Cell cycle-dependent binding of yeast heat shock factor to nucleosomes.
In the nucleus, transcription factors must contend with the presence of chromatin in order to gain access to their cognate regulatory sequences. As most nuclear DNA is assembled into nucleosomes, activators must either invade a stable, preassembled nucleosome or preempt the formation of nucleosomes on newly replicated DNA, which is transiently free of histones. We have investigated the mechanism by which heat shock factor (HSF) binds to target nucleosomal heat shock elements (HSEs), using as our model a dinucleosomal heat shock promoter (hsp82-DeltaHSE1). We find that activated HSF cannot bind a stable, sequence-positioned nucleosome in G(1)-arrested cells. It can do so readily, however, following release from G(1) arrest or after the imposition of either an early S- or late G(2)-phase arrest. Surprisingly, despite the S-phase requirement, HSF nucleosomal binding activity is restored in the absence of hsp82 replication. These results contrast with the prevailing paradigm for activator-nucleosome interactions and implicate a nonreplicative, S-phase-specific event as a prerequisite for HSF binding to nucleosomal sites in vivo.


Optogenetic neuromodulation.
Modulation of the nervous system by electrical or chemical means (neuromodulation) is becoming increasingly sophisticated, with application to a growing number of neurological diseases. However, both chemical and electrical neuromodulation are limited in their specificity. Electrical stimulation, for example, indiscriminately activates different neuronal populations within the electrical field, leading to side effects that can limit efficacy. The delivery of genes that encode proteins capable of conveying light sensitivity to neurons has provided a tool that may overcome some of the limitations of traditional neuromodulation techniques. Activation or inhibition of specific neuronal populations with different wavelengths of light opens up possibilities for modulating neural circuits with previously unimagined levels of precision. We briefly review this new technology, illustrating its advantages and potential applications.


High-throughput screening reveals a small-molecule inhibitor of the renal outer medullary potassium channel and Kir7.1.
The renal outer medullary potassium channel (ROMK) is expressed in the kidney tubule and critically regulates sodium and potassium balance. The physiological functions of other inward rectifying K(+) (Kir) channels expressed in the nephron, such as Kir7.1, are less well understood in part due to the lack of selective pharmacological probes targeting inward rectifiers. In an effort to identify Kir channel probes, we performed a fluorescence-based, high-throughput screen (HTS) of 126,009 small molecules for modulators of ROMK function. Several antagonists were identified in the screen. One compound, termed VU590, inhibits ROMK with submicromolar affinity, but has no effect on Kir2.1 or Kir4.1. Low micromolar concentrations inhibit Kir7.1, making VU590 the first small-molecule inhibitor of Kir7.1. Structure-activity relationships of VU590 were defined using small-scale parallel synthesis. Electrophysiological analysis indicates that VU590 is an intracellular pore blocker. VU590 and other compounds identified by HTS will be instrumental in defining Kir channel structure, physiology, and therapeutic potential.


IGFBP-5 enhances epithelial cell adhesion and protects epithelial cells from TGFβ1-induced mesenchymal invasion.
TGFβ1 is a major fibrotic factor and its actions involve induction of epithelial cell death, together with the stimulation and transdifferentiation of fibroblasts into collagen- and fibronectin-secreting myofibroblasts. These actions of TGFβ1 are also consistent with a pro-metastatic role, by aiding epithelial cell escape through mesenchymal tissues. Recently IGFBP-5 has been described as a pro-fibrotic (pro-metastatic?) agent and the aim of this study was to compare and contrast the actions of IGFBP-5 with TGFβ1. We used NMuMG cells and cloned stable epithelial and mesenchymal lines from the parent cells. TGFβ1 induced apoptosis and/or EMT in the epithelial cells, whereas it enhanced mesenchymal cell survival and migration. IGFBP-5, in contrast, enhanced both cell-cell and cell-ECM adhesion and also improved wound closure in epithelial cells whereas, in mesenchymal cells, IGFBP-5 decreased adhesion and migration. Furthermore, IGFBP-5 was able to antagonise the actions of TGFβ1. In a co-culture model simulating epithelial-mesenchymal boundaries, IGFBP-5 was able to antagonise the disruptive transgressions induced by TGFβ1. Overall, these findings suggest that IGFBP-5 is important in maintaining epithelial-mesenchymal boundaries and thus may limit metastasis and fibrosis by inducing an orderly repair mechanism, very distinct from the fibrotic disruption induced by TGFβ1. A role for IGFBP-5 in the inhibition of metastasis is supported by immunohistochemical studies of breast cancer microarrays, where we show that elevated IGFBP-5 expression is associated with increased disease-free survival.


When hawks give rise to doves: the evolution and transition of enforcement strategies.
The question of how altruism can evolve despite its local disadvantage to selfishness has produced a wealth of theoretical and empirical research capturing the attention of scientists across disciplines for decades. One feature that has remained consistent through this outpouring of knowledge has been that researchers have looked to the altruists themselves for mechanisms by which altruism can curtail selfishness. An alternative perspective may be that just as altruists want to limit selfishness in the population, so may the selfish individuals themselves. These alternative perspectives have been most evident in the fairly recent development of enforcement strategies. Punishment can effectively limit selfishness in the population, but it is not free. Thus, when punishment evolves among altruists, the double costs of exploitation from cheaters and punishment make the evolution of punishment problematic. Here we show that punishment can more readily invade selfish populations when associated with selfishness, whereas altruistic punishers cannot. Thereafter, the establishment of altruism because of enforcement by selfish punishers provides the ideal invasion conditions for altruistic punishment, effectively creating a transition of punishment from selfishness to altruistic. Thus, from chaotic beginnings, a little hypocrisy may go a long way in the evolution and maintenance of altruism.


The GYF domain protein CD2BP2 is critical for embryogenesis and podocyte function.
Scaffolding proteins play pivotal roles in the assembly of macromolecular machines such as the spliceosome. The adaptor protein CD2BP2, originally identified as a binding partner of the adhesion molecule CD2, is a pre-spliceosomal assembly factor that utilizes its glycine-tyrosine-phenylalanine (GYF) domain to co-localize with spliceosomal proteins. So far, its function in vertebrates is unknown. Using conditional gene targeting in mice, we show that CD2BP2 is crucial for embryogenesis, leading to growth retardation, defects in vascularization, and premature death at embryonic day 10.5 when absent. Ablation of the protein in bone marrow-derived macrophages indicates that CD2BP2 is involved in the alternative splicing of mRNA transcripts from diverse origins. At the molecular level, we identified the phosphatase PP1 to be recruited to the spliceosome via the N-terminus of CD2BP2. Given the strong expression of CD2BP2 in podocytes of the kidney, we use selective depletion of CD2BP2, in combination with next-generation sequencing, to monitor changes in exon usage of genes critical for podocyte functions, including VEGF and actin regulators. CD2BP2-depleted podocytes display foot process effacement, and cause proteinuria and ultimately lethal kidney failure in mice. Collectively, our study defines CD2BP2 as a non-redundant splicing factor essential for embryonic development and podocyte integrity.


Non-invasive biomarkers derived from the extracellular matrix associate with response to immune checkpoint blockade (anti-CTLA-4) in metastatic melanoma patients.
Excessive extracellular matrix (ECM) remodeling and a reactive stroma can affect T-cell infiltration and T-cell activity in the tumor and hereby influence response to immune checkpoint inhibitors (ICI). In the pursuit of finding biomarkers that predict treatment response, we evaluated the association between serum biomarkers of collagen and vimentin turnover and outcomes in metastatic melanoma patients treated with the anti-CTLA-4 antibody ipilimumab (IPI). Type III collagen formation (PRO-C3), MMP-degraded type I, type III and type IV collagens (C1M, C3M and C4M), and citrullinated and MMP-degraded vimentin (VICM) were measured with ELISAs in serum from metastatic melanoma patients before (n = 66) and 3 weeks after (n = 52) initiation of IPI treatment. Biomarker levels were associated with Disease Control Rate (DCR) and survival outcomes. We found that baseline levels of PRO-C3 (p = 0.011), C1M (p = 0.003), C3M (p = 0.013) and C4M (p = 0.027) were significantly elevated in patients with progressive disease (PD). Univariate Cox regression analysis identified high PRO-C3 (p = 0.021) and C4M (p = 0.008) as predictors of poor overall survival (OS) and the biomarkers remained significant when evaluated with other covariates (PRO-C3 (p = 0.049) and C4M (p = 0.046)). Multivariate analysis identified VICM as a predictor of longer OS (p = 0.026). Similarly, a high C3M/PRO-C3 ratio predicted for increased OS (p = 0.034). Only C3M (p = 0.003) and VICM (p < 0.0001) increased 3 weeks after treatment. ECM and tissue remodeling quantified in pre-treatment serum were associated with response and survival outcomes in metastatic melanoma patients treated with IPI. This highlights the importance of addressing the ECM and stromal component non-invasively in future ICI studies.


Short-term injection drug use changes following hepatitis C virus (HCV) assessment and treatment among persons who inject drugs with acute HCV infection.
It is unclear whether treatment and care for hepatitis C virus (HCV) infection can help people who inject drugs (PWID) modify their injection drug use behaviours. This study examined changes in injection drug use among PWID with acute HCV systematically referred for HCV clinical assessment and treatment and offered targeted health care services, over the course of one year. The study sample included PWID with documented acute HCV infection recruited and followed-up semi-annually at least twice in IMPACT (2007-2015), a longitudinal community-based prospective study in Montréal, Canada. Following enrolment, participants with contra-indications to treatment due to severe co-morbidity were offered targeted health care services. Pegylated interferon-alpha (12-24 weeks) was offered to all other participants who did not spontaneously resolve their infection. At each study visit, data were collected on socio-demographic factors and drug use patterns. Logistic regression was used to assess changes in injection drug use at one-year follow-up. Of the 87 eligible participants (mean age: 35.6; 78.2% male), 21.8% received treatment [(RT), Sustained virological response: 84.2%], 25.3% spontaneously resolved their infection (SR), 14.9% had contra-indication(s) (CI) and 37.9% chose not to engage in HCV care post-diagnosis (NE). In multivariate analyses adjusting for age, gender and injection drug use at baseline, the RT [Adjusted odds ratio (AOR): 0.18; 95% Confidence interval (CI): 0.04-0.76], SR (AOR: 0.34; 95% CI: 0.08-1.40), and CI (AOR: 0.24; 95% CI: 0.05-1.22) groups were less likely to report injection drug use at follow-up relative to the NE group. PWID who received treatment, spontaneously resolved their infection or presented with treatment contra-indication(s) reported reduced injection drug use at one-year follow-up relative to those who did not engage in therapy. Findings suggest that the benefits of HCV assessment and treatment may extent to helping PWID modify their injection drug use patterns.


