Production and characterization of monoclonal antibodies for the detection of the hepatitis C core antigen.
Background: Despite highly effective treatments to cure hepatitis C, almost 80% of chronically HCV-infected people are not treated, as they are unaware of their infection. Diagnostic rates and linkage to care must be substantially improved to reverse this situation. The HCV core antigen (HCVcAg) is a highly conserved protein that can be detected in the blood of HCV-infected patients and indicates active infection. Aim: To produce murine monoclonal antibodies against HCVcAg suitable for rapid and inexpensive tests to detect HCV infection. Methods: BALB/c mice were sequentially inoculated with purified recombinant HCVcAg from Gt1a, Gt3a, Gt4a, and Gt1b genotypes. Hybridomas producing the desired monoclonal antibodies were selected, and the reactivity of antibodies against HCVcAg from various genotypes was tested by Western blotting and dot blotting. The binding kinetics of the antibodies to purified HCVcAg was analyzed by surface plasmon resonance (SPR), and their ability to detect HCVcAg was tested by double antibody sandwich ELISA (DAS-ELISA). Results: Four specific monoclonal antibodies (1C, 2C, 4C, and 8C) were obtained. 1C, 2C, and 4C recognized HCVcAg of all genotypes tested (Gt1a, Gt1b, Gt2a, Gt3a, and Gt4a), while 8C did not recognize the Gt2a and Gt3a genotypes. Based on SPR data, the antibody-HCVcAg complexes formed are stable, with 2C having the strongest binding properties. DAS-ELISA with different antibody combinations easily detected HCVcAg in culture supernatants from HCV-infected cells. Conclusion: Specific and cross-reactive anti-HCVcAg monoclonal antibodies with strong binding properties were obtained that may be useful for detecting HCVcAg in HCV-infected samples.


Ghrelin-AMPK Signaling Mediates the Neuroprotective Effects of Calorie Restriction in Parkinson's Disease.
Calorie restriction (CR) is neuroprotective in Parkinson's disease (PD) although the mechanisms are unknown. In this study we hypothesized that elevated ghrelin, a gut hormone with neuroprotective properties, during CR prevents neurodegeneration in an 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of PD. CR attenuated the MPTP-induced loss of substantia nigra (SN) dopamine neurons and striatal dopamine turnover in ghrelin WT but not KO mice, demonstrating that ghrelin mediates CR's neuroprotective effect. CR elevated phosphorylated AMPK and ACC levels in the striatum of WT but not KO mice suggesting that AMPK is a target for ghrelin-induced neuroprotection. Indeed, exogenous ghrelin significantly increased pAMPK in the SN. Genetic deletion of AMPKβ1 and 2 subunits only in dopamine neurons prevented ghrelin-induced AMPK phosphorylation and neuroprotection. Hence, ghrelin signaling through AMPK in SN dopamine neurons mediates CR's neuroprotective effects. We consider targeting AMPK in dopamine neurons may recapitulate neuroprotective effects of CR without requiring dietary intervention.


Preliminary Analysis of Phytoconstituents and Evaluation of Anthelminthic Property of Cayratia auriculata (In Vitro).
Context:Phytoconstituents of medicinal plants have been playing a key role in treating various diseases all over the world since ancient times. The present study was focused on preliminary analysis of phytoconstituents and evaluation of anthelmintic property (in vitro) of different extracts of Cayratia auriculata (Family: Vitaceae) against earthworms (Pheretima posthuma). Materials and methods:Four different hexane, chloroform, ethyl acetate and methanol extracts of Cayratia auriculata were screened for their phytoconstituents. Cayratia auriculata has been shown to have various phytoconstituents such as flavonoids, phenolic compounds, tannins, alkaloids, saponins, glycosides and steroids. Hexane, chloroform, ethyl acetate and methanol extracts of Cayratia auriculata were analyzed for their anthelmintic property on earthworms (Pheretima posthuma). Each extract at three different concentrations (20 mg/mL, 40 mg/mL and 80 mg/mL) was analysed to evaluate the time taken for paralysis (P) and death (D) of adult earthworms. Albendazole was used as standard and 2% Tween 80 in distilled water as control at a concentration of 10 mg/mL. Results:Our study found that the methanolic extract of Cayratia auriculata at a concentration of 80 mg/mL [21.98±0.88 (P), 79.06±0.87(D)] expressed a stronger anthelmintic property than the standard albendazole (10 mg/mL) [26.73±0.56 (P), 87.70±0.91 (D)]. Conclusions:All extracts expressed a dose dependent anthelmintic activity, with an increasing pattern in the following order: hexane, chloroform, ethyl acetate and methanol.


Genome-wide co-localization of active EGFR and downstream ERK pathway kinases mirrors mitogen-inducible RNA polymerase 2 genomic occupancy.
Genome-wide mechanisms that coordinate expression of subsets of functionally related genes are largely unknown. Recent studies show that receptor tyrosine kinases and components of signal transduction cascades including the extracellular signal-regulated protein kinase (ERK), once thought to act predominantly in the vicinity of plasma membrane and in the cytoplasm, can be recruited to chromatin encompassing transcribed genes. Genome-wide distribution of these transducers and their relationship to transcribing RNA polymerase II (Pol2) could provide new insights about co-regulation of functionally related gene subsets. Chromatin immunoprecipitations (ChIP) followed by deep sequencing, ChIP-Seq, revealed that genome-wide binding of epidermal growth factor receptor, EGFR and ERK pathway components at EGF-responsive genes was highly correlated with characteristic mitogen-induced Pol2-profile. Endosomes play a role in intracellular trafficking of proteins including their nuclear import. Immunofluorescence revealed that EGF-activated EGFR, MEK1/2 and ERK1/2 co-localize on endosomes. Perturbation of endosome internalization process, through the depletion of AP2M1 protein, resulted in decreased number of the EGFR containing endosomes and inhibition of Pol2, EGFR/ERK recruitment to EGR1 gene. Thus, mitogen-induced co-recruitment of EGFR/ERK components to subsets of genes, a kinase module possibly pre-assembled on endosome to synchronize their nuclear import, could coordinate genome-wide transcriptional events to ensure effective cell proliferation.


Diethylstilboestrol--clinical pharmacology and alternatives in small animal practice.
Diethylstilboestrol is currently only available in Australia for oral use in dogs and cats. As an orally and systemically active non-steroidal oestrogen, DES has been widely used in small animal veterinary medicine for a variety of indications. A review of the literature reveals that many of the recommendations for use are founded on anecdotal or unreported clinical observations. While many of the uses may be valid, accurate determinations of optimum dosing regimens have not been defined. This is especially unfortunate in view of the potential toxicity of DES to small animals. Nevertheless, particularly in cases of low-dose intermittent administration, oral DES appears indicated at least until data on alternative safe and effective interventions become available.


{2-Morpholino-N-[1-(2-pyrid-yl)ethyl-idene]ethanamine-κN,N',N''}bis-(thio-cyanato-κN)zinc(II).
The asymmetric unit of the title compound, [Zn(NCS)(2)(C(13)H(19)N(3)O)], contains two crystallographically independent mol-ecules. In each mol-ecule, the Zn(II) ion is five-coordinated by the N,N',N"-tridentate Schiff base and the N atoms of two thio-cyanate ligands in a distorted square-pyramidal geometry. The two mol-ecules differ mainly in the deviations from the ideal geometry, with τ values of 0.14 and 0.33. In the crystal, inter-molecular C-H⋯S hydrogen bonds are observed. An intra-molecular C-H⋯N hydrogen bond occurs in one of the independent mol-ecules.


Strain diversity in Saccharomyces cerevisiae thiamine production capacity.
Vitamin B1 , also known as thiamine, is an important vitamin that, besides its role in human health, is converted to meat aromas upon exposure to high temperatures. Therefore, it is relevant for the production of vegan meat-like flavours. In this study, we investigated 48 Saccharomyces cerevisiae strains for their thiamine production capacity by measuring the intracellular and extracellular vitamins produced in the thiamine-free minimal medium after 72 h of growth. We found approximately an 8.2-fold difference in overall thiamine yield between the highest and lowest-producing strains. While the highest thiamine yield was 254.6 nmol/L, the highest thiamine-specific productivity was 160.9 nmol/g DW. To assess whether extracellular thiamine was due to leakage caused by cell damage, we monitored membrane permeabilization using propidium iodide (PI) staining and flow cytometry. We found a good correlation between the percentage of extracellular thiamine and PI-stained cells (Spearman's ρ = 0.85). Finally, we compared S. cerevisiae CEN.PK113-7D (wild type [WT]) to three strains evolved in a thiamine-free medium for their thiamine production capacity. On average, we saw an increase in the amount of thiamine produced. One of the evolved strains had a 49% increase in intracellular thiamine-specific productivity and a biomass increase of 20% compared with the WT. This led to a total increase in thiamine yield of 60% in this strain, reaching 208 nmol/L. This study demonstrated that it is possible to achieve thiamine overproduction in S. cerevisiae via strain selection and adaptive laboratory evolution.


Abnormalities of the central nervous system in very young children with sickle cell anemia.
To determine whether abnormalities of the CNS are present in very young children with sickle cell anemia. Thirty-nine children with hemoglobin SS between the ages of 7 and 48 months were examined with magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA). No child had a history of clinical stroke, although 3 had a history of seizures (2 neonatal). Twenty-one patients underwent developmental testing with the Bayley or McCarthy Scales. The overall prevalence of CNS abnormalities in asymptomatic children was 4 of 36 (11%, confidence interval 3, 26%). One patient had a silent infarct observed on MRI and a stenotic lesion on MRA; 3 other patients had stenotic lesions on MRA. The 3 patients who had a history of seizures all had lesions consistent with infarcts on MRI. Of the asymptomatic patients who had psychometric testing, 1 of 18 was developmentally delayed. One of 3 with a history of seizures had mild developmental delay. Very young children with sickle cell anemia (and no history of clinical stroke) have infarction in the brain and/or stenosis of major cerebral arteries, similar to those reported in older children. These findings indicate a need for larger studies to define the incidence of CNS lesions in this age group and to determine the need for early therapeutic intervention to prevent CNS sequelae of sickle cell disease.


