Non-cardiac manifestations of Marfan syndrome.
Because of the widespread distribution of fibrillin 1 in the body, Marfan syndrome (MFS) affects virtually every system. The expression of this single dominantly inherited gene is variable within a family, and between families. There is some genotype-phenotype correlation which is helpful in guiding long-term prognosis, and management. In general gene mutations have been reported in clusters, with those having mainly ocular manifestations occurring in exons 1 to 15 of this 65-exon gene; those causing cardiac problems often involving cysteine replacement in a calcium binding EGF-like sequence; the most severe mutations occurring in exons 25-32, causing neonatal MFS diagnosed at birth, and severe enough to cause death frequently before the age of 2. Other correlations will certainly be found in future. This condition is progressive, and the manifestations unfold according to age. For example, if the lens is going to dislocate this usually occurs by age 10; scoliosis usually presents itself between the ages of 8 and 15; height should be monitored carefully between the onset of puberty and cessation of growth approximately age 17 or 18. Holistic care should be offered by one doctor who oversees the patient's welfare. This should be a paediatrician, paediatric cardiologist, or general practitioner in the case of an affected child. Thereafter, the physician in charge of the most seriously affected system should be aware that other systems need to be managed through a referral network.


Development of an unsupportive social interaction scale for patients with diabetes.
The positive aspects of social support and its impact on health have been studied extensively. However, there may also be negative effects of social environments on the diabetic patient. This study developed and validated a new diabetic unsupportive social interaction scale (USIS), including two subscales: interference and insensitivity. A list of 22 items depicting unsupportive interactions associated with management of diabetes was developed. A telephone survey was administered to 764 Israelis with diabetes. The questionnaire included the USIS and questions about social networks, social support, health behaviors, and health. The characteristics, validity, and reliability of the scale were tested. A principal component analysis was performed for extraction of two factors describing unsupportive social interaction concepts: interference and insensitivity. Cronbach's alpha for the full 15-item scale was 0.84, indicating internal consistency. The two subscales were calculated to have Cronbach's alphas of 0.85 and 0.73, respectively. The USIS showed construct validity as it was associated with social support, some measures of social networks, subjective measures of health, and health behaviors. Arabs, older respondents, those defining themselves as more religious, and the less educated reported higher rates of unsupportive interactions. This study suggests a new concept of unsupportive interactions including interference and insensitivity. These unsupportive interactions may adversely affect patients' ability to adhere to treatment and may undermine their health in various ways. Identifying these problems may enable clinicians to help patients cope with their unsupportive environments.


Trends in Liver Cancer Incidence and Survival in Italy by Histologic Type, 2003-2017.
(1) Background: Liver cancer in Italy is characterised by one of the highest incidence rates worldwide outside of Asia coupled with comparatively favourable survival figures. The objective of this study was to evaluate the most recent epidemiologic trends of the disease. (2) Methods: Thirteen cancer registries covering a population of about 12,740,000 (21% of the national population) made available the records of 35,574 cases registered between 2003 and 2017. Trends in age-standardised (Europe 2013) incidence rates were analysed using the results of age-drift models. Trends in survival were analysed using 1-year, 2-year, 5-year and 10-year net survival (NS) and 5|1-year and 5|2-year conditional NS. (3) Results: Over the study period, the average annual incidence rates per 100,000 persons were 29.4 (men) and 9.4 (women) for total liver cancer; 14.6 and 3.5 for hepatocellular carcinoma (HCC); 1.8 and 1.1 for intrahepatic cholangiocarcinoma (ICC); and 13.0 and 4.8 for the 'other liver cancer types' group. The incidence of total liver cancer and HCC decreased significantly for both sexes. For total liver cancer, the estimated average annual percent change was -1.6% among men and -2.1% among women. For HCC, the change was -1.3% among men and -2.7% among women. ICC followed an opposite trend. For men, the risk of HCC had two peaks, one in the birth cohorts of 1928 and 1933 and another, more moderate peak in the cohort of 1958. Men and women exhibited comparable improvements in both early and mid-term conditional NS from HCC. In 2013-2017, 5-year NS was 28.9% (95% CI: 27.3%; 30.6%) for men and 30.1% (95% CI: 26.9%; 33.5%) for women. The uptrend in survival from ICC was less pronounced and was weakly significant, with a 5-year NS in 2013-2017 of 13.9% (95% CI: 10.8%; 17.3%) for men and 17.4% (95% CI: 13.5%; 21.7%) for women. (4) Conclusions: The opposite incidence trends of HCC and ICC confirm a pattern observed in other populations. The generalised, albeit slow, improvement in survival from HCC indicates a trend towards earlier detection coupled with improvements in treatments.


Prevalence of Neisseria meningitidis carriers in children under five years of age and teenagers in certain populations of Mexico City.
To carry out a pilot study to discover the frequency of colonization in healthy children under five years old and teenagers, as well as the distribution of the different N. meningitidis serogroups isolated from nasopharyngeal samples collected from the population under study. The population included youth between 15 and 19 years old living in social rehabilitation centers (SRC) and university teenagers (UT) as well as children under the age of five attending day care centers (DCC) in Mexico City. Nasopharyngeal exudates were processed using standard microbiological techniques in order to identify Nm, the serogroup, type and subtype of these isolates. A total of 2 310 samples (774 SRC, 800 UT and 736 DCC) were taken. Total prevalence of Nm was 1.6 per cent. In teenagers living in SRC the prevalence was 2.9%. The most frequent serogroups were Y (29.7%), C (24.3%) and B (10.8%). The prevalence of carriers in the study environment was significant (1.6%) and similar to countries in which the invasive illness is much more frequent.


Single-cell tracking reveals antibiotic-induced changes in mycobacterial energy metabolism.
ATP is a key molecule of cell physiology, but despite its importance, there are currently no methods for monitoring single-cell ATP fluctuations in live bacteria. This is a major obstacle in studies of bacterial energy metabolism, because there is a growing awareness that bacteria respond to stressors such as antibiotics in a highly individualistic manner. Here, we present a method for long-term single-cell tracking of ATP levels in Mycobacterium smegmatis based on a combination of microfluidics, time-lapse microscopy, and Förster resonance energy transfer (FRET)-based ATP biosensors. Upon treating cells with antibiotics, we observed that individual cells undergo an abrupt and irreversible switch from high to low intracellular ATP levels. The kinetics and extent of ATP switching clearly discriminate between an inhibitor of ATP synthesis and other classes of antibiotics. Cells that resume growth after 24 h of antibiotic treatment maintain high ATP levels throughout the exposure period. In contrast, antibiotic-treated cells that switch from ATP-high to ATP-low states never resume growth after antibiotic washout. Surprisingly, only a subset of these nongrowing ATP-low cells stains with propidium iodide (PI), a widely used live/dead cell marker. These experiments also reveal a cryptic subset of cells that do not resume growth after antibiotic washout despite remaining ATP high and PI negative. We conclude that ATP tracking is a more dynamic, sensitive, reliable, and discriminating marker of cell viability than staining with PI. This method could be used in studies to evaluate antimicrobial effectiveness and mechanism of action, as well as for high-throughput screening. New antimicrobials are urgently needed to stem the rising tide of antibiotic-resistant bacteria. All antibiotics are expected to affect bacterial energy metabolism, directly or indirectly, yet tools to assess the impact of antibiotics on the ATP content of individual bacterial cells are lacking. The method described here for single-cell tracking of intracellular ATP in live bacteria has many advantages compared to conventional ensemble-averaged assays. It provides a continuous real-time readout of bacterial ATP content, cell vitality, and antimicrobial mechanism of action with high temporal resolution at the single-cell level. In combination with high-throughput microfluidic devices and automated microscopy, this method also has the potential to serve as a novel screening tool in antimicrobial drug discovery.


Biosensoric potential of microbial fuel cells.
Recent progress in microbial fuel cell (MFC) technology has highlighted the potential of these devices to be used as biosensors. The advantages of MFC-based biosensors are that they are phenotypic and can function in either assay- or flow-through formats. These features make them appropriate for contiguous on-line monitoring in laboratories and for in-field applications. The selectivity of an MFC biosensor depends on the applied microorganisms in the anodic compartment where electron transfer (ET) between the artificial surface (anode) and bacterium occurs. This process strongly determines the internal resistance of the sensoric system and thus influences signal outcome and response time. Despite their beneficial characteristics, the number of MFC-based biosensoric applications has been limited until now. The aim of this mini-review is to turn attention to the biosensoric potential of MFCs by summarizing ET mechanisms on which recently established and future sensoric devices are based.


Culturomics Approaches Expand the Diagnostic Accuracy for Sexually Transmitted Infections.
Sexually transmitted infections (STIs) are a major health concern with clinical manifestations being acknowledged to cause severe reproductive impairment. Research in infectious diseases has been centered around the known major pathogens for decades. However, we have just begun to understand that the microbiota of the female genital tract is of particular importance for disease initiation, infection progression, and pathological outcome. Thus, we are now aware that many poorly described, partially not yet known, or cultured bacteria may pave the way for an infection and/or contribute to disease severity. While sequencing-based methods are an important step in diagnosing STIs, culture-based methods are still the gold-standard method in diagnostic routine, providing the opportunity to distinguish phenotypic traits of bacteria. However, current diagnostic culture routines suffer from several limitations reducing the content of information about vaginal microbiota. A detailed characterization of microbiota-associated factors is needed to assess the impact of single-bacterial isolates from the vaginal community on vaginal health and the containment of STIs. Here we provide current concepts to enable modern culture routines and create new ideas to improve diagnostic approaches with a conjunct usage of bioinformatics. We aim to enable scientists and physicians alike to overcome long-accepted limitations in culturing bacteria of interest to the human health. Eventually, this may improve the quality of culture-based diagnostics, facilitate a research interface, and lead to a broader understanding of the role of vaginal microbiota in reproductive health and STIs.


Reversible cerebral vasoconstriction syndrome.
Reversible cerebral vasoconstriction syndrome is characterised by severe headaches with or without seizures and focal neurological deficits, and constriction of cerebral arteries which resolves spontaneously in 1-3 months. It affects females slightly more than males, and mean age of onset is around 45 years. Approximately 60% of cases are secondary, mainly postpartum and after exposure to vasoactive substances. The major complications are localised cortical subarachnoid haemorrhage (22%) and parenchymal ischaemic or haemorrhagic strokes (7%) which may leave permanent sequelae. Diagnosis requires the demonstration of the "string of beads" appearance of cerebral arteries on angiography, with complete or almost complete resolution on repeat angiography 12 weeks after onset. Nimodipine seems to reduce thunderclap headaches within 48 h but has no definite effect on the haemorrhagic and ischaemic complications.


