Study of CD27 and CCR4 Markers on Specific CD4+ T-Cells as Immune Tools for Active and Latent Tuberculosis Management.
The immunological characterization of different cell markers has opened the possibility of considering them as immune tools for tuberculosis (TB) management, as they could correlate with TB latency/disease status and outcome. CD4+ T-cells producing IFN-γ+ with a low expression of CD27 have been described as an active TB marker. In addition, there are unknown homing receptors related to TB, such as CCR4, which might be useful for understanding TB pathogenesis. The aim of our study is focused on the assessment of several T-cell subsets to understand immune-mechanisms in TB. This phenotypic immune characterization is based on the study of the specific immune responses of T-cells expressing CD27 and/or CCR4 homing markers. Subjects enrolled in the study were: (i) 22 adult patients with active TB, and (ii) 26 individuals with latent TB infection (LTBI). Blood samples were drawn from each patient. The expression of CD27 and/or CCR4 markers were analyzed within CD4+ T-cells producing: (i) IFN-γ+, (ii) TNF-α+, (iii) TNF-α+IFN-γ+, and (iv) IFN-γ+ and/or TNF-α+. The percentage of CD27- within all CD4+ T-cell populations analyzed was significantly higher on active TB compared to LTBI after PPD or ESAT-6/CFP-10 stimulation. As previously reported, a ratio based on the CD27 median fluorescence intensity (MFI) was also explored (MFI of CD27 in CD4+ T-cells over MFI of CD27 in IFN-γ+CD4+ T-cells), being significantly increased during disease (p < 0.0001 after PPD or ESAT-6/CFP-10 stimulation). This ratio was also assessed on the other CD4+ T-cells functional profiles after specific stimulation, being significantly associated with active TB. Highest diagnostic accuracies for active TB (AUC ≥ 0.91) were achieved for: (i) CD27 within IFN-γ+TNF-α+CD4+ T-cells in response to ESAT-6/CFP-10, (ii) CD27 and CCR4 markers together within IFN-γ+CD4+ T-cells in response to PPD, and (iii) CD27 MFI ratio performed on IFN-γ+TNF-α+CD4+ T-cells after ESAT-6/CFP-10 stimulation. The lowest diagnostic accuracy was observed when CCR4 marker was evaluated alone (AUC ≤ 0.77). CD27 and CCR4 expression detection could serve as a good method for immunodiagnosis. Moreover, the immunological characterization of markers/subset populations could be a promising tool for understanding the biological basis of the disease.


Prevalence of myofascial temporomandibular disorder in US community women.
This study estimates the prevalence of the myofascial subtype of temporomandibular disorders (M-TMD) defined by Research Diagnostic Criteria (RDC), and relates that prevalence to the surveyed report of facial pain. From among 20 000 women selected at random in the NY metropolitan area who completed a telephone survey of facial pain, 2000 were invited for an RDC/TMD examination; 782 examinations were completed. Prevalence was estimated in analyses that were weighted to correct sampling biases. Differences among demographic strata were evaluated with logistic regression. The prevalence of M-TMD was estimated to be 10.5% (95% CL = 8.5-13.0%). Prevalence was significantly higher among younger women, among women of lower socio-economic status, among Black women, and among non-Hispanic women. The report of facial pain in the telephone survey (10.1%) had high specificity for M-TMD diagnosis (94.7%), but low sensitivity (42.7%). M-TMD is a fairly common disorder among American women. Among those reporting facial pain during the last month, half met RDC palpation criteria for M-TMD; thus, a formal physical examination is imperative to establish this diagnosis. Prevalence varies with age, socio-economic status, race and Hispanic ethnicity. A substantial number of RDC-diagnosed cases of M-TMD did not report facial pain in the survey; the reason for this requires further study.


Mechanical and molecular basis for the symmetrical division of the fission yeast nuclear envelope.
In fission yeast Schizosaccharomyces pombe, the nuclear envelope remains intact throughout mitosis and undergoes a series of symmetrical morphological changes when the spindle pole bodies (SPBs), embedded in the nuclear envelope, are pushed apart by elongating spindle microtubules. These symmetrical membrane shape transformations do not correspond to the shape behavior of an analogous system based on lipid vesicles. Here we report that the symmetry of the dividing fission yeast nucleus is ensured by SPB-chromosome attachments, as loss of kinetochore clustering in the vicinity of SPBs results in the formation of abnormal asymmetric shapes with long membrane tethers. We integrated these findings in a biophysical model, which explains the symmetry of the nuclear shapes on the basis of forces exerted by chromosomes clustered at SPBs on the extending nuclear envelope. Based on this analysis we conclude that the fission yeast nuclear envelope exhibits the same mechanical properties as simple lipid vesicles, but interactions with other cellular components, such as chromosomes, influence the nuclear shape during mitosis, allowing the formation of otherwise energetically unfavorable symmetrical dumbbell structures upon spindle elongation. The model allows us to explain the appearance of abnormal asymmetric shapes in fission yeast mutants with mis-segregated chromosomes as well as with altered nuclear membrane composition.


A registration method for improving quantitative assessment in probabilistic diffusion tractography.
Diffusion MRI-based probabilistic tractography is a powerful tool for non-invasively investigating normal brain architecture and alterations in structural connectivity associated with disease states. Both voxelwise and region-of-interest methods of analysis are capable of integrating population differences in tract amplitude (streamline count or density), given proper alignment of the tracts of interest. However, quantification of tract differences (between groups, or longitudinally within individuals) has been hampered by two related features of white matter. First, it is unknown to what extent healthy individuals differ in the precise location of white matter tracts, and to what extent experimental factors influence perceived tract location. Second, white matter lacks the gross neuroanatomical features (e.g., gyri, histological subtyping) that make parcellation of grey matter plausible - determining where tracts "should" lie within larger white matter structures is difficult. Accurately quantifying tractographic connectivity between individuals is thus inherently linked to the difficulty of identifying and aligning precise tract location. Tractography is often utilized to study neurological diseases in which the precise structural and connectivity abnormalities are unknown, underscoring the importance of accounting for individual differences in tract location when evaluating the strength of structural connectivity. We set out to quantify spatial variance in tracts aligned through a standard, whole-brain registration method, and to assess the impact of location mismatch on groupwise assessments of tract amplitude. We then developed a method for tract alignment that enhances the existing standard whole brain registration, and then tested whether this method improved the reliability of groupwise contrasts. Specifically, we conducted seed-based probabilistic diffusion tractography from primary motor, supplementary motor, and visual cortices, projecting through the corpus callosum. Streamline counts decreased rapidly with movement from the tract center (-35% per millimeter); tract misalignment of a few millimeters caused substantial compromise of amplitude comparisons. Alignment of tracts "peak-to-peak" is essential for accurate amplitude comparisons. However, for all transcallosal tracts registered through the whole-brain method, the mean separation distance between an individual subject's tract and the average tract (3.2 mm) precluded accurate comparison: at this separation, tract amplitudes were reduced by 74% from peak value. In contrast, alignment of subcortical tracts (thalamo-putaminal, pallido-rubral) was substantially better than alignment for cortical tracts; whole-brain registration was sufficient for these subcortical tracts. We demonstrated that location mismatches in cortical tractography were sufficient to produce false positive and false negative amplitude estimates in both groupwise and longitudinal comparisons. We then showed that our new tract alignment method substantially reduced location mismatch and improved both reliability and statistical power of subsequent quantitative comparisons.


Dr. Diego Gonzalez Rivas' trip to the Philippines in November 2015.
Video-assisted thoracoscopic surgery (VATS) was introduced in the Philippines in 1992. Initially, multiport VATS was utilized for lung resections in 2012. In 2015, after attending a couple of Asian uniportal VATS workshops, where one of the lecturers/surgeons was Dr. Diego Gonzalez-Rivas, Filipino surgeons were able to do uniportal VATS for a lobectomy in 2015. In November 2015, Dr. Rivas went to the Philippines to give lectures and conduct a VATS pre-congress workshop during the 25th Clinical Congress of the Association of Thoracic and Cardiovascular Surgeons of Asia (ATCSA). He then went to the Lung Center of the Philippines (LCP) where he delivered a lecture and performed a uniportal VATS lobectomy on a 64-year-old male patient who had Adenocarcinoma of the left lower lobe. Anesthetic and surgical techniques are described. Uniportal VATS is a safe procedure and oncologic results are comparable with resections done using open thoracotomy. The patients' quality of life is better because of significantly less pain after the procedure.


8-Azaguanine reporter of purine ionization states in structured RNAs.
The fluorescent nucleotide analogue 8-azaguanosine-5'-triphosphate (8azaGTP) is prepared easily by in vitro enzymatic synthesis methods. 8azaGTP is an efficient substrate for T7 RNA polymerase and is incorporated specifically opposite cytosine in the transcription template, as expected for a nucleobase analogue with the same Watson-Crick hydrogen bonding face as guanine. 8-Azaguanine (8azaG) in oligonucleotides also is recognized as guanine during ribonuclease T1 digestion. Moreover, replacement of guanine by 8azaG does not alter the melting temperature of base-paired RNAs significantly, evidence that 8azaG does not disrupt stacking and hydrogen bonding interactions. 8azaGTP displays a high fluorescent quantum yield when the N1 position is deprotonated at high pH, but fluorescence intensity decreases significantly when N1 is protonated at neutral pH. Fluorescence is quenched 10-fold to 100-fold when 8azaG is incorporated into base-paired RNA and remains pH-dependent, although apparent pKa values determined from the pH dependence of fluorescence intensity shift in the basic direction. Thus, 8azaG is a guanine analogue that does not perturb RNA structure and displays pH-dependent fluorescence that can be used to probe the ionization states of nucleobases in structured RNAs. A key application will be in determining the ionization state of active site nucleobases that have been implicated in the catalytic mechanisms of RNA enzymes.


Clinical Application of Ceramics in Anterior Cervical Discectomy and Fusion: A Review and Update.
Narrative review. Anterior cervical discectomy and fusion (ACDF) is a reliable procedure, commonly used for cervical degenerative disc disease. For interbody fusions, autograft was the gold standard for decades; however, limited availability and donor site morbidities have led to a constant search for new materials. Clinically, it has been shown that calcium phosphate ceramics, including hydroxyapatite (HA) and tricalcium phosphate (TCP), are effective as osteoconductive materials and bone grafts. In this review, we present the current findings regarding the use of ceramics in ACDF. A review of the relevant literature examining the clinical use of ceramics in anterior cervical discectomy and fusion procedures was conducted using PubMed, OVID and Cochrane. HA, coralline HA, sandwiched HA, TCP, and biphasic calcium phosphate ceramics were used in combination with osteoinductive materials such as bone marrow aspirate and various cages composed of poly-ether-ether-ketone (PEEK), fiber carbon, and titanium. Stand-alone ceramic spacers have been associated with fracture and cracks. Metallic cages such as titanium endure the risk of subsidence and migration. PEEK cages in combination with ceramics were shown to be a suitable substitute for autograft. None of the discussed options has demonstrated clear superiority over others, although direct comparisons are often difficult due to discrepancies in data collection and study methodologies. Future randomized clinical trials are warranted before definitive conclusions can be drawn.


