Analytical advances in pharmaceutical impurity profiling.
Impurities will be present in all drug substances and drug products, i.e. nothing is 100% pure if one looks in enough depth. The current regulatory guidance on impurities accepts this, and for drug products with a dose of less than 2g/day identification of impurities is set at 0.1% levels and above (ICH Q3B(R2), 2006). For some impurities, this is a simple undertaking as generally available analytical techniques can address the prevailing analytical challenges; whereas, for others this may be much more challenging requiring more sophisticated analytical approaches. The present review provides an insight into current development of analytical techniques to investigate and quantify impurities in drug substances and drug products providing discussion of progress particular within the field of chromatography to ensure separation of and quantification of those related impurities. Further, a section is devoted to the identification of classical impurities, but in addition, inorganic (metal residues) and solid state impurities are also discussed. Risk control strategies for pharmaceutical impurities aligned with several of the ICH guidelines, are also discussed.


iRegulon: from a gene list to a gene regulatory network using large motif and track collections.
Identifying master regulators of biological processes and mapping their downstream gene networks are key challenges in systems biology. We developed a computational method, called iRegulon, to reverse-engineer the transcriptional regulatory network underlying a co-expressed gene set using cis-regulatory sequence analysis. iRegulon implements a genome-wide ranking-and-recovery approach to detect enriched transcription factor motifs and their optimal sets of direct targets. We increase the accuracy of network inference by using very large motif collections of up to ten thousand position weight matrices collected from various species, and linking these to candidate human TFs via a motif2TF procedure. We validate iRegulon on gene sets derived from ENCODE ChIP-seq data with increasing levels of noise, and we compare iRegulon with existing motif discovery methods. Next, we use iRegulon on more challenging types of gene lists, including microRNA target sets, protein-protein interaction networks, and genetic perturbation data. In particular, we over-activate p53 in breast cancer cells, followed by RNA-seq and ChIP-seq, and could identify an extensive up-regulated network controlled directly by p53. Similarly we map a repressive network with no indication of direct p53 regulation but rather an indirect effect via E2F and NFY. Finally, we generalize our computational framework to include regulatory tracks such as ChIP-seq data and show how motif and track discovery can be combined to map functional regulatory interactions among co-expressed genes. iRegulon is available as a Cytoscape plugin from http://iregulon.aertslab.org.


Mitochondrial Alkbh1 localizes to mtRNA granules and its knockdown induces the mitochondrial UPR in humans and C. elegans.
The Fe(II) and 2-oxoglutarate-dependent oxygenase Alkb homologue 1 (Alkbh1) has been shown to act on a wide range of substrates, like DNA, tRNA and histones. Thereby different enzymatic activities have been identified including, among others, demethylation of N3-methylcytosine (m3C) in RNA- and single-stranded DNA oligonucleotides, demethylation of N1-methyladenosine (m1A) in tRNA or formation of 5-formyl cytosine (f5C) in tRNA. In accordance with the different substrates, Alkbh1 has also been proposed to reside in distinct cellular compartments in human and mouse cells, including the nucleus, cytoplasm and mitochondria. Here, we describe further evidence for a role of human Alkbh1 in regulation of mitochondrial protein biogenesis, including visualizing localization of Alkbh1 into mitochondrial RNA granules with super-resolution 3D SIM microscopy. Electron microscopy and high-resolution respirometry analyses revealed an impact of Alkbh1 level on mitochondrial respiration, but not on mitochondrial structure. Downregulation of Alkbh1 impacts cell growth in HeLa cells and delays development in Caenorhabditis elegans, where the mitochondrial role of Alkbh1 seems to be conserved. Alkbh1 knockdown, but not Alkbh7 knockdown, triggers the mitochondrial unfolded protein response (UPRmt) in C. elegans.


Promises of peace and development: mining and violence in Guatemala.
For Indigenous peoples of Guatemala, mining is experienced within a lingering legacy of colonialism and genocide. Here, we discuss macro-level findings of a larger study, examining the lived context of a mining-affected community in Guatemala and barriers that this poses to peace. Using an anticolonial narrative methodology, guided by participatory action research principles, we interviewed 54 participants. Their accounts pointed to intersecting and ongoing forces of poverty, dispossession, gendered oppression, genocide, and global inequity were exacerbated and triggered by local mining operations. This context posed profound threats to community well-being and signals a call to action for nurses and other global actors.


Pitting temporal against spatial integration in schizophrenic patients.
Schizophrenic patients show strong impairments in visual backward masking possibly caused by deficits on the early stages of visual processing. The underlying aberrant mechanisms are not clearly understood. Spatial as well as temporal processing deficits have been proposed. Here, by combining a spatial with a temporal integration paradigm, we show further evidence that temporal but not spatial processing is impaired in schizophrenic patients. Eleven schizophrenic patients and ten healthy controls were presented with sequences composed of Vernier stimuli. Patients needed significantly longer presentation times for sequentially presented Vernier stimuli to reach a performance level comparable to that of healthy controls (temporal integration deficit). When we added spatial contextual elements to some of the Vernier stimuli, performance changed in a complex but comparable manner in patients and controls (intact spatial integration). Hence, temporal but not spatial processing seems to be deficient in schizophrenia.


Associations of Rsf-1 overexpression with poor therapeutic response and worse survival in patients with nasopharyngeal carcinoma.
Deregulated chromatin remodelling often leads to aberrant gene expression in cells, thereby implicating tumour development and progression. As a subunit of remodelling and spacing factor complex, Rsf-1 (HBXAP), a novel nuclear protein with histone chaperon function, mediates ATPase-dependent chromatin remodelling and confers tumour aggressiveness in common carcinomas. However, the expression of Rsf-1 has never been reported in nasopharyngeal carcinoma (NPC). This study aimed at evaluating the expression status, associations with clincopathological variables and prognostic implications of Rsf-1 in a well-defined cohort of NPC. Rsf-1 immunoexpression was retrospectively assessed in biopsies of 108 consecutive NPC patients without initial distant metastasis and treated with consistent guidelines. The results were correlated with the clinicopathological features, therapeutic response, local recurrence-free survival (LRFS), distant metastasis-free survival (DMFS) and disease-specific survival (DSS). Present in 49 cases (45%), Rsf-1 overexpression was associated with N(2,3) status (p=0.016), American Joint Committee on Cancer stage 3, 4 (p=0.004), and incomplete therapeutic response (p=0.041). In multivariate analyses, Rsf-1 overexpression not only emerged as the sole independent adverse prognosticator for LRFS (p=0.0002, RR 5.287) but also independently predicted worse DMFS (p=0.0011, RR 3.185) and DSS (p<0.0001, RR 4.442), along with T(3,4) (p=0.0454) and N(2,3) (p=0.0319), respectively. Rsf-1 overexpression is common and is associated with adverse prognosticators and therapeutic response, which confers tumour aggressiveness through chromatin remodelling, and represents a potential prognostic biomarker in NPC.


Ribonomic analysis of human Pum1 reveals cis-trans conservation across species despite evolution of diverse mRNA target sets.
PUF family proteins are among the best-characterized regulatory RNA-binding proteins in nonmammalian species, but relatively little is known about mRNA targets or functions of mammalian PUF proteins. In this study, we used ribonomic analysis to identify and analyze mRNAs associated with ribonucleoproteins containing an endogenous human PUF protein, Pum1. Pum1-associated mRNAs were highly enriched for genes encoding proteins that function in transcriptional regulation and cell cycle/proliferation, results consistent with the posttranscriptional RNA regulon model and the proposed ancestral functions of PUF proteins in stem cell biology. Analysis of 3' untranslated region sequences of Pum1-associated mRNAs revealed a core Pum1 consensus sequence, UGUAHAUA. Pum1 knockdown demonstrated that Pum1 enhances decay of associated mRNAs, and relocalization of Pum1 to stress granules suggested that Pum1 functions in repression of translation. This study is the first in vivo genome-wide mRNA target identification of a mammalian PUF protein and provides direct evidence that human PUF proteins regulate stability of associated mRNAs. Comparison of Pum1-associated mRNAs to mRNA targets of PUF proteins from Saccharomyces cerevisiae and Drosophila melanogaster demonstrates how a well-conserved RNA-binding domain and cognate binding sequence have been evolutionarily rewired to regulate the collective expression of different sets of functionally related genes.


MR proton spectroscopy in multiple sclerosis.
To elucidate the natural history of visualized MR abnormalities in patients with multiple sclerosis using proton spectroscopy. MR imaging and proton spectroscopy (1H spectroscopy) were performed on 16 patients with clinically definite multiple sclerosis. All patients received gadopentetate dimeglumine (Gd-DTPA). Decreased levels of N-acetylaspartate (NAA) were demonstrated in 17 out of 21 lesions. No correlation was found between decreased NAA and Gd-DTPA enhancement. In five out of seven enhancing lesions, abnormal 1H spectra with extra peaks (termed marker peaks) at 2.1-2.6 ppm (ranging in absolute concentration from 10-50 mM protons) were observed. In nine out of 14 unenhancing lesions, no elevated marker peaks were observed. In the five other unenhancing lesions, the levels of these marker peaks were generally lower than the enhancing group. No correlation was found between the NAA levels and the levels of the marker peaks. We suggest two distinct biochemical processes: 1) decreased NAA reflecting neuronal cell loss, and 2) elevated marker peaks reflecting ongoing demyelination. Based upon these observations we infer that 1) the majority of enhancing lesions are demyelinating with extra peaks at 2.1-2.6 ppm representing a marker of this process, 2) enhancing lesions without this marker most likely represent edematous regions without significant demyelination, and 3) demyelination may be long in duration compared with transient blood-brain barrier disruption manifested by Gd-DTPA enhancement. Our results suggest that 1H spectroscopy has the ability to further categorize MR-demonstrated enhancing and unenhancing lesions in patients with multiple sclerosis and that it may be more sensitive than contrast enhancement in revealing the true time course of demyelination.