Co-ingestion of Antioxidant Drinks With an Unhealthy Challenge Meal Fails to Prevent Post-prandial Endothelial Dysfunction: An Open-Label, Crossover Study in Older Overweight Volunteers.
Eating a high calorie meal is known to induce endothelial dysfunction and it is reported that consuming drinks rich in antioxidants may be protective against this. In this study we assessed the effects of three antioxidant drinks with considerable disparity in their antioxidant content on endothelial function. Seven apparently healthy overweight and older adults (BMI 25-35; mean age 57 ± 3 years; one male, six females) completed four trials in a randomized counterbalanced design. Water (control), orange juice, green tea, or red wine were consumed with a high calorie meal (>900 kcal). Endothelial function was measured by flow-mediated dilatation immediately before (fasted, baseline) and 2 h after the meal. Blood samples were also obtained for lipid and glucose analysis, plasma nitrite ( NO 2 - ) and oxidized low-density lipoprotein (ox-LDL). Participants returned after a minimum 3 days washout to complete the remaining arms of the study. The results found that the high calorie meal induced a substantial increase in triglycerides, but not cholesterol or glucose, at 2 h after meal ingestion. FMD was significantly reduced by ∼35% at this timepoint, but the effect was not attenuated by co-ingestion of any of the antioxidant drinks. Reduced FMD was mirrored by a reduction in NO 2 - , but ox-LDL was not increased at 2 h after the meal. None of the undertaken measures were influenced by the antioxidant drinks. We conclude that co-ingestion of none of our test antioxidant drinks protected against the substantial post-prandial endothelial dysfunction induced by an unhealthy meal challenge in our sample population at a 2 h timepoint.


The impact of physiological metabolite levels on serine uptake, synthesis and utilization in cancer cells.
Serine is a non-essential amino acid that is critical for tumour proliferation and depletion of circulating serine results in reduced tumour growth and increased survival in various cancer models. While many cancer cells cultured in a standard tissue culture medium depend on exogenous serine for optimal growth, here we report that these cells are less sensitive to serine/glycine depletion in medium containing physiological levels of metabolites. The lower requirement for exogenous serine under these culture conditions reflects both increased de novo serine synthesis and the use of hypoxanthine (not present in the standard medium) to support purine synthesis. Limiting serine availability leads to increased uptake of extracellular hypoxanthine, sparing available serine for other pathways such as glutathione synthesis. Taken together these results improve our understanding of serine metabolism in physiologically relevant nutrient conditions and allow us to predict interventions that may enhance the therapeutic response to dietary serine/glycine limitation.


A 3D k-space Fourier encoding and reconstruction framework for simultaneous multi-slab acquisition.
To propose a novel 3D k-space Fourier encoding and reconstruction framework for simultaneous multi-slab (SMSlab) acquisition and demonstrate its efficacy in high-resolution imaging. First, it is illustrated in theory how the inter-slab gap interferes with the formation of the SMSlab 3D k-space. Then, joint RF and gradient encoding are applied to remove the inter-slab gap interference and form a SMSlab 3D k-space. In vivo experiments are performed to validate the proposed theory. Acceleration in the proposed SMSlab 3D k-space is also evaluated. High-resolution (1.0 mm isotropic) images can be reconstructed using the proposed SMSlab 3D framework. Controlled aliasing in parallel imaging sampling and 2D GRAPPA reconstruction can also be applied in the SMSlab 3D k-space. Compared with conventional multi-slab acquisition, SMSlab exhibits better SNR maintainability (such as lower g-factors), especially at high acceleration factors. It is demonstrated that the joint application of RF and gradient encoding enables SMSlab within a 3D Fourier encoding framework. Images with high isotropic resolution can be reconstructed, and further acceleration is also applicable. The proposed SMSlab 3D k-space can be valuable for both high-resolution and high-efficiency diffusion and functional MRI.


Effect of Sahaj Yoga on depressive disorders.
Sahaj Yoga is a meditative technique that has been found to have beneficial effects in some psycho-somatic illnesses. The study was carried out on 30 cases (19 Males, 11 females, age 18-45 years) of major depression diagnosed on the basis of DSM IV criteria. The patients were then randomly divided into two groups: Group 1: (10 Males & 5 Females) Patients who were practising Sahaj Yoga and also received conventional anti-depressants. Group 2: (9 Males & 6 Females) Patients who were only receiving conventional anti-depressants. Training in Sahaj yoga was conducted under the supervision of a trained Sahaj Yogi for 8 weeks. At start of the study, all the patients were subjected to Hamilton Rating Scale for Depression (HAM-D) and Hamilton Rating Scale for Anxiety (HAM-A). Above scales were again assessed after two months of treatment. There was significant improvement in HAM-D as well as HAM-A scores in both Group 1 and Group 2 patients (P<0.001). However, percentage improvement in HAM-D scores and HAM-A scores in patients receiving Sahaj Yoga was significantly higher than in Group 2 patients. The number of patients who went into remission after two months of intervention were also significantly higher in Group 1 patients (P=0.02). The present study demonstrates that Sahaj Yoga has got a potential role as a component in the management of depressive disorders.


Self-managed abortion: A systematic scoping review.
Self-managed abortion, when a person performs their own abortion without clinical supervision, is a model of abortion care used across a range of settings. To provide a comprehensive synthesis of the available literature on self-managed abortion, we conducted a systematic search for peer-reviewed research in April 2019 in PubMed, Embase, Web of Science, Popline, PsycINFO, Google Scholar, Scielo, and Redalyc. We included studies that had a research question focused on self-managed abortion; and were published in English or Spanish. The combined search returned 7167 studies; after screening, 99 studies were included in the analysis. Included studies reported on methods, procurement, characteristics of those who self-managed, effectiveness, safety, reasons for self-managed abortion, and emotional and physical experiences. Numerous abortion methods were reported, most frequently abortion with pills and herbs. Studies reporting on self-managed medication abortion reported high-levels of effectiveness. We identify gaps in the research, and make recommendations to address those gaps.


Recognition of tRNA bottom half by bacterial ribonuclease P.
Bacterial ribonuclease P (RNase P) contains a catalytic RNA that cleaves precursor tRNA to form the 5'-end of mature tRNA. Bacterial RNase P mainly recognizes the acceptor stem and T arm modules of tRNA molecules. The region consisting of T arm, acceptor stem and 3' CCA motif in the tRNA is generally termed "top half", and the region consisting of the others, anticodon arm, extra loop and D arm, is called "bottom half". The stems in the top half contribute to recognition, but effects of the bottom half have not been elucidated. To study the effects of the bottom half on the RNase P recognition, we have synthesized several mutant substrates that have the bottom half on different positions along the top half stem. Most of these mutants were cleaved by Escherichia coli RNase P precisely at the expected position, but the cleavage efficiencies were very different especially at low Mg2+ concentration. We also found that RNase P holoenzyme prefered somewhat mutated tRNA precursor to the wild-type tRNA precursor.


Allele-selective lowering of mutant HTT protein by HTT-LC3 linker compounds.
Accumulation of mutant proteins is a major cause of many diseases (collectively called proteopathies), and lowering the level of these proteins can be useful for treatment of these diseases. We hypothesized that compounds that interact with both the autophagosome protein microtubule-associated protein 1A/1B light chain 3 (LC3)1 and the disease-causing protein may target the latter for autophagic clearance. Mutant huntingtin protein (mHTT) contains an expanded polyglutamine (polyQ) tract and causes Huntington's disease, an incurable neurodegenerative disorder2. Here, using small-molecule-microarray-based screening, we identified four compounds that interact with both LC3 and mHTT, but not with the wild-type HTT protein. Some of these compounds targeted mHTT to autophagosomes, reduced mHTT levels in an allele-selective manner, and rescued disease-relevant phenotypes in cells and in vivo in fly and mouse models of Huntington's disease. We further show that these compounds interact with the expanded polyQ stretch and could lower the level of mutant ataxin-3 (ATXN3), another disease-causing protein with an expanded polyQ tract3. This study presents candidate compounds for lowering mHTT and potentially other disease-causing proteins with polyQ expansions, demonstrating the concept of lowering levels of disease-causing proteins using autophagosome-tethering compounds.


Dynamics of simian immunodeficiency virus SIVmac239 infection in pigtail macaques.
Pigtail macaques (PTM) are an excellent model for HIV research; however, the dynamics of simian immunodeficiency virus (SIV) SIVmac239 infection in PTM have not been fully evaluated. We studied nine PTM prior to infection, during acute and chronic SIVmac239 infections, until progression to AIDS. We found PTM manifest clinical AIDS more rapidly than rhesus macaques (RM), as AIDS-defining events occurred at an average of 42.17 weeks after infection in PTM compared to 69.56 weeks in RM (P = 0.0018). However, increased SIV progression was not associated with increased viremia, as both peak and set-point plasma viremias were similar between PTM and RM (P = 0.7953 and P = 0.1006, respectively). Moreover, this increased disease progression was not associated with rapid CD4(+) T cell depletion, as CD4(+) T cell decline resembled other SIV/human immunodeficiency virus (HIV) models. Since immune activation is the best predictor of disease progression during HIV infection, we analyzed immune activation by turnover of T cells by BrdU decay and Ki67 expression. We found increased levels of turnover prior to SIV infection of PTM compared to that observed with RM, which may contribute to their increased disease progression rate. These data evaluate the kinetics of SIVmac239-induced disease progression and highlight PTM as a model for HIV infection and the importance of immune activation in SIV disease progression.