Melanocyte stem cells express receptors for canonical Wnt-signaling pathway on their surface.
It has been reported that melanocytes play important roles in skin and hair pigmentation and are differentiated from melanocyte stem cells (MSCs) residing in the bulge area of hair follicles. Recently, interest has been growing in MSCs because regulation of the upstream of differentiated melanocytes is essential for the determination of skin and hair pigmentation; however, their precise characteristics remain to be elucidated. The aim of this study is to explore cell-surface markers expressed on MSCs in order to understand their characteristics. To explore genes specifically expressed in the bulge region, we classified a hair follicle into four areas, hair bulb, hair bulb to bulge (lower bulge), bulge, and epidermis to bulge (upper bulge), and collected these areas from back skin sections of C57BL/6 mice by laser microdissection. Real-time RT-PCR performed on these areas revealed that Frizzled (Fzd)-4, Fzd7, low density lipoprotein receptor-related protein 5 (Lrp5), and Lrp6, receptors for Wnt molecules, were expressed higher in the bulge area than other areas. Furthermore, FACS analysis showed that populations of Fzd4(+) cells and Fzd7(+) cells were different from those of Kit(+) cells (precursor of melanocytes: melanoblasts). Fzd4(+) and Fzd7(+) cells isolated by FACS required a longer culture period to differentiate into mature melanocytes than Kit(+) cells. Up-regulation of mRNA expressions of melanocyte markers (dopa chrometautomerase: Dct, tyrosinase: Tyr, tyrosinase-related protein 1: Tyrp1) was observed in Fzd4(+) and Fzd7(+) cells following Kit(+) cells during differentiation. These results suggested that Fzd4(+) and Fzd7(+) cells were more immature than melanoblasts, therefore raising the possibility that Fzd4(+) and Fzd7(+) cells are MSCs.


Monitoring coordination during bimanual movements: where is the mastermind?
One remarkable aspect of the human motor repertoire is the multitude of bimanual actions it contains. Still, the neural correlates of coordinated movements, in which the two hands share a common goal, remain debated. To address this issue, we designed two bimanual circling tasks that differed only in terms of goal conceptualization: a "coordination" task that required movements of both hands to adapt to each other to reach a common goal and an "independent" task that imposed a separate goal to each hand. fMRI allowed us to pinpoint three areas located in the right hemisphere that were more strongly activated in the coordination condition: the superior temporal gyrus (STG), the SMA, and the primary motor cortex (M1). We then used transcranial magnetic stimulation (TMS) to disrupt transiently the function of those three regions to determine their causal role in bimanual coordination. Right STG virtual lesions impaired bimanual coordination, whereas TMS to right M1 enhanced hand independence. TMS over SMA, left STG, or left M1 had no effect. The present study provides direct insight into the neural correlates of coordinated bimanual movements and highlights the role of right STG in such bimanual movements.


Comparison of 4D-microSPECT and microCT for murine cardiac function.
The objective of this study was to compare a new generation of four-dimensional micro-single photon emission computed tomography (microSPECT) with microCT for the quantitative in vivo assessment of murine cardiac function. Four-dimensional isotropic cardiac images were acquired from anesthetized normal C57BL/6 mice with either microSPECT (n = 6) or microCT (n = 6). One additional mouse with myocardial infarction (MI) was scanned with both modalities. Prior to imaging, mice were injected with either technetium tetrofosmin for microSPECT or a liposomal blood pool contrast agent for microCT. Segmentation of the left ventricle (LV) was performed using Vitrea (Vital Images) software, to derive global and regional function. Measures of global LV function between microSPECT and microCT groups were comparable (e.g., ejection fraction = 71 ± 6 % microSPECT and 68 ± 4 % microCT). Regional functional indices (wall motion, wall thickening, regional ejection fraction) were also similar for the two modalities. In the mouse with MI, microSPECT identified a large perfusion defect that was not evident with microCT. Despite lower spatial resolution, microSPECT was comparable to microCT in the quantitative evaluation of cardiac function. MicroSPECT offers an advantage over microCT in the ability to evaluate simultaneously myocardial radiotracer distribution and function, simultaneously. MicroSPECT should be considered as an alternative to microCT and magnetic resonance for preclinical cardiac imaging in the mouse.


Therapeutic Targeting of the Warburg Effect in Pancreatic Cancer Relies on an Absence of p53 Function.
The "Warburg effect" describes a peculiar metabolic feature of many solid tumors, namely their increased glucose uptake and high glycolytic rates, which allow cancer cells to accumulate building blocks for the biosynthesis of macromolecules. During aerobic glycolysis, pyruvate is preferentially metabolized to lactate by the enzyme lactate dehydrogenase-A (LDH-A), suggesting a possible vulnerability at this target for small-molecule inhibition in cancer cells. In this study, we used FX11, a small-molecule inhibitor of LDH-A, to investigate this possible vulnerability in a panel of 15 patient-derived mouse xenograft (PDX) models of pancreatic cancer. Unexpectedly, the p53 status of the PDX tumor determined the response to FX11. Tumors harboring wild-type (WT) TP53 were resistant to FX11. In contrast, tumors harboring mutant TP53 exhibited increased apoptosis, reduced proliferation indices, and attenuated tumor growth when exposed to FX11. [18F]-FDG PET-CT scans revealed a relative increase in glucose uptake in mutant TP53 versus WT TP53 tumors, with FX11 administration downregulating metabolic activity only in mutant TP53 tumors. Through a noninvasive quantitative assessment of lactate production, as determined by 13C magnetic resonance spectroscopy (MRS) of hyperpolarized pyruvate, we confirmed that FX11 administration inhibited pyruvate-to-lactate conversion only in mutant TP53 tumors, a feature associated with reduced expression of the TP53 target gene TIGAR, which is known to regulate glycolysis. Taken together, our findings highlight p53 status in pancreatic cancer as a biomarker to predict sensitivity to LDH-A inhibition, with regard to both real-time noninvasive imaging by 13C MRS as well as therapeutic response.


Initial screening of contaminated land: a comparison of US and Swedish methods.
Preliminary surveys are used to prioritize between contaminated sites to select those to be investigated more thoroughly. The data-gathering steps are almost identical between countries; however, the assessment procedures differ significantly. In this study, we have investigated 21 contaminated sites assessed as belonging to the high-risk or the very high-risk class using the Swedish Methods for Inventories of Contaminated Sites (MICS). We then applied the US Preliminary Assessment (PA) method to the same sites and compared the results and conclusions from the two screening procedures. In both cases, all sites were recommended for further investigation and the two approaches seem to corroborate one another; however, the PA assessment scores and the preliminary MICS classifications did not correlate. The results obtained with the PA method were easier to explain than the final MICS classification. The PA method also seems more transparent and easier to standardize, although objections could be made regarding the weighting scheme, because the outcome in this study was entirely dependent on the surface exposure pathway. However, to examine this in greater detail, it is necessary to include sites with less contamination: The importance of preliminary surveys in the overall risk management process gives a strong motivation for such an evaluation. Generally, the lack of research and scientific support for the various assessment procedures in use suggests that there is a need for method development, standardization, and validation.


Effects of human SAMHD1 polymorphisms on HIV-1 susceptibility.
SAMHD1 is a human restriction factor that prevents efficient infection of macrophages, dendritic cells and resting CD4+ T cells by HIV-1. Here we explored the antiviral activity and biochemical properties of human SAMHD1 polymorphisms. Our studies focused on human SAMHD1 polymorphisms that were previously identified as evolving under positive selection for rapid amino acid replacement during primate speciation. The different human SAMHD1 polymorphisms were tested for their ability to block HIV-1, HIV-2 and equine infectious anemia virus (EIAV). All studied SAMHD1 variants block HIV-1, HIV-2 and EIAV infection when compared to wild type. We found that these variants did not lose their ability to oligomerize or to bind RNA. Furthermore, all tested variants were susceptible to degradation by Vpx, and localized to the nuclear compartment. We tested the ability of human SAMHD1 polymorphisms to decrease the dNTP cellular levels. In agreement, none of the different SAMHD1 variants lost their ability to reduce cellular levels of dNTPs. Finally, we found that none of the tested human SAMHD1 polymorphisms affected the ability of the protein to block LINE-1 retrotransposition.


Restoration of Decreased T Helper 1 and CD8+ T Cell Subsets Is Associated With Regression of Lymphoproliferative Disorders Developed During Methotrexate Treatment.
Lymphoproliferative disorder (LPD), including malignant lymphoma, is a relatively rare but life-threatening complication in RA patients under methotrexate (MTX) therapy. Spontaneous regression of LPD after MTX withdrawal is regarded as a distinct characteristic in part of such LPDs. The present study aimed to investigate the immunological difference in regressive LPD and persistent LPD. We studied RA patients who developed LPD during MTX administration (n = 35) and clinically matched controls (n = 35). The time of MTX cessation was defined as week 0, and LPD patients were divided into two groups according to LPD status at week 12: regressive group (n = 22) and persistent group (n = 13). Flow cytometric analysis of whole blood samples and serum cytokine assays were conducted for LPD (n = 10) and control patients (n = 10) at weeks 0, 4, and 12. There was a significant decrease in peripheral lymphocytes and the proportion of T helper 1 cells (Th1 cells), effector memory CD8+ T cells (EMCD8+ T) and Epstein-Barr virus (EBV)-specific CD8+ T cells at the time of LPD diagnosis, and a significant increase after MTX cessation was observed in the regressive group but not in the persistent group. The expansion of Th1 cells and EMCD8+ T cells significantly correlated with an increase in serum interferon (IFN)-γ concentration. Changes in Th1 cells, EMCD8+ T cells and EBV-specific CD8+ T cells, which coincided with an increase in IFN-γ, were significantly different between regressive LPD and persistent LPD after MTX cessation.