Decreased insulin responsiveness of glucose uptake in cultured human skeletal muscle cells from insulin-resistant nondiabetic relatives of type 2 diabetic families.
To investigate the contribution of inherited biochemical defects to the peripheral insulin resistance of type 2 diabetes, we studied cultured skeletal muscle from 10 insulin-resistant nondiabetic first-degree relatives of type 2 diabetic families and 6 control subjects. Insulin stimulation of glucose uptake and glycogen synthesis was maximal in myoblasts. Insulin-stimulated glucose uptake (fold-stimulation over basal uptake) was decreased in relative compared with control myoblasts at 0.001 micromol/l (0.93 +/- 0.05 [mean +/- SE] vs. 1.15 +/- 0.06, P < 0.05) and 0.1 micromol/l (1.38 +/- 0.10 vs. 1.69 +/- 0.08, P = 0.025) insulin. Insulin responsiveness was markedly impaired in 5 of the relative myoblast cultures, and in 4 of these, there was an associated increase in basal glucose uptake (76.7 +/- 7.0 vs. 47.4 +/- 5.5 pmol x min(-1) x mg(-1) protein, relative vs. control; P < 0.02). Expression of insulin receptor substrate 1, phosphatidylinositol 3-kinase, protein kinase B, and glycogen synthase was normal in the relative cultures with impaired insulin responsiveness. Glycogen synthesis was also normal in the relative cultures. We conclude that the persistence of impaired insulin responsiveness in some of the relative cultures supports the role of inherited factors in the insulin resistance of type 2 diabetes and that the association with increased basal glucose uptake suggests that the 2 abnormalities may be linked.


Geographic cline analysis as a tool for studying genome-wide variation: a case study of pollinator-mediated divergence in a monkeyflower.
A major goal of speciation research is to reveal the genomic signatures that accompany the speciation process. Genome scans are routinely used to explore genome-wide variation and identify highly differentiated loci that may contribute to ecological divergence, but they do not incorporate spatial, phenotypic or environmental data that might enhance outlier detection. Geographic cline analysis provides a potential framework for integrating diverse forms of data in a spatially explicit framework, but has not been used to study genome-wide patterns of divergence. Aided by a first-draft genome assembly, we combined an FCT scan and geographic cline analysis to characterize patterns of genome-wide divergence between divergent pollination ecotypes of Mimulus aurantiacus. FCT analysis of 58 872 SNPs generated via RAD-seq revealed little ecotypic differentiation (mean FCT = 0.041), although a small number of loci were moderately-to-highly diverged. Consistent with our previous results from the gene MaMyb2, which contributes to differences in flower colour, 130 loci have cline shapes that recapitulate the spatial pattern of trait divergence, suggesting that they may reside in or near the genomic regions that contribute to pollinator isolation. In the narrow hybrid zone between the ecotypes, extensive admixture among individuals and low linkage disequilibrium between markers indicate that most outlier loci are scattered throughout the genome, rather than being restricted to one or a few divergent regions. In addition to revealing the genomic consequences of ecological divergence in this system, we discuss how geographic cline analysis is a powerful but under-utilized framework for studying genome-wide patterns of divergence.


A disruption in iron-sulfur center biogenesis via inhibition of mitochondrial dithiol glutaredoxin 2 may contribute to mitochondrial and cellular iron dysregulation in mammalian glutathione-depleted dopaminergic cells: implications for Parkinson's disease.
Parkinson's disease (PD) is characterized by early glutathione depletion in the substantia nigra (SN). Among its various functions in the cell, glutathione acts as a substrate for the mitochondrial enzyme glutaredoxin 2 (Grx2). Grx2 is involved in glutathionylation of protein cysteine sulfhydryl residues in the mitochondria. Although monothiol glutathione-dependent oxidoreductases (Grxs) have previously been demonstrated to be involved in iron-sulfur (Fe-S) center biogenesis, including that in yeast, here we report data suggesting the involvement of mitochondrial Grx2, a dithiol Grx, in iron-sulfur biogenesis in a mammalian dopaminergic cell line. Given that mitochondrial dysfunction and increased cellular iron levels are two important hallmarks of PD, this suggests a novel potential mechanism by which glutathione depletion may affect these processes in dopaminergic neurons. We report that depletion of glutathione as substrate results in a dose-dependent Grx2 inhibition and decreased iron incorporation into a mitochondrial complex I (CI) and aconitase (m-aconitase). Mitochondrial Grx2 inhibition through siRNA results in a corresponding decrease in CI and m-aconitase activities. It also results in significant increases in iron-regulatory protein (IRP) binding, likely as a consequence of conversion of Fe-S-containing cellular aconitase to its non-Fe-S-containing IRP1 form. This is accompanied by increased transferrin receptor, decreased ferritin, and subsequent increases in mitochondrial iron levels. This suggests that glutathione depletion may affect important pathologic cellular events associated with PD through its effects on Grx2 activity and mitochondrial Fe-S biogenesis.


Relapse according to antipsychotic treatment in schizophrenic patients: a propensity-adjusted analysis.
To compare the rate of relapse as a function of antipsychotic treatment (monotherapy vs. polypharmacy) in schizophrenic patients over a 2-year period. Using data from a multicenter cohort study conducted in France, we performed a propensity-adjusted analysis to examine the association between the rate of relapse over a 2-year period and antipsychotic treatment (monotherapy vs. polypharmacy). Our sample consisted in 183 patients; 50 patients (27.3%) had at least one period of relapse and 133 had no relapse (72.7%). Thirty-eight (37.7) percent of the patients received polypharmacy. The most severely ill patients were given polypharmacy: the age at onset of illness was lower in the polypharmacy group (p = 0.03). Patients that received polypharmacy also presented a higher general psychopathology PANSS subscore (p = 0.04) but no statistically significant difference was found in the PANSS total score or the PANSS positive or negative subscales. These patients were more likely to be given prescriptions for sedative drugs (p < 0.01) and antidepressant medications (p = 0.03). Relapse was found in 23.7% of patients given monotherapy and 33.3% given polypharmacy (p = 0.16). After stratification according to quintiles of the propensity score, which eliminated all significant differences for baseline characteristics, antipsychotic polypharmacy was not statistically associated with an increase of relapse: HR = 1.686 (0.812; 2.505). After propensity score adjustment, antipsychotic polypharmacy is not statistically associated to an increase of relapse. Future randomised studies are needed to assess the impact of antipsychotic polypharmacy in schizophrenia.


Aphid transmission studies using helper component proteins of potato virus Y expressed from a vector derived from potato virus X.
The genes encoding the helper component (HC) proteins of two strains of Potato virus Y (PVY) were cloned and the proteins expressed from a vector derived from Potato virus X (PVX). The expressed HC contained six N-terminal histidine residues to facilitate purification by metal affinity chromatography. Approximately 2-4 microg/g of purified HC was obtained from leaves of Nicotiana benthamiana plants systemically infected by recombinant PVX. Preparations of the HC protein derived from PVY ordinary strain (PVY(o)) assisted aphid transmission of purified particles of PVY(o) and PVY strain C (PVY(c); a naturally occurring non-aphid transmissible strain of PVY which contains a defective HC), as well as Potato aucuba mosaic virus. The HC derived from PVY(c) contained the Glu-Ile-Thr-Cys (EITC) motif, and mutation of Glu (E) to Lys (K) enabled the mutant PVX-expressed preparations to assist virus transmission by aphids. Expression of HC protein from the PVX vector produced biologically active protein. This approach should facilitate further studies to elucidate more precisely the molecular mechanism of virus transmission by aphids.


Subcuticular bacteria associated with two common New Zealand echinoderms: Characterization using 16S rRNA sequence analysis and fluorescence in situ hybridization.
Many echinoderms contain subcuticular bacteria (SCB), symbionts which reside in the lumen between the host's epidermal cells and outer cuticle. This relationship is common, existing in about 60% of echinoderms studied so far, yet the function of SCB remains largely unknown. In this study, phylogenetic analysis was carried out on 16S rRNA sequences obtained from echinoderm-associated bacteria, resulting in the identification of four species of putative SCB. All four bacteria were identified from the holothurian Stichopus mollis, and two of the four were also found in the asteroid Patiriella sp. Two of these bacteria belong to the Alphaproteobacteria, and two to the Gammaproteobacteria. In addition to phylogenetic analysis, fluorescence in situ hybridization (FISH) assays were carried out on Patiriella sp., S. mollis, and the asteroid Astrostole scabra. Results showed that Patiriella sp. and S. mollis contain SCB, in agreement with the phylogenetic analysis, while SCB were not detected in A. scabra. Of the bacteria detected using FISH, more than 80% were recognized as belonging to the Alphaproteobacteria in both host species. However, in S. mollis about 20% of the detected SCB successfully hybridized with the Gammaproteobacteria-specific probe, whereas bacteria belonging to this class were never observed in Patiriella sp. This is only the second study to characterize SCB by molecular means, and is the first to identify SCB in situ using FISH.


Investigating the resonance spectrum of optical frequency combs in fiber-optic cavities.
We report a detailed theoretical and experimental study of fiber-optic cavities under broadband excitation by mode-locked laser combs. We calculate the effects of fiber dispersion on the cavity transmission. For any integer ratio between the comb repetition rate and cavity free spectral range, the theoretical resonant output spectrum exhibits a narrow group of resonant teeth, surrounded by minor, unevenly spaced resonances. Also, the central resonance can be rapidly and precisely tuned over the entire comb span by only acting on its repetition rate. Experimental observations are provided by a single-mode fiber ring and a telecom-wavelength comb laser. The resulting spectral pattern agrees very well with our theoretical prediction, allowing a thorough characterization of the cavity dispersion and opening new perspectives for comb spectroscopy in dielectric resonators.


[Validation of prolonged grief disorder instrument for Portuguese population].
This study aims to validate the Portuguese population the instrument PG-13 (Prolonged Grief Disorder), created by Prigerson et al. (2007) for diagnosis of prolonged grief, whose criteria are: the experience of loss generating intense longing and yearning for the deceased that extends for a period exceeding six months; emotional, cognitive and behavioral symptoms, dysfunction and meaningful life social and occupational functioning. The population includes 102 caregivers of patients accompanied by Support Team Palliative Care, Hospital Santa Maria. The participants are mostly female (82.4%) with mean age of 58.87 (SD: 13.41) and range between 15 and 84 years. Most respondents are widowed (62.1%), and 93.2% of these people are mourning the loss of a spouse. The second largest group of subjects corresponds to married persons (29.5%) who lost one of the parental figures (64.3%) and brothers (14.3%). Deceased family members have an average age of 68.68 (SD: 11.50), with amplitude between 27 and 89 years. The gender distribution in the group of deceased patients are 57.8% male and 42.2 % female. The internal consistency in the instrument is considered very good (a=.932). We found that 22.5% of the population manifests symptoms of prolonged grief. There were no significant differences in terms of socio-demographic variables or in the circumstances of illness and death. Prolonged Grief Disorder is more prevalent in female subjects (91.3%), widowed (68.2%) and in cases where the deceased was being the spouse (65.2%).