Technical Note: A Novel Servo-Driven Dual-Roller Handrim Wheelchair Ergometer.
The measurement of handrim wheelchair propulsion characteristics and performance in the field is complicated due to the non-stationary nature of wheelchair driving. In contrast, the laboratory provides a constrained and standardisable environment to conduct measurements and experiments. Apart from wheelchair treadmills, dynamometers or ergometers for handrim wheelchairs are often custom-made, one-of-a-kind, expensive, and sparsely documented in the research literature. To facilitate standardised and comparable lab-based measurements in research, as well as in clinical settings and adapted sports, a new wheelchair ergometer was developed. The ergometer with instrumented dual rollers allows for the performance analysis of individuals in their personal handrim wheelchair and facilitates capacity assessment, training and skill acquisition in rehabilitation or adapted sports. The ergometer contains two servomotors, one for each rear wheel roller, that allow for the simulation of translational inertia and resistive forces as encountered during wheelchair propulsion based on force input and a simple mechanical model of wheelchair propulsion. A load cell configuration for left and right roller enables the measurement of effective user-generated torque and force on the handrim and the concomitant timing patterns. Preliminary results are discussed.


Chromatin Unfolding by Epigenetic Modifications Explained by Dramatic Impairment of Internucleosome Interactions: A Multiscale Computational Study.
Histone tails and their epigenetic modifications play crucial roles in gene expression regulation by altering the architecture of chromatin. However, the structural mechanisms by which histone tails influence the interconversion between active and inactive chromatin remain unknown. Given the technical challenges in obtaining detailed experimental characterizations of the structure of chromatin, multiscale computations offer a promising alternative to model the effect of histone tails on chromatin folding. Here we combine multimicrosecond atomistic molecular dynamics simulations of dinucleosomes and histone tails in explicit solvent and ions, performed with three different state-of-the-art force fields and validated by experimental NMR measurements, with coarse-grained Monte Carlo simulations of 24-nucleosome arrays to describe the conformational landscape of histone tails, their roles in chromatin compaction, and the impact of lysine acetylation, a widespread epigenetic change, on both. We find that while the wild-type tails are highly flexible and disordered, the dramatic increase of secondary-structure order by lysine acetylation unfolds chromatin by decreasing tail availability for crucial fiber-compacting internucleosome interactions. This molecular level description of the effect of histone tails and their charge modifications on chromatin folding explains the sequence sensitivity and underscores the delicate connection between local and global structural and functional effects. Our approach also opens new avenues for multiscale processes of biomolecular complexes.


Insulin resistance and adipokines serum levels in a caucasian cohort of hiv-positive patients undergoing antiretroviral therapy: a cross sectional study.
Insulin resistance is frequent in human immunodeficiency virus (HIV) infection and may be related to antiretroviral therapy. Cytokines secreted by adipose tissue (adipokines) are linked to insulin sensitivity. The present study is aimed to assess the prevalence of insulin resistance (IR) and its association with several adipokines, in a non-diabetic Romanian cohort of men and women with HIV-1 infection, undergoing combination antiretroviral therapy (cART). A cross-sectional study was conducted in an unselected sample of 89 HIV-1-positive, non-diabetic patients undergoing stable cART for at least 6 months. Metabolic parameters were measured, including fasting plasma insulin, and circulating adiponectin, leptin, resistin, tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) levels. Insulin resistance was estimated by measuring the Quantitative Insulin Sensitivity Check Index (QUICKI), using a cut-off value of 0.33. A linear regression model was fitted to QUICKI to test the association of IR and adipokines levels. A total of 89 patients (aged 18-65, median: 28 years) including 51 men (57.3%) and 38 women (42.7%) were included in the study. Fifty nine patients (66.3%) were diagnosed with IR based on QUICKI values lower than the cut-off point. IR prevalence was 72.5% in men and 57.6% in women. The presence of the IR was not influenced by either the time of the HIV diagnosis or by the duration of cART. Decreased adiponectin and increased serum triglycerides were associated with increased IR in men (R=0.43, p=0.007). Hyperleptinemia in women was demonstrated to be associated with the presence of IR (R=0.33, p=0.03). Given the significant prevalence of the IR in our young non-diabetic cohort with HIV infection undergoing antiretroviral therapy reported in our study and the consecutive risk of diabetes and cardiovascular events, we suggest that the IR management should be a central component of HIV-infection therapeutic strategy. As adipokines play major roles in regulating glucose homeostasis with levels varying according to the sex, we suggest that further studies investigating adipokines should base their analyses on gender differences.


Novel risk markers for gastric cancer screening: Present status and future prospects.
Initial identification of populations at high risk of gastric cancer (GC) is important for endoscopic screening of GC. As serum pepsinogen (PG) test-positive subjects with progression of chronic atrophic gastritis (CAG) show a high likelihood of future cancer development, this population warrants careful follow-up observation as a high-risk GC group. By combining the PG test with Helicobacter pylori (HP) antibody titers, the HP-related chronic gastritis stage can be classified, thus identifying not only a GC high-risk group but also a low-risk group. Among PG test-negative patients without CAG, those with high serum PG II levels and HP antibody titers are thought to have severe gastric mucosal inflammation and the risk of diffuse-type GC is also high. Meanwhile, in gastric mucosae obtained by endoscopic biopsy, HP infection induces aberrant DNA methylation in CpG islands in multiple gene regions and the extent of methylation clearly correlates with GC risk. By quantifying aberrant DNA methylation in suitable gene markers, we can determine the extent of the epigenetic field for cancerization. These novel concepts and risk markers will have many clinical applications in gastrointestinal endoscopy, including more efficient endoscopic GC screening and a strategic approach to metachronous multiple GCs after endoscopic treatment.


Do we produce enough fruits and vegetables to meet global health need?
Low fruit and vegetable (FV) intake is a leading risk factor for chronic disease globally, but much of the world's population does not consume the recommended servings of FV daily. It remains unknown whether global supply of FV is sufficient to meet current and growing population needs. We sought to determine whether supply of FV is sufficient to meet current and growing population needs, globally and in individual countries. We used global data on agricultural production and population size to compare supply of FV in 2009 with population need, globally and in individual countries. We found that the global supply of FV falls, on average, 22% short of population need according to nutrition recommendations (supply:need ratio: 0.78 [Range: 0.05-2.01]). This ratio varies widely by country income level, with a median supply:need ratio of 0.42 and 1.02 in low-income and high-income countries, respectively. A sensitivity analysis accounting for need-side food wastage showed similar insufficiency, to a slightly greater extent (global supply:need ratio: 0.66, varying from 0.37 [low-income countries] to 0.77 [high-income countries]). Using agricultural production and population projections, we also estimated supply and need for FV for 2025 and 2050. Assuming medium fertility and projected growth in agricultural production, the global supply:need ratio for FV increases slightly to 0.81 by 2025 and to 0.88 by 2050, with similar patterns seen across country income levels. In a sensitivity analysis assuming no change from current levels of FV production, the global supply:need ratio for FV decreases to 0.66 by 2025 and to 0.57 by 2050. The global nutrition and agricultural communities need to find innovative ways to increase FV production and consumption to meet population health needs, particularly in low-income countries.


Prognostic Value of Bone Formation and Resorption Proteins in Heterotopic Ossification in Critically-Ill Patients. a Single-Centre Study.
A potential complication in critically ill patients is the formation of bone in soft tissues, termed heterotopic ossification. The exact pathogenetic mechanisms are still undetermined. Bone morphogenetic proteins induce bone formation, while signalling through the receptor activator of nuclear factor kappa-Β (RANK) and its ligand (RANKL), regulates osteoclast formation, activation, and survival in normal bone modelling and remodelling. Osteoprotegerin protects bone from excessive bone loss by blocking RANKL from binding to RANK. The study aimed to investigate these molecules as potential prognostic biomarkers of heterotopic ossification development in critically ill patients. In this prospective observational study, BMP-2, RANKL, and osteoprotegerin were measured by ELISA in twenty-eight critically-ill, initially non-septic patients, on admission to an ICU, seven days post-admission, and thirty days after ICU discharge. In the critically-ill cohort, nine of the twenty-eight patients developed heterotopic ossification up to the 30-day follow-up time-point. The patients who developed heterotopic ossification exhibited significantly reduced BMP-2 and RANKL levels on ICU admission, compared to patients who did not; Osteoprotegerin readings were similar in both groups. Critically-ill patients who will subsequently develop heterotopic ossification, have significantly lower BMP-2 and RANKL levels at the time of ICU admission, suggesting that these proteins may be useful as prognostic markers for this debilitating condition.


Proangiogenic Tie2(+) macrophages infiltrate human and murine endometriotic lesions and dictate their growth in a mouse model of the disease.
Endometriosis affects women of reproductive age, causing infertility and pain. Although immune cells are recruited in endometriotic lesions, their role is unclear. Tie2-expressing macrophages (TEMs) have nonredundant functions in promoting angiogenesis and growth of experimental tumors. Here we show that human TEMs infiltrate areas surrounding newly formed endometriotic blood vessels. We set up an ad hoc mouse model in which TEMs, and not Tie2-expressing endothelial cells, are targeted. We transplanted in wild-type recipients bone marrow cells expressing a suicide gene (Herpes simplex virus type 1 thymidine kinase) under the Tie2 promoter/enhancer. TEMs infiltrated endometriotic lesions. TEM depletion by ganciclovir administration arrested the growth of established lesions, without toxicity. Lesion architecture was disrupted, with: i) loss of glandular organization, ii) reduced neovascularization, and iii) activation of caspase 3 in CD31(+) endothelial cells. Thus, TEMs are important for maintaining the viability of newly formed vessels and represent a potential therapeutic target in endometriosis.


In vitro and in vivo activities of dicationic diguanidino compounds against Echinococcus multilocularis metacestodes.
Alveolar echinococcosis (AE) is a disease predominantly affecting the liver, with metacestodes (larvae) of the tapeworm Echinococcus multilocularis proliferating and exhibiting tumor-like infiltrative growth. For many years, chemotherapeutical treatment against alveolar echinococcosis has relied on the benzimidazoles albendazole and mebendazole, which require long treatment durations and exhibit parasitostatic rather than parasiticidal efficacy. Although benzimidazoles have been and still are beneficial for the patients, there is clearly a demand for alternative and more efficient treatment options. Aromatic dications, more precisely a small panel of di-N-aryl-diguanidino compounds, were screened for efficacy against E. multilocularis metacestodes in vitro. Only those with a thiophene core group were active against metacestodes, while furans were not. The most active compound, DB1127, was further investigated in terms of in vivo efficacy in mice experimentally infected with E. multilocularis metacestodes. This diguanidino compound was effective against AE when administered intraperitoneally but not when applied orally. Thus, thiophene-diguanidino derivatives with improved bioavailability when administered orally could lead to treatment options against AE.