Thermoregulation in the desert iguana Dipsosaurus dorsalis.
The body temperature of desert iguanas implanted with miniature temperature-sensitive radio transmitters was continuously monitored in their natural habitat. Extensive thermoregulatory behavior occurred in retreat burrows prior to morning emergence. Such behavior permits the igluana to emerge from below ground at its preferred body temperature rather than suboptimal temperature at which activity in the burrow is initiated.


Specialty preference for cardiovascular prevention practice in the Southeast US and role of a preventive cardiologist.
Cardiovascular disease (CVD) prevention is practiced concurrently by providers from several specialties. Our goal was to understand providers' preference of specialties in CVD prevention practice and the role of preventive cardiologists. Between 11 October 2021 and 1 March 2022, we surveyed providers from internal medicine, family medicine, endocrinology, and cardiology specialties to examine their preference of specialties in managing various domains of CVD prevention. We examined categorical variables using Chi square test and continuous variables using t or analysis of variance test. Of 956 invitees, 263 from 21 health systems and 9 states responded. Majority of respondents were women (54.5%), practicing physicians (72.5%), specializing in cardiology (43.6%), and working at academic centers (51.3%). Respondents favored all specialties to prescribe statins (43.2%), ezetimibe (37.8%), sodium-glucose cotransporter-2 (SGLT2) inhibitors (30.5%), and aspirin in primary prevention (36.3%). Only 7.9% and 9.5% selected cardiologists and preventive cardiologists, respectively, to prescribe SGLT2 inhibitors. Most preferred specialists (i.e. cardiology and endocrinology) to manage advanced lipid disorders, refractory hypertension, and premature coronary heart disease. The most common conditions selected for preventive cardiologists to manage were genetic lipid disorders (17%), cardiovascular risk assessment (15%), dyslipidemia (13%), and refractory/resistant hypertension (12%). For CVD prevention practice, providers favored all specialties to manage common conditions, specialists to manage complex conditions, and preventive cardiologists to manage advanced lipid disorders. Cardiologists were least preferred to prescribe SGLT2 inhibitor. Future research should explore reasons for selected CVD prevention practice preferences to optimize care coordination and for effective use of limited expertise.


Trends in Primary Surgical Resection and Chemotherapy for Metastatic Colorectal Cancer, 2000-2016.
When, whether, and in whom primary tumor resection (PTR) for patients with metastatic colorectal cancer (CRC) is indicated remains unknown. With advances in multiagent systemic chemotherapy, PTR may be undertaken less frequently. The aim of this study was to obtain estimates of changes in the utilization of PTR and chemotherapy for metastatic CRC. Patients diagnosed with metastatic CRC between 2000 and 2016 were identified from Surveillance Epidemiology, and End Results (SEER) registry. Multivariable logistic regression defined odds of undergoing PTR. The analysis was also stratified by primary site (colon vs. rectum), age (younger than 50 vs. 50 y and older), and whether patients also underwent resection of metastatic sites (yes vs. no). The secondary endpoint of interest was the receipt of any chemotherapy, also assessed by multivariable logistic regression. Among 99,835 patients with metastatic CRC, 55,527 (55.7%) underwent PTR. The odds of undergoing PTR decreased with a later year of diagnosis, with patients diagnosed in 2016 being 61.1% less likely to undergo surgery than those diagnosed in 2000 (adjusted odds ratio=0.39, 95% confidence interval: 0.36-0.42, P<0.0001; absolute percentage: 62.3% to 43.8%). Similar trends by year for PTR were observed among each of the subgroups, although patients with colon primary, young adults (age younger than 50 y), and patients also undergoing metastasectomy were more likely to undergo PTR (P<0.001 for all). In contrast, the odds of receiving chemotherapy increased dramatically with a later year of diagnosis (adjusted odds ratio=2.21, 95% confidence interval: 2.04-2.40, P<0.0001). From 2000 to 2016, there was a sharp decline in the rate of PTR for patients with metastatic CRC, while the use of chemotherapy increased over the same period. Prospective studies are needed to define the optimal local treatment for patients with metastatic CRC.


Efficacy and tolerability of bevacizumab plus capecitabine as first-line therapy in patients with advanced hepatocellular carcinoma.
Molecularly targeted agents with anti-angiogenic activity, including bevacizumab, have demonstrated clinical activity in patients with advanced/metastatic hepatocellular carcinoma (HCC). This multicentre phase II study involving patients from several Asian countries sought to evaluate the safety and efficacy of bevacizumab plus capecitabine in this population. Histologically proven/clinically diagnosed advanced HCC patients received bevacizumab 7.5 mg kg(-1) on day 1 and capecitabine 800 mg m(-2) twice daily on days 1-14 every 3 weeks as first-line therapy. A total of 45 patients were enrolled; 44 (96%) had extrahepatic metastasis and/or major vessel invasion and 30 (67%) had hepatitis B. No grade 3/4 haematological toxicity occurred. Treatment-related grade 3/4 non-haematological toxicities included diarrhoea (n=2, 4%), nausea/vomiting (n=1, 2%), gastrointestinal bleeding (n=4, 9%) and hand-foot syndrome (n=4, 9%). The overall response rate (RECIST) was 9% and the disease control rate was 52%. Overall, median progression-free survival (PFS) and overall survival (OS) were 2.7 and 5.9 months, respectively. Median PFS and OS were 3.6 and 8.2 months, respectively, for Cancer of the Liver Italian Programme (CLIP) score < or =3 patients, and 1.4 and 3.3 months, respectively, for CLIP score 4 patients. The bevacizumab-capecitabine combination shows good tolerability and modest anti-tumour activity in patients with advanced HCC.


Characterization of calcineurin A and B genes in the abalone, Haliotis diversicolor, and their immune response role during bacterial infection.
Calcineurin (CN) is known to be involved in many biological processes, particularly, the immune response mechanism in many invertebrates. In this study, we characterized both HcCNA and HcCNB genes in Haliotis diversicolor, documented their expression in many tissues, and discerned their function as immune responsive genes against Vibrio parahaemolyticus infection. Similar to other mollusk CNs, the HcCNA gene lacked a proline-rich domain and comprised only one isoform of its catalytic unit, in contrast to CNs found in mammals. HcCNB was highly conserved in both sequence and domain architecture. Quantitative PCR and in situ hybridization revealed that the genes were broadly expressed and were not restricted to tissues traditionally associated with immune function. Upon infection of H. diversicolor with V. parahaemolyticus (a bacteria that causes serious disease in crustaceans and mollusks), both HcCNA and HcCNB genes were highly up-regulated at the early phase of bacterial infection. HcCNB was expressed significantly higher than HcCNA in response to bacterial challenge, suggesting its independent or more rapid response to bacterial infection. Together, the two CN genes are unique in their gene structure (particular HcCNA) and distribution in mollusk species and likely function as immune responsive genes along with many other genes that are enhanced in the early phase of V. parahaemolyticus infection in abalone.


Symbolic representation of hyperspectral data.
We have developed a symbolic representation of hyperspectral data using the scale space techniques of Witkin. We created a scale space image of hyperspectral data from convolution with Gaussian masks and then a fingerprint that extracts individual features from the original data. The fingerprint provides a context that pairs inflection points and assigns them to a feature, generates a measure of importance for each feature, and relates features to each other. The representation is an ordered sequence of triplets containing a measure of importance related to the area of each feature and the left and right inflection points of the feature. The description is compact, quantitative, and hierarchical, describing the hyperspectral curve by its most important structural features first, followed by features of lesser importance.


Antiviral activity of PLK1-targeting siRNA delivered by lipid nanoparticles in HBV-infected hepatocytes.
A link between HBV and PLK1 was clearly evidenced in HBV-driven carcinogenesis, and we have also recently shown that PLK1 is a proviral factor in the early phases of HBV infection. Moreover, we have shown that BI-2536, a small molecule PLK1 inhibitor, was very efficient at inhibiting HBV DNA neosynthesis, notably by affecting nucleocapsid assembly as a result of the modulation of HBc phosphorylation. Yet, as small molecule kinase inhibitors often feature poor selectivity, a more specific and safer strategy to target PLK1 would be needed for a potential development against chronic HBV infections. Here, we analysed using both freshly isolated primary human hepatocytes and differentiated HepaRG, the anti-HBV properties of an LNP-encapsulated PLK1-targeting siRNA. Standard assays were used to monitor the effect of LNP siPLK1, or controls (LNP siHBV and LNP siNon-targeting), on HBV replication and cell viability. A dose as low as 100 ng/ml of LNP-siPLK1 resulted in a >75% decrease in secreted HBV DNA (viral particles), which was comparable to that obtained with LNP siHBV or 10 µM of tenofovir (TFV), without affecting cell viability. Interestingly, and in contrast to that obtained with TFV, a strong inhibition of viral RNA and HBe/HBsAg secretions was also observed under LNP siPLK1 treatment. This correlated with a significant intracellular decrease of vRNA accumulation, which was independent of any change in cccDNA levels, thus suggesting a transcriptional or post-transcriptional modulation. Such an effect was not obtained with a biochemical approach of PLK1 inhibition, suggesting an enzymatic-independent role of PLK1. This study emphasizes that a specific PLK1 inhibition could help in achieving an improved HBsAg loss in CHB patients, likely in combination with other HBsAg-targeting strategies.


Effect of molecular weight profile of sorghum proanthocyanidins on resistant starch formation.
There is a growing interest to increase resistant starch (RS) in foods through natural modification of starch. Sorghum tannins (proanthocyanidins, PAs) were recently reported to interact with starch, increasing RS. However, there is no information about how the molecular weight profile of PAs affects RS formation. This study investigated how different-molecular-weight PAs from sorghum affected RS formation in different starch models. The levels of RS were higher (331-437 mg g(-1)) when high-amylose starch was cooked with phenolic extracts containing mostly high-molecular-weight PAs compared with extracts containing lower-molecular-weight PAs or monomeric catechin (249-285 mg g(-1)). In general, binding capacity of PAs with amylose increased proportionally with molecular weight. For example, the percentage of PAs bound to amylose increased from 45% (PAs with degree of polymerization (DP) = 6) to 94% (polymeric PAs, DP > 10). The results demonstrate that molecular weight of the PAs directly affects their interaction with starch: the higher the molecular weight, the stronger the binding to amylose and the higher the RS formation. Polymeric PAs from sorghum can naturally modify starch by interacting strongly with amylose and are thus most suitable to produce foods with higher RS.