Eating rate is a heritable phenotype related to weight in children.
There is growing interest in the heritability of behavioral phenotypes related to adiposity. One potential candidate is the speed of eating, although existing evidence for an association with weight is mixed. We aimed to assess the speed of eating in a sample of 10-12-y-old children to test the hypotheses that higher eating rate is related to greater adiposity and that eating rate is a heritable characteristic. Video data of 254 twin children eating a standard meal at home were used to record eating rate (bites/min) and changes in eating rate across the 4 quarters of the meal. Adiposity was indexed with body mass index SD scores relative to British 1990 norms; for some analyses, children were categorized into groups of overweight or obese and into 2 subgroups of normal-weight (lower normal-weight or higher normal-weight) for comparison of the eating rate within the normal range as well as between clinical and nonclinical groups. All analyses controlled for clustering in twin pairs. Heritability of eating rate was modeled by using standard twin methods. There was a significant linear association across the 3 weight groups for eating rate (P = 0.010), and regression analyses showed that eating rate increased by 0.18 bites/min for each 1-unit increase in body mass index SD score (P = 0.005). The heritability of eating rate was high (0.62; 95% CI: 0.45, 0.74). There was no association between weight group and a change (ie, deceleration) in eating rate over the mealtime. Faster eating appears to be a heritable behavioral phenotype related to higher weight.


Lumbar disc disease shows linkage to chromosome 19 overlapping with a QTL for hand OA.
Cervical and lumbar degenerative disc disease (CDD and LDD, respectively) form part of the spine osteoarthritis (OA) phenotype and are known to be influenced by genetic factors. A genome-wide linkage analysis was performed to identify new chromosomal regions of interest. Dizygotic healthy female twin volunteers (n = 348) from the TwinsUK register who had magnetic resonance imaging scans 10 years ago coded for degenerative disease, were identified. Multipoint genome-wide linkage analysis was conducted using 737 highly polymorphic markers of approximate spacing 10 cM. The mean age of the twins was 52 years. Significant linkage peaks (log of the odds (LOD) >3) were identified for LDD at three chromosomal regions. These included chromosome 1 (position 285 cM), chromosome 5 (position 175 cM) and chromosome 19 (position 80 cM). The peak on chromosome 19 had LOD = 4.06, and the empirical p = 6.7x10(-4) confirmed reliability of the linkage signal. It lies close to a linkage peak previously obtained by our group for hand OA. This genome-wide linkage study of CDD and LDD shows evidence of linkage for LDD on chromosome 19. The region of interest is likely to harbour genes that are common to LDD and hand OA.


Efficacy of the ChAdOx1 nCoV-19 Covid-19 Vaccine against the B.1.351 Variant.
Assessment of the safety and efficacy of vaccines against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in different populations is essential, as is investigation of the efficacy of the vaccines against emerging SARS-CoV-2 variants of concern, including the B.1.351 (501Y.V2) variant first identified in South Africa. We conducted a multicenter, double-blind, randomized, controlled trial to assess the safety and efficacy of the ChAdOx1 nCoV-19 vaccine (AZD1222) in people not infected with the human immunodeficiency virus (HIV) in South Africa. Participants 18 to less than 65 years of age were assigned in a 1:1 ratio to receive two doses of vaccine containing 5×1010 viral particles or placebo (0.9% sodium chloride solution) 21 to 35 days apart. Serum samples obtained from 25 participants after the second dose were tested by pseudovirus and live-virus neutralization assays against the original D614G virus and the B.1.351 variant. The primary end points were safety and efficacy of the vaccine against laboratory-confirmed symptomatic coronavirus 2019 illness (Covid-19) more than 14 days after the second dose. Between June 24 and November 9, 2020, we enrolled 2026 HIV-negative adults (median age, 30 years); 1010 and 1011 participants received at least one dose of placebo or vaccine, respectively. Both the pseudovirus and the live-virus neutralization assays showed greater resistance to the B.1.351 variant in serum samples obtained from vaccine recipients than in samples from placebo recipients. In the primary end-point analysis, mild-to-moderate Covid-19 developed in 23 of 717 placebo recipients (3.2%) and in 19 of 750 vaccine recipients (2.5%), for an efficacy of 21.9% (95% confidence interval [CI], -49.9 to 59.8). Among the 42 participants with Covid-19, 39 cases (95.1% of 41 with sequencing data) were caused by the B.1.351 variant; vaccine efficacy against this variant, analyzed as a secondary end point, was 10.4% (95% CI, -76.8 to 54.8). The incidence of serious adverse events was balanced between the vaccine and placebo groups. A two-dose regimen of the ChAdOx1 nCoV-19 vaccine did not show protection against mild-to-moderate Covid-19 due to the B.1.351 variant. (Funded by the Bill and Melinda Gates Foundation and others; ClinicalTrials.gov number, NCT04444674; Pan African Clinical Trials Registry number, PACTR202006922165132).


Contemporary results of balloon valvuloplasty and surgical valvotomy for congenital aortic stenosis.
The purpose of this study was to compare contemporary results of balloon dilatation and surgery for valvar aortic stenosis in infants and children in the five years between August 1988 and October 1993. Thirty four children underwent attempted balloon valvuloplasty (age 1 day-16 years, weight 1720 g-68 kg) (group 1), eight of whom were neonates with critical aortic stenosis. During the same period, 17 children underwent direct surgical valvotomy (group 2) (seven neonates). Successful balloon valvuloplasty was achieved in 33 (97%) with immediate reduction in the instantaneous systolic pressure gradient from 82 to 34 mm Hg (mean). There were two deaths in this group (both neonates), the second in a preterm neonate from necrotising enterocolitis. Complications requiring intervention in group 1 were aortic regurgitation in one and femoral artery injury in two. Follow up from four months to five years showed sustained results in most cases. There were two neonatal deaths in the surgical group. When the two groups were compared there was no significant difference in mortality, morbidity, or need for reintervention within 12 months. Deaths from both groups were attributed to small left ventricles. Hospital stay was significantly shorter in group 1. It is concluded that balloon dilatation for valvar aortic stenosis is effective and safe for the entire paediatric population. The results compare favourably with those of surgery.


Protein arginine methyltransferase 5 is implicated in the aggressiveness of human hepatocellular carcinoma and controls the invasive activity of cancer cells.
Protein arginine methyltransferase 5 (PRMT5) is a protein that catalyzes transfer of methyl groups to the arginine residues of proteins and is involved in diverse cellular and biological responses. While the participation of PRMT5 in cancer progression has been increasingly documented, its association with the invasive phenotype currently remains poorly understood. In the present study, we revealed that PRMT5 is overexpressed in human hepatocellular carcinoma (HCC) and in colon cancer and its depletion leads to the suppression of cell invasive activity via the reduction of the expression of MMP-2. Real-time quantitative RT-PCR analysis of 120 HCC patient tissues revealed the overexpression of PRMT5 in HCC and the association of PRMT5 with aggressive clinicopathological parameters, such as poorer differentiation (P=0.004), more frequent hepatic vein invasion (P=0.019), larger tumor size (P=0.011) and higher α-fetoprotein levels (P=0.020). Similarly to the data obtained with HCC, overexpression of PRMT5 was also displayed in colon cancer tissues, compared to matched non-tumor regions. Consistent with the significant association of the overexpression of PRMT5 with hepatic vein invasion in patient specimens, PRMT5 depletion via siRNA transfection led to a marked reduction in the invasion rate in both HCC and colon cancer cells. Reduced invasion associated with PRMT5 depletion was accompanied by a decrease in the expression of MMP-2. Collectively, our results indicated that PRMT5 overexpression in HCC and colon cancer cells contributed to their acquisition of aggressive characteristics, such as invasiveness, thus presenting a promising therapeutic target for the treatment of these diseases.


Neuronal uptake and intracellular superoxide scavenging of a fullerene (C60)-poly(2-oxazoline)s nanoformulation.
Fullerene, the third allotrope of carbon, has been referred to as a "radical sponge" because of its powerful radical scavenging activities. However, the hydrophobicity and toxicity associated with fullerene limits its application as a therapeutic antioxidant. In the present study, we sought to overcome these limitations by generating water-soluble nanoformulations of fullerene (C(60)). Fullerene (C(60)) was formulated with poly(N-vinyl pyrrolidine) (PVP) or poly(2-alkyl-2-oxazoline)s (POx) homopolymer and random copolymer to form nano-complexes. These C(60)-polymer complexes were characterized by UV-vis spectroscopy, infrared spectroscopy (IR), dynamic light scattering (DLS), atomic force microscopy (AFM) and transmission electron microscopy (TEM). Cellular uptake and intracellular distribution of the selected formulations in catecholaminergic (CATH.a) neurons were examined by UV-vis spectroscopy, immunofluorescence and immunogold labeling. Electron paramagnetic resonance (EPR) spectroscopy was used to determine the ability of these C(60)-polymer complexes to scavenge superoxide. Their cytotoxicity was evaluated in three different cell lines. C(60)-POx and C(60)-PVP complexes exhibited similar physicochemical properties and antioxidant activities. C(60)-poly(2-ethyl-2-oxazoline) (PEtOx) complex, but not C(60)-PVP complex, were efficiently taken up by CATH.a neurons and attenuated the increase in intra-neuronal superoxide induced by angiotensin II (Ang II) stimulation. These results show that C(60)-POx complexes are non-toxic, neuronal cell permeable, superoxide scavenging antioxidants that might be promising candidates for the treatment of brain-related diseases associated with increased levels of superoxide.