Subjective Sleep Quality and Sleep Architecture in Patients With Migraine: A Meta-analysis.
Sleep disturbance is often associated with migraine. However, there is a paucity of research investigating objective and subjective measures of sleep in patients with migraine. This meta-analysis aims to determine whether there are differences in subjective sleep quality measured using the Pittsburgh Sleep Quality Index (PSQI) and objective sleep architecture measured using polysomnography (PSG) between adult and pediatric patients and healthy controls. This review was preregistered on PROSPERO (CRD42020209325). A systematic search of 5 databases (Embase, MEDLINE, Global Health, APA PsycINFO, and APA PsycArticles, last searched on December 17, 2020) was conducted to find case-control studies that measured PSG or PSQI in patients with migraine. Pregnant participants and those with other headache disorders were excluded. Effect sizes (Hedges g) were entered into a random effects model meta-analysis. Study quality was evaluated with the Newcastle Ottawa Scale and publication bias with the Egger regression test. Thirty-two studies were eligible, of which 21 measured PSQI or Migraine Disability Assessment Test in adults, 6 measured PSG in adults, and 5 measured PSG in children. The overall mean study quality score was 5/9; this did not moderate any of the results and there was no risk of publication bias. Overall, adults with migraine had higher PSQI scores than healthy controls (g = 0.75, p < 0.001, 95% confidence interval [CI] 0.54-0.96). This effect was larger in those with a chronic rather than episodic condition (g = 1.03, p < 0.001, 95% CI 0.37-1.01; g = 0.63, p < 0.001, 95% CI 0.38-0.88, respectively). For polysomnographic studies, adults and children with migraine displayed a lower percentage of rapid eye movement sleep (g = -0.22, p = 0.017, 95% CI -0.41 to -0.04; g = -0.71, p = 0.025, 95% CI -1.34 to -0.10, respectively) than controls. Pediatric patients displayed less total sleep time (g = -1.37, p = 0.039, 95% CI -2.66 to -0.10), more wake (g = 0.52, p < 0.001, 95% CI 0.08-0.79), and shorter sleep onset latency (g = -0.37, p < 0.001, 95% CI -0.54 to -0.21) than controls. People with migraine have significantly poorer subjective sleep quality and altered sleep architecture compared to healthy individuals. Further longitudinal empirical studies are required to enhance our understanding of this relationship.


Open-Label Randomized Trial of Early Clinical Outcomes of Ceftaroline Fosamil Versus Vancomycin for the Treatment of Acute Bacterial Skin and Skin Structure Infections at Risk of Methicillin-Resistant Staphylococcus aureus.
Acute bacterial skin and skin structure infections (ABSSSIs) remain among the most common infectious processes seen in the clinical setting. For patients with complicated ABSSSIs deemed to require intravenous antibiotics, vancomycin remains the mainstay therapy. Ceftaroline has been shown to be non-inferior to vancomycin and may result in faster resolution of signs of infection. Multicenter, prospective, open-label, randomized trial of ceftaroline versus vancomycin for the treatment of adult patients admitted for management of ABSSSIs from April 2012 to May 2016; 166 patients in the clinically evaluable (CE) group were needed to determine a 20% difference in primary outcome of clinical response at day 2 or 3 of antibiotics. Clinical response was defined as cessation of spread of lesion and improvement in systemic signs/symptoms of infection. A secondary outcome was a ≥ 20% reduction in lesion size at day 2 or 3 of antibiotics. One hundred seventy-four patients were enrolled in the intention-to-treat (ITT) group and 108 were CE. Among CE patients, 54 were randomized to ceftaroline and 54 to vancomycin. Baseline characteristics were similar except patients in the ceftaroline arm were older and had a non-significantly higher degree of comorbidities (median Charlson score 2 vs. 4, respectively). Cellulitis was the most common type of ABSSSI (85.2% vs. 79.6%, respectively). Rapid diagnostic testing of available cultures (n = 55) demonstrated high agreement with clinical microbiology for identification of Staphylococcus aureus (100%) and MRSA (100%). There was no significant difference in primary outcome of day 2 or 3 clinical response (50.0% vs. 51.9%). Early clinical response between vancomycin- and ceftaroline-treated ABSSSIs was similar. Patients with ABSSSIs rarely remained hospitalized for > 2-3 days, thus limiting our ability to critically assess clinical outcomes. ClinicalTrials.gov identifier, NCT02582203. Allergan plc.


Cocaine-related health emergencies in Europe: a review of sources of information, trends and implications for service development.
Cocaine-related health consequences are difficult to observe. Data on drug users in health-emergency settings may be a useful source of information on consequences that are not visible via other information sources. Thirty European countries submit an annual national report on the drug situation to the EMCDDA. All reports for the period 2007-2010 were analyzed, with particular attention given to auditing cocaine-related mentions. Analysis was also performed in order to identify sources and case definitions, assess coverage, audit cases and, where possible, to identify long-term trends. Considerable heterogeneity existed between countries in their approach to recording drug-related emergencies, with only Spain and the Netherlands having established formal indicators. The highest annual numbers of cocaine-related episodes were reported by the UK (3,502), Spain (2,845) and the Netherlands (1,211). A considerable (2- to 3-fold) increase in the numbers of cocaine-related episodes has been reported since the end of the 1990s in these countries; these increases peaked in Spain and England around 2007/08. The analysis reported here suggests the need to develop more standardized approaches to monitoring drug-related emergencies. It points to the potential value of developing effective referral links between the emergency and specialized drug services working with cocaine users.


Iron fortification of infant formulas. American Academy of Pediatrics. Committee on Nutrition.
Despite the American Academy of Pediatrics' (AAP) strong endorsement for breastfeeding, most infants in the United States are fed some infant formula by the time they are 2 months old. The AAP Committee on Nutrition has strongly advocated iron fortification of infant formulas since 1969 as a way of reducing the prevalence of iron-deficiency anemia and its attendant sequelae during the first year.1 The 1976 statement titled "Iron Supplementation for Infants" delineated the rationale for iron supplementation, proposed daily dosages of iron, and summarized potential sources of iron in the infant diet.2 In 1989, the AAP Committee on Nutrition published a statement that addressed the issue of iron-fortified infant formulas3 and concluded that there was no convincing contraindication to iron-supplemented formulas and that continued use of "low-iron" formulas posed an unacceptable risk for iron deficiency during infancy. The current statement represents a scientific update and synthesis of the 1976 and 1989 statements with recommendations about the use of iron-fortified and low-iron formulas in term infants.


Multiphase modeling of geologic carbon sequestration in saline aquifers.
Geologic carbon sequestration (GCS) is being considered as a climate change mitigation option in many future energy scenarios. Mathematical modeling is routinely used to predict subsurface CO2 and resident brine migration for the design of injection operations, to demonstrate the permanence of CO2 storage, and to show that other subsurface resources will not be degraded. Many processes impact the migration of CO2 and brine, including multiphase flow dynamics, geochemistry, and geomechanics, along with the spatial distribution of parameters such as porosity and permeability. In this article, we review a set of multiphase modeling approaches with different levels of conceptual complexity that have been used to model GCS. Model complexity ranges from coupled multiprocess models to simplified vertical equilibrium (VE) models and macroscopic invasion percolation models. The goal of this article is to give a framework of conceptual model complexity, and to show the types of modeling approaches that have been used to address specific GCS questions. Application of the modeling approaches is shown using five ongoing or proposed CO2 injection sites. For the selected sites, the majority of GCS models follow a simplified multiphase approach, especially for questions related to injection and local-scale heterogeneity. Coupled multiprocess models are only applied in one case where geomechanics have a strong impact on the flow. Owing to their computational efficiency, VE models tend to be applied at large scales. A macroscopic invasion percolation approach was used to predict the CO2 migration at one site to examine details of CO2 migration under the caprock.


S. pombe placed on the prion map.
Schizosaccharomyces pombe has been used extensively as a model organism, however it is only recently that the first prion in this organism, a copper transporter protein encoded by ctr4, has been conclusively demonstrated. Prions are found in a wide range of organisms and have been implicated in a number of human neurodegenerative diseases. Research into the biology of prions has been carried out mainly in the budding yeast Saccharomyces cerevisiae, however there are many questions still to be addressed. Now, with the identification of the Ctr4 prion in S. pombe, further work in the two yeasts and comparisons of prion biology in these organisms should lead to a greater understanding of prions and their role in disease.


Real-Space Mapping of Surface-Oxygen Defect States in Photovoltaic Materials Using Low-Voltage Scanning Ultrafast Electron Microscopy.
Ultrathin layers of native oxides on the surface of photovoltaic materials may act as very efficient carrier trapping/recombination centers, thus significantly affecting their interfacial photophysical properties. How ultrathin oxide layers affect the surface and interface carrier dynamics cannot be selectively accessed by conventional ultrafast transient spectroscopic methods due to the deep penetration depth (tens to thousands of nanometers) of the pump/probe laser pulses. Herein, scanning ultrafast electron microscopy (S-UEM) at a low voltage of 1 keV electrons was recently developed at KAUST to selectively map the ultrafast charge carrier dynamics of a few layers on the top surfaces of photovoltaic materials. Unlike high-voltage 30 keV experiments, at 1 keV, the depth of detected secondary electrons (SEs) underneath the surface is significantly reduced 5 times, thus making it possible to visualize the dynamics of charge carrier from the uppermost surface of photoactive layers. More specifically, this new approach has been employed to explore and decipher the tremendous impact of native oxide layers and oxygen defect states on charge carrier dynamics in space and time simultaneously at sub-nanometer levels on several photovoltaic material surfaces, including Si, GaAs, CdTe, and CdZnTe single crystals. Interestingly, the contrast in the SEs time-resolved difference images switched from a dark "energy-loss mechanism" to a bright "energy-gain mechanism" only by removing the layers of surface oxides. Moreover, the charge carrier recombination was estimated and found to be dramatically affected by the native oxide layers. The density functional theory (DFT) calculations demonstrate that the work function of oxygen-terminated Si surface also increases slightly upon optical excitation and makes for less SE intensity, providing another piece of evidence that the origin of the dark contrast observed on these material surfaces should be assigned to the surface oxide formation, mainly oxygen defect states in the band gap and/or work function increment. Our findings provide a new method and pave the way for evaluating the effect of surface morphology and defects, including but not limited to native oxide, on charge carrier dynamics, and interfacial properties of photovoltaic absorber layers.