A neurobiological mechanism linking transportation noise to cardiovascular disease in humans.
Chronic noise exposure associates with increased cardiovascular disease (CVD) risk; however, the role of confounders and the underlying mechanism remain incompletely defined. The amygdala, a limbic centre involved in stress perception, participates in the response to noise. Higher amygdalar metabolic activity (AmygA) associates with increased CVD risk through a mechanism involving heightened arterial inflammation (ArtI). Accordingly, in this retrospective study, we tested whether greater noise exposure associates with higher: (i) AmygA, (ii) ArtI, and (iii) risk for major adverse cardiovascular disease events (MACE). Adults (N = 498) without CVD or active cancer underwent clinical 18F-fluorodeoxyglucose positron emission tomography/computed tomography imaging. Amygdalar metabolic activity and ArtI were measured, and MACE within 5 years was adjudicated. Average 24-h transportation noise and potential confounders were estimated at each individual's home address. Over a median 4.06 years, 40 individuals experienced MACE. Higher noise exposure (per 5 dBA increase) predicted MACE [hazard ratio (95% confidence interval, CI) 1.341 (1.147-1.567), P < 0.001] and remained robust to multivariable adjustments. Higher noise exposure associated with increased AmygA [standardized β (95% CI) 0.112 (0.051-0.174), P < 0.001] and ArtI [0.045 (0.001-0.090), P = 0.047]. Mediation analysis suggested that higher noise exposure associates with MACE via a serial mechanism involving heightened AmygA and ArtI that accounts for 12-26% of this relationship. Our findings suggest that noise exposure associates with MACE via a mechanism that begins with increased stress-associated limbic (amygdalar) activity and includes heightened arterial inflammation. This potential neurobiological mechanism linking noise to CVD merits further evaluation in a prospective population.


The impact of a changed legislation on reporting of adverse drug reactions in Sweden, with focus on nurses' reporting.
In March 2007, a legislative amendment was issued in Sweden compelling nurses to report all suspected adverse drug reactions (ADRs) to the national pharmacovigilance system. The aims of this study were to describe the status of ADR reporting, before and after the implementation of the legislative changes, and to describe the general characteristics of suspected ADRs reported by nurses. The Swedish pharmacovigilance system during the study period constituted six regional centres responsible for the handling of all spontaneous ADR reports within their region. In this study, we identified all individual ADR reports from 2005 and 2010, analysed in depth the ADR reports from two regional centres and collated information about the reporter and the nature of the reported ADR. From the two regional centres, a total of 898 and 1074 reports were submitted in 2005 and 2010 respectively. Nurses submitted 31% (275 reports) of the reports in 2005 and 24% (260 reports) in 2010. Nurses' reporting of serious ADRs was 3% (seven reports) in 2005 and 7% (17 reports) in 2010 with reporting of unlabelled ADRs at 4% (11 reports) in 2005 and 17% (45 reports) in 2010. Most of the serious and/or unlabelled reactions were related to vaccine administration (14 reports in 2005 and 36 reports in 2010). The overall ADR reporting by nurses did not appear to increase after the change in reporting legislation. The proportion of serious and/or unlabelled ADRs reported by nurses did however appear to increase during the same period. Taken together, our data suggests that further pro-active measures should be considered in order to involve nurses in the reporting of suspected ADRs.


Self-Monitoring vs. Implementation Intentions: a Comparison of Behaviour Change Techniques to Improve Sleep Hygiene and Sleep Outcomes in Students.
This study seeks to investigate and compare the efficacy of self-monitoring and implementation intentions-two post-intentional behaviour change techniques-for improving sleep hygiene behaviours and sleep outcomes in university students. Seventy-two undergraduate students completed baseline measures of four sleep hygiene behaviours (making the sleep environment restful, avoiding going to bed hungry/thirsty, avoiding stress/anxiety-provoking activities near bed time and avoiding caffeine in the evening), as well as the Pittsburgh sleep quality index (PSQI) and the insomnia severity index (ISI). Participants were randomly assigned to an active-control diary-keeping, self-monitoring condition or completed implementation intentions for each behaviour. Post-intervention measurement was completed 2 weeks after baseline. Repeated measures analyses of variance found significant main effects of time for improvements in making the sleep environment restful and avoiding going to bed hungry or thirsty, as well as PSQI and ISI scores. Non-significant interactions suggested no group differences on any variable, except for increasing avoidance of stress and anxiety-provoking activities before bed time, for which only implementation intentions were found to be effective. Attrition was higher amongst self-monitoring participants. Both self-monitoring and implementation intentions appear to be promising behaviour change techniques for improving sleep hygiene and sleep. Future research should examine the acceptability of the two behaviour change techniques and the relationship with differential attrition, as well as effect size variations according to behaviour and technique. Researchers should investigate potential additive or interactive effects of the techniques, as they could be utilised in a complementary manner to target different processes in effecting behaviour change.


Virus Control of Trafficking from Sorting Endosomes.
The maintenance of cell surface proteins is critical to the ability of a cell to sense and respond to information in its environment. As such, modulation of cell surface composition and receptor trafficking is a potentially important target of control in virus infection. Sorting endosomes (SEs) are control stations regulating the recycling or degradation of internalized plasma membrane proteins. Here we report that human cytomegalovirus (HCMV), a ubiquitous betaherpesvirus, alters the fate of internalized clathrin-independent endocytosis (CIE) cargo proteins, retaining them in virally reprogrammed SEs. We show that the small G protein ARF6 (ADP ribosylation factor 6), a regulator of CIE trafficking, is highly associated with SE membranes relative to uninfected cells. Combined with the observation of accumulated CIE cargo at the SE, these results suggest that infection diminishes the egress of ARF6 and its cargo from the SE. Expression of ubiquitin-specific protease 6 (USP6), also known as TRE17, was sufficient to restore ARF6 and some ARF6 cargo trafficking to the cell surface in infected cells. The USP activity of TRE17 was required to rescue both ARF6 and associated cargo from SE retention in infection. The finding that TRE17 expression does not rescue the trafficking of all CIE cargos retained at SEs in infection suggests that HCMV hijacks the normal sorting machinery and selectively sorts specific cargos into endocytic microdomains that are subject to alternative sorting fates.IMPORTANCE Cells maintain their surface composition, take up nutrients, and respond to their environment through the internalization and recycling of cargo at the cell surface through endocytic trafficking pathways. During infection with human cytomegalovirus (HCMV), host endocytic membranes are reorganized into a juxtanuclear structure associated with viral assembly and egress. Less appreciated is the effect of this reorganization on the trafficking of host proteins through the endocytic pathway. We show that HCMV retains internalized cargo and the effector of clathrin-independent endocytosis at sorting endosomes. The retention of some cargo, but not all, was reversed by overexpression of a ubiquitin-specific protease, TRE17. Our results demonstrate that HCMV induces profound reprogramming of endocytic trafficking and influences cargo sorting decisions. Further, our work suggests the presence of a novel ubiquitin-regulated checkpoint for the recycling of cargo from sorting endosome. These findings have important implications for host signaling and immune pathways in the context of HCMV infection.


Conjugated linoleic acid induces human adipocyte delipidation: autocrine/paracrine regulation of MEK/ERK signaling by adipocytokines.
Dietary conjugated linoleic acid (CLA) reduces body fat in animals and some humans. Here we show that trans-10, cis-12 CLA, but not cis-9, trans-11 CLA, when added to cultures of stromal vascular cells containing newly differentiated human adipocytes, caused a time-dependent decrease in triglyceride content, insulin-stimulated glucose and fatty acid uptake, incorporation into lipid, and oxidation compared with controls. In parallel, gene expression of peroxisome proliferator-activated receptor-gamma and many of its downstream targets were diminished by trans-10, cis-12 CLA, whereas leptin gene expression was increased. Prior to changes in gene expression and metabolism, trans-10, cis-12 CLA caused a robust and sustained activation of mitogen-activated protein kinase kinase/extracellular signal-related kinase (MEK/ERK) signaling. Furthermore, the trans-10, cis-12 CLA-mediated activation of MEK/ERK could be attenuated by pretreatment with U0126 and pertussis toxin. In parallel, pretreatment with U0126 blocked the ability of trans-10, cis-12 CLA to alter gene expression and attenuate glucose and fatty acid uptake of the cultures. Intriguingly, the induction by CLA of MEK/ERK signaling was linked to hypersecretion of adipocytokines interleukin-6 and interleukin-8. Collectively, these data demonstrate for the first time that trans-10, cis-12 CLA decreases the triglyceride content of newly differentiated human adipocytes by inducing MEK/ERK signaling through the autocrine/paracrine actions of interleukins-6 and 8.


Evaluation of electronic polarization energy in oligoacene molecular crystals using the solvated supermolecular approach.
The solvated supermolecular approach, i.e., block-localized wave function coupled with polarizable continuum model (BLW/PCM), was proposed to calculate molecular ionization potential (IP), electron affinity (EA) in the solid phase, and related electronic polarization. Via the calculations of a solvated supermolecule (5M), including four closest molecules, BLW/PCM overcomes the limitation in the calculation for the monomer PCM, that is, nearly same electronic polarization for cation (P+) and anion (P-). The solvated supermolecular approach successfully described asymmetric behaviors of P+ and P- for oligoacene crystals. In addition, we also compared two charge-localized methods, i.e., BLW and constrained density functional theory (CDFT), to calculate the molecular IP and EA in supermolecules with/without PCM. Our results demonstrate that both the BLW and CDFT correctly estimate the EA and IP values in the gas phase cluster, whereas CDFT/PCM fails to evaluate the P- value of the bulk system.


Myocyte enhancer factor 2C as a neurogenic and antiapoptotic transcription factor in murine embryonic stem cells.
Cell-based therapies require a reliable source of cells that can be easily grown, undergo directed differentiation, and remain viable after transplantation. Here, we generated stably transformed murine ES (embryonic stem) cells that express a constitutively active form of myocyte enhancer factor 2C (MEF2CA). MEF2C has been implicated as a calcium-dependent transcription factor that enhances survival and affects synapse formation of neurons as well as differentiation of cardiomyocytes. We now report that expression of MEF2CA, both in vitro and in vivo, under regulation of the nestin enhancer effectively produces "neuronal" progenitor cells that differentiate into a virtually pure population of neurons. Histological, electrophysiological, and behavioral analyses demonstrate that MEF2C-directed neuronal progenitor cells transplanted into a mouse model of cerebral ischemia can successfully differentiate into functioning neurons and ameliorate stroke-induced behavioral deficits.


