperplasia, has been performed routinely in the United States and other countries for over 20 years. Screening provides the opportunity for early detection and treatment of patients with 21OHD, preventing salt-wasting crisis during the first weeks of life. However, current first-tier screening methodologies lack specificity, leading to a large number of false positive cases, and adequate sensitivity to detect all cases of classic 21OHD that would benefit from treatment. This review summarizes the pathology of 21OHD and also the key stages of fetal hypothalamic-pituitary-adrenal axis development and adrenal steroidogenesis that contribute to limitations in screening accuracy. Factors leading to both false positive and false negative results are highlighted, along with specimen collection best practices used by laboratories in the United States and worldwide. This comprehensive review provides context and insight into the limitations of newborn screening for 21OHD for laboratorians, primary care physicians, and endocrinologists.


Complementary Medicine, Refusal of Conventional Cancer Therapy, and Survival Among Patients With Curable Cancers.
There is limited information on the association among complementary medicine (CM), adherence to conventional cancer treatment (CCT), and overall survival of patients with cancer who receive CM compared with those who do not receive CM. To compare overall survival between patients with cancer receiving CCT with or without CM and to compare adherence to treatment and characteristics of patients receiving CCT with or without CM. This retrospective observational study used data from the National Cancer Database on 1 901 815 patients from 1500 Commission on Cancer-accredited centers across the United States who were diagnosed with nonmetastatic breast, prostate, lung, or colorectal cancer between January 1, 2004, and December 31, 2013. Patients were matched on age, clinical group stage, Charlson-Deyo comorbidity score, insurance type, race/ethnicity, year of diagnosis, and cancer type. Statistical analysis was conducted from November 8, 2017, to April 9, 2018. Use of CM was defined as "Other-Unproven: Cancer treatments administered by nonmedical personnel" in addition to at least 1 CCT modality, defined as surgery, radiotherapy, chemotherapy, and/or hormone therapy. Overall survival, adherence to treatment, and patient characteristics. The entire cohort comprised 1 901 815 patients with cancer (258 patients in the CM group and 1 901 557 patients in the control group). In the main analyses following matching, 258 patients (199 women and 59 men; mean age, 56 years [interquartile range, 48-64 years]) were in the CM group, and 1032 patients (798 women and 234 men; mean age, 56 years [interquartile range, 48-64 years]) were in the control group. Patients who chose CM did not have a longer delay to initiation of CCT but had higher refusal rates of surgery (7.0% [18 of 258] vs 0.1% [1 of 1031]; P < .001), chemotherapy (34.1% [88 of 258] vs 3.2% [33 of 1032]; P < .001), radiotherapy (53.0% [106 of 200] vs 2.3% [16 of 711]; P < .001), and hormone therapy (33.7% [87 of 258] vs 2.8% [29 of 1032]; P < .001). Use of CM was associated with poorer 5-year overall survival compared with no CM (82.2% [95% CI, 76.0%-87.0%] vs 86.6% [95% CI, 84.0%-88.9%]; P = .001) and was independently associated with greater risk of death (hazard ratio, 2.08; 95% CI, 1.50-2.90) in a multivariate model that did not include treatment delay or refusal. However, there was no significant association between CM and survival once treatment delay or refusal was included in the model (hazard ratio, 1.39; 95% CI, 0.83-2.33). In this study, patients who received CM were more likely to refuse additional CCT, and had a higher risk of death. The results suggest that mortality risk associated with CM was mediated by the refusal of CCT.


Smoking intensity among male factory workers in Kunming, China.
This study investigated the intensity of cigarette consumption and its correlates in China among urban male factory workers, a cohort especially vulnerable to tobacco exposure, one that appears to have benefitted little from recent public health efforts to reduce smoking rates. Data were collected from men working in factories of Kunming city, Yunnan, China, who are current daily smokers (N = 490). A multinomial logistic regression was conducted to examine the factors in association with smoking intensity in light, moderate, and heavy levels. Light smoking correlated with social smoking, smoking the first cigarette later in the day, self-reported health condition, and quit intention. Heavy smoking was associated with purchase of lower priced cigarettes, difficulty refraining from smoking, and prehypertensive blood pressure. Even in regions where smoking is highly prevalent, even among cohorts who smoke heavily, variation exists in how cigarettes are consumed. Analyses of this consumption, with special consideration given to smoking intensity and its correlates, can help guide tobacco-control strategists in developing more effective interventions.


