Validation of an algorithm combining haemoglobin A(1c) and fasting plasma glucose for diagnosis of diabetes mellitus in UK and Australian populations.
To determine whether glycated haemoglobin (HbA(1c)) can be used in combination with fasting plasma glucose (FPG) for the diagnosis of diabetes in patients with impaired fasting glucose (IFG) and in a broader spectrum of patients. An algorithm was derived from oral glucose tolerance test (OGTT) capillary samples in 500 consecutive UK patients with IFG by World Health Organization criteria. It was validated in a further 500 UK patients and, with venous specimens, in 1175 unselected Australian patients. The derivation cohort was aged 61 years (50-69 years) (median IQ range) with 52% male and 12% South Asian. Diabetes Control and Complications Trial-aligned HbA(1c) was 6.2% (5.8-6.6%) (reference interval < 6.0%) and FPG 6.7 mmol/l (6.3-7.2 mmol/l). FPG was in the diabetes range in 36% of patients, with an OGTT identifying a further 12% with diabetes. The derived algorithm, (HbA(1c) >or= 6.0% with FPG < 7.0 mmol/l) identified those patients requiring an OGTT to diagnose diabetes. When applied to the UK validation cohort, sensitivity was 97% and specificity 100%. The algorithm was equally effective in the unselected group, aged 59 years (49-68 years) and 54% male, with sensitivity 93% and specificity 100%. HbA(1c) was 6.0% (5.6-6.6%) and FPG 6.0 mmol/l (5.3-6.8 mmol/l), with 26% having IFG. Use of the algorithm would reduce the number of OGTTs performed in the UK validation cohort by 33% and by 66% in the Australian patients studied. Use of this algorithm would simplify procedures for diagnosis of diabetes and could also be used for monitoring pre-diabetes. Validation is now required in other populations and patient groups.


The pathway from glandular fever to chronic fatigue syndrome: can the cognitive behavioural model provide the map?
The cognitive behavioural model of chronic fatigue syndrome (CFS) suggests that the illness is caused through reciprocal interactions between physiology, cognition, emotion and behaviour. The purpose of this study was to investigate whether the psychological factors operationalized in this model could predict the onset of CFS following an acute episode of infectious mononucleosis commonly known as glandular fever (GF). A total of 246 patients with GF were recruited into this prospective cohort study. Standardized self-report measures of perceived stress, perfectionism, somatization, mood, illness beliefs and behaviour were completed at the time of their acute illness. Follow-up questionnaires determined the incidence of new-onset chronic fatigue (CF) at 3 months and CFS at 6 months post-infection. Of the participants, 9.4% met the criteria for CF at 3 months and 7.8% met the criteria for CFS at 6 months. Logistic regression revealed that factors proposed to predispose people to CFS including anxiety, depression, somatization and perfectionism were associated with new-onset CFS. Negative illness beliefs including perceiving GF to be a serious, distressing condition, that will last a long time and is uncontrollable, and responding to symptoms in an all-or-nothing behavioural pattern were also significant predictors. All-or-nothing behaviour was the most significant predictor of CFS at 6 months. Perceived stress and consistently limiting activity at the time of GF were not significantly associated with CFS. The findings from this study provide support for the cognitive behavioural model and a good basis for developing prevention and early intervention strategies for CFS.


Accuracy of Different Bioinformatics Methods in Detecting Antibiotic Resistance and Virulence Factors from Staphylococcus aureus Whole-Genome Sequences.
In principle, whole-genome sequencing (WGS) can predict phenotypic resistance directly from a genotype, replacing laboratory-based tests. However, the contribution of different bioinformatics methods to genotype-phenotype discrepancies has not been systematically explored to date. We compared three WGS-based bioinformatics methods (Genefinder [read based], Mykrobe [de Bruijn graph based], and Typewriter [BLAST based]) for predicting the presence/absence of 83 different resistance determinants and virulence genes and overall antimicrobial susceptibility in 1,379 Staphylococcus aureus isolates previously characterized by standard laboratory methods (disc diffusion, broth and/or agar dilution, and PCR). In total, 99.5% (113,830/114,457) of individual resistance-determinant/virulence gene predictions were identical between all three methods, with only 627 (0.5%) discordant predictions, demonstrating high overall agreement (Fleiss' kappa = 0.98, P < 0.0001). Discrepancies when identified were in only one of the three methods for all genes except the cassette recombinase, ccrC(b). The genotypic antimicrobial susceptibility prediction matched the laboratory phenotype in 98.3% (14,224/14,464) of cases (2,720 [18.8%] resistant, 11,504 [79.5%] susceptible). There was greater disagreement between the laboratory phenotypes and the combined genotypic predictions (97 [0.7%] phenotypically susceptible, but all bioinformatic methods reported resistance; 89 [0.6%] phenotypically resistant, but all bioinformatics methods reported susceptible) than within the three bioinformatics methods (54 [0.4%] cases, 16 phenotypically resistant, 38 phenotypically susceptible). However, in 36/54 (67%) cases, the consensus genotype matched the laboratory phenotype. In this study, the choice between these three specific bioinformatic methods to identify resistance determinants or other genes in S. aureus did not prove critical, with all demonstrating high concordance with each other and phenotypic/molecular methods. However, each has some limitations; therefore, consensus methods provide some assurance.


Crystal structure of the ω-aminotransferase from Paracoccus denitrificans and its phylogenetic relationship with other class III aminotransferases that have biotechnological potential.
Apart from their crucial role in metabolism, pyridoxal 5'-phosphate (PLP)-dependent aminotransferases (ATs) constitute a class of enzymes with increasing application in industrial biotechnology. To provide better insight into the structure-function relationships of ATs with biotechnological potential we performed a fundamental bioinformatics analysis of 330 representative sequences of pro- and eukaryotic Class III ATs using a structure-guided approach. The calculated phylogenetic maximum likelihood tree revealed six distinct clades of which the first segregates with a very high bootstrap value of 92%. Most enzymes in this first clade have been functionally well characterized, whereas knowledge about the natural functions and substrates of enzymes in the other branches is sparse. Notably, in those clades 2-6 members of the peculiar class of ω-ATs prevail, many of which have proven useful for the preparation of chiral amines or artificial amino acids. One representative is the ω-AT from Paracoccus denitrificans (PD ω-AT) which catalyzes, for example, the transamination in a novel biocatalytic process for the production of L-homoalanine from L-threonine. To gain structural insight into this important enzyme, its X-ray analysis was carried out at a resolution of 2.6 Å, including the covalently bound PLP as well as 5-aminopentanoate as a putative amino donor substrate. On the basis of this crystal structure in conjunction with our phylogenetic analysis, we have identified a generic set of active site residues of ω-ATs that are associated with a strong preference for aromatic substrates, thus guiding the discovery of novel promising enzymes for the biotechnological production of corresponding chiral amines.


The effects of hypoxia on the theoretical modelling of tumour control probability.
Theoretical modelling of tumour response is increasingly used for the prediction of treatment result and has even been proposed as ranking criteria in some algorithms for treatment planning. Tumour response to radiation is greatly influenced by the details of tumour microenvironment, especially hypoxia, that unfortunately are not always taken into consideration for these simulations. This paper intends to investigate the effects of various assumptions regarding hypoxia distribution in tumours on the predictions of treatment outcome. A previously developed model for simulating theoretically the oxygenation in biologically relevant tissues, including results from oxygen diffusion, consumption and perfusion limitations in tumours, was used to investigate the effects of the different aspects of hypoxia on the predictions of treatment outcome. Thus, both the continuous distribution of values and the temporal variation of hypoxia patterns were taken into consideration and were compared with a 'black-and-white' simplification with a fully hypoxic compartment and a fully oxic one. It was found that the full distribution of oxygenation in the tissue is needed for accurate results. The 'black-and-white' simplification, while showing the same general trends for the predictions of radiation response, could lead to serious over-estimations of the tumour control probability. It was also found that the presence of some hypoxia for every treatment fraction leads to a decrease in the predicted local control, regardless of the change of the hypoxic pattern throughout the duration of the whole treatment. The results thus suggest that the assumptions regarding tumour hypoxia influence very much the predictions of treatment outcome and therefore they have to be very carefully incorporated into the theoretical modelling.


Expression of the multidrug resistance-associated protein (MRP) gene in human cancers.
We determined the expression of a newly recognized drug resistance gene, the multidrug resistance-associated protein (MRP) gene, [Cole et al., Science (Washington DC), 258: 1650-1654, 1992], in normal human tissues and in >370 human tumor biopsies using a quantitative RNase protection assay and immunohistochemistry. MRP mRNA appeared to be ubiquitously expressed at low levels in all normal tissues, including peripheral blood, the endocrine glands (adrenal and thyroid), striated muscle, the lymphoreticular system (spleen and tonsil), the digestive tract (salivary gland, esophagus, liver, gall bladder, pancreas, and colon), the respiratory tract (lung), and the urogenital tract (kidney, bladder, testis, and ovary). The human cancers analyzed could be divided into three groups with regard to MRP expression. Group 1 consists of tumors that often exhibit high to very high MRP mRNA levels (e.g., chronic lymphocytic leukemia). Group 2 comprises the tumors that often exhibit low, but occasionally exhibit high MRP mRNA expression (e.g., esophagus squamous cell carcinoma, non-small cell lung cancer, and acute myelocytic leukemia). Group 3 comprises the tumors with predominantly low levels of MRP mRNA, comparable to the levels found in normal tissues (e.g., other hematological malignancies, soft tissue sarcomas, melanoma, and cancers of the prostate, breast, kidney, bladder, testis, ovary, and colon). Using the MRP-specific mAbs MRPr1 and MRPm6, we confirmed the elevated MRP mRNA levels in tumor tissues by immunohistochemistry. We conclude that hyperexpression of MRP is observed in several human cancers, and that additional studies are needed to assess the clinical relevance of MRP.


Circadian control of chloroplast transcription by a nuclear-encoded timing signal.
Circadian timekeeping in plants increases photosynthesis and productivity. There are circadian oscillations in the abundance of many chloroplast-encoded transcripts, but it is not known how the circadian clock regulates chloroplast transcription or the photosynthetic apparatus. We show that, in Arabidopsis, nuclear-encoded SIGMA FACTOR5 (SIG5) controls circadian rhythms of transcription of several chloroplast genes, revealing one pathway by which the nuclear-encoded circadian oscillator controls rhythms of chloroplast gene expression. We also show that SIG5 mediates the circadian gating of light input to a chloroplast-encoded gene. We have identified an evolutionarily conserved mechanism that communicates circadian timing information between organelles with distinct genetic systems and have established a new level of integration between eukaryotic circadian clocks and organelles of endosymbiotic origin.


