Self-Inflicted Injury-Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP-SI): a new surveillance tool for detecting self-inflicted injury events in emergency departments.
To assess the performance of the Canadian Hospitals Injury Reporting and Prevention Program's newly developed self-harm surveillance tool (CHIRPP-SI) designed to improve emergency department (ED) hospital surveillance of youth self-inflicted injury (SI). This was a prospective, single-centre cohort study from February 2015 to September 2015. Eligible participants were aged 6-17.99 years and presented to the ED with a primary mental health complaint. The frequency of SI cases was extracted from three data sources (CHIRPP-SI, medical chart, and the National Ambulatory Care Reporting System Metadata (NACRS)). Cohen's kappa statistic was used to examine the level of agreement between data sources. Of the 250 participants who received a medical chart review, 70 completed the CHIRPP-SI. Of those who did not complete the CHIRPP-SI, 86% (n = 154) reported no SI related to their presentation, 12% (n = 22) declined to participate without specifying self-injury status, and 2% (n = 4) were unable to be interviewed prior to discharge. The three sources of surveillance data varied considerably; the medical chart captured the highest frequency of individuals reporting SI related to their ED visit (33.6%), followed by the CHIRPP-SI (28.0%), and the NACRS database (8.4%). The CHIRPP-SI captured the method of SI and the place of occurrence in 100% of individuals, and the bodily location harmed in 98.6% of individuals. Study findings highlight the disparity between different sources of data, in relation to the capture of paediatric SI, presenting to hospital EDs. If greater details of SI events are to be identified, surveillance tools such as the CHIRPP-SI should be considered.


Structural implications of hERG K+ channel block by a high-affinity minimally structured blocker.
Cardiac potassium channels encoded by human ether-à-go-go-related gene (hERG) are major targets for structurally diverse drugs associated with acquired long QT syndrome. This study characterized hERG channel inhibition by a minimally structured high-affinity hERG inhibitor, Cavalli-2, composed of three phenyl groups linked by polymethylene spacers around a central amino group, chosen to probe the spatial arrangement of side chain groups in the high-affinity drug-binding site of the hERG pore. hERG current (IhERG) recorded at physiological temperature from HEK293 cells was inhibited with an IC50 of 35.6 nm with time and voltage dependence characteristic of blockade contingent upon channel gating. Potency of Cavalli-2 action was markedly reduced for attenuated inactivation mutants located near (S620T; 54-fold) and remote from (N588K; 15-fold) the channel pore. The S6 Y652A and F656A mutations decreased inhibitory potency 17- and 75-fold, respectively, whereas T623A and S624A at the base of the selectivity filter also decreased potency (16- and 7-fold, respectively). The S5 helix F557L mutation decreased potency 10-fold, and both F557L and Y652A mutations eliminated voltage dependence of inhibition. Computational docking using the recent cryo-EM structure of an open channel hERG construct could only partially recapitulate experimental data, and the high dependence of Cavalli-2 block on Phe-656 is not readily explainable in that structure. A small clockwise rotation of the inner (S6) helix of the hERG pore from its configuration in the cryo-EM structure may be required to optimize Phe-656 side chain orientations compatible with high-affinity block.


Combining molecular data with classical morphology for uncultured phagotrophic euglenids (Excavata): a single-cell approach.
Phagotrophic euglenids are one of the most diverse and important forms of heterotrophic flagellates in sediment systems, and are key to understanding the evolution of photosynthetic euglenids and 'primary osmotrophs', yet relatively little is known about their biodiversity and phylogenetic relationships. A wealth of light microscopy-based information is available, but little progress has been made in associating this with molecular sequence data. We established a protocol to obtain light microscopy data and molecular data from single euglenid cells isolated from environmental samples. Individual cells from freshwater and marine benthic samples were isolated and rinsed by micropipetting, documented using high-resolution photomicroscopy, then subjected to single-cell nested PCR using taxon-specific primers in combination with universal eukaryotic primers, generating > 75% or full-length SSU rDNA sequences. As a proof-of-principle eight individuals were characterised and subjected to phylogenetic analyses. Many of these cells were identified as Anisonema or Dinema, and grouped with existing sequences assigned to these taxa, and with a 'Peranema sp.' sequence that we could now clearly demonstrate was misidentified or misannotated. Another cell is Heteronema c.f. exaratum, the first 'skidding heteronemid' for which sequence data are available. This is not closely related to Heteronema scaphurum, and intriguingly, branches as the sister group to primary osmotrophs. A cell similar to Ploeotia vitrea (the type of this genus), shows no particular phylogenetic affinity to Ploeotia costata, the best studied Ploeotia species. Our experimental protocol provides a useful starting point for future analyses on euglenid biodiversity (including environmental sequence surveys), and their evolution and systematics.


[Mineral-bone disorder with chronic kidney disease].
Mineral-bone disorder in chronic kidney disease is a clinical syndrome provoked by the combination of three factors: abnormal laboratory results, bone morphology disorder and extra-bone calcification. Its onset in adult age is linked with a decrease in glomerular filtration (GF < 1 ml/s). Fully developed forms occur in the course of regular dialysis treatment. The use of the traditional denomination "renal osteodystrophy" is currently restricted to the bone morphology finding. As there are two threshold types of bone turnover (low and high) and two degrees of mineralisation (low and normal), there is a total of four basic variants of mineral-bone disorder. The high turnover variants--secondary hyperparathyreosis and a combined disorder--are still the most frequent and are diagnosed in 70 to 80% of cases. Low turnover disorders include osteomalatia (OM) and adynamic bone disease (ABD). While OM is becoming increasingly rare, the occurrence of ABD is on the rise. The main reason for this may be the steady growth in the age of dialised patients and a number of risk factors, as well as treatment with inadequately high doses of vitamin D. Progressive chronic kidney disease may be linked with D-hormone deficit, negative calcium balance and with positive phosphate balance. Phosphates become a key factor in the development and progression of secondary hyperparathyreosis and extra-bone calcification in the case of D-hormone substitution. Therefore, maintaining a good phosphate balance by restricting their intake or by reducing their intestinal resorption through the use of phosphate binders is the most efficient therapeutic procedure. In patients with chronic kidney failure, adequate dialysis treatment is necessary. Hyperphosphatemia and extra-bone calcification are new independent risk factors of cardiovascular morbidity and mortality.


Stearoyl-CoA desaturase 1: A potential target for non-alcoholic fatty liver disease?-perspective on emerging experimental evidence.
Non-alcoholic fatty liver disease (NAFLD) is a progressive disease and one of the leading causes of death. An unnamed disease has become a global epidemic disease of public health concern. This spectrum of diseases manifests itself with initial accumulation of excessive triglycerides (due to de novo lipogenesis) in the hepatocytes, leading to simple steatosis. Although its aetiology is multi-factorial, lifestyle changes (diet and physical activity) are considered to be the key thriving factors. In this context, high fructose consumption is associated with an increased risk for developing NAFLD in humans, while high-fructose feeding to experimental animals results in hepatic steatosis and non-alcoholic steatohepatitis, by increasing hepatic lipogenesis. Among several lipogenic genes, the endoplasmic reticulum-bound stearoyl-CoA desaturase 1 (SCD1) is the key determinant of triglycerides biosynthesis pathway, by providing monounsaturated fatty acids, through the incorporation of a double bond at the delta-9 position of saturated fatty acids, specifically, palmitic (C16:0) and stearic (C18:0) acids, yielding palmitoleic (C16:1) and oleic (C18:1) acids, respectively. Various experimental studies involving SCD1 gene knockout and diet-induced rodent models have demonstrated that SCD1 plays a key role in the development of NAFLD, by modulating hepatic lipogenesis and thus triglyceride accumulation in the liver. Several pharmacological and dietary intervention studies have shown the benefits of inhibiting hepatic SCD1 in the pathogenesis of NAFLD. In this review, we give an overview of SCD1 in NAFLD, based on the current experimental evidence and the translational applicability of SCD1 inhibition in human NAFLD conditions, besides discussing the limitations and way-forward.