Scatter factor: molecular characteristics and effect on the invasiveness of epithelial cells.
The generation of invasiveness in transformed cells represents an essential step of tumor progression. We have previously shown that MDCK epithelial cells, which are deprived of intracellular adhesion by the addition of anti-Arc-1/uvomorulin antibodies, become invasive for collagen gels and embryonal heart tissue (Behrens, J., M. M. Mareel, F. M. Van Roy, and W. Birchmeier. 1989. J. Cell Biol. 108: 2435-2447.). Here we examined whether invasiveness is also induced by scatter factor, which is known to dissociate epithelial cells (Stoker, M., E. Gherardi, M. Perryman, and J. Gray. 1987. Nature (Lond.). 327:239-242.). Scatter factor was purified to homogeneity from conditioned medium of human fibroblasts by heparin-Sepharose chromatography, followed by cation exchange chromatography, gel filtration, or preparative SDS gel electrophoresis. We found that scatter factor represents a 92,000 mol wt glycoprotein which, apparently, is converted by limited proteolysis into disulfide-linked 62,000 and 34/32,000 mol wt subunits. Reversed phase HPLC and sequence analysis of tryptic peptides confirmed the suggested molecular structure, and revealed further that scatter factor exhibits sequence similarities to hepatocyte growth factor and to plasminogen. Purified scatter factor in fact induces the invasiveness into collagen matrices of MDCK epithelial cells, and induces or promotes the invasiveness of a number of human carcinoma cell lines. Apparently, the effect on the human cells depends on their respective degree of differentiation, i.e., cell lines with a less pronounced epithelial phenotype were more susceptible to the factor. Scatter factor does not seem to influence synthesis, steady-state level, and phosphorylation of the cell adhesion molecule Arc-1/uvomorulin. Thus, scatter factor represents a clearly defined molecular species which induces, in vitro, the progression of epithelial cells to a more motile, i.e., invasive phenotype.


Chemical control of xylem differentiation by thermospermine, xylemin, and auxin.
The xylem conducts water and minerals from the root to the shoot and provides mechanical strength to the plant body. The vascular precursor cells of the procambium differentiate to form continuous vascular strands, from which xylem and phloem cells are generated in the proper spatiotemporal pattern. Procambium formation and xylem differentiation are directed by auxin. In angiosperms, thermospermine, a structural isomer of spermine, suppresses xylem differentiation by limiting auxin signalling. However, the process of auxin-inducible xylem differentiation has not been fully elucidated and remains difficult to manipulate. Here, we found that an antagonist of spermidine can act as an inhibitor of thermospermine biosynthesis and results in excessive xylem differentiation, which is a phenocopy of a thermospermine-deficient mutant acaulis5 in Arabidopsis thaliana. We named this compound xylemin owing to its xylem-inducing effect. Application of a combination of xylemin and thermospermine to wild-type seedlings negates the effect of xylemin, whereas co-treatment with xylemin and a synthetic proauxin, which undergoes hydrolysis to release active auxin, has a synergistic inductive effect on xylem differentiation. Thus, xylemin may serve as a useful transformative chemical tool not only for the study of thermospermine function in various plant species but also for the control of xylem induction and woody biomass production.


Anthropometric profile of powerlifters: differences as a function of bodyweight class and competitive success.
This study sought to better understand the relationship between anthropometric profile and maximal strength, as assessed in the sport of powerlifting as relatively little research has examined how differences in anthropometry may contribute to bodyweight-related differences in performance or between more and less successful lifters in the same bodyweight class. To address this aim, 63 male powerlifters from an Argentine National Tournament were assessed for 31 anthropometric variables taken using ISAK (International Society for the Advancement of Kinanthropometry) protocols. Body fractionation (adipose, muscle, bone, residual and skin tissue masses) was determined using the validated Kerr & Ross five way fractionation model of body composition that has yet to be used with powerlifters. Results indicated that the powerlifters showed very elevated values of mesomorphy, muscle girths, muscle mass, bone breadths, and all this accompanied by a medium to low stature. Most of these characteristics were more pronounced in the heavier divisions. The winners had significantly larger proportional muscle mass (53.9 ± 2.2%), muscle to bone mass ratio (5.3 ± 1) and crural index (1.21 ± 0.12) than the non-winners. These comparisons reveal some potential key anthropometric determinants of high level powerlifting performance. These results further support the view that while powerlifters have unique anthropometric profiles, more successful powerlifters typically have higher degrees of muscle mass expressed per unit height and/or bone mass but similar segment lengths and segment length ratios to their less successful peers.


Cost of hospital admission for antiarrhythmic drug initiation in atrial fibrillation.
Initiation of some rhythm-control therapies for atrial fibrillation (AF) requires an inpatient hospital stay and telemetry monitoring, adding to the cost burden of AF. However, specific cost data for inpatient initiation of AF therapies are lacking. To examine costs associated with initiating sotalol or dofetilide in the inpatient setting in the US. This retrospective cohort study used data from billing/discharge records in the Premier Perspective Database for adults with a primary diagnosis of AF, hospitalized between January 2002 and September 2007. Patients had to have received 4 or more sotalol doses or 5 or more dofetilide doses starting within 2 days of admission (with >/=1 dose within 3 days of discharge). Patients admitted solely for AF drug initiation were identified by excluding patients who were admitted on an emergency basis, received care in the emergency department, or underwent major surgical procedures. The primary outcome was direct medical costs for in-hospital services during the stay. Among 7290 patients included in the analysis (4847 sotalol, 2443 dofetilide), mean total inpatient costs per patient were $3278 in the sotalol group and $3610 in the dofetilide group. The greatest costs were for room/board ($1874 sotalol, $1985 dofetilide) and cardiology/electrocardiograms ($394 sotalol, $443 dofetilide). Pharmacy costs were $230 and $201 per patient in the sotalol and dofetilide groups, respectively. The admission of patients for in-hospital initiation of AF rhythm-control therapy represents a high cost burden in the US.


Incidence and cause of fractures in European districts.
To compare fracture rates in European districts. Geographically defined areas of Wales (Swansea and Neath Port Talbot), Norway (Harstad, Trondheim, Stavanger, and Drammen), Sweden (Jamtlands), and Finland (Porvoo). Surveillance of fractures at emergency departments and hospitals and linkage with population data. Comparison of age adjusted and crude rates. Calculation of confidence intervals for ratios. A total of 4113 fractures occurred in 167 560 children during 1996. Fracture rates in south Wales (36 per 1000) were substantially higher than in Scandinavian districts (which were similar). Limiting analysis to the most severe injuries to correct for the possibility of ascertainment bias reduced some of the excess rate in Wales: the Welsh:Scandinavian fracture ratio was 1.82 (95% confidence interval: 1.64 to 2.03). Fracture rates in Welsh children are substantially higher than in Scandinavian children.


Zona pellucida mediated acrosome reaction and sperm morphology.
Sperm samples from 29 men randomly selected from the andrology laboratory, were used to evaluate acrosome reaction response to solubilized human zona pellucida. Capacitated sperm samples were exposed to a solution containing 2 zona pellucidae (ZP) per microl for 60 min, after which acrosomal status were recorded using a PSA-FITC technique. Controls included samples supplied by fertile sperm donors. After completion of acrosome reaction studies, patient samples were divided according to the percentage of morphologically normal spermatozoa. Three basic groups were identified, namely, fertile donors, teratozoospermic (normal sperm morphology 5-14%; n = 25) and severely teratozoospermic (normal sperm morphology < 4%; n = 4) groups. The mean percent normal sperm were 15.8 +/- 0.9, 10.4 +/- 0.7 and 2.7 +/- 0.7, respectively, for normozoospermic donors, teratozoospermic and severely teratozoospermic men. The mean percentage (+/-SE) ZP mediated acrosome reacted sperm among teratozoospermic and severely teratozoospermic cases was 25.8% +/- 0.9 and 19.0% +/- 0.9 (P = 0.001), compared to 36.8% +/- 0.9 for the donor controls. Results were analysed and expressed as correlations between sperm morphology and acrosomal response to human solubilized zona pellucida, spontaneous and calcium ionophore induced acrosome reaction. Predictive values for acrosome responsiveness were depicted with ROC curve analyses. Sperm morphology evaluated by strict criteria correlated positively and highly significantly with the responsiveness of the acrosome reaction (r = 0.91, P = 0.0001). At a morphology cut-off value of 4%, the ROC curve analysis showed sperm morphology to be highly predictive of zona pellucida induced acrosome responsiveness with a sensitivity of 100% and negative predictive value of 100%. Spontaneous and calcium ionophore induced acrosome reactions revealed no correlation with sperm morphology. It was concluded that (i) morphological features of human spermatozoa are indicative of specific functional characteristics; (ii) zona pellucida induction of the acrosome reaction is superior, as a predictor of sperm morphology, compared to calcium ionophore induced and spontaneous acrosome reactions.


Pharmacological characterization of the D2 dopamine receptor negatively coupled with adenylate cyclase in rat anterior pituitary.
In male and female rat anterior pituitary homogenates dopamine inhibited basal adenylate cyclase by 30% and 50%, respectively. Dopamine also inhibited vasoactive intestinal peptide-stimulated adenylate cyclase by 50% in both sexes. Sulpiride, a specific D2 antagonist, stereospecifically blocked with high affinity the dopamine inhibition in both males and females. RU 24926, a specific, non-catechol, non-ergot D2 agonist, also inhibited basal adenylate cyclase of female pituitary with a higher apparent affinity than dopamine (KDapp 20 nM and 450 nM, respectively). This effect was also stereospecifically antagonized by sulpiride. Apomorphine was also more potent (KDapp 100 nM) than dopamine, whereas norepinephrine and SKF 38393, a specific D1 agonist, were poorly active; isoproterenol and clonidine were inactive. Ergots derivatives such as CB 154, LY 14865, pergolide, and lergotrile were potent agonists. alpha-Dihydroergocryptine was a partial agonist of the dopamine receptor negatively coupled with an adenylate cyclase. Because of the slow association kinetics of this drug with the dopamine receptor, its KDapp (0.7 nM) for adenylate cyclase inhibition could be correctly determined only after a 30-min incubation period. All classical dopaminergic antagonists blocked dopamine inhibition of pituitary adenylate cyclase, pimozide (KI 1 nM) and spiperone (KI 0.8 nM) being the more potent. There were good correlations between the affinities of large series of agonists and antagonists for the anterior pituitary dopamine receptors negatively coupled with an adenylate cyclase on one hand, and for either D2 dopamine receptors labeled with [3H] dihydroergocryptine or [3H]spiroperidol in both pituitary and striatum, or D2 pituitary receptors involved in prolactin secretion on the other hand. It is concluded that the pituitary dopamine receptors negatively coupled with an adenylate cyclase are the classical D2 receptors involved in prolactin secretion.