Phase I and pharmacological study of the novel topoisomerase I inhibitor 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11) administered as a ninety-minute infusion every 3 weeks.
7-Ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11; Irinotecan), a semisynthetic analogue of camptothecin (CPT) with broad preclinical antitumor activity, has demonstrated impressive activity in phase II trials in Japan in advanced small and non-small cell lung, colorectal, cervical, and ovarian carcinomas, as well as in refractory lymphomas and leukemias. In this phase I and pharmacological study, 90-min infusions of CPT-11 were administered every 3 weeks at doses ranging from 100 to 345 mg/m2 to patients with solid malignancies. Acute, severe, and refractory vomiting, diarrhea, and/or abdominal cramps associated with flushing, warmth, and diaphoresis occurred in the immediate posttreatment period at the 240-mg/m2 dose level in several patients who were not treated with premedications. The characteristics and temporal nature of these toxicities, the prompt resolution of symptoms following treatment with diphenhydramine, and the successful use of a premedication regimen consisting of ondansetron and diphenhydramine in preventing these acute effects suggest that vasoactive substances are involved in the mediation of these acute toxicities. With the routine use of these premedications, there was no single toxicity type that limited the escalation of CPT-11 doses. Instead, a constellation of severe hematological and gastrointestinal effects precluded the repetitive administration of CPT-11 at doses above 240 mg/m2, the maximum tolerated dose and recommended phase II dose on this schedule. Major responses were observed in patients with advanced colorectal, cervical, and renal cancers. The disposition of total CPT-11 in plasma was fit by a biexponential kinetic model with renal elimination accounting for 37 +/- 4% (SE) of total drug disposition. The Cmax for the active metabolite of CPT-11, 7-ethyl-10-hydroxycamptothecin (SN-38), was achieved at 2.2 +/- 0.1 h after treatment, and mean residence times for both CPT-11 and SN-38 were long, 9.1 and 10.0 h, respectively. Compared with topotecan, another CPT analogue under development, a larger proportion of total drug exposure was accounted for by the active lactone (closed-ring) forms of CPT-11 and SN-38; areas under the time-versus concentration curve for their respective lactone were 44 and 50% of areas under the time-versus-concentration curve for total CPT-11 and SN-38. Although intermittent dosing schedules appear to be superior to single dosing schedules for CPT and some CPT analogues in preclinical tumor models, the maintenance of biologically relevant concentrations of SN-38 for relatively long durations may negate the potential pharmacological benefits of intermittent and continuous administration schedules for CPT-11.(ABSTRACT TRUNCATED AT 400 WORDS)


Screen and identification of proteins interacting with ADAM19 cytoplasmic tail.
ADAM family plays important roles in neurogenesis. The cytoplasmic tail of ADAM19 (ADAM19-CT) contains 193 residues. The presence of two putative SH3 ligand-binding sites suggests potential interactions with cytosolic proteins, which could be possibly linked to the functions of ADAM19. To address these issues, a yeast two-hybrid screen was performed in human fetal brain cDNA library to isolate proteins that interact with the cytoplasmic tail of ADAM19. Four proteins were obtained, ArgBP1, beta-cop, ubiquitin and a novel protein. GST-Pulldown assay has confirmed the interaction between AdAM19 and ArgBP1. By constructing series of deletion mutants of ADAM19-CT and ArgBP1 respectively, the interaction regions have been identified. They are the SH3 binding sites in ADAM19-CT and the P4 region in ArgBP1. And the interaction is specific. ArgBP1 does not bind to ADAM22, ADAM29 or ADAM9 (mouse). ArgBP1 may be the key protein, which accounts for the physiological function of ADAM19.


Modelling the genetic risk in age-related macular degeneration.
Late-stage age-related macular degeneration (AMD) is a common sight-threatening disease of the central retina affecting approximately 1 in 30 Caucasians. Besides age and smoking, genetic variants from several gene loci have reproducibly been associated with this condition and likely explain a large proportion of disease. Here, we developed a genetic risk score (GRS) for AMD based on 13 risk variants from eight gene loci. The model exhibited good discriminative accuracy, area-under-curve (AUC) of the receiver-operating characteristic of 0.820, which was confirmed in a cross-validation approach. Noteworthy, younger AMD patients aged below 75 had a significantly higher mean GRS (1.87, 95% CI: 1.69-2.05) than patients aged 75 and above (1.45, 95% CI: 1.36-1.54). Based on five equally sized GRS intervals, we present a risk classification with a relative AMD risk of 64.0 (95% CI: 14.11-1131.96) for individuals in the highest category (GRS 3.44-5.18, 0.5% of the general population) compared to subjects with the most common genetic background (GRS -0.05-1.70, 40.2% of general population). The highest GRS category identifies AMD patients with a sensitivity of 7.9% and a specificity of 99.9% when compared to the four lower categories. Modeling a general population around 85 years of age, 87.4% of individuals in the highest GRS category would be expected to develop AMD by that age. In contrast, only 2.2% of individuals in the two lowest GRS categories which represent almost 50% of the general population are expected to manifest AMD. Our findings underscore the large proportion of AMD cases explained by genetics particularly for younger AMD patients. The five-category risk classification could be useful for therapeutic stratification or for diagnostic testing purposes once preventive treatment is available.


Counseling about proper use of motor vehicle occupant restraints and avoidance of alcohol use while driving: U.S. Preventive Services Task Force recommendation statement.
An assessment of the independent effectiveness of primary care interventions to increase the proper use of child safety seats, booster seats, and lap-and-shoulder belts to prevent motor vehicle occupant injuries (MVOIs) and to prevent alcohol-related MVOIs in adolescents and adults. The U.S. Preventive Services Task Force reviewed evidence on the effectiveness of counseling in primary care about the proper use of child restraints in motor vehicles to prevent injury, as well as evidence on the impact of primary care counseling to prevent alcohol-related MVOIs in adolescents and adults. This included information gathered in the process of making their 1996 recommendation, as well as the accompanying systematic review of English-language articles published through 2005. Current evidence is insufficient to assess the incremental benefits, beyond the efficacy of legislation and community-based interventions, of counseling in the primary care setting to improve rates of proper use of motor vehicle occupant restraints. (I statement) Current evidence is insufficient to assess the balance of benefits and harms of routine counseling of all patients in the primary care setting to reduce driving while under the influence of alcohol or riding with drivers who are alcohol-impaired. (I statement).


Silica-Gentamicin Nanohybrids: Synthesis and Antimicrobial Action.
Orthopedic applications commonly require the administration of systemic antibiotics. Gentamicin is one of the most commonly used aminoglycosides in the treatment and prophylaxis of infections associated with orthopedic applications, but gentamicin has a short half-life. However, silica nanoparticles (SiO₂ NPs) can be used as elegant carriers for antibiotics to prolong their release. Our goal is the preparation and characterization of SiO₂-gentamicin nanohybrids for their potential antimicrobial administration in orthopedic applications. In vitro gentamicin release profile from the nanohybrids (gentamicin-conjugated SiO₂ NPs) prepared by the base-catalyzed precipitation exhibited fast release (21.4%) during the first 24 h and further extension with 43.9% release during the five-day experiment. Antimicrobial studies of the SiO₂-gentamicin nanohybrids versus native SiO₂ NPs and free gentamicin were performed against Bacillus subtilis (B. subtilis), Pseudomonas fluorescens (P. fluorescens) and Escherichia coli (E. coli). SiO₂-gentamicin nanohybrids were most effective against B. subtilis. SiO₂ NPs play no antimicrobial role. Parallel antimicrobial studies for the filter-sterilized gentamicin were performed to assess the effect of ultraviolet (UV)-irradiation on gentamicin. In summary, the initial fast gentamicin release fits the need for high concentration of antibiotics after orthopedic surgical interventions. Moreover, the extended release justifies the promising antimicrobial administration of the nanohybrids in bone applications.


Crystal structure of the toxin Msmeg_6760, the structural homolog of Mycobacterium tuberculosis Rv2035, a novel type II toxin involved in the hypoxic response.
The structure of Msmeg_6760, a protein of unknown function, has been determined. Biochemical and bioinformatics analyses determined that Msmeg_6760 interacts with a protein encoded in the same operon, Msmeg_6762, and predicted that the operon is a toxin-antitoxin (TA) system. Structural comparison of Msmeg_6760 with proteins of known function suggests that Msmeg_6760 binds a hydrophobic ligand in a buried cavity lined by large hydrophobic residues. Access to this cavity could be controlled by a gate-latch mechanism. The function of the Msmeg_6760 toxin is unknown, but structure-based predictions revealed that Msmeg_6760 and Msmeg_6762 are homologous to Rv2034 and Rv2035, a predicted novel TA system involved in Mycobacterium tuberculosis latency during macrophage infection. The Msmeg_6760 toxin fold has not been previously described for bacterial toxins and its unique structural features suggest that toxin activation is likely to be mediated by a novel mechanism.


Mechanism of the Association between Na+ Binding and Conformations at the Intracellular Gate in Neurotransmitter:Sodium Symporters.
Neurotransmitter:sodium symporters (NSSs) terminate neurotransmission by Na(+)-dependent reuptake of released neurotransmitters. Previous studies suggested that Na(+)-binding reconfigures dynamically coupled structural elements in an allosteric interaction network (AIN) responsible for function-related conformational changes, but the intramolecular pathway of this mechanism has remained uncharted. We describe a new approach for the modeling and analysis of intramolecular dynamics in the bacterial NSS homolog LeuT. From microsecond-scale molecular dynamics simulations and cognate experimental verifications in both LeuT and human dopamine transporter (hDAT), we apply the novel method to identify the composition and the dynamic properties of their conserved AIN. In LeuT, two different perturbations disrupting Na(+) binding and transport (i.e. replacing Na(+) with Li(+) or the Y268A mutation at the intracellular gate) affect the AIN in strikingly similar ways. In contrast, other mutations that affect the intracellular gate (i.e. R5A and D369A) do not significantly impair Na(+) cooperativity and transport. Our analysis shows these perturbations to have much lesser effects on the AIN, underscoring the sensitivity of this novel method to the mechanistic nature of the perturbation. Notably, this set of observations holds as well for hDAT, where the aligned Y335A, R60A, and D436A mutations also produce different impacts on Na(+) dependence. Thus, the detailed AIN generated from our method is shown to connect Na(+) binding with global conformational changes that are critical for the transport mechanism. That the AIN between the Na(+) binding sites and the intracellular gate in bacterial LeuT resembles that in eukaryotic hDAT highlights the conservation of allosteric pathways underlying NSS function.


The Automated Root Exudate System (ARES): a method to apply solutes at regular intervals to soils in the field.
Root exudation is a key component of nutrient and carbon dynamics in terrestrial ecosystems. Exudation rates vary widely by plant species and environmental conditions, but our understanding of how root exudates affect soil functioning is incomplete, in part because there are few viable methods to manipulate root exudates in situ. To address this, we devised the Automated Root Exudate System (ARES), which simulates increased root exudation by applying small amounts of labile solutes at regular intervals in the field.The ARES is a gravity-fed drip irrigation system comprising a reservoir bottle connected via a timer to a micro-hose irrigation grid covering c. 1 m2; 24 drip-tips are inserted into the soil to 4-cm depth to apply solutions into the rooting zone. We installed two ARES subplots within existing litter removal and control plots in a temperate deciduous woodland. We applied either an artificial root exudate solution (RE) or a procedural control solution (CP) to each subplot for 1 min day-1 during two growing seasons. To investigate the influence of root exudation on soil carbon dynamics, we measured soil respiration monthly and soil microbial biomass at the end of each growing season.The ARES applied the solutions at a rate of c. 2 L m-2 week-1 without significantly increasing soil water content. The application of RE solution had a clear effect on soil carbon dynamics, but the response varied by litter treatment. Across two growing seasons, soil respiration was 25% higher in RE compared to CP subplots in the litter removal treatment, but not in the control plots. By contrast, we observed a significant increase in microbial biomass carbon (33%) and nitrogen (26%) in RE subplots in the control litter treatment.The ARES is an effective, low-cost method to apply experimental solutions directly into the rooting zone in the field. The installation of the systems entails minimal disturbance to the soil and little maintenance is required. Although we used ARES to apply root exudate solution, the method can be used to apply many other treatments involving solute inputs at regular intervals in a wide range of ecosystems.