Multi-centre, multi-vendor reproducibility of 7T QSM and R2* in the human brain: Results from the UK7T study.
We present the reliability of ultra-high field T2* MRI at 7T, as part of the UK7T Network's "Travelling Heads" study. T2*-weighted MRI images can be processed to produce quantitative susceptibility maps (QSM) and R2* maps. These reflect iron and myelin concentrations, which are altered in many pathophysiological processes. The relaxation parameters of human brain tissue are such that R2* mapping and QSM show particularly strong gains in contrast-to-noise ratio at ultra-high field (7T) vs clinical field strengths (1.5-3T). We aimed to determine the inter-subject and inter-site reproducibility of QSM and R2* mapping at 7T, in readiness for future multi-site clinical studies. Ten healthy volunteers were scanned with harmonised single- and multi-echo T2*-weighted gradient echo pulse sequences. Participants were scanned five times at each "home" site and once at each of four other sites. The five sites had 1× Philips, 2× Siemens Magnetom, and 2× Siemens Terra scanners. QSM and R2* maps were computed with the Multi-Scale Dipole Inversion (MSDI) algorithm (https://github.com/fil-physics/Publication-Code). Results were assessed in relevant subcortical and cortical regions of interest (ROIs) defined manually or by the MNI152 standard space. Mean susceptibility (χ) and R2* values agreed broadly with literature values in all ROIs. The inter-site within-subject standard deviation was 0.001-0.005 ppm (χ) and 0.0005-0.001 ms-1 (R2*). For χ this is 2.1-4.8 fold better than 3T reports, and 1.1-3.4 fold better for R2*. The median ICC from within- and cross-site R2* data was 0.98 and 0.91, respectively. Multi-echo QSM had greater variability vs single-echo QSM especially in areas with large B0 inhomogeneity such as the inferior frontal cortex. Across sites, R2* values were more consistent than QSM in subcortical structures due to differences in B0-shimming. On a between-subject level, our measured χ and R2* cross-site variance is comparable to within-site variance in the literature, suggesting that it is reasonable to pool data across sites using our harmonised protocol. The harmonized UK7T protocol and pipeline delivers on average a 3-fold improvement in the coefficient of reproducibility for QSM and R2* at 7T compared to previous reports of multi-site reproducibility at 3T. These protocols are ready for use in multi-site clinical studies at 7T.


Cancer/stroma interplay via cyclooxygenase-2 and indoleamine 2,3-dioxygenase promotes breast cancer progression.
Expression of indoleamine 2,3-dioxygenase (IDO) in primary breast cancer increases tumor growth and metastasis. However, the clinical significance of stromal IDO and the regulation of stromal IDO are unclear. Metabolomics and enzyme-linked immunosorbent assay (ELISA) were used to study the effect of cyclooxygenase-2 (COX-2)-overexpressing breast cancer cells on IDO expression in co-cultured human breast fibroblasts. Biochemical inhibitors and short-hairpin RNA (shRNA) were used to clarify how prostaglandin E2 (PGE2) upregulates IDO expression. Associations of stromal IDO with clinicopathologic parameters were tested in tumor specimens. An orthotopic animal model was used to examine the effect of COX-2 and IDO inhibitors on tumor growth. Kynurenine, the metabolite generated by IDO, increases in the supernatant of fibroblasts co-cultured with COX-2-overexpressing breast cancer cells. PGE2 released by cancer cells upregulates IDO expression in fibroblasts through an EP4/signal transducer and activator of transcription 3 (STAT3)-dependent pathway. Conversely, fibroblast-secreted kynurenine promotes the formation of the E-cadherin/Aryl hydrocarbon receptor (AhR)/S-phase kinase-associated protein 2 (Skp2) complex, resulting in degradation of E-cadherin to increase breast cancer invasiveness. The enhancement of motility of breast cancer cells induced by co-culture with fibroblasts is suppressed by the IDO inhibitor 1-methyl-tryptophan. Pathological analysis demonstrates that upregulation of stromal IDO is a poor prognosis factor and is associated with of COX-2 overexpression. Co-expression of cancer COX-2 and stromal IDO predicts a worse disease-free and metastasis-free survival. Finally, COX-2 and IDO inhibitors inhibit tumor growth in vivo. Integration of metabolomics and molecular and pathological approaches reveals the interplay between cancer and stroma via COX-2, and IDO promotes tumor progression and predicts poor patient survival.


Phosphodiesterase type-5 inhibitor and oxidative stress.
Erectile dysfunction (ED) is a common medical condition that affects the sexual life of millions of men worldwide. Numerous physical and psychological factors are involved in normal erectile function, including neurological, vascular, hormonal and cavernous functions. The current therapy for the condition is pharmacological and psychotherapeutic which regulates the erectile function and amplifies the NO-mediated response. The aim of this work is to test the action of three common phosphodiesterase inhibitors: Tadalafil, Sildenafil Citrate and Vardenafil at 0.05 microM on human monocytes, analyzing the expression of iNOS protein and mRNA by Western blot and rt-PCR, and production of NO by conversion of L-(2,3,4,5)-[3H]Arginine to L-(3H) citrulline. We also tested the efficiency of the antioxidant network by spectrophotometer (SOD, CAT, GPx and Gr), under normal conditions and after stimulation with LPS. The results showed an increase in ROS levels, similar for all the molecules with regard to the antioxidant enzymes. In all cases the treatment determines a response to the limited efficiency, arriving at a situation in which phosphodiesterase inhibitors + LPS clearly show oxidative stress.


Lactobacillus plantarum HY7714 Restores TNF-α Induced Defects on Tight Junctions.
In addition to intestinal balance, probiotics are known to have beneficial effects on skin inflammation, metabolic diseases, and emotions. Previously, we have reported the skin anti-aging effects of Lactobacillus plantarum HY7714 (HY7714) in a clinical trial. To prove the protective skin effects of HY7714 through the intestinal tight junction (TJ), we investigated the effects of HY7714 on the intestines through tumor necrosis factor (TNF)-α induced TJ defects in Caco-2 cells. Specifically, 24 h treatment with HY7714 restored the decreased expression of zonula occludens-1, occludin, and claudin-1 compared to the TNF-α-treated groups (P<0.05). It also attenuated the level of pro-inflammatory cytokines interleukin-6, 8, and 1β. Further, increases in the mRNA levels of Elk-1, nuclear factor-κB, and myosin light chain kinase expression induced by TNF-α were recovered by HY7714. These findings imply that HY7714 improves intestinal barrier integrity and is a potential therapeutic agent for dysfunctions derived from TJ defects.


Power-dependent spectral shift of photoluminescence from ensembles of silicon nanocrystals.
: Nanocrystals are widely studied for their tunable optical properties, most importantly increased luminescence efficiency and emission energy. Quantum confinement effects are found for many different types of nanocrystals, and these introduce a relation between the emission wavelength and the size of nanocrystals. When ensembles of nanocrystals with a distribution of sizes are studied, this can have profound effects on their luminescence spectra. Here, we show how photoluminescence spectra of ensembles of silicon nanocrystals can shift under different excitation conditions, resulting from differences in absorption cross-section of the individual nanocrystal sizes. This effect, together with the fact that after a pulsed excitation a silicon nanocrystal can only emit a single photon, determines how the distribution of excited nanocrystals changes and leads to the spectral shift for different excitation powers. Next to this effect, the influence of different radiative rates in such ensembles is also addressed. These notions are important for the interpretation of photoluminescence data for silicon nanocrystals but can be extended to any nanoparticle system comprising size-distributed ensembles.


Transcranial Direct-Current Stimulation May Improve Discourse Production in Healthy Older Adults.
Background: The use of transcranial direct-current stimulation (tDCS) for therapeutic and neurorehabilitation purposes has become increasingly popular in recent years. Previous research has found that anodal tDCS may enhance naming ability and verbal fluency in healthy participants. However, the effect of tDCS on more functional, higher level language skills such as discourse production has yet to be understood. Aims: The present study aimed to investigate in healthy, older adults (a) the effect of anodal tDCS on discourse production vs. sham stimulation and (b) optimal electrode placement for tDCS to target language improvement at the discourse level. Methods: Fourteen healthy, older right-handed participants took part in this sham controlled, repeated measures pilot study. Each participant experienced three different experimental conditions; anodal tDCS on the left inferior frontal gyrus (IFG), anodal tDCS on the right IFG and sham stimulation while performing a story telling task. Significant changes in language performance before and after each condition were examined in three discourse production tasks: recount, procedural and narrative. Results: Left and right IFG conditions showed a greater number of significant within-group improvements (p < 0.05) in discourse production compared to sham with 6/12 for left IFG, 4/12 for right IFG and 2/12 for sham. There were no significant differences noted between tDCS conditions. No relationship was noted between language performance and physical activity, age, or gender. Conclusions: This study suggests that anodal tDCS may significantly improve discourse production in healthy, older adults. In line with previous tDCS language studies, the left IFG is highlighted as an optimal stimulation site for the modulation of language in healthy speakers. The findings support further exploration of tDCS as a rehabilitative tool for higher-level language skills in persons with aphasia.


Cyclic silicate active site and stereochemical match for apatite nucleation on pseudowollastonite bioceramic-bone interfaces.
Hydroxyapatite (Ca5(PO4)3(OH)) forms on pseudowollastonite (psW) (alpha-CaSiO3) in vitro in simulated body fluid, human parotid saliva and cell-culture medium, and in vivo in implanted rat tibias. We used crystallographic constraints with ab initio molecular orbital calculations to identify the active site and reaction mechanism for heterogeneous nucleation of the earliest calcium phosphate oligomer/phase. The active site is the planar, cyclic, silicate trimer (Si3O9) on the (001) face of psW. The trimer has three silanol groups (>SiOH) arranged at 60 degrees from each other, providing a stereochemical match for O atoms bonded to Ca2+ on the (001) face of hydroxyapatite. Calcium phosphate nucleation is modeled in steps as hydrolysis of surface Ca-O bonds with leaching of Ca2+ into solution, protonation of the surface Si-O groups to form silanols, calcium sorption as an inner-sphere surface complex and, attachment of HPO4(2-). Our model explains the experimental solution and high resolution transmission electron microscopy data for epitaxial hydroxyapatite growth on psW in vitro and in vivo. We propose that the cyclic silicate trimer is the universal active site for heterogeneous, stereochemically promoted nucleation on silicate-based bioactive ceramics. A critical active site-density and a point of zero charge of the bioceramic less than physiological pH are required for bioactivity.


A murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells.
Murine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human iPSC reprogramming in the presence of LIF yields human stem cells that display morphological, molecular, and functional properties of murine ESCs. We termed these hLR5 iPSCs because they require the expression of five ectopic reprogramming factors, Oct4, Sox2, Klf4, cMyc, and Nanog, to maintain this more naive state. The cells are "metastable" and upon ectopic factor withdrawal they revert to standard human iPSCs. Finally, we demonstrate that the hLR5 state facilitates gene targeting, and as such provides a powerful tool for the generation of recombinant human pluripotent stem cell lines.


An Orientation Dependent Size Illusion Is Underpinned by Processing in the Extrastriate Visual Area, LO1.
We use the simple, but prominent Helmholtz's squares illusion in which a vertically striped square appears wider than a horizontally striped square of identical physical dimensions to determine whether functional magnetic resonance imaging (fMRI) BOLD responses in V1 underpin illusions of size. We report that these simple stimuli which differ in only one parameter, orientation, to which V1 neurons are highly selective elicited activity in V1 that followed their physical, not perceived size. To further probe the role of V1 in the illusion and investigate plausible extrastriate visual areas responsible for eliciting the Helmholtz squares illusion, we performed a follow-up transcranial magnetic stimulation (TMS) experiment in which we compared perceptual judgments about the aspect ratio of perceptually identical Helmholtz squares when no TMS was applied against selective stimulation of V1, LO1, or LO2. In agreement with fMRI results, we report that TMS of area V1 does not compromise the strength of the illusion. Only stimulation of area LO1, and not LO2, compromised significantly the strength of the illusion, consistent with previous research that LO1 plays a role in the processing of orientation information. These results demonstrate the involvement of a specific extrastriate area in an illusory percept of size.


Disorder-immune coupled resonator optical waveguide.
We demonstrate that a photonic lattice with short- and long-range harmonic modulations of the refractive index facilitates formation of flat photonic bands and leads to slow propagation of light. The system can be considered a coupled-resonator optical waveguide (CROW): photonic bands with abnormally small dispersion are created due to the interaction of long-lived states in the cavity regions via weak coupling across tunneling barriers. Unlike previous CROW implementations, the proposed structures can be fabricated with interference photolithography (holography), sidestepping the issue of resonator-to-resonator fluctuation of the system parameters. The proposed holography-based approach enables fabrication of arrays with a large number of coupled optical resonators, which is necessary for practical applications.


Assessment of liver fibrosis and cirrhosis by transient elastography among patients with chronic HBV and HCV infection in Georgia.
The aim of the study was to evaluate liver fibrosis (LF) and cirrhosis using Transient elastography (TE) using Fibroscan in patients with chronic HBV and HCV infection. The device evaluates LF by measurement of liver stiffness (LS). 525 patients with chronic HCV infection and 105 patients with chronic HBV infection were included in the study. These patients were investigated at the Georgian-French joint hepatology clinic "Hepa" from November 2007, till November 2008. Among investigated HBV infected 105 patients 65 (61.9%) had no fibrosis (LS<5.5 kpa), 23 (21.9%) had mild fibrosis (LS-5.5-8.0 kpa), 9 (8.6%) had severe fibrosis (LS-8.0-14.0 kpa) and 8 (7.6%) had liver cirrhosis (LS>14.0 kpa). Among investigated HCV infected 525 patients 200 (38.1%) had no fibrosis (LS<5.5 kpa), 139 (26.5%) patients had mild fibrosis (LS-5.5-8.0 kpa), 87 (16.5%) patients had severe fibrosis (LS-8.0-14.0 kpa) and 99 (18.9%) patients had liver cirrhosis. It is concluded that transient elastography (TE) using Fibroscan is simple, non-invasive, reliable and easily reproducible method for assessing liver fibrosis and cirrhosis in patients with chronic HBV and HCV infection. TE is characterized with an excellent accuracy. TE results are well correlated with the clinical signs as well as with the results of laboratory and instrumental investigations. Fibrosis stages by Metavir measured using Fibroscan well corresponds with the liver biopsy results. Considering the high prevalence of fibrosis and cirrhosis among patients with chronic HBV and HCV infection, TE is a very valuable method for detecting early stages of fibrosis allowing to avoid the progression of liver damage, as well as end-stage liver disease. TE is easy to perform and therefore allows regular follow-up of the course of LF.


Extracorporeal ultrafiltration for acute heart failure: patient selection and perspectives.
Most patients presenting with acute heart failure (AHF) show signs and symptoms of fluid overload, which are closely associated with short-term and long-term outcomes. Ultrafiltration is an extremely appealing strategy for patients with AHF and concomitant overt fluid overload not fully responsive to diuretic therapy. However, although there are several theoretical beneficial effects associated with ultrafiltration, published reports have shown controversial findings. Differences in selection of the study population and in ultrafiltration indications and protocols, and high variability in the pharmacologic therapy used for the control group could explain some of these conflicting results. Here, we aimed to provide an overview on the current medical evidence supporting the use of ultrafiltration in AHF, with a special focus on the identification of potential candidates who may benefit the most from this therapeutic option.


Minimally Invasive Periodontal Treatment Using the Er,Cr: YSGG Laser. A 2-year Retrospective Preliminary Clinical Study.
MINIMALLY INVASIVE SURGERY (MIS) USING THE ERBIUM, CHROMIUM: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser (Waterlase MD, Biolase, Irvine, CA) to treat moderate to advanced periodontal disease is presented as an alternative to conventional therapies. To date, there are few short- or long-term studies to demonstrate the effects of this laser in treating and maintaining periodontal health. Electronic clinical records from 16 patients - total of 126 teeth, with pocket depths ranging from 4 mm to 9 mm - were treated with the same protocol using the Er,Cr:YSGG laser. The mean baseline probing depths (PD) were 5 mm and clinical attachment levels (CAL) were 5 mm in the 4 - 6 mm pretreated laser group. The mean baseline probing depths were 7.5 and 7.6 mm for PD and CAL respectfully in the 7 - 9 mm pretreatment laser group. At the 2 year mark, the average PD was 3.2 ± 1.1 mm for the 4-6 mm pocket group and the 7-9 mm pocket group had a mean PD of 3.7 ± 1.2 mm. mean CAL was 3.1 ± 1.1 mm for the 4-6 mm group and 3.6 ± 1.2 for the 7-9 mm group with an overall reduction of 1.9 mm and 4.0 mm respectively. At one and two years, both groups remained stable with PD comparable to the three-month gains. The CAL measurements at one and two years were also comparable to the three-month gains.


Association between liver-specific gene polymorphisms and their expression levels with nonalcoholic fatty liver disease.
Genetic factors account for a significant proportion of the phenotypic variance of nonalcoholic fatty liver disease (NAFLD); however, very few predisposing genes have been identified. We aimed to (1) identify novel genetic associations with NAFLD by performing a genome-wide association study (GWAS), and (2) examine the biological expression of the strongest genetic associations in a separate cohort. We performed GWAS of a population-based cohort (Raine Study) of 928 adolescents assessed for NAFLD by ultrasound at age 17. Expression of genes with single nucleotide polymorphisms (SNPs) that were associated with NAFLD at a significance level of P < 10(-5) was examined in adults with NAFLD and controls by quantifying hepatic messenger RNA (mRNA) expression and serum levels of protein. After adjustment for sex and degree of adiposity, SNPs in two genes expressed in liver were associated with NAFLD adolescents: group-specific component (GC) (odds ratio [OR], 2.54; P = 1.20 × 10(-6)) and lymphocyte cytosolic protein-1 (LCP1) (OR, 3.29; P = 2.96 × 10(-6)). SNPs in two genes expressed in neurons were also associated with NAFLD: lipid phosphate phosphatase-related protein type 4 (LPPR4) (OR, 2.30; P = 4.82 × 10(-6)) and solute carrier family 38 member 8 (SLC38A8) (OR, 3.14; P = 1.86 × 10(-6) ). Hepatic GC mRNA was significantly reduced (by 83%) and LCP1 mRNA was increased (by 300%) in liver biopsy samples from patients with NAFLD compared to controls (P < 0.05). Mean serum levels of GC protein were significantly lower in patients with NAFLD than controls (250 ± 90 versus 298 ± 90, respectively; P = 0.004); GC protein levels decreased with increasing severity of hepatic steatosis (P < 0.01). The association between GC and LCP1 SNPs and NAFLD as well as altered biological expression implicate these genes in the pathogenesis of NAFLD.


Imaging cAMP nanodomains in the heart.
3'-5'-cyclic adenosine monophosphate (cAMP) is a ubiquitous second messenger that modulates multiple cellular functions. It is now well established that cAMP can mediate a plethora of functional effects via a complex system of local regulatory mechanisms that result in compartmentalized signalling. The use of fluorescent probes to monitor cAMP in intact, living cells have been instrumental in furthering our appreciation of this ancestral and ubiquitous pathway and unexpected details of the nano-architecture of the cAMP signalling network are starting to emerge. Recent evidence shows that sympathetic control of cardiac contraction and relaxation is achieved via generation of multiple, distinct pools of cAMP that lead to differential phosphorylation of target proteins localized only tens of nanometres apart. The specific local control at these nanodomains is enabled by a distinct signalosome where effectors, targets, and regulators of the cAMP signal are clustered. In this review, we focus on recent advances using targeted fluorescent reporters for cAMP and how they have contributed to our current understanding of nanodomain cAMP signalling in the heart. We briefly discuss how this information can be exploited to design novel therapies and we highlight some of the questions that remain unanswered.