What is the truth? An application of the Extended Parallel Process Model to televised truth® ads.
The purpose of this study was to analyze television ads in the truth® campaign using the Extended Parallel Process Model (EPPM) as a framework. Among the ads (n = 86) analyzed, results revealed a heavy reliance on severity messages, modest attention to susceptibility messages, and no inclusion of recommended response messages in the form of self-efficacy and response efficacy. The reliance on emphasizing the health threat, without incorporating recommended response messages, is discussed with respect to the likelihood of galvanizing maladaptive responses such as psychological reactance, denial, and defensive avoidance resulting from exposure to these ads. Additionally, the unintended outcomes for secondary audiences including but not limited to stigma are considered. Implications and suggestions for practitioners and theorists are explored.


A framework for guiding health literacy research in populations with universal access to healthcare.
At least one third of the US population suffers from limited health literacy, which has been linked to poorer health status, higher costs, and individuals who are socioeconomically disadvantaged. However, research and the development of theoretical frameworks to study health literacy have only recently begun to occur. The purpose of this article is to describe theoretical frameworks that have either been used or may be used to guide health literacy research and to identify implications for nursing research and practice related to an adaptation of a health literacy framework developed specifically for conducting research in populations with universal access to healthcare.


Absolute charge calibration of scintillating screens for relativistic electron detection.
We report on new charge calibrations and linearity tests with high-dynamic range for eight different scintillating screens typically used for the detection of relativistic electrons from laser-plasma based acceleration schemes. The absolute charge calibration was done with picosecond electron bunches at the ELBE linear accelerator in Dresden. The lower detection limit in our setup for the most sensitive scintillating screen (KODAK Biomax MS) was 10 fC/mm(2). The screens showed a linear photon-to-charge dependency over several orders of magnitude. An onset of saturation effects starting around 10-100 pC/mm(2) was found for some of the screens. Additionally, a constant light source was employed as a luminosity reference to simplify the transfer of a one-time absolute calibration to different experimental setups.


Differential perinatal testosterone secretory capacity of wild house mice testes is related to aggressiveness in adulthood.
Testosterone secretory capacity of testicular Leydig cells was determined in fetal males of an aggressive and a nonaggressive genetic selection line of wild house mice. They were studied at Days 15-18 of gestation and on the first day after birth. A previously described morphometric method was used to quantify 3 beta-hydroxy steroid dehydrogenase (3 beta-HSD)-stained Leydig cells in testicular sections to determine testosterone secretory capacity, which may be considered to reflect circulating plasma testosterone in the fetus. The results of this study show that the testosterone secretory capacity of Leydig cells in the testis changes differentially during intrauterine development in males of the aggressive and nonaggressive selection lines. The peak secretory capacity is reached at Day 17 of gestation for the males of the aggressive selection line, while the peak for the nonaggressive males is reached on the first neonatal day. The larger anogenital distance observed in aggressive males suggests a higher prenatal testosterone level in these males. The importance of the difference in timing of the perinatal 3 beta-HSD peak top individual variation in adult aggressive behavior is discussed.


Diagnostic value of Doppler echocardiography for identifying hemodynamic significant pulmonary valve regurgitation in tetralogy of Fallot: comparison with cardiac MRI.
Quantification of pulmonary regurgitation (PR) is essential in the management of patients with repaired tetralogy of Fallot (TOF). We sought to evaluate the accuracy of first-line Doppler echocardiography in comparison with cardiac magnetic resonance imaging (MRI) to identify hemodynamic significant PR. Paired cardiac MRI and echocardiographic studies (n = 97) in patients with repaired TOF were retrospectively analyzed. Pressure half time (PHT) and pulmonary regurgitation index (PRi) were measured using continuous wave Doppler. The ratio of the color flow Doppler regurgitation jet width to pulmonary valve (PV) annulus (jet/annulus ratio) and diastolic to systolic time velocity integral (DSTVI; pulsed wave Doppler) were assessed. Accuracy of echocardiographic measurements was tested to identify significant PR as determined by phase-contrast MRI (PR fraction [PRF] ≥ 20%). Mean PRF was 29.4 ± 15.7%. PHT < 100 ms had a sensitivity of 93%, specificity 75%, positive predictive value (PPV) 92% and negative predictive value (NPV) 78% for identifying significant PR (C-statistic 0.82). PRi < 0.77 had sensitivity and specificity of 66% and 54%, respectively (C-statistic 0.63). Jet/annulus ratio ≥1/3 had sensitivity 96%, specificity 75%, PPV 92% and NPV 82% (C-statistic 0.87). DSTVI had sensitivity 84%, specificity 33%, PPV 84% and NPV 40%, (C-statistic 0.56). Combined jet/annulus ratio ≥1/3 and PHT < 100 ms was highly accurate in identifying PRF ≥ 20%, with sensitivity 97% and specificity 100%. PHT and jet/annulus ratio on Doppler echocardiography, especially when combined, are highly accurate in identifying significant PR and therefore seem useful in the follow-up of patients with repaired TOF.


Spinopelvic parameter changes and low back pain improvement due to femoral neck anteversion in patients with severe unilateral primary hip osteoarthritis undergoing total hip replacement.
The study of the interrelation between hip and spine disorders is gaining increasing importance in the last years, but the link between Hip Osteoarthritis (HOA) and Low Back Pain (LBP) remains still unclear. Aim of the study is to assess the relationship between Femoral Neck Anteversion (FNA), LBP, and spinopelvic parameters in patients undergoing Total Hip Replacement (THR) for unilateral severe primary HOA. 91 patients were recruited. Inclusion criteria were: grade 5 or 6 unilateral HOA, according to Turmezei, and Harris Hip score (HHS) <60. Exclusion criteria were: secondary hip osteoarthritis (dysplasia of the hip, rheumatoid arthritis, and ankylosing spondylitis); previous surgery of the spine, hip or knee; scoliosis with a Cobb angle greater than 10°; spondylolisthesis; history of spine fractures; previous bone tuberculosis or any spine infections; any contraindications to CT; BMI >30. Patients were divided into two homogeneous Groups according to the presence (Group-A) or not of concomitant LBP (Group-B). All patients underwent preoperatively a hip CT scan to evaluate FNA, Acetabular Anteversion (AA), and Combined Anteversion (CA = FNA + AA). ΔFNA, ΔAA and ΔCA were calculated as the differences between the arthritic hip and the normal hip angles in each Group. Full spinal X-rays in upstanding position were performed before (baseline) and 6 months after THR (follow-up) to calculate spinopelvic parameters. The health-related quality of life (HRQoL) was evaluated at baseline and at follow-up using Visual Analogue Scale (VAS), HHS, Oswestry Disability Index (ODI), Roland-Morris Disability Questionnaire (RM), and Short-Form Health Survey (SF-36). The intra-group and inter-group variability were assessed using, respectively, paired and unpaired t tests. At baseline, the association between HRQoL scores and ΔFNA, ΔAA, and ΔCA was analysed by the Pearson correlation test. At baseline, in Group-A, there was a significant difference between arthritic FNA and normal hip FNA, while no differences were found in AA between the two hips. A close correlation was observed between ΔFNA and Spine-VAS (r = 0.788), ODI (r = 0.824), and RM (r = 0.775). In Group-B, there was not a significant difference in FNA and AA between the two hips. At recruitment, in Group-A patients, we recorded a higher LL, SS, PI, SVA(C7), and a lower PT and T1-SPI compared with Group-B subjects. Six months after THR, in Group-A, an improvement of all clinical scores was recorded, as well as, a significant reduction of SS, LL, T1PA, and SVA(C7) and an increment of PT. In Group-B, at follow-up, an improvement of HHS, Hip-VAS, and SF-36 was recorded, while the changes in spinopelvic parameters were not significant. Patients with concomitant unilateral HOA and LBP showed a marked anteverted FNA in the arthritic hip and a spinopelvic misalignment. After THR, a relief of both hip and low back pain and a change in spinopelvic parameters is observed.


[Smoking in a psychiatric population].
The relationship between tobacco consumption and psychiatric disorders is a widely discussed topic that is generating a considerable yet controversial bibliography, that can be approached from different points of view. The authors make a revision and critical evaluation about smoking prevalence on psychiatric population, in the different psychiatric disorders and discussing the theories implicated.