High Glycemic Load Is Associated with Cognitive Decline in Apolipoprotein E ε4 Allele Carriers.
Recent evidence suggests that a high glycemic load (GL) diet is a risk factor for dementia, especially among apolipoprotein E ε4 allele (APOE4) carriers, while its association with cognitive decline is poorly known. Here, we investigated the association of high-GL meals with cognitive decline in older adults during a 12-year follow-up, according to their APOE4 carrier status. We used random-effect models and data from 2539 elderly participants from the Three-City study who completed a food frequency questionnaire (FFQ) to longitudinally assess the association of GL with changes in different cognitive domains (verbal fluency, visual memory, attention, visual motor processing speed, episodic memory). In APOE4 carriers, afternoon snack with high GL was significantly associated with cognitive decline in visual memory, episodic memory, and global cognition compared with APOE4 non-carriers. This study suggests a detrimental association between a high-GL diet and cognitive decline. The promotion of a low GL diet as a target to prevent cognitive decline in high-risk populations deserves more research.


Fourier transform infrared (FT-IR) spectromicroscopy to identify cell organelles: correlation with fluorescence staining in MCF-7 breast cancer cells.
Biomolecules display specific vibrational signatures in the infrared (IR) range, and organelles that concentrate these biomolecules can be identified by these IR signatures. Subcellular identification and location of cell organelles using IR signatures is attractive as it does not require the use of any specific trackers and is thus non-invasive and non-destructive. We show here that endogenous IR absorptions are relevant to detecting and imaging the nucleus, the cytoplasm, and the Golgi apparatus/endoplasmic reticulum in MCF-7 breast cancer cells, and we compare these results with our previous work on the HeLa cell line. We correlate maps of fixed and dried cells obtained by synchrotron radiation Fourier transform infrared (SR FT-IR) spectromicroscopy with epifluorescence images using fluorescent trackers for Golgi apparatus and nucleus, namely BODIPY TR C5-ceramide complexed to BSA and DAPI, respectively. Interestingly, the ratios of the IR bands CH2 : CH3 (both asymmetric and symmetric) and CO((ester)):amide I were shown to be reliable gauges of the lipidic character of a cellular compartment, the -CH2 and the CO((ester)) absorptions increasing with the presence of inner membranes like in the Golgi apparatus.


Thinning algorithms for gray-scale pictures.
Elongated black objects in black-and-white pictures can be ``thinned'' to arcs and curves, without changing their connectedness, by (repeatedly) deleting black border points whose deletion does not locally disconnect the black points in their neighborhoods. This technique generalizes to gray-scale pictures if we use a weighted definition of connectedness: two points are ``connected'' if there is a path joining them on which no point is lighter than either of them. We can then ``thin'' dark objects by changing each point's gray level to the minimum of its neighbors' gray levels, provided this does not disconnect any pair of points in its neighborhood. Examples illustrating the performance of this technique are given.


Adaptive radiations in natural populations of prokaryotes: innovation is key.
Prokaryote diversity makes up most of the tree of life and is crucial to the functioning of the biosphere and human health. However, the patterns and mechanisms of prokaryote diversification have received relatively little attention compared to animals and plants. Adaptive radiation, the rapid diversification of an ancestor species into multiple ecologically divergent species, is a fundamental process by which macrobiological diversity is generated. Here, we discuss whether ecological opportunity could lead to similar bursts of diversification in bacteria. We explore how adaptive radiations in prokaryotes can be kickstarted by horizontally acquired key innovations allowing lineages to invade new niche space that subsequently is partitioned among diversifying specialist descendants. We discuss how novel adaptive zones are colonized and exploited after the evolution of a key innovation and whether certain types of are more prone to adaptive radiation. Radiation into niche specialists does not necessarily lead to speciation in bacteria when barriers to recombination are absent. We propose that in this scenario, niche-specific genes could accumulate within a single lineage, leading to the evolution of an open pangenome.


Sex, gender and work segregation in the cultural industries.
This chapter addresses work 'segregation' by sex in the cultural industries. We outline some of the main forms this takes, according to our observations: the high presence of women in marketing and public relations roles; the high numbers of women in production co-ordination and similar roles; the domination of men of more prestigious creative roles; and the domination by men of technical jobs. We then turn to explanation: what gender dynamics drive such patterns of work segregation according to sex? Drawing on interviews, we claim that the following stereotypes or prevailing discourses, concerning the distinctive attributes of women and men, may influence such segregation: that women are more caring, supportive and nurturing; that women are better communicators; that women are 'better organized'; and that men are more creative because they are less bound by rules.


Using activity data to explore the influence of case-load size on care patterns.
A limited case-load size is considered crucial for some forms of intensive case management and many countries have undertaken extensive reorganisation of mental health services to achieve this. However, there has been limited empirical work to explore this specifically. To test whether there is a discrete threshold for changes in intensive case management practice determined by case-load size. "Virtual" case-load sizes were calculated for patients from their actual contacts over a 2-year period and were compared with the proportions of contacts devoted to medical and non-medical care (as a proxy for a more comprehensive service model). There were 39 025 recordings for 545 patients over 2 years, with a mean rate of contacts per full-time case manager per month of 48 (range 35-60). There was no variation in the proportion of non-medical contacts when case-load sizes were over 1:20 but there was a convincing linear relationship when sizes were between 1:10 and 1:20. Case-load size between 1:10 and 1:20 does affect the practice of case management. However, there is no support for a paradigm shift in practice at a discrete level.


Recognition of polyadenosine RNA by the zinc finger domain of nuclear poly(A) RNA-binding protein 2 (Nab2) is required for correct mRNA 3'-end formation.
Proteins bound to the poly(A) tail of mRNA transcripts, called poly(A)-binding proteins (Pabs), play critical roles in regulating RNA stability, translation, and nuclear export. Like many mRNA-binding proteins that modulate post-transcriptional processing events, assigning specific functions to Pabs is challenging because these processing events are tightly coupled to one another. To investigate the role that a novel class of zinc finger-containing Pabs plays in these coupled processes, we defined the mode of polyadenosine RNA recognition for the conserved Saccharomyces cerevisiae Nab2 protein and assessed in vivo consequences caused by disruption of RNA binding. The polyadenosine RNA recognition domain of Nab2 consists of three tandem Cys-Cys-Cys-His (CCCH) zinc fingers. Cells expressing mutant Nab2 proteins with decreased binding to polyadenosine RNA show growth defects as well as defects in poly(A) tail length but do not accumulate poly(A) RNA in the nucleus. We also demonstrate genetic interactions between mutant nab2 alleles and mutant alleles of the mRNA 3'-end processing machinery. Together, these data provide strong evidence that Nab2 binding to RNA is critical for proper control of poly(A) tail length.


Population ecology and fishery of Cittarium pica (Gastropoda: Trochidae) on the Caribbean coast of Costa Rica.
The West Indian Topshell Cittarium pica is artisanally collected on rocky shores along the Caribbean Coast of Costa Rica. There are neither data on the state of its exploitation nor exist any regulation of the fishery From October 2000 to March 2001, the population dynamics of this species were studied at an unexploited and two exploited sites to determine the present impact of the fishery on the resource. Average population density with 14 ind./m2 about three times higher at the unexploited than at the exploited sites. Length-frequency histograms showed a strong shift towards smaller specimens at the exploited sites, which is also reflected in significantly higher rates of total mortality (Z = 4.05 and 4.47) when compared to the unexploited site (Z = 1.47). Von Bertalanffy growth parameters were estimated as k = 0.19-0.28 (yr-1) and L infinity = 104 mm. No significant differences were found among sites. From these values a range of the growth performance index phi was computed (phi = 3.31-3.48) which lies at the lower end of the values reported for other tropical marine gastropods. The size at first maturity for both sexes combined was estimated as 29.20 +/- 1.14 mm. Exploitation rates > 0.6 for both exploited sites and a large fraction of small specimens (< 30 mm) in the catches suggest overexploitation and recruitment overfishing. Based on the estimated maximum sustainable yield we recommend regulative measures for the fishery such as a control of a minimum landing size of 40 mm and a closure of the fishery during its reproductive period (from July to November).


Non-Hodgkin's lymphomas, version 2.2014.
Non-Hodgkin's lymphomas (NHLs) are a heterogeneous group of lymphoproliferative disorders originating in B lymphocytes, T lymphocytes, or natural killer cells. Follicular lymphoma (FL) is the most common subtype of indolent NHL, accounting for approximately 22% of all newly diagnosed cases of NHL. The incorporation of rituximab to chemotherapy regimens has become a widely accepted standard of care for first-line therapy for patients with FL. Maintenance and consolidation therapy with rituximab and radioimmunotherapy have also been associated with improved progression-free survival in patients experiencing response to first-line therapy. Despite therapeutic advances that have improved outcomes, FL is generally considered a chronic disease characterized by multiple recurrences with current therapies. This manuscript discusses the recommendations outlined in the NCCN Guidelines for the diagnosis and management of patients with FL.


Volume image registration by cross-entropy optimization.
Cross-entropy (CE), an information-theoretic measure, quantifies the difference between two probability density functions. This measure is applied to volume image registration. When a good prior estimation of the joint distribution of the voxel values of two images in registration is available, the CE can be minimized to find an optimal registration. If such a prior estimation is not available, one seeks the registration which gives a joint distribution different from unlikely ones as much as possible, i.e., the CE is maximized to find an optimal registration. When the unlikely distribution is a uniform one, CE maximization reduces to joint entropy minimization; when the unlikely distribution is proportional to one of the marginal distributions, it reduces to conditional entropy minimization; when the unlikely distribution is the product of two marginal distributions, it degenerates to mutual-information maximization. These different CEs are added together and are used as criteria for image registration. The accuracy and robustness of this new approach are tested and compared using a likely joint distribution and various unlikely joint distributions and their combinations.