Filovirus Neutralising Antibodies: Mechanisms of Action and Therapeutic Application.
Filoviruses, especially Ebola virus, cause sporadic outbreaks of viral haemorrhagic fever with very high case fatality rates in Africa. The 2013-2016 Ebola epidemic in West Africa provided large survivor cohorts spurring a large number of human studies which showed that specific neutralising antibodies played a key role in protection following a natural Ebola virus infection, as part of the overall humoral response and in conjunction with the cellular adaptive response. This review will discuss the studies in survivors and animal models which described protective neutralising antibody response. Their mechanisms of action will be detailed. Furthermore, the importance of neutralising antibodies in antibody-based therapeutics and in vaccine-induced responses will be explained, as well as the strategies to avoid immune escape from neutralising antibodies. Understanding the neutralising antibody response in the context of filoviruses is crucial to furthering our understanding of virus structure and function, in addition to improving current vaccines & antibody-based therapeutics.


Diagnosis-dependent relationships between cytokine levels and survival in patients admitted for surgical critical care.
Death after trauma, infection, or other critical illness has been attributed to unbalanced inflammation, in which dysregulation of cytokines leads to multiple organ dysfunction and death. We hypothesized that admission cytokine profiles associated with death would differ based on admitting diagnosis. This 5-year study included patients admitted for trauma or surgical intensive care for more than 48 hours at 2 academic, tertiary care hospitals between October 2001 and May 2006. Cytokine analysis for interleukin (IL)-1, -2, -4, -6, -8, -10, -12, interferon-gamma, and tumor necrosis factor (TNF)-alpha was performed using ELISA on specimens drawn within 72 hours of admission. Mann-Whitney U test was used to compare median admission cytokine levels between alive and deceased patients. Relative risks and odds of death associated with admission cytokines were generated using univariate analysis and multivariate logistic regression models, respectively. There were 1,655 patients who had complete cytokine data: 290 infected, nontrauma; 343 noninfected, nontrauma; and 1,022 trauma. Among infected patients, nonsurvivors had higher median admission levels of IL-2, -8, -10, and granulocyte macrophage-colony stimulating factor; noninfected, nontrauma patients had higher IL-6, -8, and IL-10; and nonsurviving trauma patients had higher IL-4, -6, -8, and TNF-alpha. IL-4 was the most significant predictor of death and carried the highest relative risk of dying in trauma patients, and IL-8 in nontrauma, noninfected patients. In infected patients, no cytokine independently predicted death. Cytokine profiles of certain disease states may identify persons at risk of dying and allow for selective targeting of multiple cytokines to prevent organ dysfunction and death.


Next-generation sequencing identifies pathogenic and modifier mutations in a consanguineous Chinese family with hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a highly heterogeneous disease displaying considerable interfamilial and intrafamilial phenotypic variation, including disease severity, age of onset, and disease progression. This poorly understood variance raises the possibility of genetic modifier effects, particularly in MYBPC3-associated HCM.In a large consanguineous Chinese HCM family, we identified 8 members harboring the MYBPC3 c.3624delC (p.Lys1209Serfs) disease-causing mutation, but with very disparate phenotypes. Genotyping ruled out the modifying effect of previously described variants in renin-angiotensin-aldosterone system. Afterwards, we screened for modifying variants in all known causing genes and closely related genes for cardiomyopathy and channelopathy by performing targeted next-generation sequencing. For first time, we showed that a c.1598C>T (p.Ser533Leu) mutation in voltage-dependent l-type calcium channel subunit beta-2 (CACNB2) was present in all severely affected HCM patients, but not in those moderately affected or genotype-positive phenotype-negative patients. This CACNB2 p.Ser533Leu mutation is extremely conserved in evolution, and was not found in 550 healthy controls.Our results suggest that CACNB2 is a possible candidate genetic modifier of MYBPC3-associated familial HCM, but more genetic evidence and functional experiments are needed to confirm.


Detection of MRD may predict the outcome of patients with Philadelphia chromosome-positive ALL treated with tyrosine kinase inhibitors plus chemotherapy.
From 2001 to 2011, 122 patients with newly diagnosed Philadelphia chromosome-positive acute lymphoblastic leukemia were treated with chemotherapy + imatinib (n = 54) or + dasatinib (n = 68). One hundred fifteen (94%) achieved complete remission (CR) including 101 patients who achieved it with only 1 induction course and had at least 1 minimal residual disease (MRD) assessment; 25 patients underwent an allogeneic stem cell transplant in first CR and were excluded, leaving 76 patients as the subject of this report. MRD monitoring by multiparameter flow cytometry (MFC) and real-time quantitative polymerase chain reaction (PCR) was performed at the end of induction and at ~3-month intervals thereafter. Median age was 54 years (range, 21-84 years). There was no difference in survival by achievement of at least a major molecular response (MMR; BCR-ABL/ABL < 0.1%) at CR (P = .22). Patients achieving MMR at 3, 6, 9, and 12 months had a better survival (P = .02, .04, .05, and .01, respectively). Negative MFC at CR did not predict for improved survival (P = .2). At 3 and 12 months, negative MRD by MFC was associated with improved survival (P = .04 and .001). MRD monitoring by PCR and MFC identifies patients who benefit from treatment intensification in first CR.


Enhancing lipoaspirate efficiency by altering liposuction cannula design.
Interplay between the components of a lipoplasty system (suction pump, suction tubing, collection canister, and cannula) determines liposuction efficiency. However, in clinical practice, none of the components are more important than the cannula. Cannula design including port design, port placement, and shaft characteristics is the single most influential contributor to flow resistance and dramatically effects speed of aspiration and final contour. Many variations on port design and placement are available, yet functional enhancements to the cannula shaft have largely been ignored. We have engineered a set of novel cannulas addressing vital elements of cannula design in the effort to enhance aspiration efficiency and efficacy. Two novel cannula designs (dual- and multiport, in-line configuration), created using a unique proprietary manufacturing process, were evaluated against a popular industry standard design (tri-port, Mercedes configuration) to assess aspiration efficiency. Cannulas with shaft diameters of 3, 4, and 5 mm were attached to a standardized lipoplasty system and evaluated in real time for their ability to aspirate a viscous applesauce medium over a 5-minute time course. For each cannula, we calculated (1) the cross-sectional area of the cannula shaft, (2) single and total port area, (3) port-to-shaft ratio, and (4) theoretical resistance. The relationship between the cannula shaft and cannula port(s) directly influenced flow dynamics. Comparing medium uptake time, aspiration efficiency and the aspiration curves demonstrated a significant improvement of the 2 novel cannulas over the standard cannula in the 5- and 4-mm designations. In the 3-mm group, a difference in uptake time remained. However, a significant difference in aspiration efficiency was only seen between the dual-port novel cannula and tri-port Mercedes standard cannula. Further, differences in the aspiration curves between all 3-mm cannulas approached but did not reach significance. We have developed 2 novel cannulas that maximize port features and seek to minimize the internal shaft resistance. Both designs demonstrate enhanced aspiration and uptake compared with an industry standard design.


Self-transcendence, spiritual perspective, and sense of purpose in family caregiving relationships: a mediated model of depression symptoms in Korean older adults.
This study used structural equation modeling to test the mediated model of late-life depression to understand the mechanisms that account for the direct and indirect effects of spiritual variables and purpose in life on depression within the context of Korean family caregiving relationships. A secondary analysis study design used data from a study that tested a theory of family interdependence of 157 Korean elder-family caregiver dyads in Seoul, Korea. Both caregivers' and elders' self-transcendence was positively related to their own sense of purpose in life. However, only elders' spiritual perspective was related to purpose in life. Also, elders' purpose in life was positively associated with caregivers' purpose in life. Furthermore, there was a strong negative relationship between elders' purpose in life and their depressive symptoms, but there was not a significant negative relationship between caregivers' purpose in life and elders' depressive symptoms. Last, elders' purpose in life mediated the negative effects of elders' self-transcendence and spiritual perspective and of caregivers' self-transcendence and purpose in life on elders' depression. The findings suggest that purpose in life for both the caregiver and elder played an important role in elders' depression. Self-transcendence also was related to decreased depression in elders. It is suggested that more attention be given to caregiver and elder purpose in life in developing interventions to reduce or avoid elder depression in Korean elders.


Sub-picosecond photon-efficient 3D imaging using single-photon sensors.
Active 3D imaging systems have broad applications across disciplines, including biological imaging, remote sensing and robotics. Applications in these domains require fast acquisition times, high timing accuracy, and high detection sensitivity. Single-photon avalanche diodes (SPADs) have emerged as one of the most promising detector technologies to achieve all of these requirements. However, these detectors are plagued by measurement distortions known as pileup, which fundamentally limit their precision. In this work, we develop a probabilistic image formation model that accurately models pileup. We devise inverse methods to efficiently and robustly estimate scene depth and reflectance from recorded photon counts using the proposed model along with statistical priors. With this algorithm, we not only demonstrate improvements to timing accuracy by more than an order of magnitude compared to the state-of-the-art, but our approach is also the first to facilitate sub-picosecond-accurate, photon-efficient 3D imaging in practical scenarios where widely-varying photon counts are observed.


Reduced inter-observer and intra-observer delineation variation in esophageal cancer radiotherapy by use of fiducial markers.
Objective: Delineation variation of esophageal tumors remains a large source of geometric uncertainty. In the present study, we investigated the inter- and intra-observer variation in esophageal gross tumor volume (GTV) delineation and the impact of endoscopically implanted fiducial markers on these variations. Material/Methods: Ten esophageal cancer patients with at least two markers endoscopically implanted at the cranial and caudal tumor borders and visible on the planning computed tomography (pCT) were included in this study. Five dedicated gastrointestinal radiation oncologists independently delineated GTVs on the pCT without markers and with markers. The GTV was first delineated on pCTs where markers were digitally removed and next on the original pCT with markers. Both delineation series were executed twice to determine intra-observer variation. For both the inter- and intra-observer analyses, the generalized conformity index (CIgen), and the standard deviation (SD) of the distances between delineated surfaces (i.e., overall, longitudinal, and radial SDs) were calculated. Linear mixed-effect models were used to compare the without and with markers series (α = 0.05). Results: Both the inter- and intra-observer CIgen were significantly larger in the series with markers than in the series without markers (p < .001). For the series without markers vs. with markers, the inter-observer overall SD, longitudinal SD, and radial SD was 0.63 cm vs. 0.22 cm, 1.44 cm vs. 0.42 cm, and 0.26 cm vs. 0.18 cm, respectively (p < .05); moreover, the intra-observer overall SD, longitudinal SD, and radial SD was 0.45 cm vs. 0.26 cm, 1.10 cm vs. 0.41 cm, and 0.22 cm vs. 0.15 cm, respectively (p < .05). Conclusion: The presence of markers at the cranial and caudal tumor borders significantly reduced both inter- and intra-observer GTV delineation variation, especially in the longitudinal direction. Our results endorse the use of markers in GTV delineation for esophageal cancer patients.