Cardiovascular risk factors, radiation therapy, and myocardial infarction among lymphoma survivors.
Mediastinal radiation is associated with increased risk of myocardial infarction (MI) among non-Hodgkin lymphoma (NHL) survivors. To evaluate how preexisting cardiovascular risk factors (CVRFs) modify the association of mediastinal radiation and MI among a national population of NHL survivors with a range of CVRFs. Using Danish registries, we identified adults diagnosed with lymphoma 2000-2010. We assessed MI from one year after diagnosis through 2016. We ascertained CVRFs (hypertension, dyslipidemia, and diabetes), vascular disease, and intrinsic heart disease prevalent at lymphoma diagnosis. We used multivariable Cox regression to test the interaction between preexisting CVRFs and receipt of mediastinal radiation on subsequent MI. Among 3151 NHL survivors (median age 63, median follow-up 6.5 years), 96 were diagnosed with MI. Before lymphoma, 32% of survivors had ≥1 CVRF. 8.5% of survivors received mediastinal radiation. In multivariable analysis, we found that mediastinal radiation (HR = 1.96; 95% CI = 1.09-3.52), and presence of ≥1 CVRF (HR = 2.71; 95% CI = 1.77-4.15) were associated with an increased risk of MI. Although there was no interaction on the relative scale (p = 0.14), we saw a clinically relevant absolute increase in risk for patients with CVRF from 10-year of MI of 10.5% without radiation to 29.5% for those undergoing radiation. Patients with CVRFs have an importantly higher risk of subsequent MI if they have mediastinal radiation. Routine evaluation of CVRFs and optimal treatment of preexisting cardiovascular disease should continue after receiving cancer therapy. In patients with CVRFs, mediastinal radiation should only be given if oncologic benefit clearly outweighs cardiovascular harm.


In vitro inhibition of HBV replication by a novel compound, GLS4, and its efficacy against adefovir-dipivoxil-resistant HBV mutations.
HBV infection continues to be an important worldwide cause of morbidity and mortality. Patients with chronic hepatitis B can be successfully treated using nucleoside/nucleotide analogues. However, drug-resistant HBV mutants frequently arise, leading to treatment failure and progression to liver disease. Here, we report the effects of GLS4, a non-nucleosidic inhibitor that exhibits a novel and highly specific anti-HBV activity. The median inhibitory concentrations (IC(50)s) of GLS4 on HBV were measured by Southern blotting. HBV capsid and core protein levels were detected by immunoblotting. To determine the antiviral activity of GLS4 against adefovir dipivoxil (ADV)-resistant HBV mutants, HepG2 cells transiently transfected with PUC-HBV1.2 plasmids that contained one of three major ADV-resistant mutations (rtA181T, rtA181V and rtN236T) were treated with GLS4. Intracellular HBV replicative intermediates were detected by Southern blotting. The effect on the in vitro assembly of HBV capsid protein was examined using dynamic light scattering and electron microscopy. The IC(50) of GLS4 was 0.012 μM, which is significantly lower than that of lamivudine (0.325 μM). Immunoblot analysis of HepG2.2.15 cells and transiently transfected HepG2 cells indicated that GLS4 treatment interfered with the formation of core particles (assembly). The ADV-resistant HBV mutant strains were also sensitive to GLS4. Upon examining the in vitro assembly of HBV core protein 149 by electron microscopy, increased aberrant particles were observed after GLS4 treatment. GLS4 is a new and unique potential anti-HBV agent that possesses a different mechanism of action than existing therapeutic drugs.


The Composition of Emissions from Sanding Corian® with Different Sandpapers.
Laboratory tests were conducted to characterize the composition of emissions from sanding Corian®, a solid-surface composite material mainly composed of alumina trihydrate (ATH) and acrylic polymer. Three sandpaper materials (ceramic, silicon carbide, and aluminum oxide) were tested to distinguish the contribution of aluminum-containing dust in the emission from Corian® and sandpaper itself. The result can help identify the main cause of the pulmonary fibrosis from exposure to aluminum-containing dust while sanding Corian®. Airborne dust samples were measured using direct-reading instruments and collected using a Micro-Orifice Uniform Deposit Impactor (MOUDI) for estimating the normalized dust generation rate. The size-classified dust samples from MOUDI were analyzed for elemental aluminum content. Additionally, air samples were analyzed for characterizing methyl methacrylate (MMA). The results from the direct-reading instruments reveal that the size distribution of particulate from sanding Corian® differs from that of sawing Corian®, showing that the size distribution of dust is affected by the fabrication process. The normalized respirable dust generation rate indicates that more respirable dust was generated during sanding Corian® board. However, the use of aluminum oxide sandpaper does not result in a higher aluminum content in the respirable dust from sanding Corian®, suggesting that the aluminum content of the respirable dust is primarily originated from Corian® itself. The generation rates of MMA from sanding did not vary much among all types of sandpapers, and they were much lower than that of sawing, likely due to the higher temperature in the sawing process.


Antidepressants escitalopram and venlafaxine up-regulate BDNF promoter IV but down-regulate neurite outgrowth in differentiating SH-SY5Y neurons.
Antidepressants are used to treat depression and some anxiety disorders, including use in pregnant patients. The pharmacological actions of these drugs generally determine the uptake and metabolism of a series of neurotransmitters, such as serotonin, norepinephrine, or dopamine, along with an increase in BDNF expression. However, many aspects of antidepressant action remain unknown, particularly whether antidepressants interfere with normal neurodevelopment when taken by pregnant women. In order to reveal cellular and molecular implications crucial to the functioning of pathways related to antidepressant effects, we performed an investigation on neuronally differentiating human SH-SY5Y cells. To our knowledge, this is the first time human SH-SY5Y cells in cultures of purely neuronal cells induced by controlled differentiation with retinoic acid are followed by short-term 48-h exposure to 0.1-10 μM escitalopram or venlafaxine. Treatment with antidepressants (1 μM) did not affect the electrophysiological properties of SH-SY5Y cells. However, the percentage of mature neurons exhibiting voltage-gated sodium currents was substantially higher in cultures pre-treated with either antidepressant. After exposure to escitalopram or venlafaxine, we observed a concentration-dependent increase in activity-dependent BDNF promoter IV activation. The assessment of neurite metrics showed significant down-regulation of neurite outgrowth upon exposure to venlafaxine. Identified changes may represent links to molecular processes of importance to depression and be involved in neurodevelopmental alterations observed in postpartum children exposed to antidepressants antenatally.


Glucagon-like peptide I stimulates insulin gene expression and increases cyclic AMP levels in a rat islet cell line.
Insulin secretion is controlled by a complex set of factors. Although blood glucose levels serve as the major stimulus of insulin secretion in mammals, insulin release is also modulated by amino acids, catecholamines, glucagon, and other, intestinal hormones. The identification of factors that modulate insulin production has engendered much interest because of their potential importance in the altered dynamics of insulin secretion in response to glucose characteristic of maturity-onset diabetes mellitus. Decoding of the glucagon gene has uncovered two additional glucagon-like peptides encoded in proglucagon, the polypeptide precursor of glucagon. One of these peptides, glucagon-like peptide I, is processed from proglucagon in two forms, of 31 and 37 amino acids. We report that the smaller of the two glucagon-like peptides potently increases cAMP levels, insulin mRNA transcripts, and insulin release in cultured rat insulinoma cells. These results indicate that glucagon-like peptide I may be a physiologic modulator of insulin gene expression.


Antimicrobial guidelines in clinical practice: incorporating the ethical perspective.
Guidelines on antimicrobial therapy are subject to periodic revision to anticipate changes in the epidemiology of antimicrobial resistance and new scientific knowledge. Changing a policy to a broader spectrum has important consequences on both the individual patient level (e.g. effectiveness, toxicity) and population level (e.g. emerging resistance, costs). By combining both clinical data evaluation and an ethical analysis, we aim to propose a comprehensive framework to guide antibiotic policy dilemmas. A preliminary framework for decision-making on antimicrobial policy was constructed based on existing literature and panel discussions. Antibiotic policy themes were translated into specific elements that were fitted into this framework. The adapted framework was evaluated in two moral deliberation groups. The moral deliberation sessions were analysed using ATLAS.ti statistical software to categorize arguments and evaluate completeness of the final framework. The final framework outlines the process of data evaluation, ethical deliberation and decision-making. The first phase is a factual data exploration. In the second phase, perspectives are weighed and the policy of moral preference is formulated. Judgments are made on three levels: the individual patient, the patient population and society. In the final phase, feasibility, implementation and re-evaluation are addressed. The proposed framework facilitates decision-making on antibiotic policy by structuring existing data, identifying knowledge gaps, explicating ethical considerations and balancing interests of the individual and current and future generations.