Isolated congenital atrioventricular block diagnosed in utero: natural history and outcome.
Isolated congenital atrioventricular block (CAVB) diagnosed in utero is associated with a high morbidity and mortality. Prognosis is especially poor when heart rate drops below 55 beats per minute (bpm) and when fetal hydrops develops. We describe the natural history and outcome of 24 infants with isolated CAVB diagnosed in utero, review the literature, and assess the risk factors that could predict outcome. This was a retrospective multicenter study of 24 patients with isolated CAVB diagnosed in utero. CAVB was detected at a mean gestational age (GA) of 24.7 +/- 5.1 weeks. Ten fetuses initially presented with complete heart block. Low heart rate or incomplete heart block was the first documentation of bradyarrhythmia in the other 14 fetuses. In 11 of them, CAVB developed during pregnancy after a median time of 3 (range 1-16) weeks. Fetal hydrops developed in 10 of 24 (42%) fetuses at a mean GA of 27.6 +/- 5.1 weeks. Hydropic fetuses showed lower heart rates during pregnancy (47 +/- 10 bpm) than non-hydropic fetuses (57 +/- 10 bpm). There were three intrauterine deaths; all were hydropic and female. Nine viable females and 12 males were born at a mean GA of 37.1 +/- 6.1 weeks with an average birth weight of 3097 +/- 852 g. Fifteen CAVB patients required pacemaker (PM) intervention, 10 of them immediately after birth. Dilated cardiomyopathy (DCM) developed in three infants of whom two died of congestive heart failure, shortly after the diagnosis was made; one is still alive. Mortality before or after birth was 21%, and was associated with heart rates below 50 bpm and development of fetal hydrops. Poor outcome, defined as death, PM implantation, or development of DCM, occurred in 83% of cases and was associated with heart rates below 60 bpm during pregnancy. Isolated CAVB diagnosed in utero is associated with high morbidity and mortality. Patients who develop fetal hydrops show lower heart rates during pregnancy than patients who do not. A fetal heart rate below 50 bpm and development of fetal hydrops is associated with increased mortality. Rates below 60 bpm are associated with PM requirement and/or DCM.


Improvement in quality of life during 5 years after coronary artery bypass grafting.
To describe the changes in various aspects of quality of life (QOL) from before coronary artery bypass grafting (CABG) to 5 years after the procedure. Patients who underwent CABG in the western region of Sweden in 1988-1991 were approached with questionnaires evaluating their QOL prior to and 3 months and 1, 2, and 5 years after the operation. Three different instruments were used: the Nottingham Health Profile, the Psychological General Well-Being Index, and the Physical Activity Score. In all 2121 patients underwent CABG, of whom 310 died during 5 years of follow-up. With all three instruments QOL had improved 5 years after CABG compared with prior to the operation. However, all three instruments revealed a slight but significant deterioration in estimated QOL between 2 and 5 years after CABG. QOL 5 years after CABG is better than that prior to the operation, but between 2 and 5 years after the operation a slight deterioration in QOL is observed.


Antigenic characterization of adult Wuchereria bancrofti filarial nematodes.
Adult Wuchereria bancrofti were recovered from infected Presbytis cristatus monkeys and radio-isotope labelled extrinsically with 125I and in vitro with [35S]methionine. 125I labelling of the surface of adult W. bancrofti permitted a comparison between the major surface antigens of this species and those from the related lymphatic filariae, Brugia malayi and B. pahangi. All species bear a prominent Mr 29,000 surface antigen but among the differences observed were the strongly labelled molecules with Mr 58,000 and 67,000 in W. bancrofti which are extremely faint in the Brugia species. The [35S]methionine label was effectively incorporated into somatic parasite proteins in vitro although it was not possible to identify any secreted proteins in this way. The antigenicity of these products was investigated using a variety of sera from homologous and heterologous infections and the immunoprecipitation patterns highlighted particular differences between somatic proteins of male and female worms. One secreted antigen was detected, however, by virtue of its phosphorylcholine epitopes, in the culture medium of mixed adult worms; medium from male W. bancrofti adults was negative although homogenates of either sex of adult W. bancrofti were strongly positive in the same system.


A longitudinal study of risk factors in asthma and chronic bronchitis in childhood.
The prevalence, persistence, incidence and remission rates of reported diagnosed asthma and chronic bronchitis has been studied longitudinally over ten years in a representative community sample of children and adolescents. The objectives are to evaluate the rates of change over time, the role of family history of disease, and the social risk factors in the longitudinal course of disease. The relationship between the two reported diagnoses is very high, about half of each group having the other diagnosis. Each diagnosis has specific symptoms with which it is associated. Remission of clinical disease is associated with continuing symptoms in about half of such cases. The incidence rates and remissions of cases as they have occurred over time were associated with different symptoms and risk factors. A family history of lung disease and family social characteristics are significant risk factors affecting the presence of these diseases, and how they change. Discussion suggests these factors may affect possible precursors, such as lower respiratory tract illnesses, as well as personal habits (such as smoking), which are related to both diagnoses. Further, the interactions of these risk factors appear to significantly influence impairments of lung function at the beginning of adult life, and potential subsequent disease.


The importance of androgen receptors in breast cancer.
Breast cancer (BC) is the most common malignancy among women worldwide, and one of the leading causes of cancer-related deaths in females. For the breast malignant tumors there are numerous targeted therapies, depending on the receptors expressed. Regulating the process of epithelial-mesenchyme transcription, the steroid nuclear receptors are important in invasion and progression of BC cells. Till now, it is known that androgen receptor (AR) is present in about 60-80% of BC cells but, unfortunately, there is no targeted therapy available yet. We revised the recent literature that included the AR mechanism of action in patients diagnosed with breast cancer, the preclinical, retrospective and clinical studies and the aspects related to the prognosis of these patients, depending on the molecular subtype. A total of 12 articles were eligible for this review. AR positivity was assessed using immunohistochemistry. Herein, neither 1 nor 10% cut-points were robustly prognostic. AR was an independent prognostic marker of BC outcome, especially in triple negative BC group. AR is a potential targeted pathway which can improve the prognostic of AR positive patients with BC. Further preclinical and clinical studies are necessary to clarify the mechanism of action and to establish the drugs which can be used, either alone or in combination.


Non-invasive scoring of cellular atypia in keratinocyte cancers in 3D LC-OCT images using Deep Learning.
Diagnosis based on histopathology for skin cancer detection is today's gold standard and relies on the presence or absence of biomarkers and cellular atypia. However it suffers drawbacks: it requires a strong expertise and is time-consuming. Moreover the notion of atypia or dysplasia of the visible cells used for diagnosis is very subjective, with poor inter-rater agreement reported in the literature. Lastly, histology requires a biopsy which is an invasive procedure and only captures a small sample of the lesion, which is insufficient in the context of large fields of cancerization. Here we demonstrate that the notion of cellular atypia can be objectively defined and quantified with a non-invasive in-vivo approach in three dimensions (3D). A Deep Learning (DL) algorithm is trained to segment keratinocyte (KC) nuclei from Line-field Confocal Optical Coherence Tomography (LC-OCT) 3D images. Based on these segmentations, a series of quantitative, reproducible and biologically relevant metrics is derived to describe KC nuclei individually. We show that, using those metrics, simple and more complex definitions of atypia can be derived to discriminate between healthy and pathological skins, achieving Area Under the ROC Curve (AUC) scores superior than 0.965, largely outperforming medical experts on the same task with an AUC of 0.766. All together, our approach and findings open the door to a precise quantitative monitoring of skin lesions and treatments, offering a promising non-invasive tool for clinical studies to demonstrate the effects of a treatment and for clinicians to assess the severity of a lesion and follow the evolution of pre-cancerous lesions over time.


Glucocorticoid treatment for nasal polyps. The use of topical budesonide powder, intramuscular betamethasone, and surgical treatment.
The treatment of nasal polyps is controversial, and medical treatment alone has been little investigated to our knowledge. To examine the efficacy of therapy using only topical budesonide powder and topical budesonide powder supplemented with surgical removal or intramuscular betamethasone. Randomized, double-blind comparison of 2 dosages and additional treatment, if therapy failed. After 1 year, treatment with medication was stopped, and the demand for renewed treatment was monitored for another year. Patients with bilateral nasal polyps who demanded treatment were consecutively enrolled in a hospital outpatient clinic or specialty private practice. During a 15-month period from 1990 to 1992, a total of 126 patients entered the 2-year study. In phase 1, randomized and double-blind treatment with a topical medication, budesonide powder, 800 micrograms or 400 micrograms daily, or a placebo was given for 1 month. In phase 2, randomized and double-blind treatment with budesonide powder, 800 micrograms or 400 micrograms daily, was given. At the end of phase 1, failed therapy was supplemented by randomly assigned treatment of either surgical removal or a single injection of sustained released betamethasone. In phase 3, treatment with the medication was discontinued, and patients were monitored for another year. The time when treatment was required again was noted. The present article deals with phases 2 and 3. Patients' scores of treatment efficacy as well as symptoms and signs. Semiquantitative measurement of sense of smell and calculation of peak expiratory flow rate index based on nasal and oral peak expiratory flow. In all outcome measures, a comparison of the data before treatment with the corresponding figures during phase 2 showed statistically significant efficacy. The clinical course was described at the end of phase 2. About 85% of the patients, including those who received additional therapy because the initial therapy failed, rated total or substantial control over the symptoms. The 2 dosages investigated showed equal results. These findings were consistent with the signs recorded and the peak expiratory flow rate index. The results of phase 3 showed that 50% of patients had demanded treatment after 4 months, while 34% managed without medication after 1 year. The clinical course in this study showed that most patients with nasal polyps do well with medical treatment. Therefore, surgery was required in few patients. However, the potential of medical treatment should be further explored in future studies.