Predicting affective responses to exercise using resting EEG frontal asymmetry: does intensity matter?
Affective responses to exercise may be important for improving adherence to regular programs of exercise. The present study sought to determine whether resting frontal EEG asymmetry, an individual difference measure of affective style, is predictive of affective responses to exercise performed at distinct intensities standardized relative to a metabolic landmark (i.e., the ventilatory threshold, VT). Resting EEG was collected from 30 participants and used to predict affective responses following treadmill running at three exercise intensities: below-VT, at-VT, and above-VT. Affect was assessed [via Activation-Deactivation Adjective Check List, yielding measures of Energetic Arousal (EA) and Tense Arousal (TA)] before, immediately following exercise, after 5min cool down, and 10 and 20min post-cool down. Resting mid-frontal asymmetry (F4-F3) significantly predicted EA immediately following below-VT exercise; resting lateral frontal asymmetry (F8-F7) predicted EA at 20min post-cool down. Resting mid-frontal asymmetry predicted in EA immediately following and following cool down in above-VT exercise. As a whole, frontal asymmetry was predictive of affective responses following exercise, namely greater relative left frontal activity predicting lower EA. This was opposite to the predictions of the valenced motivation model, but may provide some support for the motivation direction model. This is based on the fact that low EA could be indicative of approach motivation, especially at higher exercise intensities.


Study of amended reports to evaluate and improve surgical pathology processes.
: Amended surgical pathology reports record defects in the process of transforming tissue specimens into diagnostic information. : Systematic study of amended reports tests 2 hypotheses: (a) that tracking amendment frequencies and the distribution of amendment types reveals relevant aspects of quality in surgical pathology's daily transformation of specimens into diagnoses and (b) that such tracking measures the effect, or lack of effect, of efforts to improve surgical pathology processes. : We applied a binary definition of altered reports as either amendments or addenda and a taxonomy of defects that caused amendments as misidentifications, specimen defects, misinterpretations, and report defects. During the introduction of a LEAN process improvement approach-the Henry Ford Productions System-we followed trends in amendment rates and defect fractions to (a) evaluate specific interventions, (b) sort case-by-case root causes of misidentifications, specimen defects, and misinterpretations, and (c) audit the ongoing accuracy of the classification of changed reports. LEAN is the management and production system of the Toyota Motor Corporation that promotes continuous improvement; it considers wasted resources expended for purposes other than creating value for end customers and targets such expenditures for elimination. : Introduction of real-time editing of amendments saw annual amendment rates increase from 4.8/1000 to 10.1/1000 and then decrease in an incremental manner to 5.6/1000 as Henry Ford Productions System-specific interventions were introduced. Before introduction of HFPS interventions, about a fifth of the amendments were due to misidentifications, a 10th were due to specimen defects, a quarter due to misinterpretation, and almost half were due to report defects. During the period of the initial application of HFPS, the fraction of amendments due to misidentifications decreased as those due to report defects increased, in a statistically linked manner. As HFPS interventions took hold, misidentifications fell from 16% to 9%, specimen defect rates remained variable, ranging between 2% and 11%, and misinterpretations fell from 18% to 3%. Reciprocally, report defects rose from 64% to 83% of all amendment-causing defects. A case-by-case study of misidentifications, specimen defects, and misinterpretations found that (a) intervention at the specimen collection level had disappointingly little effect on patient misidentifications; (b) standardization of specimen accession and gross examination reduced only specimen defects surrounding ancillary testing; but (c) a double review of breast and prostate cases was associated with drastically reduced misinterpretation defects. Finally, audit of both amendments and addenda demonstrated that 10% of the so-called addenda actually qualified as amendments. : Monitored by the consistent taxonomy, rates of amended reports first rose, then fell. Examining specific defect categories provided information for evaluating specific LEAN interventions. Tracking the downward trend of amendment rates seemed to document the overall success of surgical pathology quality improvement efforts. Process improvements modestly decreased fractions of misidentifications and markedly decreased misinterpretation fractions. Classification integrity requires real time, independent editing of both amendments (changed reports) and addenda (addition to reports).


Stroke survivors control the temporal structure of variability during reaching in dynamic environments.
Learning to control forces is known to reduce the amount of movement variability (e.g., standard deviation; SD) while also altering the temporal structure of movement variability (e.g., approximate entropy; ApEn). Such variability control has not been explored in stroke survivors during reaching movements in dynamic environments. Whether augmented feedback affects such variability control, is also unknown. Chronic stroke survivors, assigned randomly to a control/experimental group, learned reaching movements in a dynamically changing environment while receiving either true feedback of their movement (control) or augmented visual feedback (experimental). Hand movement variability was analyzed using SD and ApEn. A significant change in variability was determined for both SD and ApEn. Post hoc tests revealed that the significant decrease in SD was not retained after a week. However, the significant increase in ApEn, determined on both days of training, showed significant retention effects. In dynamically changing environments, chronic stroke survivors reduced the amount of movement variability and made their movement patterns less repeatable and possibly more flexible. These changes were not affected by augmented visual feedback. Moreover, the learning patterns characteristically involved the control of the nonlinear dynamics rather than the amount of hand movement variability. The absence of transfer effects demonstrated that variability control of hand movement after a stroke is specific to the task and the environment.


Comparative metabolic profiling to investigate the contribution of O. oeni MLF starter cultures to red wine composition.
In this research work we investigated changes in volatile aroma composition associated with four commercial Oenococcus oeni malolactic fermentation (MLF) starter cultures in South African Shiraz and Pinotage red wines. A control wine in which MLF was suppressed was included. The MLF progress was monitored by use of infrared spectroscopy. Gas chromatographic analysis and capillary electrophoresis were used to evaluate the volatile aroma composition and organic acid profiles, respectively. Significant strain-specific variations were observed in the degradation of citric acid and production of lactic acid during MLF. Subsequently, compounds directly and indirectly resulting from citric acid metabolism, namely diacetyl, acetic acid, acetoin, and ethyl lactate, were also affected depending on the bacterial strain used for MLF. Bacterial metabolic activity increased concentrations of the higher alcohols, fatty acids, and total esters, with a larger increase in ethyl esters than in acetate esters. Ethyl lactate, diethyl succinate, ethyl octanoate, ethyl 2-methylpropanoate, and ethyl propionate concentrations were increased by MLF. In contrast, levels of hexyl acetate, isoamyl acetate, 2-phenylethyl acetate, and ethyl acetate were reduced or remained unchanged, depending on the strain and cultivar evaluated. Formation of ethyl butyrate, ethyl propionate, ethyl 2-methylbutryate, and ethyl isovalerate was related to specific bacterial strains used, indicating possible differences in esterase activity. A strain-specific tendency to reduce total aldehyde concentrations was found at the completion of MLF, although further investigation is needed in this regard. This study provided insight into metabolism in O. oeni starter cultures during MLF in red wine.


Oral immunotherapy for cow's milk allergy with a weekly up-dosing regimen: a randomized single-blind controlled study.
Cow's milk allergy (CMA) in children is a important problem in medical practice. Oral desensitization has been proposed as a therapeutic approach, but current protocols are time-consuming and impractical. To establish a patient-friendly desensitization regimen with weekly up-dosing and to evaluate it in a randomized controlled trial. Thirty children with IgE-mediated CMA confirmed by double-blind placebo-controlled food challenge were equally randomized to desensitization with CM or soy milk as control. The weekly up-dosing lasted 18 weeks. The occurrence and severity of reactions after each dose was evaluated, and the desensitization was stopped if severe reactions occurred. Specific IgE and IgG4 levels to CM were measured at baseline, after 8 weeks, and at the end of the study. The double-blind food challenge was repeated once the desensitization was completed or after premature discontinuation. Two active and 1 control patient dropped out. Full tolerance to CM (200 mL) was achieved in 10 active patients and partial tolerance in 1. Two active patients discontinued the desensitization after experiencing severe reactions, whereas no reactions occurred in controls, whose sensitivity to CM remained unchanged. A significant increase in specific IgG4 levels was found only in the active group. This weekly up-dosing desensitization protocol for CMA performed under medical supervision was effective and reasonably safe and induced consistent immunologic changes.


Effect of drug-induced hyper- and hypoprolactinemia on human corpus luteum.
This study was performed on 8 patients with normal levels of PRL and ovulatory cycles. Three of these patients were treated with 5 mg/day Bromocriptine and three of them with 60 mg/day Metoclopramide from the beginning of the cycle in order to induce hypo- and hyperprolactinemia respectively. The other two women were left as control. Daily blood samples were collected for the determination of PRL, LH 17 beta-Estradiol and Progesterone. Patients were operated on between the 6th and the 8th day after the LH peak and corpora lutea enucleated and frozen for subsequent 125I-LH binding tests. An analysis of our data demonstrates that hypo- and hyper- prolactinemia induced with Bromocriptine and Metoclopramide respectively may cause luteal insufficiency, which is revealed by decreased blood levels of progesterone. Furthermore we have observed that specific binding for LH in human luteal tissue is always present (in normal, hypo- and hyperprolactinemic women) but is reduced in presence of very high or very low levels of PRL.