Spatial four-alternative forced-choice method is the preferred psychophysical method for naïve observers.
H. R. Blackwell (1952) investigated the influence of different psychophysical methods and procedures on detection thresholds. He found that the temporal two-interval forced-choice method (2-IFC) combined with feedback, blocked constant stimulus presentation with few different stimulus intensities, and highly trained observers resulted in the "best" threshold estimates. This recommendation is in current practice in many psychophysical laboratories and has entered the psychophysicists' "folk wisdom" of how to run proper psychophysical experiments. However, Blackwell's recommendations explicitly require experienced observers, whereas many psychophysical studies, particularly with children or within a clinical setting, are performed with naïve observers. In a series of psychophysical experiments, we find a striking and consistent discrepancy between naïve observers' behavior and that reported for experienced observers by Blackwell: Naïve observers show the "best" threshold estimates for the spatial four-alternative forced-choice method (4-AFC) and the worst for the commonly employed temporal 2-IFC. We repeated our study with a highly experienced psychophysical observer, and he replicated Blackwell's findings exactly, thus suggesting that it is indeed the difference in psychophysical experience that causes the discrepancy between our findings and those of Blackwell. In addition, we explore the efficiency of different methods and show 4-AFC to be more than 3.5 times more efficient than 2-IFC under realistic conditions. While we have found that 4-AFC consistently gives lower thresholds than 2-IFC in detection tasks, we have found the opposite for discrimination tasks. This discrepancy suggests that there are large extrasensory influences on thresholds--sensory memory for IFC methods and spatial attention for spatial forced-choice methods--that are critical but, alas, not part of theoretical approaches to psychophysics such as signal detection theory.


The secreted proteins of Achlya hypogyna and Thraustotheca clavata identify the ancestral oomycete secretome and reveal gene acquisitions by horizontal gene transfer.
Saprotrophic and parasitic microorganisms secrete proteins into the environment to breakdown macromolecules and obtain nutrients. The molecules secreted are collectively termed the "secretome" and the composition and function of this set of proteins varies depending on the ecology, life cycle, and environment of an organism. Beyond the function of nutrient acquisition, parasitic lineages must also secrete molecules to manipulate their host. Here, we use a combination of de novo genome and transcriptome sequencing and bioinformatic identification of signal peptides to identify the putative secreted proteome of two oomycetes, the facultative parasite Achlya hypogyna and free-living Thraustotheca clavata. By comparing the secretomes of these saprolegnialean oomycetes with that of eight other oomycetes, we were able to characterize the evolution of this protein set across the oomycete clade. These species span the last common ancestor of the two major oomycete families allowing us to identify the ancestral secretome. This putative ancestral secretome consists of at least 84 gene families. Only 11 of these gene families are conserved across all 10 secretomes analyzed and the two major branches in the oomycete radiation. Notably, we have identified expressed elicitin-like effector genes in the saprotrophic decomposer, T. clavata. Phylogenetic analyses show six novel horizontal gene transfers to the oomycete secretome from bacterial and fungal donor lineages, four of which are specific to the Saprolegnialeans. Comparisons between free-living and pathogenic taxa highlight the functional changes of oomycete secretomes associated with shifts in lifestyle.


Dissection of an antibody-catalyzed reaction.
Antibody 43C9 accelerates the hydrolysis of a p-nitroanilide by a factor of 2.5 x 10(5) over the background rate in addition to catalyzing the hydrolysis of a series of aromatic esters. Since this represents one of the largest rate accelerations achieved with an antibody, we have undertaken a series of studies aimed at uncovering the catalytic mechanism of 43C9. The immunogen, a phosphonamidate, was designed to mimic the geometric and electronic characteristics of the tetrahedral intermediate that forms upon nucleophilic attack by hydroxide on the amide substrate. Further studies, however, revealed that the catalytic mechanism is more complex and involves the fortuitous formation of a covalent acyl-antibody intermediate as a consequence of complementary side chain residues at the antibody-binding site. Several lines of evidence indicate that the catalytic mechanism involves two key residues: His-L91, which acts as a nucleophile to form the acyl-antibody intermediate, and Arg-L96, which stabilizes the anionic tetrahedral moieties. Support for this mechanism derives from the results of site-directed mutagenesis experiments and solvent deuterium isotope effects as well as direct detection of the acyl-antibody by electrospray mass spectrometry. Despite its partial recapitulation of the course of action of enzymic counterparts, the reactivity of 43C9, like other antibodies, is apparently limited by its affinity for the inducing immunogen. To go beyond this level, one must introduce additional catalytic functionality, particularly general acid-base catalysis, through either improvements in transition-state analog design or site-specific mutagenesis.


Source attribution of nontyphoid salmonellosis in new zealand using outbreak surveillance data.
In this study, 204 New Zealand outbreaks of non-typhoid salmonellosis reported from 2000 to 2009 were analyzed for information on the sources of human infection. Data were extracted from the outbreak module of EpiSurv, New Zealand's notifiable diseases database, and augmented with information from individual case reports and separate investigation reports. The outbreaks involved 1,426 cases, representing an estimated 9% of the total salmonellosis cases reported for the study period. Salmonella Typhimurium was the causative serotype in 78% of 172 outbreaks for which a serotype was available, involving 71% of outbreak cases. The most commonly reported outbreak setting was the home (47% of outbreaks), followed by commercial food operations (31%). Foodborne transmission was reported for 63% of the 123 outbreaks for which only one mode of transmission was reported, followed by person-to-person transmission (32%), waterborne transmission (3%), and zoonotic transmission (2%). However, evidence for the mode of transmission was weak or absent for 107 (63%) of the 169 outbreaks for which a mode of transmission was reported. For only 22 outbreaks was laboratory evidence successfully used to identify a potential source of infection. Of these 22 outbreaks, 7 were foodborne, 11 involved an infected food handler, 2 were attributed to contact with animals, 1 was attributed to consumption of drinking water, and 1 was attributed to multiple sources. The laboratory-confirmed contaminated foods were diverse and included imported and domestically produced foods. The results of this analysis support the hypothesis that non-typhoid salmonellosis is primarily a foodborne disease in New Zealand, but there is insufficient evidence to confirm important food vehicles.


Nonshivering thermogenesis and its adequate measurement in metabolic studies.
Alterations in nonshivering thermogenesis are presently discussed as being both potentially causative of and able to counteract obesity. However, the necessity for mammals to defend their body temperature means that the ambient temperature profoundly affects the outcome and interpretation of metabolic experiments. An adequate understanding and assessment of nonshivering thermogenesis is therefore paramount for metabolic studies. Classical nonshivering thermogenesis is facultative, i.e. it is only activated when an animal acutely requires extra heat (switched on in minutes), and adaptive, i.e. it takes weeks for an increase in capacity to develop. Nonshivering thermogenesis is fully due to brown adipose tissue activity; adaptation corresponds to the recruitment of this tissue. Diet-induced thermogenesis is probably also facultative and adaptive and due to brown adipose tissue activity. Although all mammals respond to injected/infused norepinephrine (noradrenaline) with an increase in metabolism, in non-adapted mammals this increase mainly represents the response of organs not involved in nonshivering thermogenesis; only the increase after adaptation represents nonshivering thermogenesis. Thermogenesis (metabolism) should be expressed per animal, and not per body mass [not even to any power (0.75 or 0.66)]. A 'cold tolerance test' does not examine nonshivering thermogenesis capacity; rather it tests shivering capacity and endurance. For mice, normal animal house temperatures are markedly below thermoneutrality, and the mice therefore have a metabolic rate and food consumption about 1.5 times higher than their intrinsic requirements. Housing and examining mice at normal house temperatures carries a high risk of identifying false positives for intrinsic metabolic changes; in particular, mutations/treatments that affect the animal's insulation (fur, skin) may lead to such problems. Correspondingly, true alterations in intrinsic metabolic rate remain undetected when metabolism is examined at temperatures below thermoneutrality. Thus, experiments with animals kept and examined at thermoneutrality are likely to yield an improved possibility of identifying agents and genes important for human energy balance.


Autoconfocal microscopy with a cw laser and thermionic detection.
We introduce an application of thermionic emission in a PMT photocathode. Because of the nonlinear dependence of thermionic emission on absorbed laser power, a conventional PMT is found to produce a virtual pinhole effect that rejects unfocused light at least as strongly as a physical pinhole. This virtual pinhole effect is exploited in a scanning transmission confocal microscope equipped with a cw laser source. Because the area of the PMT photocathode is large, signal descanning is not required and thermionic detection acts as a self-aligned pinhole. Our technique of thermionic-detection autoconfocal microscopy is further implemented with graded-field contrast to obtain enhanced phase-gradient sensitivity in unlabeled samples, such as rat hippocampal brain slices.


Epidemiology of pretransplant EBV and CMV serostatus in relation to posttransplant non-Hodgkin lymphoma.
BACKGROUND.: Despite the importance of non-Hodgkin lymphoma (NHL) as a posttransplant complication, the relationship between NHL and recipient seropositivity for Epstein-Barr virus (EBV) or cytomegalovirus (CMV) is incompletely understood. METHODS.: Kidney, heart, and liver transplant recipients reported to the Collaborative Transplant Study with known pretransplant EBV and CMV serostatus were analyzed in terms of clinically manifest NHL. Cox multivariate regression analysis was performed to account for a wide range of possible confounders. RESULTS.: In total, 18,682 kidney, 2042 heart, and 2616 liver transplant recipients were analyzed. Regardless of age, pretransplant EBV serostatus was significantly associated with risk of NHL in kidney transplant recipients (P<0.001). There was no significant difference in lymphoma rates according to CMV and CMV serostatus among EBV and EBV recipients (log-rank P=0.55 and P=0.57, respectively), but hospitalization for CMV disease during year 1 posttransplant was associated with subsequent NHL (hazard ratio [HR] 6.1; 95% confidence interval [CI] 2.0-18.4; P=0.001). EBV serostatus was also associated with increased risk of NHL in heart transplant patients (HR 3.6; 95% CI 1.1-11.3; P=0.031) but, contrary to expectation, not in liver recipients (HR 0.6; 95% CI 0.1-1.7; P=0.32). CONCLUSIONS.: In view of the striking increase in risk of NHL in EBV kidney transplant recipients of all ages, EBV serostatus should be determined pretransplant in all age groups. CMV serostatus was not independently associated with risk of NHL after kidney transplantation. Surprisingly, in liver transplantation, the risk of NHL was virtually unaffected by EBV serostatus.