Genome-wide meta-analysis of QTL for morphological related traits of flag leaf in bread wheat.
Flag leaf is an important organ for photosynthesis of wheat plants, and a key factor affecting wheat yield. In this study, quantitative trait loci (QTL) for flag leaf morphological traits in wheat reported since 2010 were collected to investigate the genetic mechanism of these traits. Integration of 304 QTLs from various mapping populations into a high-density consensus map composed of various types of molecular markers as well as QTL meta-analysis discovered 55 meta-QTLs (MQTL) controlling morphological traits of flag leaves, of which 10 MQTLs were confirmed by GWAS. Four high-confidence MQTLs (MQTL-1, MQTL-11, MQTL-13, and MQTL-52) were screened out from 55 MQTLs, with an average confidence interval of 0.82 cM and a physical distance of 9.4 Mb, according to the definition of hcMQTL. Ten wheat orthologs from rice (7) and Arabidopsis (3) that regulated leaf angle, development and morphogenesis traits were identified in the hcMQTL region using comparative genomics, and were speculated to be potential candidate genes regulating flag leaf morphological traits in wheat. The results from this study provides valuable information for fine mapping and molecular markers assisted selection to improve morphological characters in wheat flag leaf.


Attenuation of allergen-induced airway hyperresponsiveness is mediated by airway regulatory T cells.
Understanding the mechanisms involved in respiratory tolerance to inhaled allergens could potentially result in improved therapies for asthma and allergic diseases. Airway hyperresponsiveness (AHR) is a major feature of allergic asthma, thus the aim of the current study was to investigate mechanisms underlying suppression of allergen-induced AHR during chronic allergen exposure. Adult BALB/c mice were systemically sensitized with ovalbumin (OVA) in adjuvant and then challenged with a single 3 or 6 wk of OVA aerosols. Airway and parenchymal responses to inhaled methacholine (MCh), inflammatory cell counts, cytokines, OVA-specific IgE and IgG(1), parenchymal histology, and numbers of airway CD4(+)69(+) activated and CD4(+)25(+)FoxP3(+) regulatory T (Treg) cells were assessed 24 h after the final aerosol. Single OVA challenge resulted in AHR, eosinophilia, increased serum OVA-specific IgE, and T helper 2 (Th2) cytokines in bronchoalveolar lavage (BAL) but no difference in numbers of Treg compared with control mice. Three weeks of OVA challenges resulted in suppression of AHR and greater numbers of airway Treg cells and increased transforming growth factor-beta(1) (TGFbeta(1)) compared with control mice despite the presence of increased eosinophilia, OVA-specific IgE and IgG(1), and airway remodeling. Six weeks of OVA challenges restored AHR, whereas airway Treg numbers, TGFbeta(1), BAL eosinophilia, and Th2 cytokines returned to control levels. Partial in vivo depletion or adoptive transfer of Treg cells restored or inhibited AHR, respectively, but did not affect TGFbeta(1) or Th2 cytokine production. In conclusion, AHR suppression is mediated by airway Treg cells and potentially via a paracrine induction of TGFbeta(1) in the airways.


[Research progress on pesticide residues of Panacis Quinquefolii Radix].
Panacis Quinquefolii Radix is the dry root of Panax quinquefolium, which is a perennial plant of Araliaceae. The plant has a long growth cycle and serious growth barrier problem, which leads to the use of pesticides. As a result, the pesticide residues in Panacis Quinquefolii Radix are arousing great concern. This paper reviews the research findings on the investigation, detection methods, content analysis and risk assessment of pesticide residues in Panacis Quinquefolii Radix since 1993, and compares the pesticide residue limit standards of different countries and regions. The pesticide residues in Panacis Quinquefolii Radix have been changing from organochlorines with high toxicity to triazines and triazoles with low toxicity. The pesticide residues are generally low, while the pollution of pentachloronitrobenzene and other pesticides still exist. The detection method has evolved from chromatography to chromatography-mass spectrometry. There are no reports of health risks caused by pesticide residues of Panacis Quinquefolii Radix. Pesticide residue is a major factor restricting the sound development of Panacis Quinquefolii Radix industry in China. Therefore, we suggest to improve the registration of pesticides applied to the plant, popularize mature ecological planting mode and supporting technology, and strengthen the research on the risk assessment and limit standard of pesticide residue in Panacis Quinquefolii Radix.


Specific dermatoses of pregnancy: a review.
Some dermatoses worsen during pregnancy, some improve, yet others have unpredictable course. To conduct evidence-based search, and review of current management of specific dermatoses of pregnancy. Comprehensive literature search was conducted, with Medline and Cochrane Database regarding skin diseases in pregnancy from 1990- 2005. International pharmaceutical abstracts science search (1997- 2003) was used for search references found in the articles. All articles selected for inclusion in this review were evaluated critically with regards to their impact factor, source, and evidence based contribution on this topic as measured by their citation index and the journals they were published in. This review was limited to specific dermatoses of pregnancy generally and some of the skin disorders modified by pregnancy. Intrahepatic cholestasis of pregnancy should be managed as high risk pregnancies as several investigations have shown foetal complications. Recent randomised trials have demonstrated beneficial effects of ursodeoxycholic acid (UDCA) in intrahepatic cholestasis of pregnancy (ICP). Pruritice ruption of pregnancy is associated with multiple pregnancies. It has variable clinical features and has to be differentiated from pemphigoid gestationis, which is associated with an increased incidence of both prematurity and small for date's babies. Prurigo of pregnancy is extremely itchy with papules appearing on the extensor surfaces of the limbs and trunk. It has no maternal risk. The eruptions in pruritic folliculitis of pregnancy clear spontaneously in the postpartum period, with no associated morbidity either in the mother or baby. Some skin diseases like obstetric cholestasis may have adverse foetal outcome, while other disorders like pruritic folliculitis of pregnancy have no significant effect on either the mother or baby.


Encephalocraniocutaneous lipomatosis: a recently described neurocutaneous syndrome.
The authors report a 1-year-old Japanese boy, who was noted to have unilateral alopecia and a cloudy cornea since birth. Additional features included mild spasticity and mental retardation, DQ being 70. A non-communicating cystic lesion was found in the right middle cerebral fossa by metrizamide CT scan, and resection of the lesion by craniotomy revealed an arachnoid cyst. The case was diagnosed as encephalocraniocutaneous lipomatosis, a recently described neurocutaneous syndrome.


Peripherally inserted central catheters.
Peripherally inserted central catheters (PICCs) can meet the needs of patients requiring short- to intermediate-term intravenous therapy in a variety of settings. PICCs minimise insertion and catheter-related complications when compared to other central venous access devices (CVADs). PICCs have been introduced in the UK mainly through innovative advanced nursing practice.


Congenital disorders of glycosylation (CDG): state of the art in 2022.
Congenital disorders of glycosylation (CDG) are a complex and heterogeneous family of rare metabolic diseases. With a clinical history that dates back over 40 years, it was the recent multi-omics advances that mainly contributed to the fast-paced and encouraging developments in the field. However, much remains to be understood, with targeted therapies' discovery and approval being the most urgent unmet need. In this paper, we present the 2022 state of the art of CDG, including glycosylation pathways, phenotypes, genotypes, inheritance patterns, biomarkers, disease models, and treatments. In light of our current knowledge, it is not always clear whether a specific disease should be classified as a CDG. This can create ambiguity among professionals leading to confusion and misguidance, consequently affecting the patients and their families. This review aims to provide the CDG community with a comprehensive overview of the recent progress made in this field.