Refining the care of patients with pancreatic cancer: the AGITG Pancreatic Cancer Workshop consensus.
A meeting of the Australasian Gastro-Intestinal Trials Group (AGITG) was held to develop a consensus statement defining when a patient with pancreatic cancer has disease that is clearly operable, is borderline, or is locally advanced/inoperable. Key issues included the need for multidisciplinary team consensus for all patients considered for surgical resection. Staging investigations, to be completed within 4 weeks of presentation, should include pancreatic protocol computed tomography, endoscopic ultrasound, and, when possible, biopsy. Given marked differences in outcomes, the operability of tumours should be clearly identified by categories: those clearly resectable by standard means (group 1a), those requiring vascular resection but which are clearly operable (group 1b), and those of borderline operability requiring vascular resection (groups 2a and 2b). Patients who may require vascular reconstruction should be referred, before exploration, to a specialist unit. All patients should have a structured pathology report with standardised reporting of all seven surgical margins, which identifies an R0 (no tumour cells within a defined distance of the margin) if all surgical margins are clear from 1 mm. Neo-adjuvant therapy is increasingly recommended for borderline operable disease, while chemotherapy is recommended as initial therapy for patients with unresectable loco-regional pancreatic cancer. The value of adding radiation after initial chemotherapy remains uncertain. A small number of patients may be downstaged by chemoradiation, and trimodality therapy should only be considered as part of a clinical trial. Instituting these recommendations nationally will be an integral part of the process of improving quality of care and reducing geographic variation between centres in outcomes for patients.


[DNA polymerase mediated amplification of DNA fragments using primers with mismatches in the 3'-region].
The ability of three thermostable enzymes, Tth, Taq, and Klentaq DNA polymerases, to amplify DNA with primers containing mismatches in the 3'-terminal region was studied. It is shown that Tth polymerase, in contrast to the Taq and Klentaq enzymes, synthesizes equally well DNA with primers perfectly complementary to the template and with those containing mismatches next the 3'-end. The use of Tth DNA polymerase in the polymerase chain reaction was shown to result, in some cases, in a great number of additional, nonspecific DNA fragments as compared with Taq DNA polymerase. This may be due to the ability of Tth polymerase for DNA primer extension even if the 3'-terminal region of the primer contains nucleotides non-complementary to the template. Tth DNA polymerase and a Klentaq/Tth mixture (100:1) can be efficiently used in the amplification of DNA with degenerated primers and primers forming nonperfect duplexes with the template.


Prior deafferentation confers long term protection to CA1 against transient forebrain ischemia and sustains GluR2 expression.
Hippocampal CA1 pyramidal neurons undergo delayed neurodegeneration after transient forebrain ischemia, and the phenomenon is dependent upon hyperactivation of l-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) subtype of glutamate receptors, resulting in aberrant intracellular calcium influx. The GluR2 subunit of AMPA receptors is critical in limiting the influx of calcium. The CA1 pyramidal neurons are very sensitive to ischemic damage and attempts to achieve neuroprotection, mediated by drugs, have been unsuccessful. Moreover, receptor antagonism strategies in the past have failed to provide long-term protection against ischemic injury. Long-term protection against severe forebrain ischemia can be conferred by fimbria-fornix (FF) deafferentation, which interrupts the afferent input to CA1. Our study evaluated the long-term protective effect of FF deafferentation, 12 days prior to induction of ischemia, on vulnerable CA1 neurons. Our results indicate that at 7 and 28 days post-ischemia, prior FF deafferentation protected 60% of neurons against ischemic cell death. Furthermore, we sought to evaluate whether FF deafferentation also sustained GluR2 levels in these neurons. GluR2 protein and mRNA expression were sustained by deafferentation at 70% of control following ischemia. Correlation studies revealed a positive correlation between GluR2 protein and mRNA level. These results demonstrate that protection conferred by FF deafferentation was long-term and related to sustained GluR2 expression.


Thymine and guanine base specificity of human myeloma proteins with anti-DNA activity.
To further our understanding of the molecular basis of DNA-autoantibody interactions, we have characterized the specificities of three IgG human myeloma proteins that bind DNA. We measured their binding to synthetic single- and double-stranded homopolynucleotides, random and alternating copolymers, oligonucleotides, and nucleotides or nucleosides conjugated to non-nucleic acid carriers. All three antibodies bound single-stranded nucleic acids, including both polyribonucleotides and polydeoxyribonucleotides. They varied in relative affinities for polynucleotides of varying base composition. Polymers containing the purines guanine or hypoxanthine and/or the pyrimidine thymine were most reactive with all three proteins. A myeloma protein that reacted with poly(G), poly(I), or poly(dT) also bound to the corresponding nucleosides or nucleotides conjugated to bovine serum albumin. None of the antibodies reacted with base-paired double-helical polynucleotides (double-stranded RNA, RNA-DNA hybrid or double-stranded DNA). The results indicate that base specificity is prominent in their reactions and that the accessible epitopes in single-stranded polynucleotides become masked upon base pairing in double-stranded helices. These findings suggest a model in which positions N1 and O6 of guanine and hypoxanthine and N3 and O4 of thymine interact with amino acids of the antibody-combining site.


MO-A-213AB-08: 2D Water Equivalent Path Length Imaging Technique for Pre-Treatment Range Verification in Proton Therapy.
To assess the potentials of a novel detector for providing a transmission image of the patient in terms of the Water Equivalent Path Length (WEPL) values and to evaluate the detector potential for real-time imaging of moving targets. The method is based on the principle that for passively scattered proton beams the WEPL of any point located in the dose plateau of a spread-out Bragg-Peak can be derived from the time dependence of the dose rate function measured at this point. A flat-panel 2D detector array (Sun Nuclear Corporation, 249 diodes, pitch ∼6 mm, 2 ms time resolution) was placed distal to a range of phantoms with varying complexities. The dose rate received by all diodes were measured as functions of time and analyzed to obtain the WEPL values. To assess the potential real-time features of this imaging technique, a Lucite cube was imaged while moving with a sinusoidal pattern with the amplitude and period comparable to a typical mobile tumor. In water tanks, millimeter accuracy in the determination of the WEPL could be achieved and the geometrical shape of wedge and sphere phantoms could be reproduced. In more complex phantoms such as a Lucite step-like compensator or Medulloblastoma patient compensators, multiple Coulomb scattering and range mixing cause a slight deterioration in the reconstruction of the WEPL. We found that tracking of a moving target in the coronal plane is potentially feasible. The technique gives less than 1 cGy of dose to patients and is therefore ideal for 'range-tuning' prior to treatment. For clinical applications, the beam for 'tuning' will need a deeper range than the prescription and once the radiological path length to the dosimeter is determined and compared with that from planning, the proton range for the actual treatment can be adjusted.


Prediction of response to electroconvulsive therapy. Preliminary validation of a sign-based typology of depression.
In an open study of 35 depressed patients, psychomotor disturbance, assessed by the CORE rating system, significantly predicted response to ECT in patients with a 'retarded' but not those with an 'agitated' type of illness. In multivariate analyses, psychomotor disturbance was the sole predictor of response. This preliminary study suggests that the CORE system, which has been shown to differentiate endogenous and non-endogenous depressive disorders, has predictive validity in relation to ECT response.


Proteome analysis reveals a strong correlation between olfaction and pollen foraging preference in honeybees.
To gain a deeper understanding of the molecular basis of pollen foraging preference, we characterized the proteomes of antennae and brains of bees foraging on pear and rapeseed flowers, and the volatile compounds from nectar, anther, and inflorescence of both plants. Bees foraging on the pollen of the two plants have shaped the distinct proteome arsenals in the antenna and brain to drive olfactory and brain function. In antennae, bees foraging on pear (PA) pollen pathways associated with protein metabolism were induced to synthesize new proteins for modulation of synaptic structures via stabilizing and consolidating specific memory traces. Whereas, bees foraging on rapeseed (BA) pollen pathways implicated in energy metabolism were activated to provide metabolic fuels critical for neural activity. These findings suggest that the distinct biochemical route is functionally enhanced to consolidate the divergent olfaction in PA and BA. In brain, although the uniquely induced pathways in bees forging on both plants are likely to cement selective roles in learning and memory, pollen foraging preference in bees is mainly drived by olfaction. Furthermore, both plants have shaped different repertoires of signal odors and food rewards to attract pollinators. The suggested markers are potentially useful for selection of bees to improve their olfaction for better pollination of the plants.


Refractive errors and incident cataracts: the Beaver Dam Eye Study.
To describe the relation between refractive errors and incident age-related cataracts in a predominantly white US population. All persons aged 43 to 84 years of age in Beaver Dam, Wisconsin, were invited for a baseline examination from 1988 through 1990 and a follow-up examination 5 years later from 1993 through 1995. At both examinations, participants had refraction and photographic assessment of cataract, according to a standardized protocol. Myopia was defined as a spherical equivalent of -1.0 diopters (D) or less, hyperopia as +1.0 D or more. The relations between refractive errors at baseline and cataract at baseline (prevalent cataract), 5-year incident cataract, and incident cataract surgery were analyzed by using generalized estimating equations. When age and gender were controlled for, myopia was related to prevalent nuclear cataract (odds ratio [OR], 1.67; 95% confidence interval [CI], 1.23-2.27), but not to cortical and posterior subcapsular cataracts. Myopia was not related to 5-year incident nuclear, cortical, and posterior subcapsular cataracts, but was related to incident cataract surgery (OR 1.89; CI 1.18-3.04). Hyperopia was related to incident nuclear (OR 1.56; CI 1.25-1.95) and possibly cortical (OR 1.25; CI 0.96-1.63) cataracts, but not to posterior subcapsular cataract or cataract surgery. After further adjustment for diabetes, smoking, and education, the association between myopia and incident cataract surgery was attenuated (OR 1.60; CI 0.96-2.64), but the associations between hyperopia and incident nuclear and cortical cataracts were unchanged. These data support the cross-sectional association between myopia and nuclear cataract seen in other population-based studies, but provide no evidence of a relationship between myopia and 5-year incident cataract. Hyperopia may be related weakly to incident nuclear and cortical cataract.