Sensorimotor gating and D2 receptor signalling: evidence from a molecular genetic approach.
Converging evidence from pharmacological investigations, genetic association studies and schizophrenia research indicates an important influence of the dopamine system on sensorimotor gating as measured by prepulse inhibition (PPI) of the acoustic startle response. In particular, D2 receptor agonists have been shown to disrupt PPI in humans and rodents. In the present study, we investigated the associations of two functional DRD2 related single nucleotide polymorphisms (rs4648317 and rs1800497, the latter also known as DRD2/ANKK1 Taq1A) with PPI in two independent healthy human samples (overall n=197; Munich n=101; London n=96). Taq1A is a prominent marker of striatal D2 receptor signalling and was therefore hypothesized to impact on PPI. In line with our hypothesis, we report here reduced PPI levels in individuals with higher striatal D2 receptor signalling as indicated by the Taq1A genotype. Meta-analysis across both samples confirmed this finding. In contrast, an association between rs4648317 and PPI found in the Munich sample could not be confirmed in the London sample. Overall, the present study helps to bridge the gap between pharmacological manipulations of PPI and molecular genetics of the dopaminergic system.


Methadone treatment protects against HIV infection: two decades of experience in the Bronx, New York City.
We undertook a study of the role of methadone maintenance in protecting injecting drug users (IDUs) from human immunodeficiency virus (HIV) infection from the earliest days of the HIV epidemic in New York City to the present. The historical context of the epidemic in the Bronx is discussed. For close to two decades, we have been tracking changes in injecting drug use and HIV infection levels in a Bronx cohort study of IDUs. An initial sample of 622 IDUs was recruited from a methadone treatment program in 1985, with historical data going back to 1978. Behavioral interviews and HIV testing were performed and methadone treatment program records (urine toxicology and methadone dose history) were reviewed. We examined both prevalent and incident HIV infections. The sample included African Americans (24.3%), Latinos (50.3%), and white non-Latinos (24.4%). The average methadone dose was 64 milligrams (mg) per day with an average time in treatment of five and a half years. We found a very low rate of incident infection of 1.7 per 100 person-years observation since 1986. Because of this low rate of infection, we were unable to determine the association between methadone treatment factors and HIV seroincidence. We found that our prevalence data on the 622 IDUs enrolled from 1985 to 1988 yielded strong findings on the role of methadone maintenance in a period when most infections occurred in this population. HIV seroprevalence was 42.9%. Logistic regression analysis revealed associations of methadone dose > or = 80 mg (adjusted odds ratio = 3.07/yr, 95% confidence interval (CI): 1.23-7.68) and last year entered methadone treatment (adjusted odds ratio = 1.22/yr, 95% CI: 1.06-1.41) to HIV infection, independent of year of last cocaine injection, needle sharing in shooting galleries, number of IDU sex partners, low income, and African American of Latino ethnicity. Properly dosed, long-term methadone treatment was found to be a central protective factor in preventing HIV infection from the earliest days of the epidemic in New York City. It is crucial to have high quality drug treatment programs in place before an epidemic draws our attention to the inadequacies through excess and unnecessary morbidity and mortality.


The light enhanced removal of Bisphenol A from wastewater using cotton waste derived carbon microtubes.
The development of high performance, sustainable and inexpensive catalyst for environmental applications is a highly innovative and promising approach to meet the increasing demands from society on water treatment and pollution remediation. Carbon microtube (CMT) synthesized from cotton waste was successfully developed by direct pyrolysis of cotton bundle in argon atmosphere in different carbonization temperature (900, 1100, 1300 and 1500 °C). Carbon microtubes have been used for removal of Bisphenol A (BPA) in wastewater and showed the optimum performance for CMT11 and CMT 13. The mechanism involved in this efficient water treatment was ascribed to the strong π-π interaction and hydrogen bonds between CMT and BPA. Given the repeatability, high removal performance and cost effectiveness of the cotton based carbon microtubes when compared to other well-known catalysts such as carbon nanotubes, the carbon microtubes demonstrated great potential as low-cost, sustainable and effective catalyst for wastewater treatment.