COVID-19 in patients with autoimmune diseases: characteristics and outcomes in a multinational network of cohorts across three countries.
Patients with autoimmune diseases were advised to shield to avoid coronavirus disease 2019 (COVID-19), but information on their prognosis is lacking. We characterized 30-day outcomes and mortality after hospitalization with COVID-19 among patients with prevalent autoimmune diseases, and compared outcomes after hospital admissions among similar patients with seasonal influenza. A multinational network cohort study was conducted using electronic health records data from Columbia University Irving Medical Center [USA, Optum (USA), Department of Veterans Affairs (USA), Information System for Research in Primary Care-Hospitalization Linked Data (Spain) and claims data from IQVIA Open Claims (USA) and Health Insurance and Review Assessment (South Korea). All patients with prevalent autoimmune diseases, diagnosed and/or hospitalized between January and June 2020 with COVID-19, and similar patients hospitalized with influenza in 2017-18 were included. Outcomes were death and complications within 30 days of hospitalization. We studied 133 589 patients diagnosed and 48 418 hospitalized with COVID-19 with prevalent autoimmune diseases. Most patients were female, aged ≥50 years with previous comorbidities. The prevalence of hypertension (45.5-93.2%), chronic kidney disease (14.0-52.7%) and heart disease (29.0-83.8%) was higher in hospitalized vs diagnosed patients with COVID-19. Compared with 70 660 hospitalized with influenza, those admitted with COVID-19 had more respiratory complications including pneumonia and acute respiratory distress syndrome, and higher 30-day mortality (2.2-4.3% vs 6.32-24.6%). Compared with influenza, COVID-19 is a more severe disease, leading to more complications and higher mortality.


Glutathione S-transferase M1 and T1 Polymorphisms, Cigarette Smoking and HPV Infection in Precancerous and Cancerous Lesions of the Uterine Cervix.
Glutathione S-transferases (GSTs) play an important role in detoxification of carcinogenic electrophiles. The null genotypes in GSTM1 and GSTT1 have been implicated in carcinogenesis. Present study was planned to evaluate the influence of genetic polymorphisms of GSTM1 and GSTT1 gene loci in cervical carcinogenesis. The study was conducted in Lok Nayak hospital, New Delhi. DNA from clinical scrapes of 482 women with minor gynaecologic complaints attending Gynaecology OPD and tumor biopsies of 135 cervical cancer cases attending the cancer clinic was extracted. HPV DNA was detected by standard polymerase chain reaction (PCR) using L1 consensus primer pair. Polymorphisms of GSTM1 and GSTT1 were analysed by multiplex PCR procedures. Differences in proportions were tested using Pearson's Chi-square test with Odds ratio (OR) and 95% confidence interval (CI). The risk of cervical cancer was almost three times in women with GSTM1 homozygous null genotype (OR-2.62, 95%CI, 1.77-3.88; p<0.0001). No association of GSTM1 or GSTT1 homozygous null genotypes was observed in women with normal, precancerous and cervical cancerous lesions among ≤35 or >35 years of age groups. Smokers with null GSTT1 genotype had a higher risk of cervical cancer as compared to non-smokers (OR-3.01, 95% CI, 1.10-8.23; p=0.03). The results further showed that a significant increased risk of cervical cancer was observed in HPV positive smoker women with GSTT1 (OR-4.36, 95% CI, 1.27-15.03; p=0.02) and GSTM1T1 (OR-3.87, 95% CI, 1.05-14.23; p=0.04) homozygous null genotypes as compared to HPV positive non smokers. The results demonstrate that the GST null genotypes were alone not associated with the development of cervical cancer, but interacted with smoking and HPV to exert effects in our Delhi population.


[Pathologic change of elastic fibers with difference of microvessel density and expression of angiogenesis-related proteins in internal hemorrhoid tissues].
To investigate the pathological variations in internal hemorrhoid and evaluate the expression of nitric- oxide synthase(NOS),vascular endothelial growth factor(VEGF),matrix metalloproteinase- 2(MMP2) and MMP9. Normal anal cushion and internal hemorrhoids tissue samples were obtained from 24 patients with iii degree hemorrhoids after procedure for prolapse and hemorrhoids(PPH) procedure. The expression of NOS, VEGF, MMP2, MMP9 and CD34 were detected by immunohistochemical staining; the microvessel density (MVD) was counted by anti- CD34 antibody; the elastic fibers were detected by orcein staining. There were statistically significant differences in the expression of MVD, VEGF, MMP9 between internal hemorrhoid tissue and normal anal cushions(P< 0.05). iNOS was significantly increased in hemorrhoid tissue, but no significant difference between normal anal cushions and hemorrhoid tissue. Morphological abnormalities such as breaking, distortion, mortality, hyaline degeneration were found in elastic fibers of internal hemorrhoid tissue, but not in normal anal cushions. Angiogenesis is evident in hemorrhoid tissue, suggesting the possible mechanism in the pathogenesis of hemorrhoids. The direct degeneration effect of MMP9 on supporting structure elastic fibers in anal cushion is another important mechanism. The high expression of iNOS suggests the inflammatory factors involve in the pathogenesis of hemorrhoids, and NO may be involve in pathological effect on hemorrhoids.


Long-chain inulin increases dendritic cells in the Peyer's patches and increases ex vivo cytokine secretion in the spleen and mesenteric lymph nodes of growing female rats, independent of zinc status.
Prebiotics may increase Zn absorption, a mineral known to play a central role in the immune system. Zn-deficient states are characterised by suppressed immune function, while prebiotics may improve both gut and cell-mediated immunity. Our objective was to determine if inulin alters the number and proportion of immune cells in the spleen, mesenteric lymph nodes (MLN) and Peyer's patches (PP), ex vivo cytokine secretion, intestinal permeability and Zn status in healthy as well as Zn-deficient rats. Weanling female rats were fed diets supplemented with 5 % cellulose (CEL) or 5 % inulin (PRE) for 4 weeks. The rats received the CEL or PRE diet ad libitum (ZN) or in restricted amounts (DR), or deficient in Zn (ZD) for another 4 weeks. The PRE-fed rats had a higher number and proportion of dendritic cells in PP, and greater ex vivo secretion of IL-2, IL-10 and interferon-gamma from spleen and MLN cells compared with CEL-fed rats. PRE reduced the number and proportion of T cell receptor (TCR)-alphabeta+CD8+ cells in spleen and CD45RA+ cells in MLN compared with CEL. ZD rats had lower serum IgG2a and T cell numbers in MLN compared with ZN and DR rats. TCRgammadelta+ cell numbers in PP were higher in ZD-PRE rats compared with ZD-CEL rats. Femur Zn concentrations of DR-PRE rats were higher than those of DR-CEL rats. Intestinal permeability was unchanged. The higher proportion and number of dendritic cells in the PP of inulin-fed rats indicates a need for further research on how prebiotics and their metabolites affect immune function possibly through intestinal dendritic cells.


Successful treatment of perifolliculitis capitis abscedens et suffodiens with combined isotretinoin and dapsone.
Perifolliculitis capitis abscedens et suffodiens (PCAS) is a rare scalp disease of unknown etiology which is hard to treat. It is often accompanied by scarring alopecia, acne conglobata, and recurrent fluctuant abscesses. PCAS belongs to the family of acne inversa (hidradenitis suppurativa). A 19-year-old man presented with PCAS for 2 years; multiple systemic antibiotic therapies and surgical approaches had shown no effect. Monotherapy with isotretinoin 80 mg daily for 4 weeks had not been successful. Combination therapy with dapsone 100 mg and isotretinoin 80 mg daily produced significant improvement. During 4 weeks of treatment significant clearing was achieved. Dapsone was reduced to 50 mg daily after 6 months, while isotretinoin was discontinued gradually. Now the patient is on dapsone 50 mg every other day and has remained free of recurrences for 6 months.


Education for women undergoing HPV testing.
HPV is the main precursor to cervical cancer and is sexually transmitted. This may have psychological implications for women undergoing testing for the virus. A sensitive approach to the provision of information is required to minimise fear and stigmatisation.


Disulfide stress and its targets in acute pancreatitis.
Under physiological conditions, the balance between ROS production and removal properly maintains the intracellular redox-sensitive signaling as well as the appropriate status of protein thiols and disulfides. However, inflammation among other factors can modify this balance causing a rapid increase in intracellular ROS levels and hence thiol oxidation, eventually leading to oxidative stress. In the case of acute pancreatitis, both redox signaling and oxidative stress seem to contribute to the progression of the severe form of the disease. In this review we will focus on the reversible oxidation of protein cysteines during the course of acute pancreatitis. We describe disulfide stress in an acute inflammatory process, which is characterized by thiol oxidation in proteins, particularly protein cysteinylation, without significant changes in the glutathione redox status.