Baicalin administration attenuates hyperglycemia-induced malformation of cardiovascular system.
In this study, the effects of Baicalin on the hyperglycemia-induced cardiovascular malformation during embryo development were investigated. Using early chick embryos, an optimal concentration of Baicalin (6 μM) was identified which could prevent hyperglycemia-induced cardiovascular malformation of embryos. Hyperglycemia-enhanced cell apoptosis was reduced in embryos and HUVECs in the presence of Baicalin. Hyperglycemia-induced excessive ROS production was inhibited when Baicalin was administered. Analyses of SOD, GSH-Px, MQAE and GABAA suggested Baicalin plays an antioxidant role in chick embryos possibly through suppression of outwardly rectifying Cl(-) in the high-glucose microenvironment. In addition, hyperglycemia-enhanced autophagy fell in the presence of Baicalin, through affecting the ubiquitin of p62 and accelerating autophagy flux. Both Baicalin and Vitamin C could decrease apoptosis, but CQ did not, suggesting autophagy to be a protective function on the cell survival. In mice, Baicalin reduced the elevated blood glucose level caused by streptozotocin (STZ). Taken together, these data suggest that hyperglycemia-induced embryonic cardiovascular malformation can be attenuated by Baicalin administration through suppressing the excessive production of ROS and autophagy. Baicalin could be a potential candidate drug for women suffering from gestational diabetes mellitus.


Managing and preventing outbreaks of Gram-negative infections in UK neonatal units.
De novo guidance on the management of Gram-negative bacteria outbreaks in UK neonatal units was developed in 2012 by a Department of Health, England Antimicrobial Resistance and Healthcare Associated Infection working group. The recommendations included activation of an organisational response and establishing a control team when an outbreak is suspected; screening for the specific organism only during an outbreak; undertaking multidisciplinary reviews of cleaning routines, hand hygiene and Gram-negative bacteria transmission risks; considering deep-cleaning; cohorting colonised and infected babies preferably but not necessarily in isolation cubicles; and considering reducing beds or closing a unit to new admissions as a way of improving spacing and staff:patient ratios until the outbreak is under control. The group advised establishing mechanisms to communicate effectively across the network; informing parents of the outbreak as early as possible, and providing prewritten 'infection outbreak' information sheets. For prevention of outbreaks, the group advised meeting national staffing and cot-spacing requirements; following a Water Action Plan; using infection reduction care bundles and benchmarking; and introducing breast milk early and limiting antibiotic use.


Discrete-ordinated finite-element method for atmospheric radiative transfer and remote sensing.
Advantages and disadvantages of modern discrete-ordinates finite-element methods for the solution of radiative transfer problems in meteorology, climatology, and remote sensing applications are evaluated. After the common basis of the formulation of radiative transfer problems in the fields of neutron transport and atmospheric optics is established, the essential features of the discrete-ordinates finite-element method are described including the limitations of the method and their remedies. Numerical results are presented for 1-D and 2-D atmospheric radiative transfer problems where integral as well as angular dependent quantities are compared with published results from other calculations and with measured data. These comparisons provide a verification of the discrete-ordinates results for a wide spectrum of cases with varying degrees of absorption, scattering, and anisotropic phase functions. Accuracy and computational speed are also discussed. Since practically all discrete-ordinates codes offer a builtin adjoint capability, the general concept of the adjoint method is described and illustrated by sample problems. Our general conclusion is that the strengths of the discrete-ordinates finite-element method outweight its weaknesses. We demonstrate that existing general-purpose discrete-ordinates codes can provide a powerful tool to analyze radiative transfer problems through the atmosphere, especially when 2-D geometries must be considered.


Genome-wide association study of toxic metals and trace elements reveals novel associations.
The accumulation of toxic metals in the human body is influenced by exposure and mechanisms involved in metabolism, some of which may be under genetic control. This is the first genome-wide association study to investigate variants associated with whole blood levels of a range of toxic metals. Eleven toxic metals and trace elements (aluminium, cadmium, cobalt, copper, chromium, mercury, manganese, molybdenum, nickel, lead and zinc) were assayed in a cohort of 949 individuals using mass spectrometry. DNA samples were genotyped on the Infinium Omni Express bead microarray and imputed up to reference panels from the 1000 Genomes Project. Analyses revealed two regions associated with manganese level at genome-wide significance, mapping to 4q24 and 1q41. The lead single nucleotide polymorphism (SNP) in the 4q24 locus was rs13107325 (P-value = 5.1 × 10(-11), β = -0.77), located in an exon of SLC39A8, which encodes a protein involved in manganese and zinc transport. The lead SNP in the 1q41 locus is rs1776029 (P-value = 2.2 × 10(-14), β = -0.46). The SNP lies within the intronic region of SLC30A10, another transporter protein. Among other metals, the loci 6q14.1 and 3q26.32 were associated with cadmium and mercury levels (P = 1.4 × 10(-10), β = -1.2 and P = 1.8 × 10(-9), β = -1.8, respectively). Whole blood measurements of toxic metals are associated with genetic variants in metal transporter genes and others. This is relevant in inferring metabolic pathways of metals and identifying subsets of individuals who may be more susceptible to metal toxicity.


Phosphorescent cellular probes and uptake indicators derived from cyclometalated iridium(III) bipyridine complexes appended with a glucose or galactose entity.
A series of phosphorescent cyclometalated iridium(III) polypyridine complexes appended with a β-D-glucose moiety [Ir(N^C)2(bpy-TEG-ONCH3-β-D-glc)](PF6) [bpy-TEG-ONCH3-β-D-glc = 4-(10-N-methyl-N-(β-D-glucopyranosyl)-amino-oxy-2,5,8-trioxa-dec-1-yl)-4'-methyl-2,2'-bipyridine; HN^C = 2-((1,1'-biphenyl)-4-yl)benzothiazole) (Hbt) (1a), 2-phenylpyridine (Hppy) (2a), 2-phenylquinoline (Hpq) (3a), 7,8-benzoquinoline (Hbzq) (4a)] has been synthesized and characterized. The D-galactose counterparts [Ir(N^C)2(bpy-TEG-ONCH3-β-D-gal)](PF6) [bpy-TEG-ONCH3-β-D-gal = 4-(10-N-methyl-N-(β-D-galactopyranosyl)-amino-oxy-2,5,8-trioxa-dec-1-yl)-4'-methyl-2,2'-bipyridine; HN^C = Hbt (1b), Hppy (2b), Hpq (3b), Hbzq (4b)] and a sugar-free bt complex [Ir(bt)2(bpy-TEG-OMe)](PF6) [bpy-TEG-OMe = 4-(2,5,8,11-tetraoxa-dodec-1-yl)-4'-methyl-2,2'-bipyridine] (1c) have also been prepared. Upon photoexcitation, all the complexes displayed intense and long-lived triplet metal-to-ligand charge-transfer ((3)MLCT) [dπ(Ir) → π*(N^N)] or triplet intraligand ((3)IL) (π → π*) (N^C and N^N) emission. The lipophilicity, the cellular uptake efficiency, and cytotoxicity of the complexes toward human cervix epithelioid carcinoma cells (HeLa) have been examined. Temperature dependence and chemical inhibition experiments indicated that the transport of bt-glucose complex 1a across the cell membrane occurred through an energy-requiring process such as endocytosis, in additional to a pathway that was mediated by glucose transporters (GLUTs). Importantly, the cellular uptake efficiency of this complex was found to be strongly dependent on hormonal stimulation and inhibition, rendering it a new phosphorescent metabolic indicator. Additionally, laser-scanning confocal microscopy revealed that the complex was localized in the mitochondria and highly resistant to photobleaching compared to a fluorescent organic glucose derivative 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxy-d-glucose (2-NBDG).


Treatment of Salmonella gastroenteritis with ampicillin, amoxicillin, or placebo.
In a randomized, prospective, double-blind study infants and children with uncomplicated Salmonella gastroenteritis were treated with ampicillin (15 patients), amoxicillin (15 patients), or placebo (14 patients). The dosage of antibiotics was 100 mg/kg/day in four equally divided doses given for five days. There was no significant benefit from antibiotic therapy on the duration of diarrhea (means 8.8, 7.3, and 7.2 days, respectively) or on the duration of recovery of Salmonella from stool cultures (means 41.3, 37.0, and 20.9 days, respectively). Bacteriologic relapse was not observed in placebo-treated patients but eight patients given ampicillin (53%) and eight given amoxicillin (53%) had relapse (P = .003). Salmonella isolated in relapse were still susceptible in vitro to the antibiotics. Of the 16 patients with bacteriologic relapse six (38%) had concomitant recurrence of diarrhea. It is concluded that ampicillin or amoxicillin therapy provides no benefit to patients with uncomplicated Salmonella gastroenteritis and substantially increases the risk of bacteriologic and symptomatic relapse.


[Gradually increasing apathy and self-neglect in a 29-year-old man].
A 29-year-old man presented with progressive behavioural changes, expressed mainly in the form of increasing of apathy and self-neglect. The initial differential diagnosis included a psychotic disorder, a mood disorder or a personality disorder. After a month of medication-free observation and uncertainty regarding the results of various diagnostic approaches, we decided to use MR-imaging; this revealed a frontal meningioma which had invaded the entire frontal lobe. Although patients with some types of somatic disorders frequently present with psychiatric symptoms, there are often indications (e.g. from history or physical examination) that in fact the symptoms are of organic origin. In this case report we discuss the indications that should lead clinicians and psychiatrists to consider the possibility of organic pathology when young adults present with only psychiatric symptoms.