RDL receptors.
RDL receptors are invertebrate members of the Cys-loop family of ligand-gated ion channels. They are GABA (gamma-aminobutyric acid)-activated chloride-selective receptors that are closely related to their vertebrate orthologues, the GABA(A) receptors, as well as other Cys-loop receptors such as the ionotropic glycine, nicotinic acetylcholine and 5-HT(3) receptors. RDL receptors are widely expressed throughout the insect CNS (central nervous system) and are important in inhibitory neurotransmission. They are therefore a major insecticidal target site.


A noble metal-free proton-exchange membrane fuel cell based on bio-inspired molecular catalysts.
Hydrogen is a promising energy vector for storing renewable energies: obtained from water-splitting, in electrolysers or photoelectrochemical cells, it can be turned back to electricity on demand in fuel cells (FCs). Proton exchange membrane (PEM) devices with low internal resistance, high compactness and stability are an attractive technology optimized over decades, affording fast start-up times and low operating temperatures. However, they rely on the powerful catalytic properties of noble metals such as platinum, while lower cost, more abundant materials would be needed for economic viability. Replacing these noble metals at both electrodes has long proven to be a difficult task, so far incompatible with PEM technologies. Here we take advantage of newly developed bio-inspired molecular H2 oxidation catalysts and noble metal-free O2-reducing materials, to fabricate a noble metal-free PEMFC, with an 0.74 V open circuit voltage and a 23 μW cm-2 output power under technologically relevant conditions. X-ray absorption spectroscopy measurements confirm that the catalysts are stable and retain their structure during turnover.


Man-machine communications through brain-wave processing.
The possibility of monitoring voluntarily produced changes in the electroencephalogram (EEG) of a subject and translating these changes into a set of commands to be issued to an external device was investigated. Subjects performed five distinct tasks under both eyes-open and eyes-closed conditions. A feature set consisting of the asymmetry ratios and the power values for each lead at four frequency bands-delta (0-3 Hz) theta (4-7 Hz), alpha (8-13 Hz), and beta (14-20 Hz)-was used to characterize the EEG. The feature sets created from an estimate of the spectral density of the EEG for each task were used to test classification accuracy among the various tasks using a Bayes quadratic classifier. The results show that it is possible to distinguish, to a high degree of accuracy, among the various mental tasks studied, using only the EEG.


Depression and smoking cessation: does the evidence support psychiatric practice?
Depression and smoking are highly comorbid. The vast majority of psychiatrists treating depressed patients do not target or treat nicotine dependence, and many inpatient psychiatric facilities implicitly condone smoking by providing 'smoke breaks'. The reasons for failure to treat are unclear, but are probably linked to the notion that depressed smokers are neither willing nor able to quit, and will become more depressed if they try. We review the clinical evidence on depression and smoking cessation, and find little support for current psychiatric practice. Although quitting smoking does appear to pose a risk for the development of depression, this risk is not clearly higher in those with a past history of depression than those without. Depressed smokers are as capable as nondepressed smokers of quitting smoking, and at least one-quarter of depressed smokers is willing to try. Sustained abstinence may even lead to improvement in depressive disorders. More research is needed to understand the relationship between depression and quitting smoking, but current clinical evidence suggests more resiliency among depressed smokers than common clinical wisdom would dictate.


Ground-state destabilization in orotate phosphoribosyltransferases by binding isotope effects.
Orotate phosphoribosyltransferases (OPRTs) form and break the N-ribosidic bond to pyrimidines by way of ribocation-like transition states (TSs) and therefore exhibit large α-secondary 1'-(3)H k(cat)/K(m) kinetic isotope effects (KIEs) [Zhang, Y., and Schramm, V. L. (2010) J. Am. Chem. Soc. 132, 8787-8794]. Substrate binding isotope effects (BIEs) with OPRTs report on the degree of ground-state destabilization for these complexes and permit resolution of binding and transition-state effects from the k(cat)/K(m) KIEs. The BIEs for interactions of [1'-(3)H]orotidine 5'-monophosphate (OMP) with the catalytic sites of Plasmodium falciparum and human OPRTs are 1.104 and 1.108, respectively. These large BIEs establish altered sp(3) bond hybridization of C1' toward the sp(2) geometry of the transition states upon OMP binding. Thus, the complexes of these OPRTs distort OMP part of the way toward the transition state. As the [1'-(3)H]OMP k(cat)/K(m) KIEs are approximately 1.20, half of the intrinsic k(cat)/K(m) KIEs originate from BIEs. Orotidine, a slow substrate for these enzymes, binds to the catalytic site with no significant [1'-(3)H]orotidine BIEs. Thus, OPRTs are unable to initiate ground-state destabilization of orotidine by altered C1' hybridization because of the missing 5'-phosphate. However the k(cat)/K(m) KIEs for [1'-(3)H]orotidine are also approximately 1.20. The C1' distortion for OMP happens in two steps, half upon binding and half on going from the Michaelis complex to the TS. With orotidine as the substrate, there is no ground-state destabilization in the Michaelis complexes, but the C1' distortion at the TS is equal to that of OMP. The large single barrier for TS formation with orotidine slows the rate of barrier crossing.


QUANTITATIVE STUDIES OF SULFONAMIDE RESISTANCE.
1. In vitro experiments were performed with E. coli, using a method designed for the quantitative study of various aspects of sulfonamide resistance. 2. Resistance was found to be a gradually developing process, and was demonstrated for all four drugs tested, sulfanilamide, sulfapyridine, sulfathiazole, and sulfadiazine. 3. It was shown that the degree of resistance developed was correlated with the bacteriostatic potency of the sulfonamides, and that organisms resistant to certain bacteriostatic concentrations of one sulfonamide were equally resistant to similar bacteriostatic concentrations of the other sulfonamides. 4. These observations were interpreted as indicating that the development of sulfonamide resistance represents an interaction between the organisms and the one common structural unit of all the sulfonamides, namely, the p-amino nucleus. It is also suggested that this interaction may involve the same enzyme system (or systems) as those concerned in the antagonism of the sulfonamides by para-aminobenzoic acid. 5. The relation of these findings to the broader aspects of sulfonamide resistance is discussed, and it is postulated that, despite reports to the contrary, all organisms susceptible to the bacteriostatic action of the sulfonamides are capable of becoming resistant to all of the sulfonamides.


Evidence for bidirectional and trans-synaptic parasympathetic and sympathetic propagation of alpha-synuclein in rats.
The conversion of endogenous alpha-synuclein (asyn) to pathological asyn-enriched aggregates is a hallmark of Parkinson's disease (PD). These inclusions can be detected in the central and enteric nervous system (ENS). Moreover, gastrointestinal symptoms can appear up to 20 years before the diagnosis of PD. The dual-hit hypothesis posits that pathological asyn aggregation starts in the ENS, and retrogradely spreads to the brain. In this study, we tested this hypothesis by directly injecting preformed asyn fibrils into the duodenum wall of wild-type rats and transgenic rats with excess levels of human asyn. We provide a meticulous characterization of the bacterial artificial chromosome (BAC) transgenic rat model with respect to initial propagation of pathological asyn along the parasympathetic and sympathetic pathways to the brainstem, by performing immunohistochemistry at early time points post-injection. Induced pathology was observed in all key structures along the sympathetic and parasympathetic pathways (ENS, autonomic ganglia, intermediolateral nucleus of the spinal cord (IML), heart, dorsal motor nucleus of the vagus, and locus coeruleus (LC)) and persisted for at least 4 months post-injection. In contrast, asyn propagation was not detected in wild-type rats, nor in vehicle-injected BAC rats. The presence of pathology in the IML, LC, and heart indicate trans-synaptic spread of the pathology. Additionally, the observed asyn inclusions in the stomach and heart may indicate secondary anterograde propagation after initial retrograde spreading. In summary, trans-synaptic propagation of asyn in the BAC rat model is fully compatible with the "body-first hypothesis" of PD etiopathogenesis. To our knowledge, this is the first animal model evidence of asyn propagation to the heart, and the first indication of bidirectional asyn propagation via the vagus nerve, i.e., duodenum-to-brainstem-to-stomach. The BAC rat model could be very valuable for detailed mechanistic studies of the dual-hit hypothesis, and for studies of disease modifying therapies targeting early pathology in the gastrointestinal tract.


The spatial and temporal construction of confidence in the visual scene.
Human subjects can report many items of a cluttered field a few hundred milliseconds after stimulus presentation. This memory decays rapidly and after a second only 3 or 4 items can be stored in working memory. Here we compared the dynamics of objective performance with a measure of subjective report and we observed that 1) Objective performance beyond explicit subjective reports (blindsight) was significantly more pronounced within a short temporal interval and within specific locations of the visual field which were robust across sessions 2) High confidence errors (false beliefs) were largely confined to a small spatial window neighboring the cue. The size of this window did not change in time 3) Subjective confidence showed a moderate but consistent decrease with time, independent of all other experimental factors. Our study allowed us to asses quantitatively the temporal and spatial access to an objective response and to subjective reports.


Gene network analysis leads to functional validation of pathways linked to cancer cell growth and survival.
Hepatocellular carcinoma (HCC) represents one of the most frequently diagnosed human cancers; however, there are currently few treatment alternatives to surgical resection. In this study we performed bioinformatic analysis of previously published transcriptomic data in order to characterize liver specific networks, including biological functions, signaling pathways and transcription factors, potentially dysregulated in HCC. By incorporating specific signaling inhibitors into real-time proliferation assays using HepG2 cells, we then validated these in silico results. We found that G protein subunits Gi/G0, protein kinase C, Mek1/2, and Erk1/2 (P42/44), JAK1, PPARA and NFκB p65 subunit were the major signaling molecules required for survival and proliferation of human HCC cell lines. We also found that these pathways regulate the expression of key hepatic transcription factors involved in cell differentiation, such as CEBPA, EGR1, FOXM1 and PPARs. By combining bioinformatic and functional analyses, major signaling pathways related to tumorigenicity in HCC are revealed, thereby elucidating potential targets for drug therapies.