Customized chemotherapy in metastatic non-small cell lung cancer (NSCLC).
Metastatic non-small cell lung cancer (NSCLC) unfortunately remains a lethal disease, despite recent genetic characterization of subclasses of NSCLC, mainly adenocarcinoma, which has led to the development of targeted therapies that improve progression-free survival (PFS). Ultimately, however, patients fatally relapse. In this review we will focus on the search to improve survival for NSCLC patients deemed to be pan-negative for the common driver alterations susceptible to targeted therapy, above all those with EGFR mutations or ALK, ROS or RET translocations. Other uncommon driver mutations such as HER2 and BRAF mutations should be tested in order to rule out targeted treatment before assigning patients to chemotherapy. Chemotherapy yields short lived response with median survival still less than one year. Customized chemotherapy represents one way to attempt to prolong survival, although to date no prospective randomized customized studies have reported sufficient evidence to support this. In one attempt to demonstrate the role of tailoring chemotherapy, the Spanish Lung Cancer Group (SLCG) phase II customized chemotherapy trial (NCT00883480) showed that RAP80, a component of the BRCA1-A complex, influenced outcome in patients with low BRCA1 expression treated with cisplatin/gemcitabine, and in patients with intermediate/high BRCA1 levels receiving cisplatin/docetaxel or docetaxel alone. We are currently performing a prospective, randomized phase III trial comparing non-customized cisplatin/docetaxel with customized therapy in metastatic NSCLC patients (NCT00617656/GECP-BREC) and a parallel phase II study (ChiCTR-TRC-12001860) is being carried out in China (BREC-China) under the auspices of the SLCG.


CRK9 contributes to regulation of mitosis and cytokinesis in the procyclic form of Trypanosoma brucei.
The Trypanosoma brucei cell cycle is regulated by combinations of cyclin/CRKs (cdc2 related kinases). Recently, two additional cyclins (CYC10, CYC11) and six new CRK (CRK7-12) homologues were identified in the T. brucei genome database 12. Individual RNAi knockdowns of these new proteins in the procyclic form of T. brucei showed no apparent phenotype except for the CRK9 depletion, which enriched the cells in G2/M phase. But a similar CRK9 knockdown in the bloodstream form caused no apparent phenotype. CRK9 lacks the typical PSTAIRE motif for cyclin binding and the phenylalanine "gatekeeper" but binds to cyclin B2 in vitro and localizes to the nucleus in both forms of T. brucei. CRK9-depleted procyclic-form generated no detectable anucleate cells, suggesting an inhibition of cytokinesis by CRK9 depletion as well. The knockdown enriched cells with one nucleus, one kinetoplast and two closely associated basal bodies with an average distance of 1.08 mm in between, which was shorter than the control value of 1.36 microm, and the cells became morphologically deformed and rounded with time. CRK9 may play a role in mediating the segregation between the two kinetoplast/basal body pairs prior to cytokinetic initiation. Since such a segregation over a relatively significant distance is essential for cytokinetic initiation only in the procyclic but may not be in the bloodstream form, CRK9 could be specifically involved in regulating cytokinetic initiation in the procyclic form of T. brucei.


Clinical and neuropsychological profile of patients with dementia and chronic traumatic encephalopathy.
To determine whether subjects with chronic traumatic encephalopathy (CTE) and dementia have distinct clinical features compared to subjects with pathologically confirmed Alzheimer's disease (AD). Among 339 subjects assessed for CTE in the National Alzheimer's Coordinating Center dataset, 6 subjects with CTE and 25 subjects with AD neuropathologic change matched for age (±5 years) and sex were identified. All subjects had a clinical diagnosis of dementia. Neurological examination, neuropsychological testing and emotional/behavioural data were compared between CTE and AD subjects at the time of dementia diagnosis and last clinical visit near death. A history of traumatic brain injury with loss of consciousness (LOC) was reported in one CTE and one AD subject; information about injuries without LOC or multiple injuries was unavailable. CTE and AD subjects did not differ significantly at the time of diagnosis or last visit on the Unified Parkinson's Disease Rating Scale-Motor Exam, global measures of cognitive functioning (Mini-Mental State Exam and Clinical Dementia Rating Scale), emotional/behaviour symptoms as assessed with the Neuropsychiatric Inventory questionnaire or across neuropsychological measures. All CTE participants had co-occurring neuropathologic processes, including AD and most had TAR DNA-binding protein 43 (TDP-43) neuropathology. CTE pathology was rare in a large multicentre national dataset, and when present, was accompanied by AD and TDP-43 pathologies. CTE was not associated with a different clinical presentation from AD or with greater cognitive impairment or neurobehavioral symptoms. These findings suggest that CTE may not have a distinct clinical profile when other neuropathologic processes are coexistent with CTE pathology.


Electrocardiographic guidance for the placement of gastric feeding tubes: a pediatric case series.
The placement of nasal or oral gastric tubes is one of the most frequently performed procedures in critically ill children; tube malposition, particularly in the trachea, is an important complication. Neurally adjusted ventilatory assist (NAVA) ventilation (available only on the Servo-i ventilator, Maquet Critical Care, Solna, Sweden) requires a proprietary-design catheter (Maquet Critical Care, Solna, Sweden) with embedded electrodes that detect the electrical activity of the diaphragm (EA(di)). The EA(di) catheter has the potential benefit of confirming proper positioning of a gastric catheter, based on and the EA(di) waveforms. In a case series study, our multidisciplinary team used EA(di) guidance for immediate, real-time confirmation of proper nasal or oral gastric tube placement in 20 mechanically ventilated pediatric patients who underwent 23 oral or nasal gastric tube placements. The catheters were placed with our standard practice, with the addition of a team member monitoring the EA(di) waveforms. As the tube passes down the esophagus and posterior to the heart, a characteristic EA(di) pattern is identified and the position of the atrial signal confirms correct placement of the gastric tube. If the EA(di) waveforms indicate incorrect placement, the tube is repositioned until the proper EA(di) waveform pattern is obtained. Then proper tube placement is reconfirmed via auscultation over the stomach while air is injected into the catheter, checking the pH of fluid suctioned from the catheter (gastric pH indicates correct positioning), and/or radiograph. The group's median age was 3 years (range 4 d to 16 y). All 20 patients had successful gastric catheter placement. The EA(di) catheter provided characteristic patterns for correctly placed tubes, tubes malpositioned above or below the gastroesophageal junction, and curled tubes. Proper catheter position was confirmed via radiograph and/or gastric pH in all 20 patients. EA(di) guidance helps confirm proper gastric catheter position, is equivalent to our standard practice for confirming gastric catheter placement, and may reduce the need for radiographs and improve patient safety by avoiding catheter malpositions.


Pupylation-dependent and -independent proteasomal degradation in mycobacteria.
Bacteria make use of compartmentalizing protease complexes, similar in architecture but not homologous to the eukaryotic proteasome, for the selective and processive removal of proteins. Mycobacteria as members of the actinobacteria harbor proteasomes in addition to the canonical bacterial degradation complexes. Mycobacterial proteasomal degradation, although not essential during normal growth, becomes critical for survival under particular environmental conditions, like, for example, during persistence of the pathogenic Mycobacterium tuberculosis in host macrophages or of environmental mycobacteria under starvation. Recruitment of protein substrates for proteasomal degradation is usually mediated by pupylation, the post-translational modification of lysine side chains with the prokaryotic ubiquitin-like protein Pup. This substrate recruitment strategy is functionally reminiscent of ubiquitination in eukaryotes, but is the result of convergent evolution, relying on chemically and structurally distinct enzymes. Pupylated substrates are recognized by the ATP-dependent proteasomal regulator Mpa that associates with the 20S proteasome core. A pupylation-independent proteasome degradation pathway has recently been discovered that is mediated by the ATP-independent bacterial proteasome activator Bpa (also referred to as PafE), and that appears to play a role under stress conditions. In this review, mechanistic principles of bacterial proteasomal degradation are discussed and compared with functionally related elements of the eukaryotic ubiquitin-proteasome system. Special attention is given to an understanding on the molecular level based on structural and biochemical analysis. Wherever available, discussion of in vivo studies is included to highlight the biological significance of this unusual bacterial degradation pathway.


Metal-cluster-decorated TiO2 nanotube arrays: a composite heterostructure toward versatile photocatalytic and photoelectrochemical applications.
Recent years have witnessed increasing interest in the solution-phase synthesis of atomically precise thiolate-protected gold clusters (Aux ); nonetheless, research on the photocatalytic properties of Aux -semiconductor nanocomposites is still in its infancy. In this work, recently developed glutathione-capped gold clusters and highly ordered nanoporous layer-covered TiO2 nanotube arrays (NP-TNTAs) are employed as nanobuilding blocks for the construction of a well-defined Aux /NP-TNTA heterostructure via a facile electrostatic self-assembly strategy. Versatile photocatalytic performances of the Aux /NP-TNTA heterostructure which acts as a model catalyst, including photocatalytic oxidation of organic pollutant, photocatalytic reduction of aromatic nitro compounds and photoelectrochemical (PEC) water splitting under simulated solar light irradiation, are systematically exploited. It is found that synergistic interaction stemming from monodisperse coverage of Aux clusters on NP-TNTAs in combination with hierarchical nanostructure of NP-TNTAs reinforce light absorption of Aux /NP-TNTA heterostructure especially within visible region, hence contributing to the significantly enhanced photocatalytic and PEC water splitting performances. Moreover, photocatalytic and PEC mechanisms over Aux /NP-TNTA heterostructure are elucidated and corresponding reaction models were presented. It is anticipated that this work could boost new insight for photocatalytic properties of metal-cluster-sensitized semiconductor nanocomposites.


How Can the Arts Influence the Attitudes of Dementia Caregivers? A Mixed-Methods Longitudinal Investigation.
Arts activities can improve social connectedness and foster reciprocity between people living with dementia and their caregivers. The extent to which the arts can help shape attitudes towards dementia remains unclear. This paper explores the impact of a 12-week visual arts program "Dementia and Imagination" on the attitudes of family and professional caregivers through a mixed-methods longitudinal investigation, underpinned by a conceptual framework of the arts in dementia care. One hundred and forty-six family and professional caregivers were recruited across three settings in England and Wales (residential care homes, a county hospital, and community venues). Quantitative and qualitative data on caregivers' attitudes and perceptions of the impact of the arts program were collected through interviews at three time points. Thematic analysis identified four themes shared across the caregivers: (1) Recognizing capabilities, (2) Social connectedness, (3) Improvements to well-being, and (4) Equality and personhood. Two further themes were distinct to family caregivers: (5) Duration of the effects of the art program, and (6) Enriched perspective. The final theme Inspiring professional development was distinct to the professional caregivers. The quantitative analysis found no effect for caregivers' attitude change over time. Family caregivers scored significantly lower than professional caregivers at each of the three time points. Art programs have the potential to make visible the capabilities of the people living with dementia, enabling caregivers to see the person behind the condition. This study highlights practice implications for future implementation, such as the role of the arts in dementia care education.