Convective flow reversal in self-powered enzyme micropumps.
Surface-bound enzymes can act as pumps that drive large-scale fluid flows in the presence of their substrates or promoters. Thus, enzymatic catalysis can be harnessed for "on demand" pumping in nano- and microfluidic devices powered by an intrinsic energy source. The mechanisms controlling the pumping have not, however, been completely elucidated. Herein, we combine theory and experiments to demonstrate a previously unreported spatiotemporal variation in pumping behavior in urease-based pumps and uncover the mechanisms behind these dynamics. We developed a theoretical model for the transduction of chemical energy into mechanical fluid flow in these systems, capturing buoyancy effects due to the solution containing nonuniform concentrations of substrate and product. We find that the qualitative features of the flow depend on the ratios of diffusivities δ=D(P)/D(S) and expansion coefficients β=β(P)/β(S) of the reaction substrate (S) and product (P). If δ>1 and δ>β (or if δ<1 and δ<β ), an unexpected phenomenon arises: the flow direction reverses with time and distance from the pump. Our experimental results are in qualitative agreement with the model and show that both the speed and direction of fluid pumping (i) depend on the enzyme activity and coverage, (ii) vary with the distance from the pump, and (iii) evolve with time. These findings permit the rational design of enzymatic pumps that accurately control the direction and speed of fluid flow without external power sources, enabling effective, self-powered fluidic devices.


Reconfigurable UWB pulse generator based on pulse shaping in a nonlinear optical loop mirror and differential detection.
A reconfigurable impulse radio ultra-wideband (UWB) pulse generator for various UWB shapes (e.g., monocycle, doublet, and triplet pulses) based on a nonlinear optical loop mirror (NOLM) and differential detection is proposed and experimentally demonstrated. The proposed approach can be used with different modulation formats and may be suitable for implementation in future low-cost, high-speed, short-range UWB wireless access applications.


Improving executive function deficits by playing interactive video-games: secondary analysis of a randomized controlled trial for individuals with chronic stroke.
Executive function deficits negatively impact independence and participation in everyday life of individuals with chronic stroke. Therefore, it is important to explore therapeutic interventions to improve executive functions. The aim of this study was to determine the effectiveness of a 3-month interactive video-game group intervention compared to a traditional motor group intervention for improving executive functions in individuals with chronic stroke. This study is a secondary analysis of a single-blind randomized controlled trial for improving factors related to physical activity of individuals with chronic stroke. Assessments were administered pre and post the intervention and at 3-month follow-up by assessors blind to treatment allocation. Thirty-nine individuals with chronic stroke with executive function deficits participated in an interactive video-game group intervention (N.=20) or a traditional group intervention (N.=19). The intervention included two 1-hour group sessions per week for three months, either playing video-games or performing traditional exercises/activities. Executive function deficits were assessed using The Trail Making Test (Parts A and B) and by two performance-based assessments; the Bill Paying Task from the Executive Function Performance Test (EFPT) and the Executive Function Route-Finding Task (EFRT). Following intervention, scores for the Bill Paying Task (EFPT) decreased by 27.5% and 36.6% for the participants in the video-game and traditional intervention, respectively (F=17.3, P<0.000) and continued to decrease in the video-game group with small effect sizes. Effect size was small to medium for the TMT-B (F=0.003, P=0.954) and EFRT (F=1.2, P=0.28), without any statistical significance difference. Interactive video-games provide combined cognitive-motor stimulation and therefore have potential to improve executive functioning of individuals with chronic stroke. Further research is needed. These findings highlight the potential of utilizing interactive video-games in a small group for keeping these individuals active, while maintaining and improving executive functioning especially for individuals with chronic stroke, who have completed their formal rehabilitation.


Surface ozone response to satellite-constrained NOx emission adjustments and its implications.
Both surface and satellite observations have shown a decrease in NOx emissions in East Asian countries in recent years. In order to reflect the recent NOx emission reduction and to investigate its impact on surface O3 concentrations in Asian megacities, we adjusted two bottom-up regional emission inventories of which base years are 2006 (E2006) and 2010 (E2010), respectively. We applied direct and relative emission adjustments to both E2006 and E2010 to constrain NOx emissions using OMI NO2 vertical column densities. Except for the relative emission adjustment with E2006, modeling results with adjusted emissions exhibit that NOx emissions over East Asian megacities (Beijing, Shanghai, Seoul, and Tokyo) in the bottom-up inventories are generally overestimated. When the direct emission adjustment is applied to E2006, model biases in the Seoul Metropolitan Area (SMA), South Korea are reduced from 24 ppb to 2 ppb for NOx (=NO+NO2) and from -9 ppb to 0 ppb for O3. In addition, NO2 model biases in Beijing and Shanghai in China are reduced from 8 ppb to 18 ppb-0 ppb and 1 ppb, respectively. Daily maximum 8-h average O3 model biases over the same places are decreased by 8 ppb and 14 ppb. Further analyses suggest that the reduction in domestic South Korean NOx emissions plays a significant role in increasing O3 concentrations in SMA. We conclude that the current strong drive to reduce NOx emissions as part of the strategy to lower particulate matter concentrations in South Korea can account for increased O3 concentrations in recent years and suggest that more aggressive NOx emissions will be necessary soon.


The Effect of Speaking Valves on ICU Mobility of Individuals With Tracheostomy.
Early mobilization is part of the rehabilitation process for critically ill patients and is currently considered a means of preventing ICU-acquired muscle deterioration and worsening of physical function. We sought to determine whether the use of speaking valves in tracheostomized patients would improve their mobility. We evaluated the changes in mobility performance with the use of speaking valves in tracheostomized subjects. We performed a cohort study of a series of subjects who were tracheostomized and were being weaned from mechanical ventilation between April 2016 and May 2018. The subjects were those able to tolerate a speaking valve for a minimum of 30 min. Demographic data, comorbidities, cause of ICU admission, days in the ICU, duration of mechanical ventilation before tracheostomy, and days free from mechanical ventilation before tracheostomy were collected. Mobility status was evaluated using daily measurements of the Perme Intensive Care Unit Mobility Score. During the study period, 63 patients were tracheostomized. Patients with deficiencies in language (n = 2) or cognitive deficits (n = 36) and patients in exclusive palliative care (n = 7) were excluded. Eighteen subjects were enrolled in the study. The mean age of the subjects was 64.6 ± 14.2 y (55.5% were male), and the most common reason for ICU admission was pneumonia (n = 7; 38.8%). Perme scores increased from 11.3 (interquartile range 10.1-12.0) on the day before initiation of the speaking valve to 18.2 (IQR 16.2-20.1) immediately after the initiation of a speaking valve (P < .01). These changes were maintained during all periods of speaking valve use. The use of speaking valves in tracheostomized subjects improved mobility.


Absolute pitch: evidence for early cognitive facilitation during passive listening as revealed by reduced P3a amplitudes.
Absolute pitch (AP) is the rare ability to identify or produce different pitches without using reference tones. At least two sequential processing stages are assumed to contribute to this phenomenon. The first recruits a pitch memory mechanism at an early stage of auditory processing, whereas the second is driven by a later cognitive mechanism (pitch labeling). Several investigations have used active tasks, but it is unclear how these two mechanisms contribute to AP during passive listening. The present work investigated the temporal dynamics of tone processing in AP and non-AP (NAP) participants by using EEG. We applied a passive oddball paradigm with between- and within-tone category manipulations and analyzed the MMN reflecting the early stage of auditory processing and the P3a response reflecting the later cognitive mechanism during the second processing stage. Results did not reveal between-group differences in MMN waveforms. By contrast, the P3a response was specifically associated with AP and sensitive to the processing of different pitch types. Specifically, AP participants exhibited smaller P3a amplitudes, especially in between-tone category conditions, and P3a responses correlated significantly with the age of commencement of musical training, suggesting an influence of early musical exposure on AP. Our results reinforce the current opinion that the representation of pitches at the processing level of the auditory-related cortex is comparable among AP and NAP participants, whereas the later processing stage is critical for AP. Results are interpreted as reflecting cognitive facilitation in AP participants, possibly driven by the availability of multiple codes for tones.


Dynamical system model predicts when social learners impair collective performance.
A key question concerning collective decisions is whether a social system can settle on the best available option when some members learn from others instead of evaluating the options on their own. This question is challenging to study, and previous research has reached mixed conclusions, because collective decision outcomes depend on the insufficiently understood complex system of cognitive strategies, task properties, and social influence processes. This study integrates these complex interactions together in one general yet partially analytically tractable mathematical framework using a dynamical system model. In particular, it investigates how the interplay of the proportion of social learners, the relative merit of options, and the type of conformity response affect collective decision outcomes in a binary choice. The model predicts that, when the proportion of social learners exceeds a critical threshold, a bistable state appears in which the majority can end up favoring either the higher- or lower-merit option, depending on fluctuations and initial conditions. Below this threshold, the high-merit option is chosen by the majority. The critical threshold is determined by the conformity response function and the relative merits of the two options. The study helps reconcile disagreements about the effect of social learners on collective performance and proposes a mathematical framework that can be readily adapted to extensions investigating a wider variety of dynamics.


Consent of older children participating in BASCD coordinated dental epidemiology surveys in Wales.
New guidance on consent for England and Wales, which has positive consent at its core, has implications for the UK-wide BASCD coordinated dental epidemiology programme. This paper describes a method used in Wales for obtaining consent from older children which is believed to comply with the new guidance. The objective was to establish a more robust approach to gaining consent from 12 and 14 year olds taking part in the surveys, by building on existing "negative consent" practice and supplementing it with Gillick competent child consent. Questionnaire data from the 2002-03 survey of 6,393 13-14 year-old children and the 2004-05 survey of 6,749 11-12 year olds were used in this analysis. Questions specifically designed to establish competency to consent were asked of participating children. These ascertained whether children were happy to proceed and if so, whether they understood the nature and the purpose of the survey and whether they were happy with the outcome. Ninety-nine percent of those taking part in both survey years were happy to proceed with the examination and questionnaire. Whilst the majority of children, agreeing to take part, indicated that they had understood what was proposed and were happy with the outcome, approximately 15% of these age groups gave answers after the event which indicated that they had not understood either the nature or purpose of the survey. Use of "Gillick competent" consent in Wales did not affect participation rates adversely. The authors would suggest that indication of assent as used in Wales in these two surveys is appropriate and would only exclude 1% of children. The alternative, of examining only those children who answered questions on whether they understood the nature and purpose of what is proposed prior to assenting, would exclude 15% of children.