Role of HIV and CMV in the pathogenesis of retinitis and retinal vasculopathy in AIDS patients.
Cotton-wool spots and cytomegalovirus (CMV) retinitis are seen frequently in AIDS patients. Human immunodeficiency virus (HIV) infection of the retina has been proposed as a mechanism for the high incidence of retinal pathology. An autopsy study of the eyes from 25 consecutive cases of AIDS was performed using gross examination, light microscopy, trypsin digestion of retinal vasculatures, and immunohistochemistry to evaluate the possible role of HIV, as well as CMV, in the pathogenesis of retinitis and retinal vasculopathy. Brain tissue was studied in the first 20 of these cases to evaluate any correlation between retinal and central nervous system pathology. CMV retinitis was observed in 15 cases (60%). Cotton-wool spots were seen in nine cases (36%). CMV encephalitis was detected in four cases, whereas HIV encephalitis was noted in five cases. We were unable to demonstrate a correlation between CMV retinitis and CMV encephalitis. However, the number of cases studied was small, and the frequency of CMV encephalitis was low. On the other hand, bilateral CMV retinitis demonstrated a correlation to HIV encephalitis (P less than 0.005, Fisher's exact test). HIV infection of the retina was not detected by typical morphologic changes or immunohistochemistry. Immunohistochemistry localized CMV infection solely to areas of active retinitis. These findings suggest that bilateral CMV may serve as a marker of HIV encephalitis, possibly indicating a severely immunodepressed state.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of trabeculectomy on visual field progression in Japanese progressive normal-tension glaucoma with intraocular pressure < 15 mmHg.
We examined the effectiveness of trabeculectomy in decreasing the slope of mean deviation (MD) in Japanese patients with progressive normal-tension glaucoma (NTG) at low intraocular pressure (IOP) levels. The charts of patients who had undergone initial trabeculectomy with adjunctive mitomycin C for progressive NTG with medically controlled IOP < 15 mmHg in 2010-2013 were retrospectively reviewed. Seventeen eyes of 13 NTG patients who had undergone at least 5 times of visual field (VF) examinations in both of preoperatively and postoperatively with postoperative follow-up of ≥ 2 years were enrolled. Preoperative and postoperative MD slopes were compared to evaluate the effectiveness of trabeculectomy in slowing progression of VF. Mean IOP (8.1 ± 2.9 mmHg) and number of IOP-lowering medications (0.8 ± 1.5) were significantly lower postoperatively than preoperatively (13.9 ± 0.9 mmHg; P < 0.001 and 3.0± 0.4; P < 0.0001). In total, 91.7% of eyes with single-digit IOP postoperatively showed improvement in MD slope, whereas only 20.0% of eyes with IOP ≥ 10 mmHg postoperatively showed the improvement. Three eyes (17.6%) showed a decrease in visual acuity (VA) of ≥ 0.1 unit; this group had a lower mean postoperative IOP (6.0 ± 1.0 vs. 8.6 ± 3.0 mmHg; P = 0.1717) and a higher mean IOP reduction rate (56.2 vs. 38.5%; P = 0.8296) than eyes with a VA decrease of < 0.1 unit or no change. Thus, in this analysis of Japanese NTG patients with medically controlled IOP < 15 mmHg, achieving an IOP < 10 mmHg with trabeculectomy was beneficial for reducing the VF progression rate in progressive NTG at low IOP levels. However, an IOP < 7 mmHg by surgery would be required careful attention to VA decline.


A highly selective, colorimetric, and environment-sensitive optical potassium ion sensor.
Potassium ions (K+) play vital roles in many biological processes and thus highly selective sensors for K+ are critical for disease diagnosis and health monitoring. Herein, we report a colorimetric K+ sensor (KS7) in which a hemicyanine dye was used as a fluorophore and phenylaza-[18]crown-6 lariat ether (ACLE) was utilized as a K+ ligand. The maximum absorption peak of KS7 shifted hypsochromically by 77 nm (from 515 to 438 nm) with an isosbestic point at 452 nm upon the addition of K+ to its aqueous solution accompanied by a color change from red to yellow. This sensor exhibited a linear response range to K+ from 1 to 200 mM, indicating its wide detection range for cellular, urinary, and environmental potassium ions. Further, this sensor is solvent-sensitive, implying its environmental sensitivity. For the demonstration of its applications, we prepared filter paper-based K+ test strips, which were used to detect K+ in urine conveniently.


Frankenrigs: building character rigs from multiple sources.
We present a new rigging and skinning method which uses a database of partial rigs extracted from a set of source characters. Given a target mesh and a set of joint locations, our system can automatically scan through the database to find the best-fitting body parts, tailor them to match the target mesh, and transfer their skinning information onto the new character. For the cases where our automatic procedure fails, we provide an intuitive set of tools to fix the problems. When used fully automatically, the system can generate results of much higher quality than a standard smooth bind, and with some user interaction, it can create rigs approaching the quality of artist-created manual rigs in a small fraction of the time.


Laparoscopic excision of a large retroperitoneal lymphovascular malformation in an adult.
Retroperitoneal lymphangioma is a rare benign tumour of the retroperitoneal lymphatics that usually manifests in infancy. It is worth reporting of an unexpected presentation, especially in an adult. They frequently affect the neck (75%) and the axilla (20%). Intra-abdominal lymphangiomas (<5%) have been reported in the mesentery, gastrointestinal tract, spleen, liver and pancreas. Retroperitoneal lymphangiomas account for nearly 1% of all lymphangiomas and are uncommon incidental findings usually at surgery, autopsy or lymphography. Differentiating cystic lymphangiomas from other cystic growths by imaging studies alone is often inconclusive, and surgery is frequently required for definitive diagnosis. An interesting and rare case of a retroperitoneal lymphangioma in an adult patient which was removed laparoscopically is described here.


Considerations for the mitigation of nitrate contamination: stable isotopes and insights into the importance of soil processes.
Nutrient management is widely promoted to minimize the impact of intensive fertilizer use on groundwater quality, however watershed-scale stable isotope studies in eastern North America suggest nitrogen transport to groundwater is dominated by non-growing season fluxes derived principally from the mineralization and nitrification of soil organic matter. In the current field scale study, delta15N ratios of nitrate in tile drain effluents from experimental potato plots treated with 300 kg/ha ammonia nitrate and those with no fertilizer both average +4.7 per thousand, close to the +4.0 per thousand ratios observed in soils of the same plots, and distinct from values near 0 per thousand for inorganic fertilizer. A source apportionment model using delta15N and delta18O in nitrate suggests that even with heavy fertilizer application, less than 10% of non-growing season N flux is derived from direct leaching of fertilizer, the remainder representing N from various sources, including residual fertilizer that has been assimilated into the broader soil organic matter pool and subsequently released via mineralization and nitrification. Factors controlling these losses could be as closely related to cropping practices as initial N application rates, providing potential opportunities for more efficiently utilizing N available in the soil profile and reducing initial N application rates.


The application of network label propagation to rank biomarkers in genome-wide Alzheimer's data.
Ranking and identifying biomarkers that are associated with disease from genome-wide measurements holds significant promise for understanding the genetic basis of common diseases. The large number of single nucleotide polymorphisms (SNPs) in genome-wide studies (GWAS), however, makes this task computationally challenging when the ranking is to be done in a multivariate fashion. This paper evaluates the performance of a multivariate graph-based method called label propagation (LP) that efficiently ranks SNPs in genome-wide data. The performance of LP was evaluated on a synthetic dataset and two late onset Alzheimer's disease (LOAD) genome-wide datasets, and the performance was compared to that of three control methods. The control methods included chi squared, which is a commonly used univariate method, as well as a Relief method called SWRF and a sparse logistic regression (SLR) method, which are both multivariate ranking methods. Performance was measured by evaluating the top-ranked SNPs in terms of classification performance, reproducibility between the two datasets, and prior evidence of being associated with LOAD.On the synthetic data LP performed comparably to the control methods. On GWAS data, LP performed significantly better than chi squared and SWRF in classification performance in the range from 10 to 1000 top-ranked SNPs for both datasets, and not significantly different from SLR. LP also had greater ranking reproducibility than chi squared, SWRF, and SLR. Among the 25 top-ranked SNPs that were identified by LP, there were 14 SNPs in one dataset that had evidence in the literature of being associated with LOAD, and 10 SNPs in the other, which was higher than for the other methods. LP performed considerably better in ranking SNPs in two high-dimensional genome-wide datasets when compared to three control methods. It had better performance in the evaluation measures we used, and is computationally efficient to be applied practically to data from genome-wide studies. These results provide support for including LP in the methods that are used to rank SNPs in genome-wide datasets.


Water dynamics--the effects of ions and nanoconfinement.
Hydrogen bond dynamics of water in highly concentrated NaBr salt solutions and reverse micelles are studied using ultrafast 2D-IR vibrational echo spectroscopy and polarization-selective IR pump-probe experiments performed on the OD hydroxyl stretch of dilute HOD in H(2)O. The vibrational echo experiments measure spectral diffusion, and the pump-probe experiments measure orientational relaxation. Both experimental observables are directly related to the structural dynamics of water's hydrogen bond network. The measurements performed on NaBr solutions as a function of concentration show that the hydrogen bond dynamics slow as the NaBr concentration increases. The most pronounced change is in the longest time scale dynamics which are related to the global rearrangement of the hydrogen bond structure. Complete hydrogen bond network randomization slows by a factor of approximately 3 in approximately 6 M NaBr solution compared to that in bulk water. The hydrogen bond dynamics of water in nanoscopically confined environments are studied by encapsulating water molecules in ionic head group (AOT) and nonionic head group (Igepal CO 520) reverse micelles. Water dynamics in the nanopools of AOT reverse micelles are studied as a function of size by observing orientational relaxation. Orientational relaxation dynamics deviate significantly from bulk water when the size of the reverse micelles is smaller than several nm and become nonexponential and slower as the size of the reverse micelles decreases. In the smallest reverse micelles, orientational relaxation (hydrogen bond structural randomization) is almost 20 times slower than that in bulk water. To determine if the changes in dynamics from bulk water are caused by the influence of the ionic head groups of AOT or the nanoconfinement, the water dynamics in 4 nm nanopools in AOT reverse micelles (ionic) and Igepal reverse micelles (nonionic) are compared. It is found that the water orientational relaxation in the 4 nm diameter nanopools of the two types of reverse micelles is almost identical, which indicates that confinement by an interface to form a nanoscopic water pool is a primary factor governing the dynamics of nanoscopic water rather than the presence of charged groups at the interface.