Authoring Data-Driven Videos with DataClips.
Data videos, or short data-driven motion graphics, are an increasingly popular medium for storytelling. However, creating data videos is difficult as it involves pulling together a unique combination of skills. We introduce DataClips, an authoring tool aimed at lowering the barriers to crafting data videos. DataClips allows non-experts to assemble data-driven "clips" together to form longer sequences. We constructed the library of data clips by analyzing the composition of over 70 data videos produced by reputable sources such as The New York Times and The Guardian. We demonstrate that DataClips can reproduce over 90% of our data videos corpus. We also report on a qualitative study comparing the authoring process and outcome achieved by (1) non-experts using DataClips, and (2) experts using Adobe Illustrator and After Effects to create data-driven clips. Results indicated that non-experts are able to learn and use DataClips with a short training period. In the span of one hour, they were able to produce more videos than experts using a professional editing tool, and their clips were rated similarly by an independent audience.


Factors Influencing the Prevalence of Animal Cruelty During Adolescence.
Adolescents' interactions with animals are of increasing interest and their beneficial developmental outcomes are well known. However, negative interactions such as perpetrating cruelty toward animals during childhood and adolescence have been related with child abuse, domestic violence, and later interpersonal violence. Cruelty toward animals by adolescents has been reported predominately in criminal and clinical samples, and links have been made between animal cruelty and interpersonal violence. However, studies often lack a clear definition of animal cruelty and the animal involved. The present study addresses methodological shortcomings by providing a clear definition of the cruelty acts and the animals involved and the time frame within which cruelty acts have been taken place. Therefore, we investigated the prevalence of animal cruelty of 979 British adolescents (419 male, 497 female, Mage = 15.1 ± 1.57 years) by means of a survey questionnaire administered in school. Animal cruelty was investigated encompassing deliberate and nondeliberate cruelty acts, a clear definition of the target animals was included and a time frame was provided. Furthermore, acceptability of animal cruelty, engaging in antisocial behavior, and family affluence were investigated. Results show high reliabilities for the measures applied. Exploratory factor analysis reveals different types of animal cruelty. Gender differences were observed for deliberate and accidental cruelty acts, with boys reporting higher levels than girls. Younger adolescents reported higher accidental cruelty acts than older ones. Acceptance of animal cruelty played a significant role in predicting animal cruelty, together with antisocial behaviors and place of living. The present study shows for the first time the importance of distinguishing between different types of animal cruelty and defining the animals involved.


Correlation of structure and mechanical response in solid-like polymers.
Employing large scale molecular dynamics simulations, we measure the uniaxial tensile response of amorphous and semicrystalline states of a coarse-grained PVA bead-spring model. The response beyond the elastic limit encompasses strain-softening and strain-hardening regimes. To understand the underlying mechanisms of plastic deformation, we analyse conformational and structural changes of polymers. In particular, we characterise the volume distribution of crystalline domains along the stress-strain curve. The strain-softening regime in semicrystalline samples is dominated by deformation of crystalline parts, while strain-hardening involves unfolding and alignment of chains in both amorphous and crystalline parts. Comparing the tensile response of semicrystalline and amorphous polymers, we find similar conformations of polymers for both systems in the strain-hardening regime.


Chrysanthemum indicum L. extract induces apoptosis through suppression of constitutive STAT3 activation in human prostate cancer DU145 cells.
Chrysanthemum indicum L. has been shown to possess antiinflammatory and anticancer activities, but its molecular targets/pathways are not yet fully understood in tumor cells. In the present study, the potential effects of C. indicum on signal transducer and activator of transcription 3 (STAT3) signaling pathway in different tumor cells were examined. The solvent fractions (hexane, CH₂Cl₂, EtOAc, and BuOH,) were obtained from a crude extract (80% EOH extract) of C. indicum. The methylene chloride fraction of C. indicum (MCI) exhibited strong cytotoxic activity as compared with the other fractions and clearly suppressed constitutive STAT3 activation against both DU145 and U266 cells, but not MDA-MB-231 cells. The suppression of constitutive STAT3 activation by MCI is associated with blocking upstream JAK1 and JAK2, but not Src. MCI downregulated the expression of STAT3-regulated gene products; this is correlated with the accumulation of the cell cycle at sub-G1 phase, the induction of caspase-3 activation, and apoptosis. Moreover, the major components of the MCI were bioactive compounds such as sudachitin, hesperetin, chrysoeriol, and acacetin. Sudachitin, chrysoeriol, and acacetin also exerted significantly cytotoxicity, clearly suppressed constitutive STAT3 activation, and induced apoptosis, although hesperetin did not show any significant effect in DU145 cells. Overall, our results demonstrate that MCI could induce apoptosis through inhibition of the JAK1/2 and STAT3 signaling pathways.


Locating the nucleation sites for protein encapsulated gold nanoclusters: a molecular dynamics and fluorescence study.
Fluorescent gold nanoclusters encapsulated by proteins have attracted considerable attention in recent years for their unique properties as new fluorescence probes for biological sensing and imaging. However, fundamental questions, such as the nucleation sites of gold nanoclusters within proteins and the fluorescence mechanism remain unsolved. Here we present a study of the location of gold nanoclusters within bovine serum albumin (BSA) combining both fully atomistic molecular dynamic (MD) simulations and fluorescence spectroscopic studies. The MD simulations show gold clusters growing close to a number of cysteine sites across all three domains of BSA, although just two major sites in domains IIB and IA were found to accommodate large clusters comprising more than 12 atoms. The dependence of the fluorescence on pH is found to be compatible with possible nucleation sites in domains IIB and IA. Furthermore, the energy transfer between tryptophan and gold nanoclusters reveals a separation of 29.7 Å, further indicating that gold nanoclusters were most likely located in the major nucleation site in domain IIB. The disclosure of the precise location of the gold nanoclusters and their surrounding amino acid residues should help better understanding of their fluorescence mechanism and aid their optimization as fluorescent nanoprobes.


Rheumotologitsts' view on the use of hydroxychloroquine to treat COVID-19.
The current pandemic coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) calls urgently for effective therapies. Anti-malarial medicine chloroquine (CQ) and particularly its chemical analogue hydroxychloroquine (HCQ) have been recommended as promising candidate therapeutics that are now under either compassionate off-label use or clinical trials for the treatment of COVID-19 patients. However, there are public concerns and disputes about both the safety and efficacy of CQ and HCQ for this new application. Given the fact that for decades HCQ has been approved as an immunomodulatory drug for the long term treatment of chronic rheumatic diseases, as experienced rheumatologists, we would like to share our thoughts in this regard and trigger a brainstorm among clinical care providers for exchanging their diverse opinions on this urgent topic.


Structural determinants of PINK1 topology and dual subcellular distribution.
PINK1 is a mitochondria-targeted kinase that constitutively localizes to both the mitochondria and the cytosol. The mechanism of how PINK1 achieves cytosolic localization following mitochondrial processing remains unknown. Understanding PINK1 subcellular localization will give us insights into PINK1 functions and how mutations in PINK1 lead to Parkinson's disease. We asked how the mitochondrial localization signal, the transmembrane domain, and the kinase domain participate in PINK1 localization. We confirmed that PINK1 mitochondrial targeting signal is responsible for mitochondrial localization. Once inside the mitochondria, we found that both PINK1 transmembrane and kinase domain are important for membrane tethering and cytosolic-facing topology. We also showed that PINK1 dual subcellular distribution requires both Hsp90 interaction with the kinase domain and the proteolysis at a cleavage site downstream of the transmembrane domain because removal of this cleavage site completely abolished cytosolic PINK1. In addition, the disruption of the Hsp90-PINK1 interaction increased mitochondrial PINK1 level. Together, we believe that once PINK1 enters the mitochondria, PINK1 adopts a tethered topology because the transmembrane domain and the kinase domain prevent PINK1 forward movement into the mitochondria. Subsequent proteolysis downstream of the transmembrane domain then releases PINK1 for retrograde movement while PINK1 kinase domain interacts with Hsp90 chaperone. The significance of this dual localization could mean that PINK1 has compartmental-specific functions.


Discordant neutralizing antibody and T cell responses in asymptomatic and mild SARS-CoV-2 infection.
Understanding the nature of immunity following mild/asymptomatic infection with SARS-CoV-2 is crucial to controlling the pandemic. We analyzed T cell and neutralizing antibody responses in 136 healthcare workers (HCW) 16-18 weeks after United Kingdom lockdown, 76 of whom had mild/asymptomatic SARS-CoV-2 infection captured by serial sampling. Neutralizing antibodies (nAb) were present in 89% of previously infected HCW. T cell responses tended to be lower following asymptomatic infection than in those reporting case-definition symptoms of COVID-19, while nAb titers were maintained irrespective of symptoms. T cell and antibody responses were sometimes discordant. Eleven percent lacked nAb and had undetectable T cell responses to spike protein but had T cells reactive with other SARS-CoV-2 antigens. Our findings suggest that the majority of individuals with mild or asymptomatic SARS-CoV-2 infection carry nAb complemented by multispecific T cell responses at 16-18 weeks after mild or asymptomatic SARS-CoV-2 infection.


A single-component light sensor system allows highly tunable and direct activation of gene expression in bacterial cells.
Light-regulated modules offer unprecedented new ways to control cellular behaviour with precise spatial and temporal resolution. Among a variety of bacterial light-switchable gene expression systems, single-component systems consisting of single transcription factors would be more useful due to the advantages of speed, simplicity, and versatility. In the present study, we developed a single-component light-activated bacterial gene expression system (eLightOn) based on a novel LOV domain from Rhodobacter sphaeroides (RsLOV). The eLightOn system showed significant improvements over the existing single-component bacterial light-activated expression systems, with benefits including a high ON/OFF ratio of >500-fold, a high activation level, fast activation kinetics, and/or good adaptability. Additionally, the induction characteristics, including regulatory windows, activation kinetics and light sensitivities, were highly tunable by altering the expression level of LexRO. We demonstrated the usefulness of the eLightOn system in regulating cell division and swimming by controlling the expression of the FtsZ and CheZ genes, respectively, as well as constructing synthetic Boolean logic gates using light and arabinose as the two inputs. Taken together, our data indicate that the eLightOn system is a robust and highly tunable tool for quantitative and spatiotemporal control of bacterial gene expression.