Time-free spiking neural P systems.
Different biological processes take different times to be completed, which can also be influenced by many environmental factors. In this work, a realistic definition of nonsynchronized spiking neural P systems (SN P systems, for short) is considered: during the work of an SN P system, the execution times of spiking rules cannot be known exactly (i.e., they are arbitrary). In order to establish robust systems against the environmental factors, a special class of SN P systems, called time-free SN P systems, is introduced, which always produce the same computation result independent of the execution times of the rules. The universality of time-free SN P systems is investigated. It is proved that these P systems with extended rules (several spikes can be produced by a rule) are equivalent to register machines. However, if the number of spikes present in the system is bounded, then the power of time-free SN P systems falls, and in this case, a characterization of semilinear sets of natural numbers is obtained.


Trace amounts of sporadically reappearing HCV RNA can cause infection.
Successful hepatitis C virus (HCV) treatment is defined as the absence of viremia 6 months after therapy cessation. We previously reported that trace amounts of HCV RNA, below the sensitivity of the standard clinical assay, can reappear sporadically in treatment responders. Here, we assessed the infectivity of this RNA and infused 3 chimpanzees sequentially at 9-week intervals with plasma or PBMCs from patients who tested positive for trace amounts of HCV RNA more than 6 months after completing pegylated IFN-α/ribavirin therapy. A fourth chimpanzee received HCV RNA-negative plasma and PBMCs from healthy blood donors. The 3 experimental chimpanzees, but not the control chimpanzee, generated HCV-specific T cell responses against nonstructural and structural HCV sequences 6-10 weeks after the first infusion of patient plasma and during subsequent infusions. In 1 chimpanzee, T cell responses declined, and this animal developed high-level viremia at week 27. Deep sequencing of HCV demonstrated transmission of a minor HCV variant from the first infusion donor that persisted in the chimpanzee for more than 6 months despite undetectable systemic viremia. Collectively, these results demonstrate that trace amounts of HCV RNA, which appear sporadically in successfully treated patients, can be infectious; furthermore, transmission can be masked in the recipient by an extended eclipse phase prior to establishing high-level viremia.


Traumatic subarachnoid-pleural fistula in a dog.
Subarachnoid-pleural fistula is a rare occurrence in humans as a result of trauma or spinal surgery. Such fistulas commonly remain undiagnosed until sufficient cerebrospinal fluid accumulates in the pleural space to cause respiratory distress. We describe a subarachnoid-pleural fistula in a dog that occurred subsequent to blunt trauma sustained during a fall, with concurrent acute, traumatic intervertebral disc rupture. The extruded disc material penetrated the dura mater, allowing communication between the subarachnoid space and the extrapleural thoracic cavity. Radiographic, myelographic, and computed tomographic (CT) findings are reviewed. Abnormalities noted during myelography included an intradural-extramedullary lesion at T11-T12, with epidural leakage of contrast medium from the region of T12 extending cranially. In images from myelography and CT there was extravasation of contrast medium extending from the subarachnoid and epidural space into the extrapleural thoracic cavity.


Phosphoproteomic analysis of differentiating Leishmania parasites reveals a unique stage-specific phosphorylation motif.
Protists of the genus Leishmania are obligatory intracellular parasites that cause a wide range of cutaneous, mucocutaneous, and visceral diseases in humans. They cycle between phagolysosomes of mammalian macrophages and the sand fly midgut, proliferating as intracellular amastigotes and extracellular promastigotes, respectively. Exposure to a lysosomal environment, i.e. acidic pH and body temperature, signals promastigotes to differentiate into amastigotes. Time course analyses indicated that Leishmania differentiation is a highly regulated and coordinated process. However, the role of posttranslational events such as protein phosphorylation in this process is still unknown. Herein, we analyzed and compared the phosphoproteomes of L. donovani amastigotes and promastigotes using an axenic host-free system that simulates parasite differentiation. Shotgun phosphopeptide analysis revealed 1614 phosphorylation residues (p-sites) corresponding to 627 proteins. The analysis indicated that the majority of the p-sites are stage-specific. Serine phosphorylation in a previously identified trypanosomatid-specific "SF" motif was significantly enriched in amastigotes. We identified a few phosophotyrosines (pY), mostly in proteins known to participate in signal transduction pathways. The analysis indicated that Leishmania contains proteins with multiple p-sites that are phosphorylated at distinct stages of the life cycle. For over half of the phosphorylation events, changes in phosphoprotein abundance did not positively correlate with changes in protein abundance, suggesting functional regulation. This study compares, for the first time, the phosphoproteins of L. donovani axenic promastigotes and amastigotes and provides the largest data set of the Leishmania phosphoproteome to date.


MR imaging of intradural inflammatory diseases of the spine.
Twenty-eight patients with intradural inflammatory disease of the spine were studied in order to characterize the MR imaging findings of infectious and inflammatory conditions. Patients were categorized according to the spinal compartment involved. Among the 12 patients in the intradural extramedullary group, unenhanced scans were either normal or nonspecific while contrast-enhanced scans were helpful in visualizing and localizing the lesion. Nevertheless, contrast-enhanced MR studies were unable to differentiate infection and inflammation from tumor in this compartment. Among 16 patients with intramedullary lesions, four had granulomatous disease and 12 had nongranulomatous disease. The granulomatous lesions resembled tumors and displayed MR characteristics of a focal lesion with large nodular enhancement. The patients with nongranulomatous intramedullary lesions exhibited two subsets of MR findings. In the first subset of nine patients, diffuse cord swelling and high signal were seen on long TR images, combined with either no enhancement or peripheral, diffuse, or speckled enhancement of the spinal cord on contrast-enhanced short TR images. In the second subset of three patients, minimal or no spinal cord swelling was displayed despite the visualization of high signal on long TR scans and nodular enhancement with contrast administration on short TR scans. Both subsets were sufficiently unique that nongranulomatous myelitis could usually be differentiated from spinal cord tumors.


The role of endogenous molecules in modulating pain through transient receptor potential vanilloid 1 (TRPV1).
Pain is a physiological response to a noxious stimulus that decreases the quality of life of those sufferring from it. Research aimed at finding new therapeutic targets for the treatment of several maladies, including pain, has led to the discovery of numerous molecular regulators of ion channels in primary afferent nociceptive neurons. Among these receptors is TRPV1 (transient receptor potential vanilloid 1), a member of the TRP family of ion channels. TRPV1 is a calcium-permeable channel, which is activated or modulated by diverse exogenous noxious stimuli such as high temperatures, changes in pH, and irritant and pungent compounds, and by selected molecules released during tissue damage and inflammatory processes. During the last decade the number of endogenous regulators of TRPV1's activity has increased to include lipids that can negatively regulate TRPV1, as is the case for cholesterol and PIP2 (phosphatidylinositol 4,5-biphosphate) while, in contrast, other lipids produced in response to tissue injury and ischaemic processes are known to positively regulate TRPV1. Among the latter, lysophosphatidic acid activates TRPV1 while amines such as N-acyl-ethanolamines and N-acyl-dopamines can sensitize or directly activate TRPV1. It has also been found that nucleotides such as ATP act as mediators of chemically induced nociception and pain and gases, such as hydrogen sulphide and nitric oxide, lead to TRPV1 activation. Finally, the products of lipoxygenases and omega-3 fatty acids among other molecules, such as divalent cations, have also been shown to endogenously regulate TRPV1 activity. Here we provide a comprehensive review of endogenous small molecules that regulate the function of TRPV1. Acting through mechanisms that lead to sensitization and desensitization of TRPV1, these molecules regulate pathways involved in pain and nociception. Understanding how these compounds modify TRPV1 activity will allow us to comprehend how some pathologies are associated with its deregulation.


Failure and success of hydrodynamic interaction models.
In suspensions with charged particles, electrostatic forces and hydrodynamic interactions are both important to describe the system. We study different models of hydrodynamic interaction for monopolarly charged particles in a non-polar liquid. In this case, there is no screening of the Coulomb repulsion, so the repulsion between all pairs must be taken into account. The particles are expected to drift away from each other, however at a lower rate when hydrodynamic interaction between the particles is taken into account. Existing, frequently used models of hydrodynamic interactions tend to overestimate the slowing down of the charged particles, even to the extent that the particles effectively attract each other. This is demonstrated for some selected particle setups. We find that these anomalies even occur in dilute systems, if they contain sufficiently many particles. We explain why these anomalies can be avoided by an approach, in which the superposition of interactions is done in the friction tensor instead of the mobility tensor.


Effectiveness of combined nifedipine and propranolol treatment in hypertension.
The long-term antihypertensive effect of combined nifedipine and propranolol therapy was assessed in an open trial in 26 hypertensive patients (19 men, seven women, mean age 53 years). On propranolol alone (160 to 240 mg/day), the patients' average sitting blood pressure was 192 +/- 5/114 +/- 2 mm Hg. Propranolol was continued in a fixed dose and nifedipine was added in a dose that was gradually increased from 30 to 90 mg/day to achieve blood pressure (BP) values below 160/95 mm Hg. Twenty-two patients remained on the combined regimen for 14 to 30 weeks. Their BP decreased to 136 +/- 3/84 +/- 2 mm Hg on an average daily dose of 59.5 mg nifedipine. Seventeen of the 22 subjects were subsequently treated sequentially with propranolol alone, combined therapy, and nifedipine alone, to assess the relative efficacy of each mode of therapy. The combined regimen was found to be more effective than either drug alone. Side effects occurred in 13 of 26 patients. Four dropped out 4 to 11 weeks after starting nifedipine because of either intolerable flushing (2), allergic rash (1), or headache (1). Nine subjects experienced mild reactions that were well tolerated. It is concluded that the combined use of propranolol and nifedipine is effective in the long-term treatment of moderately severe hypertension and offers an alternative therapeutic approach that deserves further evaluation.