Thinking nuclear medicine--PET activation.
PET activation, although restricted to a limited number of research centers, is currently the gold standard for mapping functional areas of the brain. This paper outlines how and why activation studies are performed and reviews the major uses of this technique. Special emphasis is on cognitive neuropsychology including the results of a project designed to map the areas of the brain responsible for controlling automatic counting and simple calculation. These fascinating studies are, by definition, a part of nuclear medicine. As such, we argue that knowledge of PET activation is an essential component of professional development and that, given opportunity, commitment and the will to learn, nuclear medicine technologists have the potential for involvement, collaboration or leadership in this area of research.


Impact of molecular residual disease post allografting in myelofibrosis patients.
We screened 136 patients with myelofibrosis and a median age of 58 years who underwent allogeneic stem cell transplantation (AHSCT) for molecular residual disease for JAKV617F (n=101), thrombopoietin receptor gene (MPL) (n=4) or calreticulin (CALR) (n=31) mutation in peripheral blood on day +100 and +180 after AHSCT. After a median follow-up of 78 months, the 5-year estimated overall survival was 60% (95% confidence interval (CI): 50-70%) and the cumulative incidence of relapse at 5 years was 26% (95% CI: 18-34%) for the entire study population. The percentage of molecular clearance on day 100 was higher in CALR-mutated patients (92%) in comparison with MPL- (75%) and JAKV617F-mutated patients (67%). Patients with detectable mutation at day +100 or at day +180 had a significant higher risk of clinical relapse at 5 years than molecular-negative patients (62% vs 10%, P<0.001) and 70% vs 10%, P<0.001, respectively) irrespectively of the underlying mutation. In a multivariate analysis, high-risk diseases status (hazard ratio (HR) 2.5; 95% CI: 1.18-5.25, P=0.016) and detectable MRD at day 180 (HR 8.36, 95% CI: 2.76-25.30, P<0.001) were significant factors for a higher risk of relapse.


ECG-related fasciculation potential.
We report the observation of an unusual fasciculation potential (FP). This FP was time-locked to the electrocardiogram (ECG) in 87% of the discharges. The motor unit action potential (MUAP) of the FP was also elicited as H-reflex. As mechanism we propose spindle afferent discharges mechanically triggered by the arterial pulse, i.e. a reflex fasciculation. We suggest that such a heartbeat-related mechanism could explain synchronous fasciculations described by Norris (Arch. Neurol., 1965, 13: 495-500).


Permeability of surface-modified polyamidoamine (PAMAM) dendrimers across Caco-2 cell monolayers.
Aim of this study was to prepare polyamine-conjugated PAMAM dendrimers and study their permeability across Caco-2 cell monolayers. Polyamines, namely, arginine and ornithine were conjugated to the amine terminals of the G4 PAMAM dendrimers by Fmoc synthesis. The apical-to-basolateral (AB) and basolateral-to-apical (BA) apparent permeability coefficients (P(app)) for the PAMAM dendrimers increased by conjugating the dendrimers with both of the polyamines. The enhancement in permeability was dependent on the dendrimer concentration and duration of incubation. The correlation between monolayer permeability and the decrease in transepithelial electrical resistance (TEER) with both the PAMAM dendrimers and the polyamine-conjugated dendrimers suggests that paracellular transport is one of the mechanisms of transport across the epithelial cells. Cytotoxicity of the polyamine-conjugated dendrimers was evaluated in Caco-2 cells by MTT (methylthiazoletetrazolium) assay. Arginine-conjugated dendrimers were slightly more toxic than PAMAM dendrimer as well as ornithine-conjugated dendrimers. Though investigations on the possible involvement of other transport mechanisms are in progress, results of the present study suggest the potential of dendrimer-polyamine conjugates as drug carriers to increase the oral absorption of drugs.


E. coli DNA Polymerase I and the Klenow Fragment.
Escherichia coli DNA Pol I can carry out three enzymatic reactions: It possesses 5' → 3' DNA polymerase activity and 3' → 5' and 5' → 3' exonuclease activity. Pol I can be cleaved by mild treatment with subtilisin into two fragments; the larger fragment is known as the Klenow fragment. The Klenow fragment retains the polymerizing activity and the 3' → 5' exonuclease of the holo-enzyme but lacks its powerful 5' → 3' exonuclease activity. These enzymes and their applications in molecular cloning are introduced here.


Are cognitive control and stimulus-driven processes differentially linked to inattention and hyperactivity in preschoolers?
Temperament and attention-deficit/hyperactivity disorder (ADHD) are both typically viewed as biologically based behavioural constructs. There is substantial overlap between ADHD symptoms and specific temperamental traits, such as effortful control, especially in young children. Recent work by Martel and colleagues ( 2009 , 2011 ) suggests that cognitive control temperamental processes are more closely related to inattention symptoms, whereas stimulus-driven temperamental processes are linked to hyperactivity-impulsivity. The present study tested a model of temperament and ADHD symptoms in typically developing preschoolers and those at risk for ADHD using structural equation modelling. Data were from larger study on ADHD in a short-term longitudinal sample with parent/teacher reports and neurocognitive testing. Participants included 214 preschool children (72.9% male) from diverse ethnic/racial backgrounds and a wide range of socioeconomic status from a large metropolitan center. Cognitive control processes, such as effortful control, but not stimulus-driven processes, are related to inattention and hyperactivity. In contrast, stimulus-driven processes, such as emotional reactivity, were related only to hyperactivity symptoms longitudinally. These results suggest that early temperament behaviours and cognitive processes may be indicators of later childhood behavioural difficulties with lasting consequences.


Phylogenetically Structured Differences in rRNA Gene Sequence Variation among Species of Arbuscular Mycorrhizal Fungi and Their Implications for Sequence Clustering.
Arbuscular mycorrhizal (AM) fungi form mutualisms with plant roots that increase plant growth and shape plant communities. Each AM fungal cell contains a large amount of genetic diversity, but it is unclear if this diversity varies across evolutionary lineages. We found that sequence variation in the nuclear large-subunit (LSU) rRNA gene from 29 isolates representing 21 AM fungal species generally assorted into genus- and species-level clades, with the exception of species of the genera Claroideoglomus and Entrophospora However, there were significant differences in the levels of sequence variation across the phylogeny and between genera, indicating that it is an evolutionarily constrained trait in AM fungi. These consistent patterns of sequence variation across both phylogenetic and taxonomic groups pose challenges to interpreting operational taxonomic units (OTUs) as approximations of species-level groups of AM fungi. We demonstrate that the OTUs produced by five sequence clustering methods using 97% or equivalent sequence similarity thresholds failed to match the expected species of AM fungi, although OTUs from AbundantOTU, CD-HIT-OTU, and CROP corresponded better to species than did OTUs from mothur or UPARSE. This lack of OTU-to-species correspondence resulted both from sequences of one species being split into multiple OTUs and from sequences of multiple species being lumped into the same OTU. The OTU richness therefore will not reliably correspond to the AM fungal species richness in environmental samples. Conservatively, this error can overestimate species richness by 4-fold or underestimate richness by one-half, and the direction of this error will depend on the genera represented in the sample. Arbuscular mycorrhizal (AM) fungi form important mutualisms with the roots of most plant species. Individual AM fungi are genetically diverse, but it is unclear whether the level of this diversity differs among evolutionary lineages. We found that the amount of sequence variation in an rRNA gene that is commonly used to identify AM fungal species varied significantly between evolutionary groups that correspond to different genera, with the exception of two genera that are genetically indistinguishable from each other. When we clustered groups of similar sequences into operational taxonomic units (OTUs) using five different clustering methods, these patterns of sequence variation caused the number of OTUs to either over- or underestimate the actual number of AM fungal species, depending on the genus. Our results indicate that OTU-based inferences about AM fungal species composition from environmental sequences can be improved if they take these taxonomically structured patterns of sequence variation into account.