Drastic population fluctuations explain the rapid extinction of the passenger pigeon.
To assess the role of human disturbances in species' extinction requires an understanding of the species population history before human impact. The passenger pigeon was once the most abundant bird in the world, with a population size estimated at 3-5 billion in the 1800s; its abrupt extinction in 1914 raises the question of how such an abundant bird could have been driven to extinction in mere decades. Although human exploitation is often blamed, the role of natural population dynamics in the passenger pigeon's extinction remains unexplored. Applying high-throughput sequencing technologies to obtain sequences from most of the genome, we calculated that the passenger pigeon's effective population size throughout the last million years was persistently about 1/10,000 of the 1800's estimated number of individuals, a ratio 1,000-times lower than typically found. This result suggests that the passenger pigeon was not always super abundant but experienced dramatic population fluctuations, resembling those of an "outbreak" species. Ecological niche models supported inference of drastic changes in the extent of its breeding range over the last glacial-interglacial cycle. An estimate of acorn-based carrying capacity during the past 21,000 y showed great year-to-year variations. Based on our results, we hypothesize that ecological conditions that dramatically reduced population size under natural conditions could have interacted with human exploitation in causing the passenger pigeon's rapid demise. Our study illustrates that even species as abundant as the passenger pigeon can be vulnerable to human threats if they are subject to dramatic population fluctuations, and provides a new perspective on the greatest human-caused extinction in recorded history.


A dated phylogeny and collection records reveal repeated biome shifts in the African genus Coccinia (Cucurbitaceae).
Conservatism in climatic tolerance may limit geographic range expansion and should enhance the effects of habitat fragmentation on population subdivision. Here we study the effects of historical climate change, and the associated habitat fragmentation, on diversification in the mostly sub-Saharan cucurbit genus Coccinia, which has 27 species in a broad range of biota from semi-arid habitats to mist forests. Species limits were inferred from morphology, and nuclear and plastid DNA sequence data, using multiple individuals for the widespread species. Climatic tolerances were assessed from the occurrences of 1189 geo-referenced collections and WorldClim variables. Nuclear and plastid gene trees included 35 or 65 accessions, representing up to 25 species. The data revealed four species groups, one in southern Africa, one in Central and West African rain forest, one widespread but absent from Central and West African rain forest, and one that occurs from East Africa to southern Africa. A few individuals are differently placed in the plastid and nuclear (LFY) trees or contain two ITS sequence types, indicating hybridization. A molecular clock suggests that the diversification of Coccinia began about 6.9 Ma ago, with most of the extant species diversity dating to the Pliocene. Ancestral biome reconstruction reveals six switches between semi-arid habitats, woodland, and forest, and members of several species pairs differ significantly in their tolerance of different precipitation regimes. The most surprising findings of this study are the frequent biome shifts (in a relatively small clade) over just 6 - 7 million years and the limited diversification during and since the Pleistocene. Pleistocene climate oscillations may have been too rapid or too shallow for full reproductive barriers to develop among fragmented populations of Coccinia, which would explain the apparently still ongoing hybridization between certain species. Steeper ecological gradients in East Africa and South Africa appear to have resulted in more advanced allopatric speciation there.


Templating multiple length scales by combining phase separation, self-assembly and photopatterning in porous films.
Patternable nanoporous silica thin films with pore sizes on multiple length scales are fabricated using preformed block copolymer/homopolymer blend films as templates. Previous work by Tirumala et al. [V.R. Tirumala, R.A. Pai, S. Agarwal, J.J. Testa, G. Bhatnagar, A.H. Romang, C. Chandler, B.P. Gorman, R.L. Jones, E.K. Lin, J.J. Watkins, Adv. Mater. 18 (8) (2008) 1603] has demonstrated that hydrogen bonding between an amphiphilic copolymer and a homopolymer leads to significant enhancements in the long range order of the template self assembly. However if the copolymer template is simply changed from Pluronic F127 to Brij78, a well ordered template is no longer always obtained; the blend phase separates with apparent selective partitioning of the photoacid generator (PAG) at the interface of the polymer phases. UV exposure selectively generates a photoacid, which is utilized to catalyze tetraethylorthosilicate (TEOS) condensation. The large disparity in diffusivity of the photoacid between the glassy poly(hydroxystyrene) (PHOSt) and rubbery Brij78 phases results in selective templating of the Brij mesostructure and limited reaction into the PHOSt. Calcination yields relatively monodisperse mesopores from the Brij phase and macropores from the PHOSt phase. Simple variations in processing parameters allow the macropore morphology to be tuned to create high surface area materials with structures on the order of 1 nm, 100 nm and microms from self assembly, phase separation and lithographic patterning respectively.


Responsive parenting: establishing early foundations for social, communication, and independent problem-solving skills.
Mothers whose infants varied in early biological characteristics (born at term, n = 120; born at very low birth weight [VLBW], n = 144) were randomized to a target group (n = 133) or developmental feedback comparison group (n = 131) to determine whether learning responsive behaviors would facilitate infant development. The target condition included videotaped examples, problem-solving activities, and mothers' critique of their own behaviors through video procedures across 10 home visits. All target versus comparison mothers showed greater increases across multiple responsiveness behaviors observed in 4 assessments conducted across 6-13 months of age; changes in emotionally supportive behaviors were strongest for target mothers of infants born at VLBW. Increased maternal responsiveness facilitated greater growth in target infants' social, emotional, communication, and cognitive competence, supporting a causal role for responsiveness on infant development. Although benefits were generally comparable across risk groups, aspects of social and emotional skills showed greater change for those born at VLBW. Evidence for responsiveness as a multidimensional construct was provided as well as the importance of different aspects of responsiveness mediating the effect of the intervention on different infant skill domains.


Medical and social costs after using financial incentives to improve medication adherence: results of a 1 year randomised controlled trial.
Offering a financial incentive ('Money for Medication') is effective in improving adherence to treatment with depot antipsychotic medications. We investigated the cost-effectiveness in terms of medical costs and judicial expenses of using financial incentives to improve adherence. The effects of financial incentives on depot medication adherence were evaluated in a randomised controlled trial. Patients in the intervention group received €30 a month over 12 months if antipsychotic depot medication was accepted. The control group received mental health care as usual. For 133 patients outcomes were calculated based on self-reported service use and delinquent behaviour and expressed as standard unit costs to value resource use. The financial incentive resulted in higher average costs related to mental health care (€449.6 versus €355.7). and lower medical costs related to other healthcare services (€52.0 versus €78.4). Relevant differences in social costs related to delinquent behaviour were not found. Although wide confidence intervals indicate uncertainty, incremental cost-effectiveness ratio's (ICER) indicate that it costs €2080 for achieving a 20% increase in adherence or €3332 for achieving over 80% adherence. In sum, offering money as financial incentive for increasing compliance did not lead to an overall cost reduction as compared to care as usual. Trial registration NTR2350, 01 June 2010.


Antimicrobial activity of essential oils against Helicobacter pylori.
Helicobacter pylori is an important pathogen responsible for gastroduodenal diseases in humans. Although the eradication of H. pylori using antibiotics often improves gastroduodenal diseases, resistance to the antibiotics is emerging. The antimicrobial effect of essential oils and the development of resistance to the essential oils were evaluated in vitro and in vivo. Thirteen essential oils used in this study completely inhibited the growth of H. pylori in vitro at a concentration of 0.1% (v/v). Cymbopogon citratus (lemongrass) and Lippia citriodora (lemon verbena) were bactericidal against H. pylori at 0.01% at pH 4.0 and 5.0. Resistance to lemongrass did not develop even after 10 sequential passages, whereas resistance to clarithromycin developed under the same conditions. In in vivo studies, the density of H. pylori in the stomach of mice treated with lemongrass was significantly reduced compared with untreated mice. These results demonstrate that the essential oils are bactericidal against H. pylori without the development of acquired resistance, suggesting that essential oils may have potential as new and safe agents for inclusion in anti-H. pylori regimens.


The Evidence Base for an Ideal Care Pathway for Frail Multimorbid Elderly: Combined Scoping and Systematic Intervention Review.
There is a call for bold and innovative action to transform the current care systems to meet the needs of an increasing population of frail multimorbid elderly. International health organizations propose complex transformations toward digitally supported (1) Person-centered, (2) Integrated, and (3) Proactive care (Digi-PIP care). However, uncertainty regarding both the design and effects of such care transformations remain. Previous reviews have found favorable but unstable impacts of each key element, but the maturity and synergies of the combination of elements are unexplored. This study aimed to describe how the literature on whole system complex transformations directed at frail multimorbid elderly reflects (1) operationalization of intervention, (2) maturity, (3) evaluation methodology, and (4) effect on outcomes. We performed a systematic health service and electronic health literature review of care transformations targeting frail multimorbid elderly. Papers including (1) Person-centered, integrated, and proactive (PIP) care; (2) at least 1 digital support element; and (3) an effect evaluation of patient health and/ or cost outcomes were eligible. We used a previously published ideal for the quality of care to structure descriptions of each intervention. In a secondary deductive-inductive analysis, we collated the descriptions to create an outline of the generic elements of a Digi-PIP care model. The authors then reviewed each intervention regarding the presence of critical elements, study design quality, and intervention effects. Out of 927 potentially eligible papers, 10 papers fulfilled the inclusion criteria. All interventions idealized Person-centered care, but only one intervention made what mattered to the person visible in the care plan. Care coordinators responsible for a whole-person care plan, shared electronically in some instances, was the primary integrated care strategy. Digitally supported risk stratification and management were the main proactive strategies. No intervention included workflow optimization, monitoring of care delivery, or patient-reported outcomes. All interventions had gaps in the chain of care that threatened desired outcomes. After evaluation of study quality, 4 studies remained. They included outcome analyses on patient satisfaction, quality of life, function, disease process quality, health care utilization, mortality, and staff burnout. Only 2 of 24 analyses showed significant effects. Despite a strong common-sense belief that the Digi-PIP ingredients are key to sustainable care in the face of the silver tsunami, research has failed to produce evidence for this. We found that interventions reflect a reductionist paradigm, which forces care workers into standardized narrowly focused interventions for complex problems. There is a paucity of studies that meet complex needs with digitally supported flexible and adaptive teamwork. We predict that consistent results from care transformations for frail multimorbid elderly hinges on an individual care pathway, which reflects a synergetic PIP approach enabled by digital support.