Signaling of high mobility group box 1 (HMGB1) through toll-like receptor 4 in macrophages requires CD14.
High mobility group box 1 (HMGB1) is a DNA-binding protein that possesses cytokinelike, proinflammatory properties when released extracellularly in the C23-C45 disulfide form. HMGB1 also plays a key role as a mediator of acute and chronic inflammation in models of sterile injury. Although HMGB1 interacts with multiple pattern recognition receptors (PRRs), many of its effects in injury models occur through an interaction with toll-like receptor 4 (TLR4). HMGB1 interacts directly with the TLR4/myeloid differentiation protein 2 (MD2) complex, although the nature of this interaction remains unclear. We demonstrate that optimal HMGB1-dependent TLR4 activation in vitro requires the coreceptor CD14. TLR4 and MD2 are recruited into CD14-containing lipid rafts of RAW264.7 macrophages after stimulation with HMGB1, and TLR4 interacts closely with the lipid raft protein GM1. Furthermore, we show that HMGB1 stimulates tumor necrosis factor (TNF)-α release in WT but not in TLR4(-/-), CD14(-/-), TIR domain-containing adapter-inducing interferon-β (TRIF)(-/-) or myeloid differentiation primary response protein 88 (MyD88)(-/-) macrophages. HMGB1 induces the release of monocyte chemotactic protein 1 (MCP-1), interferon gamma-induced protein 10 (IP-10) and macrophage inflammatory protein 1α (MIP-1α) in a TLR4- and CD14-dependent manner. Thus, efficient recognition of HMGB1 by the TLR4/MD2 complex requires CD14.


Bacteriophage Tailspikes and Bacterial O-Antigens as a Model System to Study Weak-Affinity Protein-Polysaccharide Interactions.
Understanding interactions of bacterial surface polysaccharides with receptor protein scaffolds is important for the development of antibiotic therapies. The corresponding protein recognition domains frequently form low-affinity complexes with polysaccharides that are difficult to address with experimental techniques due to the conformational flexibility of the polysaccharide. In this work, we studied the tailspike protein (TSP) of the bacteriophage Sf6. Sf6TSP binds and hydrolyzes the high-rhamnose, serotype Y O-antigen polysaccharide of the Gram-negative bacterium Shigella flexneri (S. flexneri) as a first step of bacteriophage infection. Spectroscopic analyses and enzymatic cleavage assays confirmed that Sf6TSP binds long stretches of this polysaccharide. Crystal structure analysis and saturation transfer difference (STD) NMR spectroscopy using an enhanced method to interpret the data permitted the detailed description of affinity contributions and flexibility in an Sf6TSP-octasaccharide complex. Dodecasaccharide fragments corresponding to three repeating units of the O-antigen in complex with Sf6TSP were studied computationally by molecular dynamics simulations. They showed that distortion away from the low-energy solution conformation found in the octasaccharide complex is necessary for ligand binding. This is in agreement with a weak-affinity functional polysaccharide-protein contact that facilitates correct placement and thus hydrolysis of the polysaccharide close to the catalytic residues. Our simulations stress that the flexibility of glycan epitopes together with a small number of specific protein contacts provide the driving force for Sf6TSP-polysaccharide complex formation in an overall weak-affinity interaction system.