Experimental Evaluation of the Optimal Suture Pattern With a Flexible Endoscopic Suturing System.
The ability to perform reliable, secure endoluminal closure of the gastrointestinal tract wall, is a prerequisite to support the progress of the emerging field of endoluminal surgery. Along with advanced clipping systems, flexible endoscopic suturing devices are commercially available. Current systems can replicate traditional surgical suturing patterns in the endoluminal environment. The aim of this study was to evaluate the optimal endoluminal suturing technique using a flexible endoscopic suturing device. Procedures were performed on bench-top simulators containing 20 explanted porcine stomachs. A standardized 3-cm full-thickness incision was created on the anterior wall of each stomach using monopolar cautery. The gastrotomy was closed endoscopically using an over-the-scope suturing device (OverStitch, Apollo Endosurgery; Austin, TX). Three different techniques were used: single stitches, figure-of-8 pattern, and running suture. Material consumption and operation time were recorded and bursting pressure measurement of the closure was performed. No statistically significant differences were identified in suturing time. Suturing time (minutes) was slightly shorter with the figure-of-8 technique (41.14 ± 4.6) versus interrupted (45.75 ± 1.1) versus continuous (51.44 ± 10.0), but the difference was not statistically significant. The number of sutures required was greater in the interrupted group. No significant difference was found in the burst pressure (mm Hg): figure-of-8 (45.85 ± 26.2) versus interrupted (30.5 ± 22.89) versus continuous (32.0 ± 26.5). In the figure-of-8 group, 85.5% of cases were leakproof above 30 mm Hg, while in the other groups only 50% of cases were so. A figure-of-8 suturing pattern seems to be the preferable suturing technique with the endoscopic suturing device.


Heterologous promoter recognition leading to high-level expression of cloned foreign genes in Bombyx mori cell lines and larvae.
The baculovirus expression system using the Autographa californica nuclear polyhedrosis virus (AcNPV) has been extensively utilized for high-level expression of cloned foreign genes, driven by the strong viral promoters of polyhedrin (polh) and p10 encoding genes. A parallel system using Bombyx mori nuclear polyhedrosis virus (BmNPV) is much less exploited because the choice and variety of BmNPV-based transfer vectors are limited. Using a transient expression assay, we have demonstrated here that the heterologous promoters of the very late genes polh and p10 from AcNPV function as efficiently in BmN cells as the BmNPV promoters. The location of the cloned foreign gene with respect to the promoter sequences was critical for achieving the highest levels of expression, following the order + 35 > + 1 > -3 > -8 nucleotides (nt) with respect to the polh or p10 start codons. We have successfully generated recombinant BmNPV harboring AcNPV promoters by homeologous recombination between AcNPV-based transfer vectors and BmNPV genomic DNA. Infection of BmN cell lines with recombinant BmNPV showed a temporal expression pattern, reaching very high levels in 60-72 h post infection. The recombinant BmNPV harboring the firefly luciferase-encoding gene under the control of AcNPV polh or p10 promoters, on infection of the silkworm larvae led to the synthesis of large quantities of luciferase. Such larvae emanated significant luminiscence instantaneously on administration of the substrate luciferin resulting in 'glowing silkworms'. The virus-infected larvae continued to glow for several hours and revealed the most abundant distribution of virus in the fat bodies. In larval expression also, the highest levels were achieved when the reporter gene was located at + 35 nt of the polh.


Genetic Diversification and Dispersal of Taro (Colocasia esculenta (L.) Schott).
Taro (Colocasia esculenta (L.) Schott) is widely distributed in tropical and sub-tropical areas. However, its origin, diversification and dispersal remain unclear. While taro genetic diversity has been documented at the country and regional levels in Asia and the Pacific, few reports are available from Americas and Africa where it has been introduced through human migrations. We used eleven microsatellite markers to investigate the diversity and diversification of taro accessions from nineteen countries in Asia, the Pacific, Africa and America. The highest genetic diversity and number of private alleles were observed in Asian accessions, mainly from India. While taro has been diversified in Asia and the Pacific mostly via sexual reproduction, clonal reproduction with mutation appeared predominant in African and American countries investigated. Bayesian clustering revealed a first genetic group of diploids from the Asia-Pacific region and to a second diploid-triploid group mainly from India. Admixed cultivars between the two genetic pools were also found. In West Africa, most cultivars were found to have originated from India. Only one multi-locus lineage was assigned to the Asian pool, while cultivars in Madagascar originated from India and Indonesia. The South African cultivars shared lineages with Japan. The Caribbean Islands cultivars were found to have originated from the Pacific, while in Costa Rica they were from India or admixed between Indian and Asian groups. Taro dispersal in the different areas of Africa and America is thus discussed in the light of available records of voyages and settlements.


Exchanging Appointment Data Among Healthcare Institutions.
The introduction of national electronic patient records such as the electronic patient dossier EPD in Switzerland provides a new basis for digitizing healthcare processes at a national level. One process however, that is currently neglected within the Swiss EPD, is the scheduling process in healthcare. The objective of this work is to analyze the appointment scheduling process and the involved IT systems in order to develop an appointment data structure and a concept for cross-institutional exchange of appointment data. The analysis showed that various outpatient and inpatient information systems support appointment booking through proprietary solutions. A true standard for appointment data exchange is missing. We suggest an appointment data structure and a corresponding data exchange process based on the FHIR standard. In its current implementation, the Swiss EPD does not support this proposed appointment scheduling process. We discuss how potential additions such as the IHE Care Services Discovery (CSD) profile can provide better compatibility.


Reproducibility of right ventricular volumes and ejection fraction using real-time three-dimensional echocardiography: comparison with cardiac MRI.
The nongeometric nature of the right ventricle (RV) makes it difficult to measure. We sought to determine whether real-time three-dimensional echocardiography (RT3DE) is superior to two-dimensional echocardiography (2DE) for the follow-up of RV function by validation vs cardiac MRI. RV volumes and ejection fraction (EF) were studied with 2DE (including area-length [A-L], the modified two-dimensional subtraction [2DS] method, and the Simpson method of discs), RT3DE, and MRI in 50 patients with left ventricular wall motion abnormalities, the results of which suggested possible RV infarction. Test-retest variation was performed by a complete restudy using a separate sonographer within 24 h without the alteration of hemodynamics or therapy. Interobserver and intraobserver variations were noted in a subgroup of 20 patients. EF estimations were similar using each technique. The mean (+/- SD) MRI end-diastolic volume (87 +/- 22 mL) was only slightly underestimated by RT3DE (mean difference, -3 +/- 10; p < 0.05), with a greater mean difference for 2DE A-L (-29 +/- 10; p < 0.05), and the Simpson method of discs (-29 +/- 23; p < 0.05), and was greatly overestimated by 2DS (mean difference, 26 +/- 23; p < 0.05). Similarly, the mean MRI end-systolic volume (46 +/- 17 mL) was only slightly underestimated by RT3DE (-4 +/- 7; p < 0.05), compared with 2DE A-L (-16 +/- 8; p < 0.05) and the Simpson method of discs (-16 +/- 8; p < 0.05), and was overestimated by 2DS (14 +/- 13; p < 0.05). RT3DE findings had a higher correlation with each parameter than any 2DE technique. There was also good intraobserver and interobserver correlation between RT3DE by two sonographers. RT3DE had less test-retest variation of RV volumes and EF than any 2DE measure. RT3DE is more accurate than two-dimensional approaches and reduces the test-retest variation of RV volumes and EF measurements in follow-up RV assessment.


An evaluation of analytical approaches for understanding change in cognition in the context of aging and health.
In this article, we discuss the importance of studying the relationship between health and cognitive function, and some of the methods with which this relationship has been studied. We consider the challenges involved, in particular operationalization of the health construct and causal inference in the context of observational data. We contrast the approaches taken, and review the questions addressed: whether health and cognition are associated, whether changes in health are associated with changes in cognition, and the degree of interdependency among their respective trajectories. A variety of approaches for understanding the association between cognition and health in aging individuals have been used. Much of the literature on cognitive change and health has relied on methods that are based at least in part on the reorganization of between-person differences (e.g., cross-lag analysis) rather than relying more fully on analysis of within-person change and joint analysis of individual differences in within-person change in cognition and health. We make the case for focusing on the interdependency between within-person changes in health and cognition and suggest methods that would support this.


Introduction and executive summary: care of the critically ill and injured during pandemics and disasters: CHEST consensus statement.
Natural disasters, industrial accidents, terrorism attacks, and pandemics all have the capacity to result in large numbers of critically ill or injured patients. This supplement provides suggestions for all of those involved in a disaster or pandemic with multiple critically ill patients, including front-line clinicians, hospital administrators, professional societies, and public health or government officials. The current Task Force included a total of 100 participants from nine countries, comprised of clinicians and experts from a wide variety of disciplines. Comprehensive literature searches were conducted to identify studies upon which evidence-based recommendations could be made. No studies of sufficient quality were identified. Therefore, the panel developed expert-opinion-based suggestions that are presented in this supplement using a modified Delphi process. The ultimate aim of the supplement is to expand the focus beyond the walls of ICUs to provide recommendations for the management of all critically ill or injured adults and children resulting from a pandemic or disaster wherever that care may be provided. Considerations for the management of critically ill patients include clinical priorities and logistics (supplies, evacuation, and triage) as well as the key enablers (systems planning, business continuity, legal framework, and ethical considerations) that facilitate the provision of this care. The supplement also aims to illustrate how the concepts of mass critical care are integrated across the spectrum of surge events from conventional through contingency to crisis standards of care.


A clinician's guide to a woman's heart.
Contrary to the perception of most women that breast cancer is the leading cause of death in women, it is heart disease and stroke that claim the most lives of women older than 35 years in the United States. The prevalence of cardiovascular disease is likely to increase as more women-notably the "baby boomers"- enter the menopausal period, a time of increased cardiac vulnerability. Primary care physicians should be on the forefront in taking an inventory of risk factors in their female patients and in educating them about the reality of heart disease in their gender and about the risk factors that these patients can modify to decrease their risk. To aid in their efforts, this article provides an overview of those risk factors that are modifiable, such as weight reduction, exercise, and smoking cessation. It also offers an evaluation of the benefit and nonbenefit of vitamin and herbal supplements in reduction of cardiac events.


The impact of Thatcherism on health and well-being in Britain.
Margaret Thatcher (1925-2013) was the United Kingdom's prime minister from 1979 to 1990. Her informal transatlantic alliance with U.S. President Ronald Reagan from 1981 to 1989 played an important role in the promotion of an international neoliberal policy agenda that remains influential today. Her critique of UK social democracy during the 1970s and her adoption of key neoliberal strategies, such as financial deregulation, trade liberalization, and the privatization of public goods and services, were popularly labeled Thatcherism. In this article, we consider the nature of Thatcherism and its impact on health and well-being during her period as prime minister and, to a lesser extent, in the years that follow; we focus mainly on Great Britain (England, Scotland, and Wales). Thatcher's policies were associated with substantial increases in socioeconomic and health inequalities: these issues were actively marginalized and ignored by her governments. In addition, her public-sector reforms applied business principles to the welfare state and prepared the National Health Service for subsequent privatization.