Optical configuration for TV holography measurement of in-plane and out-of-plane deformations.
A novel TV holography method is proposed for parallel evaluation of in-plane and out-of-plane deformation fields. The method permits a trade-off between in-plane and out-of-plane measuring sensitivity. A four-exposure, four-frame phase shifting technique is used in the experiments; the experimental results for an aluminum specimen subjected to both rotation in its own plane and a bending couple load at the center are presented.


Centrifugal force stretcher a new of in vitro mechanical cell stimulator.
A number of mechanical cell stimulators have been used to study the effect of mechanical stimulation on cells in vitro. But the efficiency of these devices is not fully desirable. We recently developed a new device for mechanical cell stimulation, the centrifugal force stretcher, and compared its efficacy with that of the traditional Flexercell Strain Unit. When the mechanical stretcher circumrotates with certain speed, cardiac myocytes attached on the plate are stretched and elongated by centrifugal force. Neonatal rat cardiac myocytes were isolated by enzymatic dissociation from the hearts of 3~5 d old Sprague Dawley rats, and were mechanically stimulated by traditional 20% stretch and 180 r/min centrifugal force for 12 and 24 h. The effects of mechanical stimulation on the hypertrophic response of neonatal rat cardiac myocytes and production of angiotensin II (Ang II) were examined. Compared with the non-stretch group, the radioactivity of (3)H-leucine incorporated into the stretch-stimulated cardiac myocytes in the centrifugal force stretch group was significantly higher [(1295.17+/-51.19) vs (1122.67+/-51.63) in 12 h; (1447.5+/-35.96) vs (1210.67+/-90.92) in 24 h, P<0.05]. Ang II was also dramatically increased by 128% in 12 h (P<0.05) and 139% in 24 h (P<0.01). After the myocytes was stretched for 24 h, the LDH level in the medium in the Flexercell Strain Unit group was significantly higher than that in the centrifugal force group [(14.5+/-8.7) U/L vs (7.8+/-4.3) U/L, P<0.05]. The centrifugal force stretcher is a new and improved mechanical cell stimulator with the same effects on the protein synthesis and Ang II secretion of the cardiac myocytes, and the damage to the cells bronght by this stimulator is relatively slighter in comparison with the Flexercell Strain Unit.


Assimilation and depuration of microcystin-LR by the zebra mussel, Dreissena polymorpha.
Zebra mussels (Dreissena polymorpha) are an important component of the foodweb of shallow lakes in the Netherlands, amongst others in Lake IJsselmeer, an international important wetland. Large numbers of ducks feed on these mussels in autumn and winter. The mussels are filter feeders and are exposed to high densities of cyanobacteria in summer and autumn. Mussels and cyanobacteria both thrive in Lake IJsselmeer. Apparently the mussels are somehow protected against accumulation of harmful quantities of cyanobacterial toxins. In this study, we investigated the assimilation of the cyanobacterial toxin microcystin-LR (MC-LR) in zebra mussels when fed the toxic cyanobacterium Microcystis aeruginosa as sole food or in a mixture with the eustigmatophyte Nannochloropsis limnetica. After 3 weeks of assimilation we studied the depuration of MC-LR during 3 weeks when the food of the mussels was free of cyanobacteria. These assimilation/depuration experiments were combined with grazing experiments, using the same food treatments. Microcystins were analyzed using liquid chromatography-mass spectrometry (LC-MS); in addition, covalently bound MC were analyzed using the MMPB method. The mussels showed higher clearance rates on Microcystis than on Nannochloropsis. No selective rejection of either phytoplankton species was observed in the excretion products of the mussels. Zebra mussels fed Microcystis as single food, assimilated microcystin-LR relatively fast, and after 1 week the maximum value of free unbound microcystin assimilation (ca. 11 microg g DW(-1)) was attained. For mussels, fed with the mixed food, a maximum of only 3.9 microg g DW(-1) was recorded after 3 weeks. Covalently bound MC never reached high values, with a maximum of approximately 62% of free MC in the 2nd week of the experiment. In the depuration period microcystin decreased rapidly to low values and after 3 weeks only very low amounts of microcystin were detectable. The amount of toxin that accumulated in the mussels would appear to be high enough to cause (liver) damage in diving ducks. However, death by exposure to microcystin seems unlikely. Mussels seem efficient in minimizing the assimilation of microcystin. If it were not for this, mass mortalities of ducks in shallow lakes in the Netherlands would presumably occur on a much more widespread scale than is currently observed.


INCOG guidelines for cognitive rehabilitation following traumatic brain injury: methods and overview.
Traumatic brain injury results in complex cognitive sequelae. However, clinicians have difficulty implementing the available evidence. An international group of researchers and clinicians (known as INCOG) convened to develop clinical practice guidelines for cognitive rehabilitation posttraumatic brain injury. The Guidelines Adaptation and Development cycle was used to derive the recommendations. Previously published cognitive rehabilitation recommendations were identified and tabulated. An expert panel met to select appropriate recommendations. Afterward, the team enhanced the recommendations by reviewing available literature. To address shortfalls of previous guidelines, the team developed decision algorithms incorporating the recommendations based on inclusion and exclusion criteria of published trials and expert opinion. The team then prioritized the recommendations for implementation and developed audit criteria to evaluate adherence to best practice. The team recommends that individuals have detailed assessments of cognition after resolution of posttraumatic amnesia. Cognitive assessment and rehabilitation should be tailored to the patient's neuropsychological profile, premorbid cognitive characteristics, and goals for life activities and participation. Clinical algorithms and audit tools to evaluate current practice are provided. Cognitive rehabilitation should be offered to select individuals with traumatic brain injury. These guidelines provide assistance to clinicians who want to provide evidence-based care.


Formation and drying of colloidal crystals using nanosized silica particles.
Much interest has been generated in the fabrication of colloidal crystals from suspensions because of the promise of photonic band gap applications. However, since the case of small, nonsedimenting colloidal particles indeed remains rather rarely treated, spherical silica particles with diameters varying from 75 down to 20 nm have been used in the present work to fabricate colloidal crystals by drying the suspending liquid. Typical events that take place during the drying process of a particulate film, such as cracking, compaction and penetration of air into a porous network, have been evaluated using existing theories, and the maximum stress in the drying film could be approximated. Investigation on the dry film structure by scanning electron microscopy showed the arrangement of particles in a close-packed system. To interpret the formation of such crystals, the amplitudes of the interparticle and capillary forces have been estimated from existing models. The repulsive interparticle forces allow the particles to remain stable and thus rearrange up to fairly high particle concentration. These modeling results showed the dominance of the capillary contribution at the end of the drying process. Nitrogen adsorption/desorption measurements gave very coherent results regarding both pore volume and pore size of the dry particulate films when compared to the expected ordered packing arrangements.


Anxiolytic-like effect of inhalation of essential oil from Lavandula officinalis: investigation of changes in 5-HT turnover and involvement of olfactory stimulation.
Essential oil extracted from Lavandula officinalis (LvEO) has a long history of usage in anxiety alleviation with good evidence to support its use. However, findings and information regarding the exact pathway involved and mechanism of action remain inconclusive. Therefore, we aimed to (1) reveal the influence of olfactory stimulation, and (2) determine whether the serotonergic system is involved in the anxiolytic effect of LvEO when it is inhaled. To this end, we first compared the anxiety-related behaviors of normosmic and anosmic mice. LvEO inhalation caused notable elevation in anxiety-related parameters with or without olfactory perception, indicating that olfactory stimulation is not necessarily required for LvEO to be effective. Neurochemical analysis of the serotonin (5-HT) turnover rate, accompanied by EPM testing, was then performed. LvEO significantly increased the striatal and hippocampal levels of 5-HT and decreased turnover rates in accordance with the anxiolytic behavioral changes. These results, together with previous findings, support the hypothesis that serotonergic neurotransmission plays a certain role in the anxiolytic properties of LvEO.


18F-FDG-PET partial volume effect correction using a modified recovery coefficient approach based on functional volume and local contrast: physical validation and clinical feasibility in oncology.
2-deoxy-2-[18F]fluoro-D-glucose 18F-FDG uptake within tumors reflects the glucose consumption of malignant tumors, i.e., the tumor activity. Thus, 18F-FDG uptake measurements enable improved therapeutic monitoring of patients in chemo- or radiotherapy treatment through the detection of changes in tumor uptake via quantitative measurements of the lesion standard uptake values (SUVs) or activity concentrations. A major bias that affects positron emission tomography (PET) image quantitation is the partial volume effect (PVE), which most strongly affects the smallest structures due to the poor spatial resolution of PET. Thus, PVE corrections are important when 18F-FDG-PET images are used as a quantitative tool for monitoring responses to therapy. The aim of this paper was to propose a PVE correction based on a modified recovery coefficient method (termed FARCAS) that considers the functional volumes and local contrasts of lesions that are automatically determined using a semi-automatic iterative segmentation algorithm. The FARCAS method consists of establishing a set of calibration curves based on the mathematical fitting of the RC values as a function of the automatically determined functional lesion volume and local lesion contrast. We set up our method using data from a cylindrical phantom that included spheres of different volumes (range: 0.43 to 97.8 mL) and contrasts (range: 1.7 to 22.9), and we assessed the method using both cylindrical and anthropomorphic phantom data that also included spheres of different volumes and contrasts. FARCAS was also compared with conventional RC methods that only considered the lesion functional volume, either automatically determined (RCVa) or using the ground truth volume (RCVgt). Finally, the clinical feasibility of FARCAS and its evaluation on tumor classification were also assessed on 24 NSCLC lesions. Whatever the phantom considered, for the spheres with contrast <5, FARCAS obtained comparable results to RCVgt and better than RCVa. For the spheres with contrast >5, FARCAS and RCVa were not statistically different, neither for the cylindrical and nor the anthropomorphic phantom. For the cylindrical phantom FARCAS yielded corrections that were not statistically different to those of RCVa for the smallest spheres (V<2 mL), but statistically superior for the larger spheres (V≥2 mL). RCVgt maintained a non-statistically superior accuracy. Regarding the anthropomorphic data, FARCAS was statistically more accurate than RCVa but not RCVgt. As main findings regarding the clinical data, FARCAS modified the classifications of five of 24 NSCLC lesions based on quantitative PERCIST criteria. The PVE correction proposed in this paper allows the accurate quantification of the PVE-corrected SUV, allowing also an automatic definition of the Metabolic Target Volume (MTV). Our results revealed that the PVE correction based on FARCAS is a better approach than conventional RC to significantly reduce the impact of PVE on lesion quantification, thus improving the evaluation of tumor response to treatment based on PET-CT images.