[Management of postoperative chyle leak after surgery for digestive malignancies].
To investigate the treatment of postoperative chyle leak after surgery for digestive malignancies. From December 2008 to February 2012, in the Sun Yat-sen Memorial Hospital of Sun Yat-sen University, clinical data of 19 patients with chyle leak after digestive system cancer surgery were retrospective analyzed. Nineteen cases of chyle leak were all identified between the second and the fourth postoperative day and were all initially managed with conservative treatment including early fasting, parenteral nutrition(PN), 24-hour continuous infusion of somatostatin, and low pressure suction drainage. Eight patients were treated successfully for 6 to 10 days with a significant reduction of the daily drainage volume. Ten patients had enteral nutrition(EN) and their drain tubes were repeatedly washed with 30 ml of compound meglumine diatrizoate injection every day until the drainage volume decreased to 200 ml/day. The time to resolution of chyle leak in these ten patients ranged from 12 to 24 days. One patient had no significant decrease in fluid drainage and developed abdominal distension after one week of conservative treatment. Surgical closure of chyle leak was performed on the 11th postoperative day, abdominal cavity drainage tube was removed on the 4th postoperative day. The patient was discharged home in good condition. Most postoperative chyle leak after surgery for digestive malignancies can be successfully managed with conservative treatment. Somatostatin and the drainage are the main therapeutic approaches. When chyle leak is not resolved with conservative treatment, surgical treatment should be considered to prevent serious complications.


Examination of the Biomark assay as an alternative to Oncotype DX for defining chemotherapy benefit.
Currently the 21-gene recurrence score (RS) assay called Oncotype DX is recommended by the National Comprehensive Cancer Network guideline for defining the benefit of chemotherapy. To overcome the cost disadvantages of the Oncotype DX assay and the turnaround time, a multigene assay was examined to compare the correlation of the RS and the predicted score (PS) of the present study. Paraffin-embedded tissues of 50 cases with early-stage estrogen receptor (ER)-positive breast cancer, who underwent the Oncotype DX test were used. A total of 149 candidate genes with high correlation to the RS were identified, in another project (Lee et al, unpublished data). Reverse transcription-quantitative polymerase chain reaction biomark assays were conducted using the dynamic array integrated fluidic circuit and the correlation analysis was performed with BRB ArrayTools. A predictive model was developed by the coefficient and gene expression, and 41 genes were identified. If the cut-off was ≥18, the predicted model was 18/50 cases, and the RS was 19, indicating that the differential rate of predicted response against RS was 2%. If the cutoff was ≥11, the predicted model was 38/50 cases and the RS was 34, indicating a difference of 8%. Genes common to the Oncotype DX and the Biomark assay include marker of proliferation Ki-67, aurora kinase A, Erb-B2 receptor tyrosine kinase 2, glutathione S-transferase Mu 1, estrogen receptor 1, progesterone receptor, B-cell lymphoma 2, signal peptide CUB domain EGF-like 2 and 5 reference genes. The remaining 28 genes are involved in various pathways and functions. This result indicates that there is a significant correlation between PS and RS scores, although validation of results is required to accurately determine the risk of distant recurrence. The Biomark assay is an easy and inexpensive way to measure mRNA expression. The present study demonstrates the possibility of the Biomark assay as an alternative for defining chemotherapy benefit in individual patients with ER-positive early-stage breast cancer.


Rendering Dynamic Source Motion in Surface Haptics via Wave Focusing.
Emerging surface haptic technologies can display localized haptic feedback anywhere on a touch surface by focusing mechanical waves generated via sparse arrays of actuators. However, rendering complex haptic scenes with such displays is challenging due to the infinite number of physical degrees of freedom intrinsic to such continuum mechanical systems. Here, we present computational focusing methods for rendering dynamic tactile sources. They can be applied to a variety of surface haptic devices and media, including those that exploit flexural waves in thin plates and solid waves in elastic media. We describe an efficient rendering technique based on time-reversal of waves emitted from a moving source, and motion path discretization. We combine these with intensity regularization methods that reduce focusing artifacts, improve power output, and increase dynamic range. We demonstrate the utility of this approach in experiments with a surface display that uses elastic wave focusing to render dynamic sources, achieving millimeter-scale resolution in experiments. Results of a behavioral experiment show that participants could readily feel and interpret rendered source motion, attaining 99% accuracy across a wide range of motion speeds.