Topical Use of NaCl Solution with Different Concentration Affects Lens Transparency in Anesthetized Mice.
To study the influence of NaCl solution with different concentration on lens transparency in anesthetized mice. Four kinds of NaCl solution with different concentration were prepared as eye drops to imply graded osmolarity (100, 300, 500 and 1000 mOsmol/kg). Five groups of anesthetized mice were set-up to induce lens opacity, in which four groups were treated with NaCl solution and another group naturally exposed to air. The lens opacity was graded as no opacity, mild, medium and severe opacity at 0, 10, 20, 30, 45 and 60 min after the start of the experiment. A numerical value from 0 to 3 was assigned to each grade for the cataract index (CI) calculation and data analysis. The same procedure was repeated in all groups 48 h later. The reversion process of lens opacity was explored using a hypotonic NaCl solution (100 mOsmol/kg) in another pair of groups, a 500 mOsmol/kg NaCl solution group and natural exposure group. The gross appearance and time course of development and reversion of lens opacity were assessed. Lens opacity primarily developed in a hypertonic NaCl solution-treated and naturally exposed eyes, and the gross anatomical appearance were similar. The speed of lens opacity development and CI changes were osmolarity-dependent, and the higher NaCl concentration solution used, the faster and more severe the formation of opacification. Both hypertonic NaCl-solution-induced lens opacity and natural exposure induced lens opacity could be resolved by hypotonic NaCl solution prior to anesthesia recovery. This study indicates a crucial effect of NaCl concentration on the development and reversion of lens opacity in the anesthetized mice, and support the osmolarity theory in the reversible lens opacification phenomenon. It is also of practical significance to mouse eye studies that require lens transparency.


Clinical Trials and Clinical Research: A Comprehensive Review.
Clinical research is an alternative terminology used to describe medical research. Clinical research involves people, and it is generally carried out to evaluate the efficacy of a therapeutic drug, a medical/surgical procedure, or a device as a part of treatment and patient management. Moreover, any research that evaluates the aspects of a disease like the symptoms, risk factors, and pathophysiology, among others may be termed clinical research. However, clinical trials are those studies that assess the potential of a therapeutic drug/device in the management, control, and prevention of disease. In view of the increasing incidences of both communicable and non-communicable diseases, and especially after the effects that Coronavirus Disease-19 (COVID-19) had on public health worldwide, the emphasis on clinical research assumes extremely essential. The knowledge of clinical research will facilitate the discovery of drugs, devices, and vaccines, thereby improving preparedness during public health emergencies. Therefore, in this review, we comprehensively describe the critical elements of clinical research that include clinical trial phases, types, and designs of clinical trials, operations of trial, audit, and management, and ethical concerns.


Cardiovascular effects of peptide kinin B2 receptor antagonists in rats.
Bradykinin (BK) is a vasoactive peptide reputed to play an important role in cardiovascular homeostasis. In this study, we describe the cardiovascular changes (mean blood pressure (BP) and heart rate (HR)) induced by the i.v. administration (left jugular vein) of two selective kinin B2 receptor antagonist, namely icatibant (0.1-1 micromol/kg as a bolus) and MEN1 1270 (0.1-1 micromol/kg as a bolus or 1 micromol/kg infused in 15 or 60 min), in urethane-anaesthetized or conscious rats with an indwelling catheter implanted in the right carotid artery for BP measurements. In conscious rats, icatibant at 0.1 or 0.3 micromol/kg did not change BP but at 0.1 micromol/kg increased HR at 30 min from administration. MEN1 1270 at 0.1 or 0.3 micromol/kg induced a dose-related increase in BP and a concomitant bradycardia (significant at 0.3 micromol/kg) lasting for 5 or 30 min, respectively. Icatibant at 1 micromol/kg induced a slight (P < 0.05) increase in BP that resolved in 5 min and a biphasic tachycardia (peaks at 30 and 90 min from administration). MEN1 1270 at 1 micromol/kg induced a triphasic change in HR (tachycardia in the first 5 min, bradycardia at 30 min, and tachycardia at 90 and 120 min) and a biphasic change in BP (hypotension at 15 min and hypertension at 30 min). The i.v. infusion of MEN1 1270 (1 micromol/kg in 15 or 60 min) produced hypertension, whereas HR was increased only following the 15-min infusion. In urethane-anaesthetized rats, both icatibant and MEN1 1270 (0.1 micromol/kg as a bolus) increased BP and the onset for this effect was correlated with the time course of the antagonism of BK-induced hypotension, where the effect of MEN1 1270 was more rapid than that of icatibant. These results indicate that kinin B2 receptor antagonists can induce acute cardiovascular effects, and the reason for the different haemodynamic profile between icatibant and MEN1 1270 could be putatively attributed to kinetic characteristics.


Metal-support interaction in platinum and palladium nanoparticles loaded on nitrogen-doped mesoporous carbon for oxygen reduction reaction.
Mesoporous carbons are highly porous materials, which show large surface area, chemical inertness and electrochemical performances superior to traditional carbon material. In this study, we report the preparation of nitrogen-doped and undoped mesoporous carbons by an optimized hard template procedure employing silica as template, sucrose and ammonia as carbon and nitrogen source, respectively. Surface area measurements assert a value of 900 and 600 m(2) g(-1) for the best doped and undoped samples, respectively. Such supports were then thoroughly characterized by surface science and electron microscopy tools. Afterward, they were decorated with Pt and Pd nanoparticles, and it was found that the presence of nitrogen defects plays a significant role in improving the metal particles dimension and dispersion. In fact, when doped supports are used, the resulting metal nanoparticles are smaller (2-4 nm) and less prone to aggregation. Photoemission measurements give evidence of a binding energy shift, which is consistent with the presence of an electronic interaction between nitrogen atoms and the metal nanoparticles, especially in the case of Pd. The catalytic properties of electrodes decorated with such catalyst/support systems were investigated by linear sweep voltammetry and by rotating disk electrode measurements, revealing excellent stability and good activity toward oxygen reduction reaction (ORR). In particular, although Pd nanoparticles always result in lower activity than Pt ones, both Pt and Pd electrodes based on the N-doped supports show an increased activity toward ORR with respect to the undoped ones. At the same mass loading, the Tafel slope and the stability test of the Pt@N-doped electrocatalysts indicate superior performances to that of a commercial Pt@C catalysts (30 wt % Pt on Vulcan XC-72, Johnson Matthey).


Tocolysis of preterm contractions does not improve preterm delivery rate or perinatal outcomes.
To determine whether hospital admission and parenteral tocolysis of patients with threatened preterm labor results in a decreased preterm delivery rate and improved perinatal outcome, we performed a prospective cohort study comparing tocolytic treatment versus no tocolytic treatment. Women between 20 and 37 weeks' gestation who presented with at least eight uterine contractions per hour and a cervical examination <2 cm dilated, <80% effaced, and no cervical change over a 2-hr period were entered into the study. Exclusion criteria included cervical change, multiple gestation, chorioamnionitis, rupture of membranes, placenta previa or abruption, undiagnosed vaginal bleeding, and prior tocolytic therapy in the current pregnancy. Seventy-five patients were admitted to the hospital for tocolysis (tocolysis cohort) and 81 were observed and discharged to home (observation cohort). There was no significant difference in mean gestational age at delivery, mean birth weight, or in preterm delivery rate between the two cohorts. Four of the eight women in the observation cohort who delivered preterm had a history of preterm delivery. We conclude that discharging women with preterm contractions without objective evidence of labor does not result in an increase in preterm births. More importantly, tocolysis did not decrease the preterm delivery rate in this cohort.


The role of interaural differences on speech intelligibility in complex multi-talker environments.
Interaural differences in time (ITDs) and interaural differences in level (ILDs) contribute to a listener's ability to achieve spatial release from masking (SRM), and help to improve speech intelligibility in noisy environments. In this study, the extent to which ITDs and ILDs contribute to SRM and the relationships with aging and hearing loss were examined. SRM was greatest when stimuli were presented with consistent ITD and ILD, relative to ITD or ILD alone, all of which produced greater SRM than when ITD and ILD cues were in conflict with each other. This pattern was independent of age and hearing loss.


A practical overview of pediatric atopic dermatitis, part 2: triggers and grading.
In part 2 of this 3-part series on atopic dermatitis (AD) in children, triggers for the appearance and flaring of AD are reviewed. The role of AD in the atopic march is explored. Furthermore, the usage of grading systems in the development of therapeutics and in clinical care is discussed. The natural history of AD has changed from improvement to 50% persistence and therefore it is important to council guardians and patients accordingly.