Genetic and environmental effects on vocal symptoms and their intercorrelations.
Recently, Simberg et al. (2009) found genetic effects on a composite variable consisting of 6 vocal symptom items measuring dysphonia. The purpose of the present study was to determine genetic and environmental effects on the individual vocal symptoms in a population-based sample of Finnish twins. The sample comprised 1,728 twins (125 monozygotic and 108 dizygotic twin pairs) born between 1961 and 1989, who completed a questionnaire concerning 6 vocal symptoms. Values for additive genetic, dominant genetic, shared environmental, and nonshared environmental components were computed separately for all symptoms. Multivariate analyses to determine genetic and environmental associations between the vocal symptoms were also performed. Variance was explained by significant additive genetic effects (27%) in only one of the vocal symptoms, namely, voice gets low or hoarse, whereas the variance of one of the vocal symptoms, voice gets strained or tires, could be explained by nonshared environmental influence alone. Multivariate analyses showed that the correlations for most of the symptom combinations were significant. Both genetic and environmental components influence vocal symptoms. Genetic and environmental influences seem to be differently balanced in different vocal symptoms. Genetic effects are moderate, whereas environmental effects seem to be the most important factor contributing to the presence of vocal symptoms.


Primary care REFerral for EchocaRdiogram (REFER) in heart failure: a diagnostic accuracy study.
Symptoms of breathlessness, fatigue, and ankle swelling are common in general practice but deciding which patients are likely to have heart failure is challenging. To evaluate the performance of a clinical decision rule (CDR), with or without N-Terminal pro-B type natriuretic peptide (NT-proBNP) assay, for identifying heart failure. Prospective, observational, diagnostic validation study of patients aged >55 years, presenting with shortness of breath, lethargy, or ankle oedema, from 28 general practices in England. The outcome was test performance of the CDR and natriuretic peptide test in determining a diagnosis of heart failure. The reference standard was an expert consensus panel of three cardiologists. Three hundred and four participants were recruited, with 104 (34.2%; 95% confidence interval [CI] = 28.9 to 39.8) having a confirmed diagnosis of heart failure. The CDR+NT-proBNP had a sensitivity of 90.4% (95% CI = 83.0 to 95.3) and specificity 45.5% (95% CI = 38.5 to 52.7). NT-proBNP level alone with a cut-off <400 pg/ml had sensitivity 76.9% (95% CI = 67.6 to 84.6) and specificity 91.5% (95% CI = 86.7 to 95.0). At the lower cut-off of NT-proBNP <125 pg/ml, sensitivity was 94.2% (95% CI = 87.9 to 97.9) and specificity 49.0% (95% CI = 41.9 to 56.1). At the low threshold of NT-proBNP <125 pg/ml, natriuretic peptide testing alone was better than a validated CDR+NT-proBNP in determining which patients presenting with symptoms went on to have a diagnosis of heart failure. The higher NT-proBNP threshold of 400 pg/ml may mean more than one in five patients with heart failure are not appropriately referred. Guideline natriuretic peptide thresholds may need to be revised.


Cryopreservation affects bovine sperm intracellular parameters associated with capacitation and acrosome exocytosis.
Although semen cryopreservation is widely and commonly used in the bovine breeding industry, half the spermatozoa do not survive and most of those that do survive undergo numerous physiological changes that affect their fertilising ability. The aim of the present study was to determine how cryopreservation affects the intracellular events involved in sperm capacitation and acrosome reaction. Immediately after thawing and washing, almost 50% of spermatozoa were capacitated and more than 20% had lost their acrosome. The sperm cAMP concentration was lower than that in freshly ejaculated spermatozoa, but the cytosolic pH (pHcyt) was in the expected range. The free cytosolic Ca2+ concentration ([Ca2+]cyt) was higher than in fresh spermatozoa and cryopreserved spermatozoa had internally stored Ca2+. Phenylarsine oxide increased pHcyt and both cytosolic and stored Ca2+ concentrations, whereas orthovanadate enhanced acrosome loss and protein tyrosine phosphorylation (P-Tyr). Heparin increased the percentage of spermatozoa expressing the B (capacitated) chlortetracycline binding pattern, pHcyt, P-Tyr and Ca2+ storage. Moreover, positive correlations exist between capacitation, cAMP, P-Tyr and stored Ca2+, whereas the acrosome reaction is positively correlated with pHcyt and [Ca2+]cyt. These results demonstrate that sperm regulatory mechanisms may be affected by the cryopreservation procedure, but frozen-thawed sperm can still regulate their capacitation and acrosome reaction signalling pathways.


Methylthioadenosine Phosphorylase and Breast Cancer 1 Protein-Associated Protein 1 as Biomarkers for the Peritoneal Mesothelioma.
Combination of Breast Cancer 1 protein-associated protein 1 (BAP1) and methylthioadenosine phosphorylase (MTAP) in the peritoneal mesothelioma (PeM) has yet to be explored. We aim to assess the diagnostic value of combined BAP1 and MTAP to distinguish biphasic mesothelioma (BM) from epithelioid mesothelioma (EM) with reactive stroma in peritoneum, as well as its prognostic value in PeM. This is a retrospective study from June 2014 to December 2021. This study included 18 cases of BM and 27 cases of EM with reactive stroma, excluded sarcomatoid, and EM without reactive stroma cases, and clinicopathological information was collected. The associations between MTAP and BAP1 levels and clinicopathological features or prognosis were analyzed. Clinical follow-up data were reviewed to correlate with pathological prognostic factors using Kaplan-Meier estimator and univariate/multivariate Cox proportional hazards regression models. Loss/decrease of BAP1/MTAP was observed in 6 (33.3%) BM cases and 12 (44.4%) EM cases. In 5 (27.8%) cases, loss of or decreased BAP1/MTAP expression was observed in both EC and SC of BM. BAP1/MTAP loss/decrease was observed in 12 (44.4%) cases of only EC of EM but not in reactive stroma. Compared with histology alone, a combination of BAP1 and MTAP immunohistochemistry (IHC) in spindled PeM provides a more objective mean to distinguish BM from EM with reactive stroma. Loss/decrease of BAP1/MTAP was associated with peritoneal cancer index (PCI) score (P = 0.047) and completeness of cytoreduction (CC) score (P = 0.038). BM patients have worse overall survival (OS) than EM with reactive stroma (P = 0 .007). Combination of BAP1/MTAP by IHC is helpful for differential diagnosis of peritoneal BM from EM with reactive stroma. Nevertheless, BAP1/MTAP may help to evaluate the biological behavior of PeM.


Efficient stochastic sampling of first-passage times with applications to self-assembly simulations.
Models of reaction chemistry based on the stochastic simulation algorithm (SSA) have become a crucial tool for simulating complicated biological reaction networks due to their ability to handle extremely complicated networks and to represent noise in small-scale chemistry. These methods can, however, become highly inefficient for stiff reaction systems, those in which different reaction channels operate on widely varying time scales. In this paper, we develop two methods for accelerating sampling in SSA models: an exact method and a scheme allowing for sampling accuracy up to any arbitrary error bound. Both methods depend on the analysis of the eigenvalues of continuous time Markov models that define the behavior of the SSA. We show how each can be applied to accelerate sampling within known Markov models or to subgraphs discovered automatically during execution. We demonstrate these methods for two applications of sampling in stiff SSAs that are important for modeling self-assembly reactions: sampling breakage times for multiply connected bond networks and sampling assembly times for multisubunit nucleation reactions. We show theoretically and empirically that our eigenvalue methods provide substantially reduced sampling times for a large class of models used in simulating self-assembly. These techniques are also likely to have broader use in accelerating SSA models so as to apply them to systems and parameter ranges that are currently computationally intractable.


Monitoring neuronal activity with voltage-sensitive fluorophores.
Voltage imaging in living cells offers the tantalizing possibility of combining the temporal resolution of electrode-based methods with the spatial resolution of imaging techniques. Our lab has been developing voltage-sensitive fluorophores, or VoltageFluors, that respond to changes in cellular and neuronal membrane potential via a photoinduced electron transfer (PeT)-based mechanism. This unique mechanism enables both the fast response kinetics and high sensitivity required to record action potentials in single trials, across multiple cells without the need for stimuli-triggered averaging. In this chapter, we present a methodology for imaging membrane potential dynamics from dozens of neurons simultaneously in vitro. Using simple, commercially available cameras, illumination sources, and microscope optics in combination with the far-red synthetic voltage-sensitive fluorophore BeRST-1 (Berkeley Red Sensor of Transmembrane potential) provides a readily applied method for monitoring neuronal activity in cultured neurons. We discuss different types of voltage-sensitive dyes, considerations for selecting imaging modalities, and outline procedures for the culture of rat hippocampal neurons and performing voltage imaging experiments with these samples. Finally, we provide an example of how changes to the metabolic input to cultured hippocampal neurons can alter their activity profile.


Influence of prehospital airway management on neurological outcome in patients transferred to a heart attack centre following out-of-hospital cardiac arrest.
To describe the association between prehospital airway management and neurological outcomes in patients transferred by the ambulance service directly to a heart attack centre (HAC) post-return of spontaneous circulation (ROSC). A retrospective observational cohort study in which ambulance records were reviewed to determine prehospital airway management strategy and collect physiological and demographic data. HAC notes were obtained to determine in-hospital management and quantify neurological outcome via the cerebral performance category (CPC) scale. Statistical analyses were performed via χ2 -test, Mann-Whitney U-test, odds ratios and binomial logistic regression. Two hundred and twenty patients were included between August 2013 and August 2014, with complete outcome data obtained for 209. Median age of patients with complete outcome data was 67 years and 71.3% were male (n = 149). Airway management was provided using a supraglottic airway (SGA) in 72.7% of cases (n = 152) with the remainder undergoing endotracheal intubation (ETI). There was no significant difference in the proportion of patients who had a good neurological outcome (CPC 1 and 2) at discharge between the SGA and ETI groups (P = 0.29). Binomial logistic regression incorporating factors known to influence outcome demonstrated no significant difference in neurological outcomes between the SGA and ETI groups (adjusted OR 0.73, 95% CI 0.34-1.56). In this observational study, there was no significant difference in the proportion of good neurological outcomes in patients managed with SGA versus ETI during cardiac arrest and in the post-ROSC transfer phase. Further research is required to provide more definitive evidence in relation to the optimal airway management strategy in out-of-hospital cardiac arrest.