Survey of academic field experiences (SAFE): trainees report harassment and assault.
Little is known about the climate of the scientific fieldwork setting as it relates to gendered experiences, sexual harassment, and sexual assault. We conducted an internet-based survey of field scientists (N = 666) to characterize these experiences. Codes of conduct and sexual harassment policies were not regularly encountered by respondents, while harassment and assault were commonly experienced by respondents during trainee career stages. Women trainees were the primary targets; their perpetrators were predominantly senior to them professionally within the research team. Male trainees were more often targeted by their peers at the research site. Few respondents were aware of mechanisms to report incidents; most who did report were unsatisfied with the outcome. These findings suggest that policies emphasizing safety, inclusivity, and collegiality have the potential to improve field experiences of a diversity of researchers, especially during early career stages. These include better awareness of mechanisms for direct and oblique reporting of harassment and assault and, the implementation of productive response mechanisms when such behaviors are reported. Principal investigators are particularly well positioned to influence workplace culture at their field sites.


Phorbol ester increases the dihydropyridine-sensitive calcium conductance in a vascular smooth muscle cell line.
In vascular smooth muscle, phorbol esters cause a slowly developing contraction and an associated transmembrane calcium flux, both of which are inhibited by dihydropyridine calcium channel antagonists. In the A7r5 cultured vascular cell line, we used the whole-cell voltage-clamp technique to identify voltage-dependent calcium conductances and investigate the effect of phorbol esters on that conductance having characteristic dihydropyridine sensitivity (slowly inactivating, high-threshold, "L-type"). With barium as the charge carrier, large-amplitude (100-800 pA) inward currents of two types were characterized by their kinetics and voltage dependence. With holding potential--80 mV, a rapidly inactivating, low-threshold current ("T-type") was activated by depolarizations above-40 mV and was maximal at -10 mV. With holding potential -30 mV, this component was inactivated, and a second slowly inactivating, high-threshold current was activated above -10 mV and was maximal at +10 to +20 mV. These currents are similar to the T-type and L-type currents previously described in vascular smooth muscle cells. When added to the bath, the active phorbol ester, 12-O-tetradecanoyl phorbol-13-acetate (100 nM) increased the slowly inactivating (L-type) current by 32 +/- 20% (n = 8, +/- SD). Phorbol-12,13-dibutyrate (100 nM) caused a similar effect, but the inactive phorbol, 4-alpha-phorbol (100 nM), did not. We conclude that at least two distinct calcium conductances are expressed in A7r5 vascular smooth muscle cells, and that the dihydropyridine-sensitive calcium conductance is acutely modulated by phorbol esters, presumably acting through stimulation of protein kinase C. Such modulation may play a role in increasing transmembrane calcium influx mediated by agonist-receptor interactions that lead to activation of protein kinase C and may help to sustain or amplify calcium-dependent cell responses.


What is the effect of age on pancreatic resection?
Both morbidity and mortality rates following pancreatic resection increase with advanced age. The reported mortality rates following pancreatic surgery arc underestimated in single-institution studies. There is a significant publication bias where only centers with good results report their outcomes. The population-based data are critical to provide a more realistic view of mortality rates following pancreatic resection. It is essential in counseling elderly patients that they understand that mortality rates are increased, morbidity rates are increased, and the effect of complications often leads to a prolonged convalescence. They will have a longer length of stay and up to a 30% to 40% chance that they will not be able to go home but will need to recover in an extended care facility following hospital discharge. Although the morbidity and mortality rates are increased for elderly patients, they are well within the acceptable range for major abdominal surgery when performed at experienced centers. Patients also need to be aware that surgical resection is the only curative option for pancreatic cancer. In reasonable risk elderly patients the benefit of surgical resection does not decrease with age and these patients can experience long-term survival and good quality of life. Once patients over 80 get beyond the 2-year survival mark without cancer recurrence their survival parallels that of their age-matched counterparts. One should also keep in mind that the reported survival rates are mostly for pancreatic cancer, but patients with other periampullary cancers have improved long-term survival when compared with those with pancreatic cancer. Elderly patients also need to be aware of the fact that hospital volume and surgeon experience significantly impact outcomes. The mortality rates following surgery in the oldest patients, those over 80, are nearly twice as high at low-volume facilities compared with high-volume facilities. The overall mortality rate and the difference decrease with decreasing age. This likely represents improved processes of care at experienced centers and better ability to manage the complications of pancreatic surgery, which occur more commonly in elderly patients. It is important to educate both physicians and elderly patients about this difference. Currently, elderly patients are less likely to be resected at high-volume centers than younger patients. The reasons for this are unclear but include lack of awareness of the importance of hospital volume and surgeon experience and reluctance of patients in this age group to travel long distances from home for their care. When reviewing the data, one must be aware that these studies (both population-based and single-institution) are retrospective and subject to significant selection bias. The elderly patients undergoing resection were dearly carefully chosen. There is still nihilism, however, toward aggressive care in these patients, with fewer than 10% of patients over 80 with locoregional disease and no comorbidities being resected, whereas 40% of patients 66 to 70 in the same category are resected. These data provide an excellent foundation to guide informed decision-making in the elderly population with pancreatic and periampullary cancer. Patients need to know that surgical resection offers the only hope for cure and that the benefit of surgical resection does not diminish with age. The diagnosis (pancreatic versus other periampullary cancers versus benign disease or premalignant lesions) needs to be taken into account to balance fully the risks and benefits. Older patients need to be aware of the increased morbidity, mortality, and prolonged convalescence they may experience. They also need to be advised to have their surgery done by experienced surgeons at experienced centers where these complications can be best managed. Further studies are needed to guide patient selection. The effect of patient comorbidities, cognitive status, preoperative functional status, and frailty need to be more formally assessed to select patients, maximize surgical resection in appropriate candidates, and improve short-term outcomes. Once better characterized, specialized geriatric pathways may optimize surgical resection rates, streamline care, and improve outcomes in this challenging population. Age alone, however, should not be a contraindication to pancreatic resection in elderly patients with pancreatic cancer.


Antibody-mediated early death in vivo after infection with yellow fever virus.
The phenomenon known as antibody-dependent enhancement (ADE) has been demonstrated in vitro but its significance in viral pathogenesis is uncertain even though it has been associated with dengue shock syndrome. Here we report for the first time the enhancement of virus virulence in mice using monoclonal antibodies (MAbs) prepared against yellow fever (YF) viruses. Our results show that the average survival time of mice was reduced by up to 33% (i.e. 6.7 to 4.5 days) and that ADE is both antibody dose-dependent and antibody- and virus strain-specific. A total of 12 YF viruses and 11 MAbs were examined and of these only three YF viruses (FNV, Asibi and B11) could be enhanced in vivo by only two MAbs (427 and 126). A particular combination of virus and antibody is required for ADE to take place.


Effects of age on metacognitive efficiency.
Humans have a capacity to become aware of thoughts and behaviours known as metacognition. Metacognitive efficiency refers to the relationship between subjective reports and objective behaviour. Understanding how this efficiency changes as we age is important because poor metacognition can lead to negative consequences, such as believing one is a good driver despite a recent spate of accidents. We quantified metacognition in two cognitive domains, perception and memory, in healthy adults between 18 and 84years old, employing measures that dissociate objective task performance from metacognitive efficiency. We identified a marked decrease in perceptual metacognitive efficiency with age and a non-significant decrease in memory metacognitive efficiency. No significant relationship was identified between executive function and metacognition in either domain. Annual decline in metacognitive efficiency after controlling for executive function was ∼0.6%. Decreases in metacognitive efficiency may explain why dissociations between behaviour and beliefs become more marked as we age.


Recent progress on single-molecule localization microscopy.
Super-resolution imaging based on single-molecule localization has been developed for more than a decade. These techniques can break through diffraction limit of fluorescent microscopy and initially improve the resolution by an order of magnitude to ~20 nm, by introducing photoactivatable/photoswitching probes and centroid fitting method. As the demand of biological research, the localization precision of single-molecules was further improved by several state-of-the-art methods in the past several years. This review focuses on the latest developed techniques which have greatly improved the performance of single-molecule localization microscopy, from measurement principle to hardware design. These methods are essential for the study of nanostructures and biomacromolecule dynamics inside of cells.


Compensatory growth of healthy cardiac cells in the presence of diseased cells restores tissue homeostasis during heart development.
Energy generation by mitochondrial respiration is an absolute requirement for cardiac function. Here, we used a heart-specific conditional knockout approach to inactivate the X-linked gene encoding Holocytochrome c synthase (Hccs), an enzyme responsible for activation of respiratory cytochromes c and c1. Heterozygous knockout female mice were thus mosaic for Hccs function due to random X chromosome inactivation. In contrast to midgestational lethality of Hccs knockout males, heterozygous females appeared normal after birth. Analyses of heterozygous embryos revealed the expected 50:50 ratio of Hccs deficient to normal cardiac cells at midgestation; however, diseased tissue contributed progressively less over time and by birth represented only 10% of cardiac tissue volume. This change is accounted for by increased proliferation of remaining healthy cardiac cells resulting in a fully functional heart. These data reveal an impressive regenerative capacity of the fetal heart that can compensate for an effective loss of 50% of cardiac tissue.


In vivo Detection of Alzheimer's Disease.
Recent revisions to the diagnostic criteria for Alzheimer's disease (AD) incorporated conceptual advances in the field. Specifically, AD is now recognized to encompass a continuum, spanning from preclinical (accruing brain pathology in the absence of symptoms) through symptomatic predementia (prodromal AD, mild cognitive impairment) and dementia phases. The role of biological markers (biomarkers) of both the underlying molecular pathologies and related neurodegenerative changes has also been acknowledged. In this abridged review, we provide an overview of fluid (cerebrospinal fluid and blood) and molecular imaging-based biomarkers used within the field and discuss the potential role of computer driven artificial intelligence approaches for both the early and accurate identification of AD and as a tool for population enrichment in clinical trials testing candidate disease modifying therapies.