Genetic variation in the endangered wild apple (Malus sylvestris (L.) Mill.) in Belgium as revealed by amplified fragment length polymorphism and microsatellite markers.
The genetic variation within and between wild apple samples (Malus sylvestris) and cultivated apple trees was investigated with amplified fragment length polymorphisms (AFLP) and microsatellite markers to develop a conservation genetics programme for the endangered wild apple in Belgium. In total, 76 putative wild apples (originating from Belgium and Germany), six presumed hybrids and 39 cultivars were typed at 12 simple sequence repeats (SSR) and 139 amplified fragment length polymorphism (AFLP) loci. Principal co-ordinate analysis and a model-based clustering method classified the apples into three major gene pools: wild Malus sylvestris genotypes, edible cultivars and ornamental cultivars. All presumed hybrids and two individuals (one Belgian, one German) sampled as M. sylvestris were assigned completely to the edible cultivar gene pool, revealing that cultivated genotypes are present in the wild. However, gene flow between wild and cultivated gene pools is shown to be almost absent, with only three genotypes that showed evidence of admixture between the wild and edible cultivar gene pools. Wild apples sampled in Belgium and Germany constitute gene pools that are clearly differentiated from cultivars and although some geographical pattern of genetic differentiation among wild apple populations exists, most variation is concentrated within samples. Concordant conclusions were obtained from AFLP and SSR markers, which showed highly significant correlations in both among-genotypes and among-samples genetic distances.


Alopecia and platelet-derived therapies.
Platelet rich plasma (PRP) injections have emerged as a promising regenerative therapy for androgenetic alopecia (AGA). To date, injections of both autologous native and activated PRP have been administered to hair loss patients, and positive results have been observed. However, little to no work has yet to be seen wherein PRP treatments are combined with hair restoration surgeries. Furthermore, the PRP activation protocol in the hair restoration setting employs compounds with potentially deleterious side effects, namely thrombin or calcium gluconate. Therefore, the objectives of this work are to evaluate the effectiveness of platelet and platelet-derived products as augmented graft therapies in hair restoration surgeries and to compare the follicular regeneration rate of follicles transplanted in the presence of platelet lysate (PL) versus activated PRP (AA-PRP). PL was administered to the frontal scalp of three male AGA patients. Three treatment zones measuring 4 cm-2 were mapped in the midline scalp region of each patient and equal number of follicular grafts were placed in each box along with PL, AA-PRP, or normal saline. The transplanted follicular grafts of a fourth patient were placed solely with PL. Hair checks in which the surface area of hair coverage was quantified were performed at follow-up appointments ranging from 3.5 to 7 months post-surgery. In these appointments, the number of follicular units with hairs measuring 50 mm or more were counted to determine the percentage of graft hair regeneration. Growth factor (GF) concentrations [vascular endothelial growth factor (VEGF), transforming growth factor beta 1 (TGF-β1), PDGF-BB, IGF-1] in PL and AA-PRP were also measured for an independent subject set. Follicular regeneration in transplanted grafts was found to be superior for those placed with PL rather than AA-PRP or saline at all follow-up dates. Specifically, at 3.5 months post-op, 89%±9%, 74%±7%, and 57%±10% of follicular units had regenerated hair in the PL, AA-PRP, and saline treatment zones, respectively. At 4 months post-op, 99%, 75%, and 71% of follicle regeneration had occurred in the PL, AA-PRP, and saline treatment areas, respectively. Impressively, when PL was injected alone, the patient experienced a 50% increase in follicular unit density and a 122% increase in hair density 7 months post-injection. When GF concentrations were measured, PL generated from a 30-min sonication of PRP was found to have significantly higher levels of VEGF, PDGF-BB, and TGF-β1 than AA-PRP. PRP remains a promising hair loss therapy and should be evaluated further for use not only as an independent therapeutic tool, but also as a treatment to augment surgical procedures. PL in particular affords an effective and efficacious therapeutic product given that the lysate may be obtained by mechanical rather than chemical means. Ultrasonic waves provide sufficient energy to rupture platelet cell walls, and centrifugation may be used to separate the lysate from cell fragments prior to delivery.


Clavicles, interclavicles, gastralia, and sternal ribs in sauropod dinosaurs: new reports from diplodocidae and their morphological, functional and evolutionary implications.
Ossified gastralia, clavicles and sternal ribs are known in a variety of reptilians, including dinosaurs. In sauropods, however, the identity of these bones is controversial. The peculiar shapes of these bones complicate their identification, which led to various differing interpretations in the past. Here we describe different elements from the chest region of diplodocids, found near Shell, Wyoming, USA. Five morphotypes are easily distinguishable: (A) elongated, relatively stout, curved elements with a spatulate and a bifurcate end resemble much the previously reported sauropod clavicles, but might actually represent interclavicles; (B) short, L-shaped elements, mostly preserved as a symmetrical pair, probably are the real clavicles, as indicated by new findings in diplodocids; (C) slender, rod-like bones with rugose ends are highly similar to elements identified as sauropod sternal ribs; (D) curved bones with wide, probably medial ends constitute the fourth morphotype, herein interpreted as gastralia; and (E) irregularly shaped elements, often with extended rugosities, are included into the fifth morphotype, tentatively identified as sternal ribs and/or intercostal elements. To our knowledge, the bones previously interpreted as sauropod clavicles were always found as single bones, which sheds doubt on the validity of their identification. Various lines of evidence presented herein suggest they might actually be interclavicles - which are single elements. This would be the first definitive evidence of interclavicles in dinosauromorphs. Previously supposed interclavicles in the early sauropodomorph Massospondylus or the theropods Oviraptor and Velociraptor were later reinterpreted as clavicles or furculae. Independent from their identification, the existence of the reported bones has both phylogenetic and functional significance. Their presence in non-neosauropod Eusauropoda and Flagellicaudata and probable absence in rebbachisaurs and Titanosauriformes shows a clear character polarity. This implicates that the ossification of these bones can be considered plesiomorphic for Sauropoda. The proposed presence of interclavicles in sauropods may give further support to a recent study, which finds a homology of the avian furcula with the interclavicle to be equally parsimonious to the traditional theory that furcula were formed by the fusion of the clavicles. Functional implications are the stabilizing of the chest region, which coincides with the development of elongated cervical and caudal vertebral columns or the use of the tail as defensive weapon. The loss of ossified chest bones coincides with more widely spaced limbs, and the evolution of a wide-gauge locomotor style.


Multiple exemplar-based facial image retrieval using independent component analysis.
In this paper, we design a content-based image retrieval system where multiple query examples can be used to indicate the need to retrieve not only images similar to the individual examples, but also those images which actually represent a combination of the content of query images. We propose a scheme for representing content of an image as a combination of features from multiple examples. This scheme is exploited for developing a multiple example-based retrieval engine. We have explored the use of machine learning techniques for generating the most appropriate feature combination scheme for a given class of images. The combination scheme can be used for developing purposive query engines for specialized image databases. Here, we have considered facial image databases. The effectiveness of the image retrieval system is experimentally demonstrated on different databases.


Response of the cerebral circulation in baboons to changing perfusion pressure after indomethacin.
An earlier study has demonstrated that indomethacin, a prostaglandin synthesis inhibitor, blocks the cerebrovascular response to hypercapnia. This response is believed to be mediated by a lowering of pH in the cerebral interstitial fluid. Should autoregulation of cerebral blood flow (CBF) to changing perfusion pressure also be mediated by a changing interstitial pH (the "metabolic" theory), then indomethacin should impair autoregulation. This hypothesis was tested in anesthetized baboons. CBF was measured by the intracarotid 133Xe clearance technique; the preparation and the indomethacin protocol were identical to those of our previous investigation. Arterial pressure was increased by the intravenous infusion of angiotensin and decreased by controlled hemorrhage. Indomethacin was given by continuous infusion into the internal carotid artery. Although it reduced resting CBF, the cerebrovascular response to changing perfusion pressure was unchanged. Because indomethacin affects the response to changing CO2 but not that to changing perfusion pressure, the mechanisms for these two reactions presumably are different and it is improbable that changing interstitial pH is responsible for autoregulation in the cerebral circulation.


Perceived friends' use as a risk factor for marijuana use across young adulthood.
Perceived social norms of substance use are commonly identified as a risk factor for use. How the strength of association between perceived friends' use and substance use may change across development has not yet been documented. The current analysis considers how the associations between perceived friends' marijuana use and participants' own use of any marijuana in the past year changes from ages 18 to 30 using longitudinal data from the United States national Monitoring the Future study from 1976 to 2014 (N = 30,794 people). Time-varying effect modeling (TVEM) was used to examine the associations between perceived friends' use of marijuana and participants' own annual marijuana use by age, as well as the extent to which these time-varying associations were moderated by sex, race/ethnicity, and parental education. Associations between perceived friends' use and own marijuana use increased with age. In addition, the association between perceived friends' use and own marijuana use significantly varied by demographic groups, such that it was significantly greater for men from ages 19 to 24 and from ages 27 to 30, compared with women; for Whites, compared with other race/ethnicities, across all ages; and for individuals whose parents attended college, compared with those whose parents had a high school education or less, across all ages. Results suggest that perceived friends' marijuana use becomes an even more important marker for increased marijuana use as people age through young adulthood. Therefore, the role of peers in substance use remains crucial beyond adolescence and should be incorporated into intervention strategies for young adults. (PsycINFO Database Record


Graft materials and bone marrow stromal cells in bone tissue engineering.
Bone augmentation procedures rely on osteogenic/osteoconductive properties of bone graft material (BGM). A further improvement is represented by use of autologous bone marrow stromal cells (BMSC), expanded in vitro and seeded on BGM before implantation in the bone defect. The effect of different BGMs on BMSC osteogenic differentiation was evaluated. BMSC were cultured in vitro in the presence of different BGM (natural, synthetic, and mixed origins). Cellular morphology was analyzed with scanning electron microscopy. The capability of BMSC to differentiate was determined in vitro by alkaline phosphatase gene expression and enzyme activity at different time points (7, 14, and 28 days) and in vivo by ectopic bone formation of implanted tissue constructs in an immunodeficient murine model. BGM supports the cell adhesion and osteogenic differentiation of BMSC developing a useful tool in the bone tissue engineering.