Study protocol to investigate the effects of testosterone therapy as an adjunct to exercise rehabilitation in hypogonadal males with chronic heart failure.
Testosterone deficiency is a common occurrence in men with chronic heart failure (CHF) and may underpin features of advanced disease, including reduced skeletal muscle mass and fatigue. It is positively correlated with cardiac output and exercise capacity in patients with CHF, whereas a significant improvement in both these parameters has been observed following testosterone replacement therapy. Testosterone therapy has also been shown to reduce circulating levels of inflammatory markers, (TNF-alpha, sICAM-1 and sVCAM-1) in patients with established coronary artery disease and testosterone deficiency. This pilot study will assess the feasibility of a combined exercise rehabilitation and adjunctive testosterone therapy intervention for evoking improvements in exercise capacity, circulating inflammatory markers, cardiac and skeletal muscle function, indices of psychological health status and quality of life in hypogonadal males with chronic heart failure. Following ethical approval, 36 patients will be randomly allocated to one of two groups: testosterone or placebo therapy during exercise rehabilitation. A combined programme of moderate intensity aerobic exercise and resistance (strength) training will be used. The primary outcome measure is exercise capacity, assessed using an incremental shuttle walk test. Secondary outcome measures include measures of peak oxygen uptake, cardiac function, lower-limb skeletal muscle contractile function and oxygenation during exercise, circulating inflammatory markers, psychological health status and quality of life. Exercise rehabilitation can safely increase exercise capacity in stable CHF patients but there is a need for studies which are aimed at evaluating the long-term effects of physical training on functional status, morbidity and mortality. This pilot study will provide valuable preliminary data on the efficacy of testosterone therapy as an adjunct to exercise rehabilitation on a range of functional, physiological and health-related outcomes in this patient population. Preliminary data will be used in the design of a large-scale randomised controlled trial, aimed at informing clinical practice with respect to optimisation of exercise rehabilitation in this patient group.


Protrusio acetabuli in Marfan syndrome: age-related prevalence and associated hip function.
Protrusio acetabuli is known to occur in patients with Marfan syndrome, but its prevalence, its effects on hip function, and its possible association with the subsequent development of degenerative hip disease have not been studied in a large population. Nevertheless, some clinicians have recommended prophylactic hip surgery for preadolescents with Marfan syndrome and protrusio acetabuli. We performed a cross-sectional study of 173 patients (346 hips) with Marfan syndrome who were interviewed and examined for calculation of the Iowa hip score. Anteroposterior radiographs of the pelvis were made, and two radiographic indices of acetabular depth were measured: (1) the center-edge angle of Wiberg and (2) the acetabular-ilioischial distance. The presence of protrusio was defined with use of two extant definitions: (1) a center-edge angle of >50 degrees or (2) an acetabular-ilioischial distance of >/=3 mm in male patients or >/=6 mm in female patients. Linear regression analyses were performed between these radiographic indices of acetabular depth and patient age, Iowa hip scores, the magnitude of the radiographic joint space, and range of motion. The prevalence of protrusio acetabuli was 27% according to the center-edge angle criterion and 16% according to the acetabular-ilioischial distance criterion. The prevalence of protrusio increased until the age of twenty years and remained stable after the age of twenty years. Slight but significant negative correlations were detected between the two radiographic indices of acetabular depth and both the Iowa hip score and the summed range of motion (p < 0.02 for all). No significant relationship was found between the two radiographic indices and pain scores. In patients with Marfan syndrome who were more than forty years old, the Iowa hip scores for hips with protrusio were not significantly lower than those for hips without protrusio. In patients with Marfan syndrome, the prevalence of protrusio acetabuli increases during the first two decades of life and then plateaus in terms of both population-wide prevalence and radiographic severity. In this population, protrusio generally is not associated with severely problematic hip function but it is associated with slightly decreased range of motion of the hip. We concluded that prophylactic surgical intervention is not indicated for most patients with Marfan syndrome who have a radiographic diagnosis of protrusio.


[The regional budget for mental health care: a new approach to combine inpatient and outpatient care].
Due to increasing health care expenditures the discussion about advantages and disadvantages of new methods for resource allocation in mental health care has been intensified. A promising model is the Regional Budget for Mental Health Care, which is currently being examined in Schleswig-Holstein. The present paper describes first experiences with the new resource allocation model. BASIC CONDITIONS: An annual budget, provided for the treatment of a fixed number of patients, makes it possible to reduce inpatient capacity in favour of improved community-integrated approaches for the treatment of acute psychiatric illness. In a first step inpatient capacity will be reduced by 8 percent. By the end of 2007 capacity for hospital day care shall be increased by 87 percent and a home treatment will be implemented. The previous working method, orientated to treatment setting, will be replaced by an approach specialized in diagnostic groups. The Regional Budget could improve the continuity and flexibility of patient care. Service providers become motivated to treat in a way, which with little resource consumption achieves a long lasting health status improvement. For health insurances the Regional Budget is an opportunity to limit cost increases.


Single-port for laparoscopic gastric resection with a novel platform.
Laparoscopic gastrointestinal resections using single-port are possible, but triangulation problems and the need of articulated instruments difficult the procedures. To present a surgical alternative using single-port laparoscopic device on gastric resection. The patient is placed in a supine and reverse Trendelenburg position with surgeon between patient's legs. First assistant was on the right side of the patient with the monitor placed on the patient's cranial side. With the patient under general anesthesia, a transumbilical 3 cm skin incision is performed. A single-incision advanced access platform with gelatin cap, self-retaining sleeve and wound protector is introduced through this incision. Three 5-12 mm operating ports were introduced through the single-port device. Due to the gel cap and sleeves, no articulated instruments are necessary. CO2 pneumoperitoneum is established at 12 mmHg. A rigid 30 degree 10 mm laparoscope is introduced. Operation begins with access to the lesser sac by opening the omentum along the greater curvature of the stomach using harmonic scalpel. Once the stomach is fully exposed and a stay suture is place around the tumor. Gastric wall is divided with cautery 1 cm away from the tumor. Tumor is excised. Gastric wall is sutured with two-layer running suture. No drain was used. Umbilical incision was closed. This procedure was used in one patient with gastric duplication. Operative time was 200 minutes. Blood loss was minimal. Recovery was uneventful and patient discharged on postoperative day 2. Final aspect of the umbilical incision was good. Gastric resection with single-port laparoscopic platform is feasible and may be safely performed in selected patients.


Motor cortex stimulation for phantom limb pain: comprehensive therapy with spinal cord and thalamic stimulation.
The effects of spinal cord stimulation (SCS), deep brain stimulation (DBS) of the thalamic nucleus ventralis caudalis (VC) and motor cortex stimulation (MCS) were analyzed in 19 patients with phantom limb pain. All of the patients underwent SCS and, if the SCS failed to reduce the pain, the patients were considered for DBS and/or MCS. Satisfactory pain control for the long-term was achieved in 6 of 19 (32%) by SCS, 6 of 10 (60%) by DBS and 1 (20%) of 5 by MCS. SCS and DBS of the VC sometimes produced a dramatic effect on the pain, leading to a long pain-free interval and infrequent use of stimulation. The effects of both DBS of the VC and MCS were tested in four. One patient of them reported better pain control by MCS than by DBS, whereas two reported the opposite results. There is no evidence at present for an advantage of MCS over SCS and DBS of the VC in controlling phantom limb pain.


Replication-compromised cells require the mitotic checkpoint to prevent tetraploidization.
Replication stress often induces chromosome instability. In this study, we explore which factors in replication-compromised cells promote abnormal chromosome ploidy. We expressed mutant forms of either polymerase α (Polα) or polymerase δ (Polδ) in normal human fibroblasts to compromise DNA replication. Cells expressing the mutant Polα-protein failed to sustain mitotic arrest and, when propagated progressively, down-regulated Mad2 and BubR1 and accumulated 4N-DNA from the 2N-DNA cells. Significantly, a population of these cells became tetraploids. The Polα mutant expressing cells also exhibited elevated cellular senescence markers, suggesting as a mechanism to limit proliferation of the tetraploids. Expression of the Polδ mutant also caused cells to accumulate 4N-DNA. In contrast to the Polα mutant expressing cells, the Polδ mutant expressing cells expressed sufficient levels of Mad2, BubR1, and cyclin B1 to sustain mitotic arrest, and these cells had normal chromosome ploidy. Together, these results suggest that replication-compromised cells depend on the mitotic checkpoint to prevent mitotic slippage that could result in tetraploidization.


Current progress in non-invasive imaging of beta cell mass of the endocrine pancreas.
The increasing incidence of diabetes requires a better understanding of the pathogenesis of the clinical disease. Studies in prevention and treatment have been hampered by the single end-point of diagnosis of diabetes and hyperglycemia. The common pathology in both type 1 and type 2 diabetes is insufficient beta-cell mass to meet the metabolic demand. Unfortunately, current diagnostic methods rely on metabolic responses that do not accurately reflect true beta-cell mass. Recent advances in beta-cell imaging have utilized multiple modalities in experimental and clinical settings. While no "gold-standard" exists to measure beta-cell mass, modalities such as single photon emission computed tomography, optical and fluorescent imaging, magnetic resonance imaging, and positron emission tomography have been used with mixed success. Many of the methods are limited by the inability to translate to the clinical setting, poor discrimination between the exocrine and endocrine pancreas, or a poor measurement of beta-cell mass. However, promising new "neurofunctional imaging" approaches have emerged as improved measures of beta-cell mass. We review the current understanding of the pathogenesis and evaluation of diabetes, as well as experimental approaches to assessing beta-cell mass.


Methylation and inactivation of estrogen, progesterone, and androgen receptors in prostate cancer.
Prostate cancer development is initially steroid hormone dependent. Estrogen receptors (ERs), androgen receptors (ARs), and progesterone receptors (PRs) have been identified in normal and cancerous prostate tissues. We investigated whether the promoter regions of these steroid receptor genes are methylated and inactivated in prostate cancer cells and tissues. The expression and promoter methylation status of three ERalpha isoforms (ERalpha-A, ERalpha-B, and ERalpha-C), ERbeta, two PR isoforms (PR-A and PR-B), and AR were investigated in five prostate cancer cell lines (ND1, DU145, PC3, LNCaP, and DUPro) and in pairs of normal and cancerous prostate tissues from 38 patients with prostate cancer. Methylation-specific polymerase chain reaction, reverse transcription--polymerase chain reaction, and 5' rapid amplification of complementary DNA ends were used. All statistical tests were two-sided. ERalpha-C was expressed in all cell lines, but ERalpha-A and ERalpha-B were not expressed in any cell line. ERalpha-A and ERalpha-B promoters were methylated, but ERalpha-C was unmethylated. Promoters for ERbeta, AR, PR-A, and PR-B were methylated and thus inactivated in some cell lines but not in others. Treating cells with the demethylating reagent 5-aza-2'-deoxycytidine restored expression of all steroid receptor genes with previously methylated promoters. All 38 pairs of cancer and normal tissues had unmethylated ERalpha-C promoters. Thirty-six (95%) of 38 cancers had methylated ERalpha-A, 35 (92%) of 38 cancers had methylated ERalpha-B, but all normal tissues had unmethylated ERalpha-A and ERalpha-B (both P<.001). ERbeta was methylated in 30 (79%) of 38 cancers but unmethylated in all normal tissues. AR was methylated in three (8%) of 38 cancers but unmethylated in all normal tissues. PR-A and PR-B were unmethylated in all tissues. Certain steroid receptor genes appear to be inactivated by CpG methylation in prostate cancer tissue and cell lines.