Does problem-based learning work? A meta-analysis of evaluative research.
The purpose of this review is to synthesize all available evaluative research from 1970 through 1992 that compares problem-based learning (PBL) with more traditional methods of medical education. Five separate meta-analyses were performed on 35 studies representing 19 institutions. For 22 of the studies (representing 14 institutions), both effect-size and supplementary vote-count analyses could be performed; otherwise, only supplementary analyses were performed. PBL was found to be significantly superior with respect to students' program evaluations (i.e., students' attitudes and opinions about their programs)--dw (standardized differences between means, weighted by sample size) = +.55, CI.95 = +.40 to +.70 - and measures of students' clinical performance (dw = +.28, CI.95 = +.16 to +.40). PBL and traditional methods did not differ on miscellaneous tests of factual knowledge (dw = -.09, CI.95 = +.06 to -.24) and tests of clinical knowledge (dw = +.08, CI.95 = -.05 to +.21). Traditional students performed significantly better than their PBL counterparts on the National Board of Medical Examiners Part I examination--NBME I (dw = -.18, CI.95 = -.10 to -.26). However, the NBME I data displayed significant overall heterogeneity (Qt = 192.23, p < .001) and significant differences among programs (Qb = 59.09, p < .001), which casts doubt on the generality of the findings across programs. The comparative value of PBL is also supported by data on outcomes that have been studied less frequently, i.e., faculty attitudes, student mood, class attendance, academic process variables, and measures of humanism. In conclusion, the results generally support the superiority of the PBL approach over more traditional methods. Acad. Med. 68 (1993):550-563.


Determination of eight carbonyl compounds in aerosols trapped in phosphate buffer saline solutions to support in vitro assessment studies.
When investigating the toxicological impact of aerosols using in vitro systems like cell cultures, it is essential to have a quantitative measurement of the chemicals that the cells are exposed to. Carbonyl compounds represent an important class of marker compounds for in vitro and in vivo exposure to different toxicological agents, including cigarette smoke (CS). A new LC-MS/MS method that quantifies eight of these analytes in aerosols trapped in phosphate-buffered saline solutions has been developed to measure exposure. During the method development phase, particular attention has been paid to the efficient derivatization of the target compounds in the trapped aerosols and to avoid the formation of poly-derivatized molecules, which could lead to inaccurate quantifications. The method has been successively validated using the accuracy profile procedure. Selectivity, detection limits, precision, and accuracy have been evaluated for Vitrocell®, Gas Vapor Phase (GVP), and Whole Smoke (WS) matrices of smoke generated by 3R4F cigarettes and aerosol generated by the Tobacco Heating System (THS) 2.2, a heat-not-burn tobacco product developed by Philip Morris International (Smith et al., 2016) [1]. Validation results confirmed that the established working ranges also allow the analysis of THS aerosols, where the concentrations of carbonyl compounds are substantially lower than those generated by 3R4F cigarettes. Moreover, data gathered on 3R4F aerosol samples trapped with DNPH in acetonitrile solutions have been compared to the quantification given by an in-house UHPLC-MS/MS and reference values from the literature.


Inducible nitric oxide synthase expression in cerebrovascular smooth muscle and neutrophils after traumatic brain injury in immature rats.
The inflammatory response after traumatic brain injury (TBI) includes cytokine production, leukocyte infiltration, and microglial activation. Production of nitric oxide by inducible nitric oxide synthase (iNOS) occurs during acute inflammation outside of the CNS and in models of cerebral ischemia, and therefore may contribute to the inflammatory response after TBI. The purpose of this study was to localize and define the time course of iNOS expression after TBI in the immature rat. Immature Wistar rats (age 3.5-4.5 wk) were anesthetized and subjected to percussive trauma to the right parietal cortex. Nontraumatized rats were used as controls (n = 7). At 2, 24, 48, or 168 h (n = 3/group) posttrauma rats were killed by perfusion fixation. Brains were removed, frozen, sectioned, immunostained with antibodies against iNOS and glial fibrillary acidic protein (GFAP, a marker specific for astrocytes), and imaged using fluorescent detection systems. There was no detectable expression of iNOS in control brains. At 2h, minimal cerebrovascular iNOS expression was seen in the peritrauma area. At 24 and 48 h, there was marked peritrauma cerebrovascular iNOS expression that appeared to be restricted to vascular smooth muscle cells and infiltrated leukocytes. Further dual-immunolabeling showed that the leukocytes expressing iNOS were predominantly neutrophils. At 168 h, iNOS expression was no longer detectable. iNOS was not detectable in GFAP-positive cells. The prominent expression of iNOS protein after TBI in cerebrovascular smooth muscle cells and infiltrated neutrophils suggests that iNOS may play a role in cerebrovascular disturbances and secondary brain injury after trauma.


Pyrantel resistance in two herds of donkey in the UK.
Resistance to currently available anthelmintics is a serious phenomenon which is prevalent globally. Cyathostomins are one of the major parasites, and are of primary concern in donkeys. There have been reports of emerging resistance to pyrantel, but the status of pyrantel resistance in donkey populations in the UK is largely unknown. This report investigates pyrantel resistance in two geographically isolated donkey herds in the South West of England. The first herd had suspected pyrantel resistance, with already established resistance to other anthelmintics. In the second herd the efficacy of pyrantel was not suspected at the time the study took place. Faecal Egg Count Reduction Test (FECRT) was carried out, revealing large scale resistance. Eighty one percent of the first herd and 73% of the second herd had a FEC of less than 95% after treatment, and anthelmintic resistance was confirmed using the World Association for the Advancement of Veterinary Parasitology guidelines. These findings indicate that anthelmintic resistance to pyrantel exists in both tested donkey populations and illustrate the continuing development of resistance through different classes of chemotherapeutics.


The creep and wear of highly cross-linked polyethylene: a three-year randomised, controlled trial using radiostereometric analysis.
The creep and wear behaviour of highly cross-linked polyethylene and standard polyethylene liners were examined in a prospective, double-blind randomised, controlled trial using radiostereometric analysis. We randomised 54 patients to receive hip replacements with either highly cross-linked polyethylene or standard liners and determined the three-dimensional penetration of the liners over three years. After three years the mean total penetration was 0.35 mm (SD 0.14) for the highly cross-linked polyethylene group and 0.45 mm (SD 0.19) for the standard group. The difference was statistically significant (p = 0.0184). From the pattern of penetration it was possible to discriminate creep from wear. Most (95%) of the creep occurred within six months of implantation and nearly all within the first year. There was no difference in the mean degree of creep between the two types of polyethylene (highly cross-linked polyethylene 0.26 mm, SD 0.17; standard 0.27 mm, SD 0.2; p = 0.83). There was, however, a significant difference (p = 0.012) in the mean wear rate (highly cross-linked polyethylene 0.03 mm/yr, SD 0.06; standard 0.07 mm/yr, SD 0.05). Creep and wear occurred in significantly different directions (p = 0.01); creep was predominantly proximal whereas wear was anterior, proximal and medial. We conclude that penetration in the first six months is creep-dominated, but after one year virtually all penetration is due to wear. Highly cross-linked polyethylene has a 60% lower rate of wear than standard polyethylene and therefore will probably perform better in the long term.


The effect of dietary sodium modification on blood pressure in adults with systolic blood pressure less than 140 mmHg: a systematic review.
Modifying dietary sodium intake is a cornerstone of diet advice for lowering blood pressure (BP) under the assumption that it is protective against cardiovascular disease. Previous meta-analyses of normotensive participants have not excluded all studies that recruited participants with systolic blood pressure (SBP) > 140 mmHg, which greatly hinders generalization to the wider normotensive population. The objective of this review was to identify the effectiveness of reducing or increasing sodium intake on BP in normotensive participants with SBP ≤ 140 mmHg. This review considered studies on adult participants (≥18 years) with SBP ≤ 140 mmHg. Studies on pregnant women or patients prescribed antihypertensive or vasoactive medications were excluded. Interventions that quantitatively evaluated dietary sodium intake for equal to or greater than four weeks duration were considered. Only studies that included two study arms comprising different levels of sodium intake were included. Studies that reported SBP, diastolic blood pressure (DBP), pulse wave velocity (PWV), pulse wave analysis or flow mediated dilatation were considered. Experimental study designs including randomized controlled trials and non-randomized controlled trials were considered. An initial search strategy was conducted on databases MEDLINE and CINAHL before an extensive search of all relevant published and gray literature databases, and clinical trial registries were searched. Potential papers were assessed for methodological validity using the standardized critical appraisal instrument from the Joanna Briggs Institute Meta-Analysis of Statistics Assessment and Review Instrument (JBI-MAStARI). Quantitative data were extracted from papers using the standardized data extraction tool from JBI-MAStARI. Quantitative data were pooled in statistical meta-analysis. Effect sizes were expressed as weighted mean differences and 95% confidence intervals. Meta-analysis was conducted using a random-effect model, and heterogeneity assessed statistically using the standard Chi-square test and the I index. A priori sub-group analysis was undertaken on studies achieving ≥40 mmol versus <40 mmol in urinary sodium excretion and post hoc on studies with a mean body mass index (BMI) ≥ 30 versus less than 30. Five trials were included with a total of 1214 participants. The overall reduction in SBP was -0.71 mmHg (95% CI: -2.62, 1.20, P = 0.47) and DBP -0.57 mmHg (95% CI: -1.26, 0.12, P = 0.10). There was no significant change in PWV following reduction of dietary sodium over a four to six-week period. Sub-group analysis did not find a significant effect of urinary sodium excretion or BMI on outcomes; however, a trend toward a greater reduction in BP was observed in those with a higher BMI (MD -2.41, 95% CI -5.72, +0.91, P = 0.16). Blood pressure in normotensive participants was not significantly affected by sodium modification and was controlled to within 1% of baseline values. Reducing dietary sodium in normotensive participants may still be of importance for cardiovascular risk management; however, good quality interventional research is limited.


Mathematical modeling of the hypothalamic-pituitary-adrenal gland (HPA) axis, including hippocampal mechanisms.
This paper presents a mathematical model of the HPA axis. The HPA axis consists of the hypothalamus, the pituitary and the adrenal glands in which the three hormones CRH, ACTH and cortisol interact through receptor dynamics. Furthermore, it has been suggested that receptors in the hippocampus have an influence on the axis. A model is presented with three coupled, non-linear differential equations, with the hormones CRH, ACTH and cortisol as variables. The model includes the known features of the HPA axis, and includes the effects from the hippocampus through its impact on CRH in the hypothalamus. The model is investigated both analytically and numerically for oscillating solutions, related to the ultradian rhythm seen in data, and for multiple fixed points related to hypercortisolemic and hypocortisolemic depression. The existence of an attracting trapping region guarantees that solution curves stay non-negative and bounded, which can be interpreted as a mathematical formulation of homeostasis. No oscillating solutions are present when using physiologically reasonable parameter values. This indicates that the ultradian rhythm originate from different mechanisms. Using physiologically reasonable parameters, the system has a unique fixed point, and the system is globally stable. Therefore, solutions converge to the fixed point for all initial conditions. This is in agreement with cortisol levels returning to normal, after periods of mild stress, in healthy individuals. Perturbing parameters lead to a bifurcation, where two additional fixed points emerge. Thus, the system changes from having a unique stable fixed point into having three fixed points. Of the three fixed points, two are stable and one is unstable. Further investigations show that solutions converge to one of the two stable fixed points depending on the initial conditions. This could explain why healthy people becoming depressed usually fall into one of two groups: a hypercortisolemic depressive group or a hypocortisolemic depressive group.