Access to firearms: When and how do mental health clients become prohibited from owning guns?
Psychologists and other mental health clinicians are often called upon to assess violence and suicide risk. Access to firearms has important implications for client safety, but most psychologists report receiving little or no training on discussing firearms with clients in the context of delivering psychological services such as screening, assessment, or psychotherapy. Understanding the extent to which firearm legislation limits legal firearm ownership among people with mental illness can help clinicians identify and respond to situations in which people at high risk for violence may legally own or acquire a firearm. In this article, we discuss three clinical cases and review firearm legislation as it pertains to people with mental illness. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Risk-neutral density extraction from option prices: improved pricing with mixture density networks.
One of the central goals in finance is to find better models for pricing and hedging financial derivatives such as call and put options. We present a new semi-nonparametric approach to risk-neutral density extraction from option prices, which is based on an extension of the concept of mixture density networks. The central idea is to model the shape of the risk-neutral density in a flexible, nonlinear way as a function of the time horizon. Thereby, stylized facts such as negative skewness and excess kurtosis are captured. The approach is applied to a very large set of intraday options data on the FTSE 100 recorded at LIFFE. It is shown to yield significantly better results in terms of out-of-sample pricing accuracy in comparison to the basic and an extended Black-Scholes model. It is also significantly better than a more elaborate GARCH option pricing model which includes a time-dependent volatility process. From the perspective of risk management, the extracted risk-neutral densities provide valuable information for value-at-risk estimations.


[Training in social skills in patients with acquired brain damage].
Brain damaged patients have cognitive deficits, behaviour disorders and personality changes which affect socio-familial behaviour. Frequently, these changes generate considerable disturbance between family members and make it very difficult to return to work. We describe a pilot study which led to the development and application of a focal rehabilitation programme aimed at retraining this group in social skills. The programme was intensive (3 months) and included 6 outpatients (5 men and 1 woman) who took part in programmes of neuropsychological rehabilitation after traumatic or vascular cerebral lesions (X = 19.2 months after the lesion occurred). The average age of the patients was 27.2 years and their average IQ 109.2. Individual and group interventions were combined (role-playing, 'make-believe' work, video filming) emphasizing particularly the processes of learning without making mistakes and the extension of achievements to the natural setting (controlled trials). We state and describe the relationship between the nature of neuropsychological and psychopathological deficits, and the characteristics of the limitations in social behaviour. The patients showed a significant reduction in the level of anxiety and aggressive behaviour and improvement in consciousness of their defects. Some improvement was seen in expressing opinions and emotions, and in ability to adapt social conduct to different situations (flexible behaviour). Finally we describe the conclusions drawn as to this treatment for future review and improvement of the programme.


Double minute amplification of mutant PDGF receptor α in a mouse glioma model.
In primary brain tumors, oncogenes are frequently amplified and maintained on extrachromosomal DNA as double minutes (DM), but the underlying mechanisms remain poorly understood. We have generated a mouse model of malignant glioma based on knock-in of a mutant PDGF receptor α (PDGFRα) that is expressed in oligodendrocyte precursor cells (OPCs) after activation by a Cre recombinase. In the tumor suppressor INK4/Arf(-/-) background, mutant animals frequently developed brain tumors resembling anaplastic human gliomas (WHO grade III). Besides brain tumors, most animals also developed aggressive fibrosarcomas, likely triggered by Cre activation of mutant PDGFRα in fibroblastic cell lineages. Importantly, in the brain tumors and cell lines derived from brain tumor tissues, we identified a high prevalence of DM Pdgfra gene amplification, suggesting its occurrence as an early mutational event contributing to the malignant transformation of OPCs. Amplicons extended beyond the Pdgfra locus and included in some cases neighboring genes Kit and Kdr. Our genetically defined mouse brain tumor model therefore supports OPC as a cell of origin for malignant glioma and offers an example of a defined temporal sequence of mutational events, thus providing an entry point for a mechanistic understanding of DM gene amplification and its functionality in gliomagenesis.


[Post-orthodontic rehabilitation of the sequellae of alveolar clefts: implantation or periodontal plastic surgery?].
Just as they do for many cranio-facial deformities, the treatment team begins the care of cleft palate patients early and continues treatment over a long period. Throughout the growth period, they perform well-timed interventions and re-evaluations according to the individual plan they have prepared. When growth is completed and the orthopedic and orthodontic corrections have contributed as much as possible to an esthetic and functional equilibrium, a final prosthetic replacement will frequently be required. When the correct edentulous space between the teeth on each side of the defect has been created, or maintained, practitioners can choose between two alternatives: implants or traditional fixed bridges. They should analyze any secondary anatomic discrepancies, severe or superficial, that may have developed, and prepare the best therapeutic pathway for a reconstruction of crestal gingival morphology by means of periodontal surgery.


Nascent RNA sequencing reveals mechanisms of gene regulation in the human malaria parasite Plasmodium falciparum.
Gene expression in Plasmodium falciparum is tightly regulated to ensure successful propagation of the parasite throughout its complex life cycle. The earliest transcriptomics studies in P. falciparum suggested a cascade of transcriptional activity over the course of the 48-hour intraerythrocytic developmental cycle (IDC); however, the just-in-time transcriptional model has recently been challenged by findings that show the importance of post-transcriptional regulation. To further explore the role of transcriptional regulation, we performed the first genome-wide nascent RNA profiling in P. falciparum. Our findings indicate that the majority of genes are transcribed simultaneously during the trophozoite stage of the IDC and that only a small subset of genes is subject to differential transcriptional timing. RNA polymerase II is engaged with promoter regions prior to this transcriptional burst, suggesting that Pol II pausing plays a dominant role in gene regulation. In addition, we found that the overall transcriptional program during gametocyte differentiation is surprisingly similar to the IDC, with the exception of relatively small subsets of genes. Results from this study suggest that further characterization of the molecular players that regulate stage-specific gene expression and Pol II pausing will contribute to our continuous search for novel antimalarial drug targets.


The effect of phenotypic traits and external cues on natal dispersal movements.
1. Natal dispersal has the potential to affect most ecological and evolutionary processes. However, despite its importance, this complex ecological process still represents a significant gap in our understanding of animal ecology due to both the lack of empirical data and the intrinsic complexity of dispersal dynamics. 2. By studying natal dispersal of 74 radiotagged juvenile eagle owls Bubo bubo (Linnaeus), in both the wandering and the settlement phases, we empirically addressed the complex interactions by which individual phenotypic traits and external cues jointly shape individual heterogeneity through the different phases of dispersal, both at nightly and weekly temporal scales. 3. Owls in poorer physical conditions travelled shorter total distances during the wandering phase, describing straighter paths and moving slower, especially when crossing heterogeneous habitats. In general, the owls in worse condition started dispersal later and took longer times to find further settlement areas. Net distances were also sex biased, with females settling at further distances. Dispersing individuals did not seem to explore wandering and settlement areas by using a search image of their natal surroundings. Eagle owls showed a heterogeneous pattern of patch occupancy, where few patches were highly visited by different owls whereas the majority were visited by just one individual. During dispersal, the routes followed by owls were an intermediate solution between optimized and randomized ones. Finally, dispersal direction had a marked directionality, largely influenced by dominant winds. These results suggest an asymmetric and anisotropic dispersal pattern, where not only the number of patches but also their functions can affect population viability. 4. The combination of the information coming from the relationships among a large set of factors acting and integrating at different spatial and temporal scales, under the perspective of heterogeneous life histories, are a fruitful ground for future understanding of natal dispersal.


Caffeine induces neurobehavioral effects through modulating neurotransmitters.
Evidence demonstrates that chronic caffeine exposure, primarily through consumption of coffee or tea, leads to increased alertness and anxiety. Preclinical and clinical studies showed that caffeine induced beneficial effects on mood and cognition. Other studies using molecular techniques have reported that caffeine exhibited neuroprotective effects in animal models by protecting dopaminergic neurons. Moreover, caffeine interacts with dopaminergic system, which leads to improvements in neurobehavioral measures in animal models of depression or attention deficit hyperactivity disorder (ADHD). Glutamatergic receptors have been found to be involved on the neurobiological effects of caffeine. Additionally, caffeine has been found to suppress the inhibitory (GABAergic) activity and modulate GABA receptors. Studies have also found that modulating these neurotransmitters leads to neurobehavioral effects. The linkage between the modulatory role of caffeine on neurotransmitters and neurobehavioral effects has not been fully discussed. The purpose of this review is to discuss in detail the role of neurotransmitters in the effects of caffeine on neurobehavioral disorders.


Functional analyses of thymic CD5+ B cells. Responsiveness to major histocompatibility complex class II-restricted T blasts but not to lipopolysaccharide or anti-IgM plus interleukin 4.
The function of thymic B cells in several standard in vitro assays was investigated. Thymic B cells, 75% of which were CD5+, showed a poor responsiveness to the mitogens LPS or anti-mu plus IL-4. Both proliferation and antibody formation were much lower in thymic than splenic B cell cultures. However, CD5- B cells purified using a cell sorter responded well to B cell stimulants, whereas purified CD5+ thymic B cells did not, indicating that CD5+ thymic B cells were unresponsive to B cell growth factor or LPS. Thymic B cells could be activated polyclonally by direct interaction with alloreactive T blasts, as manifested by DNA synthesis and antibody formation. These findings indicate that CD5+ thymic B cells may not be stimulated via sIg and IL-4, but require instead direct interaction with T blasts.


Reduction of trimethylamine N-oxide by Escherichia coli as anaerobic respiration.
E. coli was found to grow anaerobically on lactate in the presence of trimethylamine N-oxide (TMANO), reducing it to trimethylamine. Anaerobic growth on glucose was promoted in the presence of TMANO. When a culture grown in complex medium was transferred to defined medium, growth on glucose and ammonia took place in the presence of TMANO after consumption of complex nutrients introduced with the preculture, in contrast to growth in nitrate respiration. The amounts of ethanol, succinate, and lactate among the fermentation products were decreased and that of acetate was increased in the presence of TMANO. Formate generation was much reduced at pH 7.4, whereas stoichiometric formation of formate was observed in the absence of TMANO. Cells grown anaerobically in the presence of TMANO had a higher activity of amine N-oxide reductase than cells grown under other conditions. The content of cytochrome-558 was elevated in the presence of TMANO during growth in complex medium. Cytochrome c-552 found in cells grown in diluted complex medium or defined medium in the presence of TMANO was oxidized by TMANO in cell extracts. The molar growth yield on glucose was higher in the presence of TMANO than in its absence and lower than that in the presence of nitrate.