Treatment of petroleum refinery wastewater by distillation-assisted catalytic oxidation under low temperature and low pressure.
A distillation-assisted catalytic oxidation (DACO) process under low temperature (100 degrees C) and atmospheric pressure was investigated to treat heavily contaminated wastewater from oil refining industry. The DACO experiments were carried out in a distillation batch reactor, using CuO/gamma-A1(2)O3 as catalyst. The experimental temperature was kept at 100 degrees C and H2O2 oxidant was supplied into the reactive system with 200 mL/L. The results demonstrated that more than 92.2% of chemical oxygen demand removal was obtained and the absorbance of the refinery wastewater after treatment was zero, indicating significant decolorization efficiency for the solution. The research of life and stability showed that the catalyst had a good stability. The present study indicates that this DACO approach may have a significant application potential for industrial wastewater treatment.


Expression of 5-lipoxygenase and 15-lipoxygenase in rheumatoid arthritis synovium and effects of intraarticular glucocorticoids.
It was previously shown that lipoxygenase (LO) pathways are important in the rheumatoid arthritis (RA) inflammatory process and that synovial fluid from RA patients contains high amounts of leukotrienes. We therefore aimed to investigate the 5-LO and 15-LO-1 expression pattern in RA and ostheoarthritis (OA) synovial tissue and to study the effect of intraarticular glucocorticoid (GC) therapy on enzyme expression. Expression of LOs was evaluated by immunohistochemistry in RA and OA synovial biopsies. Cellular localization of these enzymes was analyzed by double immunofluorescence. In synovial biopsies from 11 RA patients, 5-LO and 15-LO-1 expression was evaluated before and after triamcinolone hexacetonide knee injection and assessed by image analysis to quantify their expression. We also investigated the presence of 15-LO-1 by immunohistochemistry in synovial fluid (SF) cells as well as their ability to form 15-hydroxyeicosatetraenoic acid (15-HETE) following treatment with arachidonic acid (AA). 5-LO and 15-LO-1 are present in RA and OA synovium, with 5-LO being mostly expressed in lining and sublining macrophages, neutrophils and mast cells and 15-LO-1 mainly in lining macrophages, fibroblasts and sublining endothelial cells. Intraarticular GC treatment resulted in a significant suppression of 5-LO expression, but did not influence the 15-LO-1 enzyme significantly. Also, SF cells express a functional 15-LO-1 and produce 15-HETE when challenged with AA. These data demonstrate that local therapy with GC decreases 5-LO expression in RA synovium and offer an additional possible mechanism for the efficiency of intraarticular adjuvant therapy in RA.


Case Study: Symptomatic Exercise-Associated Hyponatremia in an Endurance Runner Despite Sodium Supplementation.
Symptomatic exercise-associated hyponatremia (EAH) is known to be a potential complication from overhydration during exercise, but there remains a general belief that sodium supplementation will prevent EAH. We present a case in which a runner with a prior history of EAH consulted a sports nutritionist who advised him to consume considerable supplemental sodium, which did not prevent him from developing symptomatic EAH during a subsequent long run. Emergency medical services were requested for this runner shortly after he finished a 17-hr, 72-km run and hike in Grand Canyon National Park during which he reported having consumed 9.2-10.6 L of water and >6,500 mg of sodium. First responders determined his serum sodium concentration with point-of-care testing was 122 mEq/L. His hyponatremia was documented to have improved from field treatment with an oral hypertonic solution of 800 mg of sodium in 200 ml of water, and it improved further after significant aquaresis despite in-hospital treatment with isotonic fluids (lactated Ringer's). He was discharged about 5 hr after admission in good condition. This case demonstrates that while oral sodium supplementation does not necessarily prevent symptomatic EAH associated with overhydration, early recognition and field management with oral hypertonic saline in combination with fluid restriction can be effective treatment for mild EAH. There continues to be a lack of universal understanding of the underlying pathophysiology and appropriate hospital management of EAH.


A detailed description of the phenotypic spectrum of North Sea Progressive Myoclonus Epilepsy in a large cohort of seventeen patients.
In 2011, a homozygous mutation in GOSR2 (c.430G > T; p. Gly144Trp) was reported as a novel cause of Progressive Myoclonus Epilepsy (PME) with early-onset ataxia. Interestingly, the ancestors of patients originate from countries bound to the North Sea, hence the condition was termed North Sea PME (NSPME). Until now, only 20 patients have been reported in literature. Here, we provide a detailed description of clinical and neurophysiological data of seventeen patients. We collected clinical and neurophysiological data from the medical records of seventeen NSPME patients (5-46 years). In addition, we conducted an interview focused on factors influencing myoclonus severity. The core clinical features of NSPME are early-onset ataxia, myoclonus and seizures, with additionally areflexia and scoliosis. Factors such as fever, illness, heat, emotions, stress, noise and light (flashes) all exacerbated myoclonic jerks. Epilepsy severity ranged from the absence of or incidental clinical seizures to frequent daily seizures and status epilepticus. Some patients made use of a wheelchair during their first decade, whereas others still walked independently during their third decade. Neurophysiological features suggesting neuromuscular involvement in NSPME were variable, with findings ranging from indicative of sensory neuronopathy and anterior horn cell involvement to an isolated absent H-reflex. Although the sequence of symptoms is rather homogeneous, the severity of symptoms and rate of progression varied considerably among individual patients. Common triggers for myoclonus can be identified and myoclonus is difficult to treat; to what extent neuromuscular involvement contributes to the phenotype remains to be further elucidated.


Construction and validation of a polycompetitor construct (SWITCH) for use in competitive RT-PCR to assess tachyzoite-bradyzoite interconversion in Toxoplasma gondii.
The obligate intracellular protozoan parasite, Toxoplasma gondii exists as 2 life-cycle forms in intermediate hosts. The rapidly dividing tachyzoites responsible for acute disease, present in the first 14 days of infection, give rise to slowly dividing bradyzoites that reside in tissue cysts. Reactivation of disease is associated with conversion of bradyzoites to tachyzoites. A sensitive method for detection and assessment of the number of each life-cycle stage would be useful for following these events. Herein we describe the construction and validation of a plasmid (pSWITCH) containing a polycompetitor construct (SWITCH) for use in competitive reverse transcriptase-PCR (cRT-PCR). pSWITCH contains competitors for SAG2A and LDH2 genes, which are exclusively expressed by tachyzoite and bradyzoite stages respectively, and for beta-tubulin, a gene expressed by both stages. Using cRT-PCR, samples can first be accurately normalized for expression of the housekeeping gene, beta-tubulin and then the relative levels of SAG2A and LDH2 expression compared to follow stage conversion. The abundance of transcripts for other genes of interest can then be followed during this process as demonstrated here for the SAG2-related family of genes. This technique offers a powerful tool for studying the processes involved in tachyzoite and bradyzoite interconversion.


Antimicrobial drugs for treating methicillin-resistant Staphylococcus aureus colonization.
Eradication strategies for methicillin-resistant Staphylococcus aureus (MRSA) are variable. We sought to summarize the evidence for use of antimicrobial agents to eradicate MRSA. To describe the effects of topical and systemic antimicrobial agents on nasal and extra-nasal MRSA carriage, adverse events, and incidence of subsequent MRSA infections. We searched the Cochrane Infectious Diseases Group's trials register (August 2003), the Cochrane Central Register of Controlled Trials (Issue 3, 2003), MEDLINE (1966 to 2003), EMBASE (1988 to 2003), handsearched relevant literature, and contacted MRSA experts and the manufacturer of mupirocin. Randomized controlled trials of patients colonized with MRSA comparing topical or systemic antimicrobials to placebo or no treatment, and trials comparing various combinations of topical or systemic agents to no treatment, placebo, or to topical or systemic agents. Two reviewers independently applied inclusion criteria to potentially relevant trials, assessed trial methodological quality, and extracted data. Primary outcomes included eradication of MRSA, infection due to MRSA, and adverse events. Six trials (384 participants) met the inclusion criteria. No difference in MRSA eradication was detected in four studies: one that compared mupirocin to placebo, two that compared one systemic agent to no treatment (fusidic acid in one and rifampin or minocycline in the other) and one that compared mupirocin to topical fusidic acid and oral trimethoprim-sulfamethoxazole, examining nasal MRSA eradication as an outcome. One study compared minocycline to rifampin, with rifampicin being more effective in relation to eradication of MRSA from all sites at day 30 (relative risk 0.16; 95% confidence intervals 0.02 to 1.00), but the difference at 90 days was not statistically significant (n = 18). Two studies (one testing novobiocin and rifampin, the other ciprofloxacin and rifampin, versus trimethoprim-sulfamethoxazole and rifampin) did not demonstrate a difference in eradication of MRSA at all sites (n = 94). Adverse events with systemic agents occurred in up to 20% of participants, however reporting was sporadic and denominators small. All trials reported development of resistance to antimicrobial agents used. There is insufficient evidence to support use of topical or systemic antimicrobial therapy for eradicating nasal or extra-nasal MRSA. There is no demonstrated superiority of either topical or systemic therapy, or of combinations of these agents. Potentially serious adverse events and development of antimicrobial resistance can result from therapy.


Neuroimmunotherapy of untreatable metastatic solid tumors with subcutaneous low-dose interleukin-2, melatonin and naltrexone: modulation of interleukin-2-induced antitumor immunity by blocking the opioid system.
The preliminary applications of the psychoneuroimmunological knowledges to the treatment of human diseases have confirmed the possibility to amplify IL-2-dependent anticancer immunity by the pineal hormone melatonin (MLT) or by opioid antagonist, such as naltrexone (NTX), which act by activating TH1 lymphocytes or suppressing TH2 lymphocytes, respectively. At present, however, there are no data about the immunobiological effects of a concomitant administration of both MLT and NTX on IL-2-induced anticancer immunity. This preliminary study was carried out to evaluate whether the association of NTX may further enhance the lymphocytosis induced by the neuroimmunotherapy with IL-2 plus MLT. The study included 14 consecutive untreatable metastatic solid tumor patients. According to a cross-over randomized study, the patients were treated during two consecutive immunotherapeutic cycles at 21-day intervals with IL-2 plus MLT alone or with IL-2 plus MLT plus NTX. IL-2 was injected subcutaneously at 3 MIU/day for 6 days/week for 4 weeks, MLT was given orally at 20 mg /day in the evening every day, and NTX was given orally at 100 mg in the morning every next day. For the immune evaluation, venous blood samples were drawn before the onset of treatment and at weekly intervals. Lymphocyte mean number significantly increased after both IL-2 plus MLT and IL-2 plus MLT plus NTX. However, the concomitant administration of NTX induced a significantly higher increase in lymphocyte mean number with respect to that achieved with IL-2 plus MLT alone. In contrast, the increase in eosinophil mean number was significantly higher on IL-2 plus MLT alone. This preliminary study shows that the association of NTX further amplifies the lymphocytosis obtained by IL-2 plus MLT. Since the lymphocytosis represents the most important favourable prognostic variable predicting the anticancer efficacy of IL-2 immunotherapy, it is probable that a cancer neuroimmunotherapy with IL-2 plus both MLT and NTX to activate TH1 and suppress TH2 cells respectively, may deserve more promising results in the treatment of human neoplasms according to the psychoneuroimnunological knowledge.