The preterm birth syndrome: issues to consider in creating a classification system.
A comprehensive classification system for preterm birth requires expanded gestational boundaries that recognize the early origins of preterm parturition and emphasize fetal maturity over fetal age. Exclusion of stillbirths, pregnancy terminations, and multifetal gestations prevents comprehensive consideration of the potential causes and presentations of preterm birth. Any step in parturition (cervical softening and ripening, decidual-membrane activation, and/or myometrial contractions) may initiate preterm parturition, and should be recorded for every preterm birth, as should the condition of the mother, fetus, newborn, and placenta, before a phenotype is assigned.


Epidemiology of coronary heart disease and acute coronary syndrome.
The aim of this review is to summarize the incidence, prevalence, trend in mortality, and general prognosis of coronary heart disease (CHD) and a related condition, acute coronary syndrome (ACS). Although CHD mortality has gradually declined over the last decades in western countries, this condition still causes about one-third of all deaths in people older than 35 years. This evidence, along with the fact that mortality from CHD is expected to continue increasing in developing countries, illustrates the need for implementing effective primary prevention approaches worldwide and identifying risk groups and areas for possible improvement.


Shrink Wrapping Cells in a Defined Extracellular Matrix to Modulate the Chemo-Mechanical Microenvironment.
Cell-matrix interactions are important for the physical integration of cells into tissues and the function of insoluble, mechanosensitive signaling networks. Studying these interactions in vitro can be difficult because the extracellular matrix (ECM) proteins that adsorb to in vitro cell culture surfaces do not fully recapitulate the ECM-dense basement membranes to which cells such as cardiomyocytes and endothelial cells adhere to in vivo. Towards addressing this limitation, we have developed a surface-initiated assembly process to engineer ECM proteins into nanostructured, microscale sheets that can be shrink wrapped around single cells and small cell ensembles to provide a functional and instructive matrix niche. Unlike current cell encapsulation technology using alginate, fibrin or other hydrogels, our engineered ECM is similar in density and thickness to native basal lamina and can be tailored in structure and composition using the proteins fibronectin, laminin, fibrinogen, and/or collagen type IV. A range of cells including C2C12 myoblasts, bovine corneal endothelial cells and cardiomyocytes survive the shrink wrapping process with high viability. Further, we demonstrate that, compared to non-encapsulated controls, the engineered ECM modulates cytoskeletal structure, stability of cell-matrix adhesions and cell behavior in 2D and 3D microenvironments.


Baseline Frontoparietal Task-Related BOLD Activity as a Predictor of Improvement in Clinical Symptoms at 1-Year Follow-Up in Recent-Onset Psychosis.
The early course of illness in psychotic disorders is highly variable, and predictive biomarkers of treatment response have been lacking. Trial and error remains the basis for care in early psychosis, and poor outcomes are common. Early prediction of nonimprovement in response to treatment could help identify those who would benefit from alternative and/or supplemental interventions. The goal of this study was to evaluate the ability of functional MRI (fMRI) measures of cognitive control-related brain circuitry collected at baseline to predict symptomatic response in patients after 1 year. Patients with recent-onset (<2 years) psychotic disorders (N=82) in early psychosis specialty care were classified as improvers (>20% improvement in total score on the Brief Psychiatric Rating Scale [BPRS] at 1-year follow-up compared with baseline) or as nonimprovers. Behavioral (d' context) and fMRI (proactive control-associated activation in a priori frontoparietal regions of interest) measures of cognitive control were then evaluated on their ability to predict BPRS improvement using linear and logistic regression. Cognitive control-associated measures significantly predicted BPRS improvement and improver status, with 70% positive predictive value, 60% negative predictive value, and 66% accuracy. Only the fMRI-based measure (and not the behavioral measure) significantly predicted status. These results suggest that frontoparietal activation during cognitive control performance at baseline significantly predicts subsequent symptomatic improvement during early psychosis specialty care. Potential implications for fMRI-based personalized patient treatment are discussed.


Effects of NSAIDs and hydroxyapatite coating on osseointegration.
The aim of our study is to investigate the bone ongrowth of two different alternative surfaces and the effect of nonsteroidal anti-inflammatory drugs (NSAIDs) on osseointegration. Hips of 40 New Zealand white rabbits were operated bilaterally. Hydroxyapatite (HA)-coated titanium rods were implanted into the right femur, and grit-blasted titanium rods were implanted into the left femur. They were divided into three groups. At the end of 8 weeks, both femora of the rabbits were removed and investigated biomechanically and histologically. HA-coated implants had a significantly better failure load and "percentage of bone-implant contact" than grit-blasted implants. There was no significant difference between the medication groups as a result of the biomechanical and histologic investigations. Our results indicate that NSAIDs did not have any negative effect on the osseointegration. HA-coated implants may provide more tensile strength and greater bone-implant contact rate in comparison with grit-blasted implants.


Varicella seroprevalence in a random sample of the Turkish population.
Chicken pox highly contagious and common throughout the world, is an infectious disease caused by varicella zoster virus (VZV). This study was conducted to determine the seroprevalence of VZV in a population under age 30 and to identify the relationship of VZV seroprevalence and several characteristics of the study subjects in nine provinces of Turkey. The sampling method of 30 clusters recommended for field studies was used for selecting subjects of a pre-determined number in the rural and urban areas in each province. For this, a total of 60 groups, 30 clusters in the rural and 30 in the urban areas were determined. It was planned that a total of 4800 subjects, including 600 subjects from five big provinces (Istanbul, Ankara, Izmir, Adana, Diyarbakir) and 450 subjects from the remaining smaller provinces (Samsun, Erzurum, Trabzon, Edirne), be included in the study. ELISA method was used to examine the blood samples for VZV seropositivity. Positive VZV seroprevalence was detected in 77.8% of 4387 subjects under age 30 in nine provinces of Turkey. There was no difference in seroprevalence rate between rural and urban areas. Seroprevalence was found to be 79.0% in urban areas and 76.3% in rural areas. Seroprevalence increased with age. Seroprevalence was 20% at the age of 1 year, subsequently increased to 40% at the age of 4 years, 60% at the age 6 years, 80% at the age of 8 years, 85% at the age of 10 years, and then remained at 85-90% in subjects over the age of 10 years. In order to develop vaccination protocols and take appropriate preventive health care measures against diseases in different countries, it is very important to know the seroprevalence of any disease for an individual country.


Ultrasensitive responses and specificity in cell signaling.
Interconnected cell signaling pathways are able to efficiently and accurately transmit a multitude of different signals, despite an inherent potential for undesirable levels of cross-talk. To ensure that an appropriate response is produced, biological systems have evolved network-level mechanisms that insulate pathways from crosstalk and prevent 'leaking' or 'spillover' between pathways. Many signaling pathways have been shown to respond in an ultrasensitive (switch-like) fashion to graded input, and this behavior may influence specificity. The relationship of ultrasensitivity to signaling specificity has not been extensively explored. We studied the behavior of simple mathematical models of signaling networks composed of two interconnected pathways that share an intermediate component, asking if the two pathways in the network could exhibit both output specificity (preferentially activate their own output) and input fidelity (preferentially respond to their own input). Previous results with weakly-activated pathways indicated that neither mutual specificity nor mutual fidelity were obtainable in the absence of an insulating mechanism, such as cross-pathway inhibition, combinatorial signaling or scaffolding/compartmentalization. Here we found that mutual specificity is obtainable for hyperbolic or ultrasensitive pathways, even in the absence of an insulating mechanism. However, mutual fidelity is impossible at steady-state, even if pathways are hyperbolic or ultrasensitive. Nevertheless, ultrasensitivity does provide advantages in attaining specificity and fidelity to networks that contain an insulating mechanism. For networks featuring cross-pathway inhibition or combinatorial signaling, ultrasensitive activation can increase specificity in a limited way, and can only be utilized by one of the two pathways. In contrast, for networks featuring scaffolding/compartmentalization, ultrasensitive activation of both pathways can dramatically improve network specificity. There are constraints to obtaining performance objectives associated with signaling specificity; such constraints may have influenced the evolution of signal transduction networks. Notably, input fidelity (preferential response to an authentic input) is a more difficult objective to achieve than output specificity (preferential targeting to an authentic output). Indeed, mutual fidelity is impossible in the absence of an insulating mechanism, even if pathways are ultrasensitive. Ultrasensitivity does, however, significantly enhance the performance of several insulating mechanisms. In particular, the ultrasensitive activation of both pathways can provide substantial improvement to networks containing scaffolding/compartmentalization.


Do you want to convince me or to be understood?: preference-consistent information sharing and its motivational determinants.
In two experiments, we provide evidence for a fundamental discussion asymmetry, namely, preference-consistent information sharing. Despite being in a dyadic situation requiring open information exchange and being given no incentive to do so, participants communicated more information that supported their individually preferred decision alternative than information that contradicted it. Preference-consistent information sharing was not caused by biased recall and occurred in written as well as in face-to-face communication. Moreover, we tested whether preference-consistent information sharing was influenced by statements by bogus discussion partners indicating that they held a congruent versus incongruent preference to the participants' preference and that they understood versus did not understand the participants' preference. We found that when partners stated that they understood the participants' preference, subsequent preference-consistent information sharing was considerably reduced. This indicates that a motivation to be understood by others might be an important driving force underlying preference-consistent information sharing.


Inspiring Life in Frozen Communities: Supporting Migrant Women in Brussels to Regain Control over their Lives.
In Brussels, many migrant women without legal status have no or limited access to health care and other basic services. Their access to descent care is mainly hampered by a lack of information, limited financial resources and poor experiences in the past. Three non-governmental organisations joint efforts to help migrant women without legal status to come out of their isolation. Action research during the implementation process was conducted in order to know which elements contributed to increased feelings of trust and reinforced autonomy among the target group and more willingness to support migrants among a larger population. Our major conclusion is that mental health and well-being is largely defined by (the quality of) social relations and interactions - an aspect that is too often forgotten as a result of the medicalization of mental health related problems.