Ganglionic Eminence Anomalies and Coexisting Cerebral Developmental Anomalies on Fetal MR Imaging: Multicenter-Based Review of 60 Cases.
The ganglionic eminences are transient fetal brain structures that produce a range of neuron types. Ganglionic eminence anomalies have been recognized on fetal MR imaging and anecdotally found in association with a number of neurodevelopmental anomalies. The aim of this exploratory study was to describe and analyze the associations between ganglionic eminence anomalies and coexisting neurodevelopmental anomalies. This retrospective study includes cases of ganglionic eminence anomalies diagnosed on fetal MR imaging during a 20-year period from 7 centers in Italy and England. Inclusion criteria were cavitation or increased volume of ganglionic eminences on fetal MR imaging. The studies were analyzed for associated cerebral developmental anomalies: abnormal head size and ventriculomegaly, reduced opercularization or gyration, and abnormal transient layering of the developing brain mantle. The results were analyzed using χ2 and Fisher exact tests. Sixty fetuses met the inclusion criteria (21 females, 24 males, 15 sex unknown). Thirty-four had ganglionic eminence cavitations (29 bilateral and 5 unilateral), and 26 had increased volume of the ganglionic eminences (19 bilateral, 7 unilateral). Bilateral ganglionic eminence cavitations were associated with microcephaly (P = .01), reduced opercularization, (P < .001), reduced gyration (P < .001), and cerebellar anomalies (P = .01). Unilateral ganglionic eminence cavitations were not significantly associated with any particular feature. Bilateral increased volume of the ganglionic eminences showed an association with macrocephaly (P = .03). Unilateral increased volume was associated with macrocephaly (P = .002), abnormal transient layering (P = .001), unilateral polymicrogyria (P = .001), and hemimegalencephaly (P < .001). Ganglionic eminence anomalies are associated with specific neurodevelopmental anomalies with ganglionic eminence cavitations and increased ganglionic eminence volume apparently having different associated abnormalities.


Trichomonas vaginalis interactions with fibronectin and laminin.
The sexually transmitted protozoan Trichomonas vaginalis cytoadheres to vaginal epithelial cells and causes contact-dependent cytotoxicity which, when combined with the normal exfoliation process, leads to erosion of the epithelium, which may allow trichomonads into extracellular matrix and basement membrane sites. Therefore, the association of T. vaginalis with immobilized fibronectin (FN) and laminin (LM) on cover-slips was examined. Binding of live parasites to coated cover-slips was time- and parasite-density-dependent. Coincubation with an inhibitor of trichomonad cysteine proteinases resulted in an increased attachment of parasites to FN but had no effect on binding to LM, denoting that protease activity influenced optimal FN associations. Further, 20 h mid-exponential phase trichomonads placed in fresh culture medium for 3 h gave higher levels of binding to FN, suggesting that changes during growth in vitro to T. vaginalis organisms affect maximal levels of binding to FN. Extended incubation with substrates diminished the capacity of parasites to bind FN and LM. Treatment of live organisms with periodate reduced binding to LM but not FN, suggesting a role for carbohydrates. In addition, trypsinization of live parasites decreased numbers bound to both substrates. Placement of trypsinized parasites in medium for 2 h fully regenerated binding to FN but not LM. Incubation of trypsinized parasites with cycloheximide abrogated regeneration of attachment to FN, affirming a role for synthesized surface proteins in FN binding. Importantly, the T. vaginalis adhesin proteins that mediate cytoadherence, and iron, a factor that regulates adhesin synthesis, were not involved in FN and LM recognition. These results suggest a role for surface proteins and carbohydrates in trichomonal associations with FN and LM, respectively.


Which Are the Key Kinematic and Kinetic Components to Distinguish Recovery Strategies for Overground Slips Among Community-Dwelling Older Adults?
Slip outcomes are categorized as either a backward loss of balance (LOB) or a no loss of balance (no-LOB) in which an individual does not take a backward step to regain their stability. LOB includes falls and nonfalls, while no-LOB includes skate overs and walkovers. Researchers are uncertain about which factors determine slip outcomes and at which critical instants they do so. The purpose of the study was to investigate factors affecting slip outcomes in proactive and early reactive phases by analyzing 136 slip trials from 68 participants (age: 72.2 [5.3] y, female: 22). Segment angles and average joint moments in the sagittal plane of the slipping limb were compared for different slip outcomes. The results showed that knee flexor, hip extensor, and plantar flexor moments were significantly larger for no-LOB than for LOB in the midproactive phase, leading to smaller shank-ground and foot-ground angles at the slip onset, based on forward dynamics. In the early reactive phase, the hip extensor and plantar flexor moments were larger for no-LOB than for LOB, and all segment angles were smaller for no-LOB. Our findings indicate that the shank angle and knee moment were the major determinants of slip outcomes in both proactive and reactive phases.


Phonetic-phonological performance of typical younger and aged adults from Brazil's capital city.
Given the sociodemographic diversity in Brazil, it is fundamental to understand the speech performance of a sample from the Brazilian capital. The repetition task can assess phonological and motor-phonetic planning. Previous studies found phonological-phonetic performance of speakers to be associated with education, age, and other demographic factors. To compare the phonetic-phonological performance for speech of younger and aged adults in the capital of Brazil, Federal District (FD); to compare FD performance against national normative means based on São Paulo; to determine the association of phonetic-phonological agility with sociodemographic, cognitive, and neuropsychiatric variables for the sample. Cross-sectional study. A total of 60 volunteers from the FD, comprising 30 older adults and 30 younger ones, were stratified by education into two subgroups: 2‒7 years and ≥8 years of education. Data on age, educational level, and socioeconomic status were collected. The Verbal Agility subtest of the Boston Diagnostic Aphasia Examination was applied to assess phonetic-phonological performance. No statistically significant difference in performance for verbal agility was found between aged and younger adults from the FD. There was a statistically significant difference in the phonetic-phonological performance of the FD sample compared with the Brazilian normative mean values. Cognitive and socioeconomic variables were associated with verbal agility. In the capital of Brazil, economic status, age, education, and cognitive variables were associated with verbal agility performance, despite there being no difference in phonetic-phonological performance between younger and aged adult groups. Regional differences in phonetic-phonological performance were also evident.


Multicenter study using paraffin-embedded tumor tissue testing PITX2 DNA methylation as a marker for outcome prediction in tamoxifen-treated, node-negative breast cancer patients.
We recently reported DNA methylation of the paired-like homeodomain transcription factor 2 (PITX2) gene to be strongly correlated with increased risk of recurrence in node-negative, hormone receptor-positive, tamoxifen-treated breast cancer patients using fresh frozen specimens. Aims of the present study were to establish determination of PITX2 methylation for routine analysis in formalin-fixed paraffin-embedded (FFPE) breast cancer tissue and to test PITX2 DNA methylation as a biomarker for outcome prediction in an independent patient cohort. Real-time polymerase chain reaction (PCR) technology was validated for FFPE tissue by comparing methylation measurements in FFPE specimens with those in fresh frozen specimens from the same tumor. The impact of PITX2 methylation on time to distant metastasis was then evaluated in FFPE specimens from hormone receptor-positive, node-negative breast cancer patients (n = 399, adjuvant tamoxifen monotherapy). Reproducibility of the PCR assay in replicate measurements (r(s) > or = 0.95; n = 150) and concordant measurements between fresh frozen and FFPE tissues (r(s) = 0.81; n = 89) were demonstrated. In a multivariate model, PITX2 methylation added significant information (hazard ratio = 2.35; 95% CI, 1.20 to 4.60) to established prognostic factors (tumor size, grade, and age). PITX2 methylation can be reliably assessed by real-time PCR technology in FFPE tissue. Together with our earlier studies, we have accumulated substantial evidence that PITX2 methylation analysis holds promise as a practical assay for routine clinical use to predict outcome in node-negative, tamoxifen-treated breast cancer, which might allow, based on future validation studies, the identification of low-risk patients who may be treated by tamoxifen alone.


Time trends in smoking in Russia in the light of recent tobacco control measures: synthesis of evidence from multiple sources.
The study aims at identifying long-term trends and patterns of current smoking by age, gender, and education in Russia, including the most recent period from 2008 during which tobacco control policies were implemented, and to estimate the impact on mortality of any reductions in prevalence. We present an in-depth analysis based on an unprecedentedly large array of survey data. We examined pooled micro-data on smoking from 17 rounds of the Russian Longitudinal Monitoring Study of 1996-2016, 11 other surveys conducted in Russia in 1975-2017, and two comparator surveys from England and the USA. Standardization by age and education, regression and meta-analysis were used to estimate trends in the prevalence of current smoking by gender, age, and educational patterns. From the mid-1970s to the mid-2000s smoking prevalence among men was relatively stable at around 60%, after which time prevalence declined in every age and educational group. Among women, trends in smoking were more heterogeneous. Prevalence more than doubled above the age of 55 years from very low levels (< 5%). At younger ages, there were steep increases until the mid-2000s after which prevalence has declined. Trends differed by educational level, with women in the lowest educational category accounting for most of the long-term increase. We estimate that the decline in male smoking may have contributed 6.2% of the observed reduction in cardiovascular deaths among men in the period 2008-16. The implementation of an effective tobacco control strategy in Russia starting in 2008 coincided with a decline in smoking prevalence among men from what had been stable, high levels over many decades regardless of age and education. Among women, the declines have been more uneven, with young women showing recent downturns, while the smoking prevalence in middle age has increased, particularly among those with minimal education. Among men, these positive changes will have made a small contribution to the reduction in mortality seen in Russia since 2005.


Preferences for rural specialist health care in the treatment of Parkinson's disease: exploring the role of community-based nursing specialists.
People living with Parkinson's disease (PWPD) in rural areas have limited access to local condition-specific care. This paper examines the healthcare preferences of PWPD living in rural areas and how a community-driven initiative to employ a movement disorder nurse (MDN) functioned to address barriers to health services access. A qualitative design facilitated an understanding of how interactions with the health system shaped PWPD and their carer's experiences of living in a regional community. A total of 42 semi-structured interviews were conducted 6-9 months apart; 19 interviews with PWPD and 23 dyadic interviews. The findings support the contention that specialist care can be effectively delivered through allied health professionals in some settings. In particular, having access to a specialist MDN can cushion the effects of living with Parkinson's disease in regional and rural areas where continuity of care and access to timely support is often difficult for people to find. The quality of social support provided by the MDN may increase people's ability to cope in the face of an unpredictable disease course. This is consistent with prior research, which identified that a specialist nurse or allied health services for people living with chronic conditions is enhances quality of life.