Cranberry proanthocyanidins induce apoptosis and inhibit acid-induced proliferation of human esophageal adenocarcinoma cells.
The occurrence of esophageal adenocarcinoma and its only recognized precursor lesion, Barrett's esophagus, has rapidly increased during the past three decades. The precise reason for the rise remains to be elucidated, but increasing rates have been linked to multiple nutritional factors. Plant-based diets have generally been associated with a reduction of risk for esophageal adenocarcinoma and those of animal origin with risk escalation. Moreover, a number of recent in vitro and limited in vivo investigations have reported that cranberry extracts affect multiple cancer-associated processes in breast, colon, prostate, and other cancer cell lines of epithelial origin. Thus, this study sought to investigate the chemopreventive potential of a cranberry proanthocyanidin rich extract (PAC) in SEG-1 human esophageal adenocarcinoma (EAC) cells. PAC pretreatment significantly inhibited the viability and proliferation of EAC cells in a time- and dose-dependent manner. Moreover, PAC (50 microg/mL) significantly inhibited acid-induced cell proliferation of SEG-1 cells. PAC treatment induced cell cycle arrest at the G1 checkpoint and significantly reduced the percentage of SEG-1 cells in S-phase following 24 and 48 h of exposure. PAC treatment also resulted in significant induction of apoptosis. Thus, PAC modulates cell cycle regulation, aberrant proliferation, and apoptosis, all key biological processes altered during progression to esophageal adenocarcinoma. These findings support that further mechanistic studies are warranted to more fully elucidate the inhibitory potential of PAC against esophageal cancer.


New ataxic tottering-6j mouse allele containing a Cacna1a gene mutation.
Voltage-gated Ca(2+) (Ca(v)) channels control neuronal functions including neurotransmitter release and gene expression. The Cacna1a gene encodes the α1 subunit of the pore-forming Ca(v)2.1 channel. Mice with mutations in this gene form useful tools for defining channel functions. The recessive ataxic tottering-6j strain that was generated in the Neuroscience Mutagenesis Facility at The Jackson Laboratory has a mutation in the Cacna1a gene. However, the effect of this mutation has not been investigated in detail. In this study, mutation analysis shows a base substitution (C-to-A) in the consensus splice acceptor sequence linked to exon 5, which results in the skipping of exon 5 and the splicing of exon 4 directly to exon 6. The effect of this mutation is expected to be severe as the expressed α1 subunit protein lacks a significant part of the S4-S5 linker, S5, and part of S5-S6 linker in domain I. Tottering-6j mice display motor dysfunctions in the footprint, rotating rod, and hind-limb extension tests. Although cytoarchitecture of the mutant brains appears normal, tyrosine hydroxylase was persistently expressed in cerebellar Purkinje cells in the adult mutant mice. These results indicate that tottering-6j is a useful model for functional studies of the Ca(v)2.1 channel.


Effects of high-fiber rice Dodamssal (Oryza sativa L.) on glucose and lipid metabolism in mice fed a high-fat diet.
We investigated the effects of high amylose rice variety, Dodamssal (DO) (Oryza sativa L.), on glucose homeostasis and lipid metabolism in mice. Experiment 1: Oral administration of DO for 1 week significantly improved glucose and insulin tolerance (p < .001) and reduced plasma triglyceride and low-density lipoprotein cholesterol concentrations. Experiment 2: Administration of DO-containing diet for 5 weeks also significantly reduced fasting glucose concentrations and hepatic lipid accumulation. DO induced GLP-1, adiponectin, and PYY levels. In the liver, DO suppressed the gene expression of G6pc, key gene in gluconeogenesis and induced AKT phosphorylation. DO increased fecal bile acid excretion regulating the expression in key genes in bile acid metabolism. DO suppressed plasma Trimethylamine N-oxide and intestinal lipopolysaccharide concentrations. DO may be achieved the hypolipidemic effects by direct activation of hepatic Pparα expression and its responsive genes regulating hepatic fatty acid uptake and β-oxidation, while downregulating the hepatic fatty acid synthesis Our results demonstrate that high-fiber rice, DO, might be a potential supplement for the amelioration of insulin resistance and hyperlipidemia. PRACTICAL APPLICATIONS: The results from the present study suggest that newly developed DO (Oryza sativa L.) high amylose rice strain may improve insulin sensitivity and activates the Akt pathway. DO consumption tends to counteract the deleterious effects characterized during the intake of high-fat-diet related to plasma TG, ALT, and AST concentrations. Therefore, DO supplementation might be a potential adjuvant to ameliorate dyslipidemia and adiposity.


The extent and degree of utterance-final word lengthening in spontaneous speech from 10 languages.
Words in utterance-final positions are often pronounced more slowly than utterance-medial words, as previous studies on individual languages have shown. This paper provides a systematic cross-linguistic comparison of relative durations of final and penultimate words in utterances in terms of the degree to which such words are lengthened. The study uses time-aligned corpora from 10 genealogically, areally, and culturally diverse languages, including eight small, under-resourced, and mostly endangered languages, as well as English and Dutch. Clear effects of lengthening words at the end of utterances are found in all 10 languages, but the degrees of lengthening vary. Languages also differ in the relative durations of words that precede utterance-final words. In languages with on average short words in terms of number of segments, these penultimate words are also lengthened. This suggests that lengthening extends backwards beyond the final word in these languages, but not in languages with on average longer words. Such typological patterns highlight the importance of examining prosodic phenomena in diverse language samples beyond the small set of majority languages most commonly investigated so far.


Research on cognitive impairment in community and home settings.
Mounting evidence demonstrates the significant impact of cognitive impairment on the lives of older adult patients and their caregivers. In addition, cognitive impairment presents challenging conditions for healthcare providers who are frequently unprepared to manage the care of these patients. These developments have spawned a wide range of research to more effectively identify and diagnose individuals with cognitive impairments, to develop better strategies to differentiate the types of cognitive impairment, and more accurately estimate the incidence and prevalence of the different types of impairments within different treatment settings. The 4 articles presented in this article represent a small sampling of recent research on these topics. Each article possesses methodological and other limitations that limit the generalizability of their findings. Nonetheless, there are important lessons to be learned from each. The first article examines the effectiveness of using a single-item measure to screen for delirium, one form of cognitive impairment. The second article presents the findings of a systematic review of research articles that estimate the prevalence of missed and delayed diagnoses of dementia. Efforts to understand factors that contribute to missed or delayed diagnoses may result in more robust strategies to improve timely diagnoses and effective interventions. The third article attempts to differentiate the prevalence of cognitive impairment that does not progress to a diagnosis of dementia. The findings suggest that, contrary to conventional thought, not every patient with cognitive impairment will ultimately have dementia. The final article illustrates the difficulties of identifying community-dwelling older adults with dementia. Once identified, the authors determined that patients and their families have wide range of unmet needs that may be addressed with better preparation of healthcare providers. Future research may produce more robust evidence on these important topics.


Public funding of pharmaceuticals in The Netherlands: investigating the effect of evidence, process and context on CVZ decision-making.
The College Voor Zorgverzekeringen (CVZ) provides guidance to the Dutch healthcare system on funding and use of new pharmaceutical technologies. This study examined the impact of evidence, process and context factors on CVZ decisions in 2004-2009. A data set of CVZ decisions pertaining to pharmaceutical technologies was created, including 29 variables extracted from published information. A three-category outcome variable was used, defined as the decision to 'recommend', 'restrict' or 'not recommend' a technology. Technologies included in list 1A/1B or on the expensive drug list were considered recommended; those included in list 2 or for which patient co-payment is required were considered restricted; technologies not included on any reimbursement list were classified as 'not recommended'. Using multinomial logistic regression, the relative contribution of explanatory variables on CVZ decisions was assessed. In all, 244 technology appraisals (256 technologies) were analysed, with 51%, of technologies recommended, 33% restricted and 16% not recommended by CVZ for funding. The multinomial model showed significant associations (p ≤ 0.10) between CVZ outcome and several variables, including: (1) use of an active comparator and demonstration of statistical superiority of the primary endpoint in clinical trials, (2) pharmaceutical budget impact associated with introduction of the technology, (3) therapeutic indication and (4) prevalence of the target population. Results confirm the value of a comprehensive and multivariate approach to understanding CVZ decision-making.


Duodenal follicular lymphoma: comprehensive gene expression analysis with insights into pathogenesis.
Follicular lymphoma (FL) of the gastrointestinal tract, particularly duodenal follicular lymphoma (DFL), is a rare variant of FL with indolent clinical behavior, and this disease is included in the 2008 World Health Organization classification system. In contrast to nodal follicular lymphoma (NFL), DFL occurs most frequently in the second part of the duodenum, lacks follicular dendritic cell meshworks and has memory B-cell characteristics. However, its molecular pathogenesis is still unclear. In the present study, we examined 10 DFL, 18 NFL and 10 gastric MALT lymphoma samples using gene expression analysis. Quantitative RT-PCR experiments and immunohistochemical analysis for 72 formalin-fixed, paraffin-embedded tissues from an independent series, including 32 DFL, 19 gastric MALT lymphoma and 27 NFL samples, were performed for validation of microarray data. Gene expression profiles of the three lymphoma types were compared using 2918 differentially expressed genes (DEG) and results suggested that DFL shares characteristics of MALT lymphoma. Among these DEG, CCL20 and MAdCAM-1 were upregulated in DFL and MALT but downregulated in NFL. In contrast, protocadherin gamma subfamily genes were upregulated in DFL and NFL. Quantitative RT-PCR and immunohistochemical studies demonstrated concordant results. Double immunofluorescence studies revealed that CCL20 and CCR6 were co-expressed in both DFL and MALT. We hypothesize that increased expression of CCL20 and MAdCAM-1 and co-expression of CCL20 and CCR6 may play an important role in tumorigenesis.