Plasma prolactin levels in maturing intact and cryptorchid male rats: development of stress response.
In order to determine the possible role of the seminiferous tubules in the regulation of prolactin secretion during sexual development, male rats were rendered cryptorchid at 22 days of age, and thereafter different groups of animals were decapitated at 8-10 day intervals between Day 32 and Day 70. Cryptorchid rats showed destruction of the germinal epithelium accompanied by increased plasma FSH and, to a much lesser extent, increased plasma LH titers. Nevertheless, plasma prolactin levels were similar to those of intact controls throughout the entire period studied. Plasma prolactin titers in intact controls remained uniformly low from Day 20 to Day 70, contrasting with previous reports in which increasing prolactin levels have been observed during sexual development. To determine the reason for this apparent discrepancy, a longitudinal experiment was conducted in which intact and cryptorchid male rats were bled every 10 days from Day 30 to Day 70, following a 3-min period of exposure to ether fumes. The prolactin response to this stress increased markedly with age. A similar pattern of prolactin was observed in a cross-sectional study in which different groups of intact animals were bled following a 3-min period of ether exposure, at ages ranging from 20 to 70 daysmthe results indicate that unlike FSH secretion, prolactin secretion is not controlled by the seminiferous tubules. In addition, they suggest that the pattern of increasing plasma prolactin previously described in the developing male rat is at least in part caused by an age-dependent increase in responsiveness of prolactin to stress.


The VanS-VanR two-component regulatory system controls synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147.
Plasmid pIP816 of Enterococcus faecium BM4147 confers inducible resistance to vancomycin and encodes the VanH dehydrogenase and the VanA ligase for synthesis of depsipeptide-containing peptidoglycan precursors which bind the antibiotic with reduced affinity. We have characterized a cluster of five genes of pIP816 sufficient for peptidoglycan synthesis in the presence of vancomycin. The distal part of the van cluster encodes VanH, VanA, and a third enzyme, VanX, all of which are necessary for resistance. Synthesis of these enzymes was regulated at the transcriptional level by the VanS-VanR two-component regulatory system encoded by the proximal part of the cluster. VanR was a transcriptional activator related to response regulators of the OmpR subclass. VanS stimulated VanR-dependent transcription and was related to membrane-associated histidine protein kinases which control the level of phosphorylation of response regulators. Analysis of transcriptional fusions with a reporter gene and RNA mapping indicated that the VanR-VanS two-component regulatory system activates a promoter used for cotranscription of the vanH, vanA, and vanX resistance genes.


Adaptation of Francisella tularensis to the mammalian environment is governed by cues which can be mimicked in vitro.
The intracellular bacterium Francisella tularensis survives in mammals, arthropods, and freshwater amoeba. It was previously established that the conventional media used for in vitro propagation of this microbe do not yield bacteria that mimic those harvested from infected mammals; whether these in vitro-cultivated bacteria resemble arthropod- or amoeba-adapted Francisella is unknown. As a foundation for our goal of identifying F. tularensis outer membrane proteins which are expressed during mammalian infection, we first sought to identify in vitro cultivation conditions that induce the bacterium's infection-derived phenotype. We compared Francisella LVS grown in brain heart infusion broth (BHI; a standard microbiological medium rarely used in Francisella research) to that grown in Mueller-Hinton broth (MHB; the most widely used F. tularensis medium, used here as a negative control) and macrophages (a natural host cell, used here as a positive control). BHI- and macrophage-grown F. tularensis cells showed similar expression of MglA-dependent and MglA-independent proteins; expression of the MglA-dependent proteins was repressed by the supraphysiological levels of free amino acids present in MHB. We observed that during macrophage infection, protein expression by intracellular bacteria differed from that by extracellular bacteria; BHI-grown bacteria mirrored the latter, while MHB-grown bacteria resembled neither. Naïve macrophages responding to BHI- and macrophage-grown bacteria produced markedly lower levels of proinflammatory mediators than those in cells exposed to MHB-grown bacteria. In contrast to MHB-grown bacteria, BHI-grown bacteria showed minimal delay during intracellular replication. Cumulatively, our findings provide compelling evidence that growth in BHI yields bacteria which recapitulate the phenotype of Francisella organisms that have emerged from macrophages.


Poly (I:C)- and doxorubicin-loaded magnetic dendrimeric nanoparticles affect the apoptosis-related gene expressions in MCF-7 cells.
Use of nanoparticles as drug carrier vectors has great potential to circumvent the limitations associated with chemotherapy, including drug resistance and destructive side effects. For this purpose, magnetic generation 4 dendrimeric nanoparticles were prepared to carry chemotherapeutic agent doxorubicin (G4-DOX) and immune modulator polyinosinic:polycytidylic acid [Poly(I:C)]. As previously reported, DOX and Poly(I:C) was loaded onto G4 nanoparticles (PIC-G4-DOX). Cellular internalization study using confocal microscopy demonstrated high levels of cellular internalization of PIC-G4-DOX nanoparticles by MCF-7 cells. This resulted in higher efficacy of PIC-G4-DOX nanoparticles in killing MCF-7 breast cancer cells. Alteration in the expression levels of selected genes was determined by RT-qPCR analyses. Proapoptotic NOXA, PUMA, and BAX genes were upregulated, and SURVIVIN, APOLLON, and BCL-2 genes were downregulated, indicating the cell-killing effectiveness of PIC-G4-DOX nanoparticles. Gene expression analysis provided some insights into the possible molecular mechanisms on cytotoxicity of DOX and Poly(I:C) delivered through G4 magnetic nanoparticles. The results demonstrated that PIC-G4-DOX can be useful for targeted delivery affecting apoptotic pathways, resulting in an advanced degree of cancer-cell-killing. They are promising for targeting cancer-cells because of their stability, biocompatibility, higher internalization, and toxicity.


TIME (Tumor Immunity in the MicroEnvironment) classification based on tumor CD274 (PD-L1) expression status and tumor-infiltrating lymphocytes in colorectal carcinomas.
Inhibitors targeting the PDCD1 (programmed cell death 1, PD-1) immune checkpoint pathway have revolutionized cancer treatment strategies. The TIME (Tumor Immunity in the MicroEnvironment) classification based on tumor CD274 (PDCD1 ligand 1, PD-L1) expression and tumor-infiltrating lymphocytes (TIL) has been proposed to predict response to immunotherapy. It remains to be determined clinical, pathological, and molecular features of TIME subtypes of colorectal cancer. Using 812 colon and rectal carcinoma cases from the Nurses' Health Study and Health Professionals Follow-up Study, we examined the association of tumor characteristics and survival outcomes with four TIME subtypes (TIME 1, CD274low/TILabsent; TIME 2, CD274high/TILpresent; TIME 3, CD274low/TILpresent; and TIME 4, CD274high/TILabsent). In survival analyses, Cox proportional hazards models were adjusted for potential confounders, including microsatellite instability (MSI) status, CpG island methylator phenotype (CIMP) status, LINE-1 methylation level, and KRAS, BRAF, and PIK3CA mutation status. TIME subtypes 1, 2, 3 and 4 had 218 (27%), 117 (14%), 103 (13%), and 374 (46%) colorectal cancer cases, respectively. Compared with TIL-absent subtypes (TIME 1 and 4), TIL-present subtypes (TIME 2 and 3) were associated with high-level MSI, high-degree CIMP, BRAF mutation, and higher amounts of neoantigens (p < 0.001). TIME subtypes were not significantly associated with colorectal cancer-specific or overall survival. In conclusion, TIL-present TIME subtypes of colorectal cancer are associated with high levels of MSI and neoantigen load, supporting better responsiveness to cancer immunotherapy. Further studies examining tumor molecular alterations and additional factors in the tumor microenvironment may inform development of immunoprevention and immunotherapy strategies.


Prioritizing research for integrated implementation of early childhood development and maternal, newborn, child and adolescent health and nutrition platforms.
Existing health and nutrition services present potential platforms for scaling up delivery of early childhood development (ECD) interventions within sensitive windows across the life course, especially in the first 1000 days from conception to age 2 years. However, there is insufficient knowledge on how to optimize implementation for such strategies in an integrated manner. In light of this knowledge gap, we aimed to systematically identify a set of integrated implementation research priorities for health, nutrition and early child development within the 2015 to 2030 timeframe of the Sustainable Development Goals (SDGs). We applied the Child Health and Nutrition Research Initiative method, and consulted a diverse group of global health experts to develop and score 57 research questions against five criteria: answerability, effectiveness, deliverability, impact, and effect on equity. These questions were ranked using a research priority score, and the average expert agreement score was calculated for each question. The research priority scores ranged from 61.01 to 93.52, with a median of 82.87. The average expert agreement scores ranged from 0.50 to 0.90, with a median of 0.75. The top-ranked research question were: i) "How can interventions and packages to reduce neonatal mortality be expanded to include ECD and stimulation interventions?"; ii) "How does the integration of ECD and MNCAH&N interventions affect human resource requirements and capacity development in resource-poor settings?"; and iii) "How can integrated interventions be tailored to vulnerable refugee and migrant populations to protect against poor ECD and MNCAH&N outcomes?". Most highly-ranked research priorities varied across the life course and highlighted key aspects of scaling up coverage of integrated interventions in resource-limited settings, including: workforce and capacity development, cost-effectiveness and strategies to reduce financial barriers, and quality assessment of programs. Investing in ECD is critical to achieving several of the SDGs, including SDG 2 on ending all forms of malnutrition, SDG 3 on ensuring health and well-being for all, and SDG 4 on ensuring inclusive and equitable quality education and promotion of life-long learning opportunities for all. The generated research agenda is expected to drive action and investment on priority approaches to integrating ECD interventions within existing health and nutrition services.