Molecular mechanism for repression of 17alpha-hydroxylase expression and androstenedione production in granulosa cells.
According to the traditional two-cell two-gonadotropin model of follicular steroidogenesis, androgen production arises exclusively from theca cells. The granulosa cells, in turn, utilize androstenedione and testosterone, which are aromatized into estrone and estradiol, respectively. Differential expression of the activator protein-1 (AP-1) transcription factor, c-fos, has been postulated to result in distinct patterns of steroidogenesis in the theca and granulosa cell compartments. We hypothesize that c-fos functions to inhibit the production of 17alpha-hydroxylase 17,20 lyase (CYP17) in granulosa cells, thereby suppressing androgen synthesis. Our objective was to define the role of c-fos in the regulation of CYP17 production in granulosa cells. Human luteinized granulosa (HGL5) cells were utilized for all experiments. The following techniques were used: mRNA extraction, steroid quantification, small interfering RNA silencing, microarray analysis, and immunohistochemistry. Immunohistochemistry studies demonstrated significant staining of c-fos in the granulosa cell layer, but absent staining for CYP17. Conversely, the theca cell layer did not stain for c-fos, but staining was evident for CYP17. Treatment of HGL5 cells with the MAPK kinase inhibitor PD98059 resulted in an 11-fold increase in CYP17 mRNA levels. In c-fos gene silenced cells, CYP17 mRNA levels increased 8-fold. Androstenedione production was increased 13-fold after treatment with PD98059. These results suggest that the AP-1 transcription factor, c-fos, may be one of the factors responsible for CYP17 repression and hence suppression of androstenedione production in granulosa cells. This may provide an explanation for the lack of CYP17 in granulosa cells.


Lake Tanganyika--a 'melting pot' of ancient and young cichlid lineages (Teleostei: Cichlidae)?
A long history of research focused on the East Africa cichlid radiations (EAR) revealed discrepancies between mtDNA and nuclear phylogenies, suggesting that interspecific hybridisation may have been significant during the radiation of these fishes. The approximately 250 cichlid species of Lake Tanganyika have their roots in a monophyletic African cichlid assemblage, but controversies remain about the precise phylogenetic origin and placement of different lineages and consequently about L. Tanganyika colonization scenarios. 3312 AFLP loci and the mitochondrial ND2 gene were genotyped for 91 species representing almost all major lacustrine and riverine haplotilapiine east African cichlid lineages with a focus on L. Tanganyika endemics. Explicitly testing for the possibility of ancient hybridisation events, a comprehensive phylogenetic network hypothesis is proposed for the origin and diversification of L. Tanganyika cichlids. Inference of discordant phylogenetic signal strongly suggests that the genomes of two endemic L. Tanganyika tribes, Eretmodini and Tropheini, are composed of an ancient mixture of riverine and lacustrine lineages. For the first time a strong monophyly signal of all non-haplochromine mouthbrooding species endemic to L. Tanganyika ("ancient mouthbrooders") was detected. Further, in the genomes of early diverging L. Tanganyika endemics Trematocarini, Bathybatini, Hemibatini and Boulengerochromis genetic components of other lineages belonging to the East African Radiation appear to be present. In combination with recent palaeo-geological results showing that tectonic activity in the L. Tanganyika region resulted in highly dynamic and heterogeneous landscape evolution over the Neogene and Pleistocene, the novel phylogenetic data render a single lacustrine basin as the geographical cradle of the endemic L. Tanganyika cichlid lineages unlikely. Instead a scenario of a pre-rift origin of several independent L. Tanganyika precursor lineages which diversified in ancient rivers and precursor lakes and then amalgamated in the extant L. Tanganyika basin is put forward as an alternative: the 'melting pot Tanganyika' hypothesis.


Cerebral Small Vessel Disease Biomarkers Detection on MRI-Sensor-Based Image and Deep Learning.
Magnetic resonance imaging (MRI) offers the most detailed brain structure image available today; it can identify tiny lesions or cerebral cortical abnormalities. The primary purpose of the procedure is to confirm whether there is structural variation that causes epilepsy, such as hippocampal sclerotherapy, local cerebral cortical dysplasia, and cavernous hemangioma. Cerebrovascular disease, the second most common factor of death in the world, is also the fourth leading cause of death in Taiwan, with cerebrovascular disease having the highest rate of stroke. Among the most common are large vascular atherosclerotic lesions, small vascular lesions, and cardiac emboli. The purpose of this thesis is to establish a computer-aided diagnosis system based on small blood vessel lesions in MRI images, using the method of Convolutional Neural Network and deep learning to analyze brain vascular occlusion by analyzing brain MRI images. Blocks can help clinicians more quickly determine the probability and severity of stroke in patients. We analyzed MRI data from 50 patients, including 30 patients with stroke, 17 patients with occlusion but no stroke, and 3 patients with dementia. This system mainly helps doctors find out whether there are cerebral small vessel lesions in the brain MRI images, and to output the found results into labeled images. The marked contents include the position coordinates of the small blood vessel blockage, the block range, the area size, and if it may cause a stroke. Finally, all the MRI images of the patient are synthesized, showing a 3D display of the small blood vessels in the brain to assist the doctor in making a diagnosis or to provide accurate lesion location for the patient.


A partially linear tree-based regression model for multivariate outcomes.
In the genetic study of complex traits, especially behavior related ones, such as smoking and alcoholism, usually several phenotypic measurements are obtained for the description of the complex trait, but no single measurement can quantify fully the complicated characteristics of the symptom because of our lack of understanding of the underlying etiology. If those phenotypes share a common genetic mechanism, rather than studying each individual phenotype separately, it is more advantageous to analyze them jointly as a multivariate trait to enhance the power to identify associated genes. We propose a multilocus association test for the study of multivariate traits. The test is derived from a partially linear tree-based regression model for multiple outcomes. This novel tree-based model provides a formal statistical testing framework for the evaluation of the association between a multivariate outcome and a set of candidate predictors, such as markers within a gene or pathway, while accommodating adjustment for other covariates. Through simulation studies we show that the proposed method has an acceptable type I error rate and improved power over the univariate outcome analysis, which studies each component of the complex trait separately with multiple-comparison adjustment. A candidate gene association study of multiple smoking-related phenotypes is used to demonstrate the application and advantages of this new method. The proposed method is general enough to be used for the assessment of the joint effect of a set of multiple risk factors on a multivariate outcome in other biomedical research settings.


MMP-9-induced increase in intestinal epithelial tight permeability is mediated by p38 kinase signaling pathway activation of MLCK gene.
Matrix metalloproteinase-9 (MMP-9) has been implicated as being an important pathogenic factor in inflammatory bowel disease (IBD). MMP-9 is markedly elevated in intestinal tissue of patients with IBD, and IBD patients have a defective intestinal tight-junction (TJ) barrier manifested by an increase in intestinal permeability. The loss of intestinal epithelial barrier function is an important contributing factor in the development and prolongation of intestinal inflammation; however, the role of MMP-9 in intestinal barrier function remains unclear. The purpose of this study was to investigate the effect of MMP-9 on the intestinal epithelial TJ barrier and to delineate the intracellular mechanisms involved by using in vitro (filter-grown Caco-2 monolayers) and in vivo (mouse small intestine recycling perfusion) systems. MMP-9 caused a time- and dose-dependent increase in Caco-2 TJ permeability. MMP-9 also caused an increase in myosin light-chain kinase (MLCK) gene activity, protein expression, and enzymatic activity. The pharmacological MLCK inhibition and siRNA-induced knockdown of MLCK inhibited the MMP-9-induced increase in Caco-2 TJ permeability. MMP-9 caused a rapid activation of the p38 kinase signaling pathway and inhibition of p38 kinase activity prevented the MMP-9-induced increase in MLCK gene activity and the increase in Caco-2 TJ permeability. MMP-9 also caused an increase in mouse intestinal permeability in vivo, which was accompanied by an increase in MLCK expression. The MMP-9-induced increase in mouse intestinal permeability was inhibited in MLCK-deficient mice. These data show for the first time that the MMP-9-induced increase in intestinal TJ permeability in vitro and in vivo was mediated by the p38 kinase signal transduction pathway upregulation of MLCK gene activity and that therapeutic targeting of these pathways can prevent the MMP-9-induced increase in intestinal TJ permeability. NEW & NOTEWORTHY MMP-9 is highly elevated in patients with IBD. IBD patients have compromised intestinal TJ barrier function manifested by an increase in intestinal permeability and intestinal inflammation. This study shows that MMP-9, at clinically achievable concentrations, causes an increase in intestinal TJ permeability in vitro and in vivo. In addition, a MMP-9-induced increase in intestinal TJ permeability was mediated by an increase in MLCK gene and protein expression via the p38 kinase pathway.


Autologous Cord Blood Infusions Are Safe and Feasible in Young Children with Autism Spectrum Disorder: Results of a Single-Center Phase I Open-Label Trial.
Despite advances in early diagnosis and behavioral therapies, more effective treatments for children with autism spectrum disorder (ASD) are needed. We hypothesized that umbilical cord blood-derived cell therapies may have potential in alleviating ASD symptoms by modulating inflammatory processes in the brain. Accordingly, we conducted a phase I, open-label trial to assess the safety and feasibility of a single intravenous infusion of autologous umbilical cord blood, as well as sensitivity to change in several ASD assessment tools, to determine suitable endpoints for future trials. Twenty-five children, median age 4.6 years (range 2.26-5.97), with a confirmed diagnosis of ASD and a qualified banked autologous umbilical cord blood unit, were enrolled. Children were evaluated with a battery of behavioral and functional tests immediately prior to cord blood infusion (baseline) and 6 and 12 months later. Assessment of adverse events across the 12-month period indicated that the treatment was safe and well tolerated. Significant improvements in children's behavior were observed on parent-report measures of social communication skills and autism symptoms, clinician ratings of overall autism symptom severity and degree of improvement, standardized measures of expressive vocabulary, and objective eye-tracking measures of children's attention to social stimuli, indicating that these measures may be useful endpoints in future studies. Behavioral improvements were observed during the first 6 months after infusion and were greater in children with higher baseline nonverbal intelligence quotients. These data will serve as the basis for future studies to determine the efficacy of umbilical cord blood infusions in children with ASD. Stem Cells Translational Medicine 2017;6:1332-1339.