Efficacy, safety and clinical pharmacology of macitentan in comparison to other endothelin receptor antagonists in the treatment of pulmonary arterial hypertension.
Macitentan is a novel dual endothelin receptor antagonist (ERA) showing sustained receptor occupancy. In vitro and in vivo animal studies have demonstrated its potency in antagonizing endothelin-induced disorders. A large morbidity/mortality study in patients with pulmonary arterial hypertension (PAH) taking macitentan has been completed recently. This drug evaluation reviews the efficacy, safety and clinical pharmacology of macitentan in the treatment of PAH. The large Phase III study (SERAPHIN) tested macitentan in more than 700 PAH patients and has provided unique long-term outcome data for this ERA, not available for other members of this class. The effect on a composite clinically relevant morbidity/mortality end point was highly significant at a 10 mg/day dose. The safety profile of macitentan appears to be superior with respect to hepatic safety and edema/fluid retention than bosentan and ambrisentan, respectively, and is similar when considering decrease in hemoglobin concentration. The drug has a low propensity for drug-drug interactions and has one circulating pharmacologically active metabolite. The pharmacokinetics of macitentan in patients with renal or hepatic impairment does not require dose adjustments. Based on its characteristics, macitentan is an important addition to the therapeutic armamentarium in the long-term treatment of PAH. Its potential use in other disorders is under investigation.


Off-clamp vs on-clamp robotic partial nephrectomy: Perioperative, functional and oncological outcomes from a propensity-score matching between two high-volume centers.
Aim of the study was to compare perioperative, functional and oncological outcomes after off-clamp vs on-clamp robotic partial nephrectomy (RPN). Patients who underwent off-clamp or on-clamp (warm ischemia) RPN were extracted from 2 institutional prospectively-maintained databases. 123 patients who underwent off-clamp RPN at one institution were excluded, so that each institution contributed with unselected patients (institution 1:on-clamp RPN vs institution 2:off-clamp). 2:1 propensity-score matching (age, sex, smoking, diabetes, hypertension, ASA score, solitary kidney, preoperative eGFR, tumor size and R.E.N.A.L.score). Perioperative outcomes were compared. A linear mixed model was fitted to eGFR as the outcome regressed on fixed effects for 1) management of clamping (on-clamp/off-clamp), 2) time (at baseline, at discharge, at 12 and 24 months postoperatively), and 3) clamp/time interaction. Survival events were compared between groups. 1983 patients were pooled. After matching, 400 on-clamp vs 200 off-clamp patients were analyzed. No significant differences were found in key perioperative outcomes. The effect of on-clamp on eGFR changed over time. At discharge, groups had similar drop in eGFR. The difference between groups was greatest at 12-months postoperatively, with on-clamp patients showing a deficit of 5 ml/min. At 24-months follow-up, this gap shrunk to 2 ml/min. There were no significant differences in overall survival (p = 0.1), recurrence (χ2 = 0.008, p = 0.9), or metastasis free survival (χ2 = 0.962 p = 0.3). Only one cancer-specific death occurred in off-clamp group. We confirm no significant differences in the perioperative and oncological outcomes between off-clamp and on-clamp RPN. Avoided ischemia benefits renal function within 1-year follow-up after surgery. At longer follow-up, difference with on-clamp is softened.


[Macrophages in asthma].
Every time they exist more demonstrations of the paper than performs the line monocytes-macrophage in the patogenesis of the bronchial asthma. The mononuclear phagocytes cells, as the alveolar macrophages, also they can be activated during allergic methods. The monocytes macrophages are possible efficient inductors of the inflammation; this due to the fact that they can secrete inflammatory mediators, between those which are counted the pre-forming granules of peptides, metabolites of oxidation activation, activator of platelets activator and metabolites of the arachidonic acid. The identification of IL-1 in the liquidate of the bronchial ablution of sick asthmatic, as well as the identification of IL-1 in the I bronchioalveolar washing of places of allergens cutaneous prick, supports the activation concept mononuclear of phagocytic cells in allergic sufferings.


Relationship between Pressure and Output Parameters in Belt Grinding of Steels and Nickel Alloy.
Belt grinding of flat surfaces of typical parts made of steel and alloys, such as grooves, shoulders, ends, and long workpieces, is a good alternative to milling. Several factors can influence the belt grinding process of flat surfaces of metals, such as cutting speed and pressure. In this work, the importance of pressure in the belt grinding was investigated in terms of technological and experimental aspects. The grinding experiments were performed on structural alloy steel 30KhGSN2/30KhGSNA, structural carbon steel AISI 1045, corrosion-resistant and heat-resistant stainless steel AISI 321, and heat-resistant nickel alloy KHN77TYuR. The performance of the grinding belt was investigated in terms of surface roughness, material removal rate (MRR), grinding belt wear, performance index. Estimated indicators of the belt grinding process were developed: cutting ability; reduced cutting ability for belt grinding of steels and heat-resistant alloy. It was found that with an increase in pressure p, the surface roughness of the processed surface Ra decreased while the tool wear VB and MRR increased. With a decrease in plasticity and difficulty of machinability, the roughness, material removal rate, reduced cutting capacity (Performance index) qper, material removal Q decreased, and the tool wear VB increased. The obtained research results can be used by technologists when creating belt grinding operations for steels and alloys to ensure the required performance is met.


A high-fat diet induces rapid changes in the mouse hypothalamic proteome.
Prolonged over-consumption of a high-fat diet (HFD) commonly leads to obesity and insulin resistance. However, even 3 days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus. Mice were fed either a low-fat diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from fat for 3 days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with palmitic acid (PA) and oleic acid (OA). Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3 days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose metabolism and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer's disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA. These results demonstrate that within 3 days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.


Avian metapneumovirus subtype B experimental infection and tissue distribution in chickens, sparrows, and pigeons.
Avian metapneumovirus (aMPV) is a respiratory virus that infects a range of avian hosts, including chickens and turkeys. Migratory and local wild birds are implicated in aMPV spread among farms, countries, and seasonal outbreaks of the disease. A subtype B aMPV isolate from commercial chicken flocks suffering from respiratory disease was experimentally inoculated oculonasally into 7-week old chickens, young pigeons, and sparrows. Chickens showed minimal tracheal rales, whereas pigeons and sparrows were asymptomatic. Shedding of aMPV was detected by reverse transcription polymerase chain reaction on homogenates from nasal turbinates. At 5 days postinfection, 5 of 5 chickens, 2 of 5 pigeons, and 1 of 5 sparrows were positive; at 10 or 15 days, none were positive. At 2 and 5 days, aMPV antigens were localized at the ciliated boarder of respiratory epithelium in nasal cavity and trachea of chickens, as well as to the conjunctival epithelium. Pigeons had detectable viral antigens in only the trachea at 2 and 5 days; sparrow tissues did not show any positive staining. At the end of the experiment, at 21 days postinfection, 14 of 15 inoculated chickens seroconverted against aMPV, but none of the inoculated pigeons or sparrows did. The authors believe that pigeons and sparrows have the ability to transmit the virus between chicken farms, although they do not consider pigeons and sparrows as natural hosts for aMPV, given that they failed to seroconvert. In conclusion, pigeons and sparrows are partially susceptible to aMPV infection, probably acting more as mechanical vectors because infection is only temporary and short-lived.


Structure of the cell surface.
The cell surface is the locus for many important biochemical functions of cells and for the interactions of cells with one another and with their environment. The structure of the cell surface may be thought of as three-layered, with a central plasma membrane to which certain macromolecular components are attached on the outer face (the exoskeleton) and other components on the inner face (the membrane cytoskeleton). In the last decade, the basic molecular structure of the plasma membrane has been elucidated and can be represented by the fluid mosaic model as a first approximation. The binding of specific integral proteins of the membrane to individual peripheral proteins outside or inside the cell is most likely the basis for the three-layered structure of the cell surface. Studies of the last several years on the molecular structures of these three-layered cell surfaces of cultured normal fibroblasts and of fibroblasts transformed by oncogenic viruses are beginning to shed light on the molecular mechanisms responsible for changes in cell shape, adhesiveness, and in contact inhibition of motility associated with neoplastic transformation.


Effect of nebivolol on endothelial dysfunction in patients with Behçet's disease; a prospective single-arm controlled study.
Behçet's disease (BD) is a systemic vasculitis, capable of involving all types of vessels. Endothelial dysfunction (ED) has been previously documented in BD. Previous studies showed that nebivolol might improve endothelial functions in endothelial dysfunction. The aim of our study is to assess the effects of nebivolol on endothelial dysfunction in patients with Behçet's disease. This study was designed as prospective single-arm controlled study. We prospectively studied 35 Behçet's patients who were diagnosed according to the International Study Group criteria. Patients received 5 mg nebivolol per day for 3 months. Endothelial dysfunction was evaluated by brachial artery flow-mediated dilatation (FMD) method using high-resolution vascular ultrasound device at baseline and after for 3-month therapy. The paired samples t test, Wilcoxon test, Pearson, Spearman correlation analyses were used for statistical analysis. A significant improvement was observed in FMD after therapy period (4.23±1.19 vs 7.95±2.21%, p<0.001). The correlation analysis showed a negative correlation between post-treatment high-sensitive C-reactive protein and FMD (r=-0.435, p=0.009). No adverse effects were observed in treatment period. Nebivolol improved endothelial dysfunction in Behçet's patients. However, further comprehensive studies are needed to determine the long-term effects of nebivolol.


Validity of the Walsall Community Pressure Sore Risk Calculator.
The Walsall Community Risk Score Calculator has been in use for the past 8 years. Outcomes from a trust-wide prevalence audit identified that this assessment tool may benefit from re-evaluation. This study examined the tool's validity and reliability and has resulted in some changes being made to the original score. To assess validity, a cross-sectional study of 720 patients was undertaken looking at the incidence of pressure damage, despite intervention, over a 12-week period. Using logistic regression analysis new weightings for each category were developed. This also led to the removal of one factor (bodily pain) which did not appear to contribute to increased risk of pressure damage. The updated score shows an improved prediction of pressure sore development compared with the existing method.


An innovative influenza vaccination policy: targeting last season's patients.
Influenza vaccination is the primary approach to prevent influenza annually. WHO/CDC recommendations prioritize vaccinations mainly on the basis of age and co-morbidities, but have never considered influenza infection history of individuals for vaccination targeting. We evaluated such influenza vaccination policies through small-world contact networks simulations. Further, to verify our findings we analyzed, independently, large-scale empirical data of influenza diagnosis from the two largest Health Maintenance Organizations in Israel, together covering more than 74% of the Israeli population. These longitudinal individual-level data include about nine million cases of influenza diagnosed over a decade. Through contact network epidemiology simulations, we found that individuals previously infected with influenza have a disproportionate probability of being highly connected within networks and transmitting to others. Therefore, we showed that prioritizing those previously infected for vaccination would be more effective than a random vaccination policy in reducing infection. The effectiveness of such a policy is robust over a range of epidemiological assumptions, including cross-reactivity between influenza strains conferring partial protection as high as 55%. Empirically, our analysis of the medical records confirms that in every age group, case definition for influenza, clinical diagnosis, and year tested, patients infected in the year prior had a substantially higher risk of becoming infected in the subsequent year. Accordingly, considering individual infection history in targeting and promoting influenza vaccination is predicted to be a highly effective supplement to the current policy. Our approach can also be generalized for other infectious disease, computer viruses, or ecological networks.


Primary phonological planning units in spoken word production are language-specific: Evidence from an ERP study.
It is widely acknowledged in Germanic languages that segments are the primary planning units at the phonological encoding stage of spoken word production. Mixed results, however, have been found in Chinese, and it is still unclear what roles syllables and segments play in planning Chinese spoken word production. In the current study, participants were asked to first prepare and later produce disyllabic Mandarin words upon picture prompts and a response cue while electroencephalogram (EEG) signals were recorded. Each two consecutive pictures implicitly formed a pair of prime and target, whose names shared the same word-initial atonal syllable or the same word-initial segments, or were unrelated in the control conditions. Only syllable repetition induced significant effects on event-related brain potentials (ERPs) after target onset: a widely distributed positivity in the 200- to 400-ms interval and an anterior positivity in the 400- to 600-ms interval. We interpret these to reflect syllable-size representations at the phonological encoding and phonetic encoding stages. Our results provide the first electrophysiological evidence for the distinct role of syllables in producing Mandarin spoken words, supporting a language specificity hypothesis about the primary phonological units in spoken word production.


Matrix-binding proteins of Staphylococcus aureus: functional analysis of mutant and hybrid molecules.
The fibrinogen-binding protein ClfA and the collagen-binding protein Cna are surface-associated adhesins of Staphylococcus aureus. ClfA has a dipeptide repeat region R composed mainly of serine and aspartate residues, more than 40 of which are required along with the 28-residue region W, the LPXTG motif and region M to display the ligand-binding region A on the cell surface in a functional form. Cna has a 61-residue region W and at least one 187-residue region B linking the collagen-binding region A to peptidoglycan. A cna mutant of S. aureus lacking region B was shown to bind collagen at the same level as wild-type Cna+ cells, indicating that region B is not necessary for ligand binding. Furthermore, altering the number of B repeats did not influence the level of collagen binding. In order to study the ability of C-terminal domains of Cna and ClfA to support functional ligand-binding activity of different adhesins, chimeric proteins were constructed and expressed in S. aureus. Surprisingly, the presence of a single Cna B domain and a nonapeptide linker located between ClfA region A and Cna region WM failed to support fibrinogen binding by S. aureus cells, despite the fact that ClfA region A was detected on the bacterial surface by immunoblotting. In contrast, the ClfA region A-Cna region B hybrid expressed as a recombinant protein in Escherichia coli did bind fibrinogen in Western ligand blots and in an ELISA-type assay. It is concluded that Cna region B cannot support functional display of ClfA region A on the bacterial cell surface. However, the ClfA dipeptide repeat region R and region WM did promote functional surface expression of the Cna collagen-binding domain in a hybrid Cna-ClfA protein.


Clusterin glycopeptide variant characterization reveals significant site-specific glycan changes in the plasma of clear cell renal cell carcinoma.
Cancer-related alterations in protein glycosylation may serve as diagnostic or prognostic biomarkers or may be used for monitoring disease progression. Clusterin is a medium abundance, yet heavily glycosylated, glycoprotein that is upregulated in clear cell renal cell carcinoma (ccRCC) tumors. We recently reported that the N-glycan profile of clusterin is altered in the plasma of ccRCC patients. Here, we characterized the occupancy and the degree of heterogeneity of individual N-glycosylation sites of clusterin in the plasma of patients diagnosed with localized ccRCC, before and after curative nephrectomy (n = 40). To this end, we used tandem mass spectrometry of immunoaffinity-enriched plasma samples to analyze the individual glycosylation sites in clusterin. We determined the levels of targeted clusterin glycoforms containing either a biantennary digalactosylated disialylated (A2G2S2) glycan or a core fucosylated biantennary digalactosylated disialylated (FA2G2S2) glycan at N-glycosite N374. We showed that the presence of these two clusterin glycoforms differed significantly in the plasma of patients prior to and after curative nephrectomy for localized ccRCC. Removal of ccRCC led to a significant increase in the levels of both FA2G2S2 and A2G2S2 glycans in plasma clusterin. These changes were further confirmed by lectin blotting of plasma clusterin. It is envisioned that these identified glycan alterations may provide an additional level of therapeutic or biomarker sensitivity than levels currently achievable by monitoring expression differences alone.


Vitreous levels of bevacizumab and vascular endothelial growth factor-A in patients with choroidal neovascularization.
To investigate the vitreous levels of bevacizumab and vascular endothelial growth factor-A (VEGF-A) after intravitreal injection of the drug in patients with choroidal neovascularization (CNV). Interventional case series. Eleven eyes of 11 patients with submacular hemorrhage and CNV due to age-related macular degeneration (n = 10) or angioid streaks (n = 1). All patients were treatment naïve except for a single dose of intravitreal injection of bevacizumab (1.25 mg/50 muL dose) and subsequent vitrectomy after various intervals (1-101 days) because of active and progressive lesion. Intravitreal free bevacizumab and VEGF-A levels were measured using enzyme-linked immunosorbent assay and microsphere-based immunoassay, respectively. Vitreous VEGF-A isoforms were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting. Intravitreal bevacizumab and VEGF-A levels were measured and pharmacokinetic parameters were calculated. Pharmacokinetics of intravitreal bevacizumab followed a 2-compartment model with initial and terminal half-lives of 0.5 and 6.7 days, respectively. Bevacizumab could be detected in all cases, ranging from 2.63 ng/ml to 165 microg/ml. The peak concentration was observed on the second day after intravitreal bevacizumab injection. Vitreous free VEGF-A levels ranged from 0.2 to 33.9 pg/ml and showed a negative correlation with the bevacizumab concentration (P<0.001; r = -0.955) and a positive correlation with time (P<0.001; r = 0.964). However, the percentage expression of VEGF-A(165) exhibited a positive correlation with the bevacizumab concentration (P = 0.032, r = 0.645) and a negative correlation with time (P = 0.007, r = -0.755). A time-dependent increase was found for the percentage expression of VEGF-A(189) (P = 0.023, r = 0.673). Neither bevacizumab- nor time-related alterations were found for VEGF-A(121). Based on pharmacokinetics, the interval of 6-7 weeks would be appropriate for efficacy, although clinical trials should guide dosing recommendations. Vitreous levels of free VEGF-A showed a negative correlation with the bevacizumab concentration, which confirmed the in vivo binding affinity of bevacizumab to VEGF-A. The analysis of the VEGF-A isoforms suggests differences of interaction between bevacizumab and individual VEGF-A isoforms.


A prospective multicentre phase II study of cisplatin and weekly docetaxel as first-line treatment for recurrent or metastatic nasopharyngeal cancer (KCSG HN07-01).
The purpose of this phase II study was to determine the efficacy and toxicity of cisplatin and weekly docetaxel combination chemotherapy as a first-line treatment in patients with recurrent or metastatic nasopharyngeal cancer. Recurrent or metastatic nasopharyngeal cancer patients were enrolled and received a combination of weekly docetaxel (35 mg/m(2) on Day1 and Day8) and cisplatin (70 mg/m(2) D1) every 21 days, for up to a maximum of 6 cycles. The primary endpoint was objective response rate, and the secondary endpoints included toxicity of combination chemotherapy, progression-free survival, overall survival and 1-year survival rate. In total, 47 patients were enrolled and analysed, and 46 patients (97.9%) completed the planned protocol. In an intent-to-treat analysis, 6 patients (12.8%) achieved complete response (CR) and 27 patients (57.4%) showed partial response (PR), with an objective response rate of 70.2%. The median progression-free survival and overall survival were 9.6 months (95% C.I. 5.7-13.5 months) and 28.5 months (95% C.I. 16.9-40.1 months), respectively, and the 1-year survival rate was 89.9%. The common grade 3 adverse events were stomatitis (1.2%), neutropenia (0.8%), anaemia (0.8%), infection (0.8%) and diarrhoea (0.8%). Grade 4 adverse events were not observed in this study. The combination chemotherapy of cisplatin and weekly docetaxel is highly effective and shows favourable toxicity as a first-line chemotherapy in patients with recurrent or metastatic nasopharyngeal cancer.


[Democracy and conflict in pluralist contexts: an interview with Chantal Mouffe].
Chantal Mouffe, along with Argentinian political theorist Ernesto Laclau (1935-2014), laid down the bases of discourse theory in 1985. She later developed her work by exploring in more detail how discourse theory formulations influence the analysis of contemporary democracies. Approaching conflict as a product of the encounter with difference, Mouffe sees it as an indelible part of the constitution of social relationships. In this encounter with the author, we seek to reflect upon certain themes and problematics that are central to her work, and upon the implications of her theory for the field of contemporary education.