The gut takes nearly all: threonine kinetics in infants.
Threonine is an essential amino acid that is abundantly present in intestinally produced glycoproteins. Animal studies show that intestinal first-pass threonine metabolism is high, particularly during a restricted enteral protein intake. The objective of the study was to quantify intestinal first-pass threonine metabolism in preterm infants during full enteral feeding and during restricted enteral intake. Eight preterm infants (x +/- SD birth weight: 1.1 +/- 0.1 kg; gestational age: 29 +/- 2 wk) were studied during 2 periods. During period A, 40% of total intake was administered enterally and 60% was administered parenterally. Total threonine intake was 58 +/- 6 micromol kg(-1) h(-1). During period B, the infants received full enteral feeding, and the total threonine intake was 63 +/- 6 micromol kg(-1) h(-1). Dual stable-isotope tracer techniques were used to assess splanchnic and whole-body threonine kinetics. The fractional first-pass threonine uptake by the intestine was remarkably high in both periods: 82 +/- 6% during partial enteral feeding and 70 +/- 6% during full enteral feeding. Net threonine retention was not affected by the route of feeding. In preterm infants, the splanchnic tissues extract a very large amount of the dietary threonine intake, which indicates a high obligatory visceral need for threonine, presumably for the purposes of synthesis.


Multiple post-translational modifications affect heterologous protein synthesis.
Post-translational modifications (PTMs) are required for proper folding of many proteins. The low capacity for PTMs hinders the production of heterologous proteins in the widely used prokaryotic systems of protein synthesis. Until now, a systematic and comprehensive study concerning the specific effects of individual PTMs on heterologous protein synthesis has not been presented. To address this issue, we expressed 1488 human proteins and their domains in a bacterial cell-free system, and we examined the correlation of the expression yields with the presence of multiple PTM sites bioinformatically predicted in these proteins. This approach revealed a number of previously unknown statistically significant correlations. Prediction of some PTMs, such as myristoylation, glycosylation, palmitoylation, and disulfide bond formation, was found to significantly worsen protein amenability to soluble expression. The presence of other PTMs, such as aspartyl hydroxylation, C-terminal amidation, and Tyr sulfation, did not correlate with the yield of heterologous protein expression. Surprisingly, the predicted presence of several PTMs, such as phosphorylation, ubiquitination, SUMOylation, and prenylation, was associated with the increased production of properly folded soluble proteins. The plausible rationales for the existence of the observed correlations are presented. Our findings suggest that identification of potential PTMs in polypeptide sequences can be of practical use for predicting expression success and optimizing heterologous protein synthesis. In sum, this study provides the most compelling evidence so far for the role of multiple PTMs in the stability and solubility of heterologously expressed recombinant proteins.


[The importance of nutrition in inflammatory bowel disease in children].
Many children with inflammatory bowel disease, mainly Crohn's disease, suffer from malnutrition and growth retardation. Maintenance of adequate nutrition is extremely important in the management of pediatric patients with Crohn's disease. This can usually be achieved by exclusive elemental formulae and polymeric formulae and rarely necessitates the use of parenteral nutrition. This review presents an update on the nutritional aspects of inflammatory bowel disease in children.


Metabolite amyloids: a new paradigm for inborn error of metabolism disorders.
The formation of ordered amyloid assemblies is associated with major human degenerative disorders, including Alzheimer's disease, Parkinson's disease, and type 2 diabetes. Amyloid fibrils are elongated nanoscale structures that bind to specific dyes (including thioflavin T and Congo red). Amyloid fibrillar assemblies or their early intermediates are known to induce apoptotic cytotoxic effect. Until recently, amyloid fiber formation was observed only with proteins and peptides. We reported in 2012 that a single amino acid, phenylalanine, could also form typical amyloid fibrils with the same morphology, dye-binding specificity, and electron diffraction pattern as protein amyloids. X-ray crystallography demonstrated the formation of supramolecular β-sheet-like organization by phenylalanine at its zwitterionic form. Metabolite amyloids had pronounced cytotoxicity that could be depleted by treatment with antibodies raised against the phenylalanine structures. We suggested that the observed amyloid formation could explain some of the symptoms observed in phenylketonuria (PKU) upon the accumulation of phenylalanine. Follow-up studies by other groups revealed the ability of phenylalanine amyloids to bind to membranes, as observed with protein amyloids. Furthermore, the doxycycline amyloid formation inhibitor was shown to also affect the formation of phenylalanine amyloids. In 2015, it was reported that other metabolites involved in metabolic disorders, including adenine, uracil, tyrosine, and orotic acid, could form amyloid-like assemblies. It was further demonstrated that the assemblies induce apoptotic cell death. Taken together, we suggest a new hypothesis to understand the etiology of degenerative processes observed in inborn error of metabolism disorders and indicate new avenues for treatment.


Resonant escape over an oscillating barrier in underdamped Josephson tunnel junctions.
The escape from a metastable state over an oscillating barrier of an underdamped Josephson tunnel junction has been experimentally investigated with oscillation frequency well separated from the plasma frequency of the junction. The resonant escape, namely, a minimum of the average escape time as a function of the oscillation frequency, was observed. For the oscillation frequency much smaller than the "resonant frequency," the average escape time is the average of the times required to cross over each of the barriers. On the other hand, for the oscillation frequency much greater than the "resonant frequency," the average escape time is that required to cross the average barrier.


A Holistic and Community-Based Service Program Promoting Mental Health among Latina Breast Cancer Survivors and Informal Caregivers.
This report discusses the components and uptake of a holistic-health community-based service program designed to promote mental health among Latina breast cancer survivors and their caregivers. Salud Holística para la Mujer's (SAHOM) strategic plan tackled all phases of the mental health prevention continuum and of the cancer care continuum.


Intercellular adhesion molecule-1 (ICAM-1) in ulcerative colitis: presence, visualization, and significance.
The intensive research of recent years has suggested that the cause of ulcerative colitis (UC) involves a genetic predisposition to an uncontrolled or unbalanced immune response to luminal or epithelial antigens or against other external factors. Intercellular adhesion molecule-1 (ICAM-1) is pivotal for the influx of neutrophil granulocytes into colonic mucosa, and gene analyses have found polymorphisms in the gene encoding ICAM-1, indicating that changes in ICAM-1 function may be involved in the pathogenesis of UC. Clinical trials of the ICAM-1 antisense oligonucleotide Alicaforsen, which inhibits the synthesis of ICAM-1, have shown positive results in the treatment of patients with left-sided (distal) UC. In addition to emphasizing the central role of ICAM-1 in active stages of UC, the results provide hope for the development and introduction of a more specific and efficient treatment for UC than those currently available. This review discusses the results from studies on the expression of ICAM-1 in colonic tissue from patients with UC.


Mean nuclear area of fine needle aspirates of primary preoperative palpable breast carcinoma using image cytometry.
Early, operable breast cancers in appropriate patients are increasingly being treated preoperatively using neoadjuvant chemotherapy. A good response rate is seen with high grade tumors. Nuclear size, which may reflect the grade of the tumor, is also of possible prognostic value in breast cancer. We measured the mean nuclear area (MNA) of 114 consecutive preoperative fine needle aspirates of palpable, operable breast cancers. We used computerized image cytometry to measure nuclear area to determine tumor biology and possible grade prior to treatment. Histologic grade distribution was as follows: low grade, 15%; moderate grade, 40%; and high grade, 45%. Mann-Whitney test for trend on tumor size and histologic grade between MNA showed a significant relationship between MNA and tumor size (P=.016) but no significance between MNA and histologic grade (P =.22). The chi2 and Fisher Exact Test between MNA and node-positive or -negative patients showed no significance. When correlating MNA with tumor size and histologic grade, high MNA is present at a higher frequency as tumor size and histologic grade increase.


Osteoclast differentiation independent of the TRANCE-RANK-TRAF6 axis.
Osteoclasts are derived from myeloid lineage cells, and their differentiation is supported by various osteotropic factors, including the tumor necrosis factor (TNF) family member TNF-related activation-induced cytokine (TRANCE). Genetic deletion of TRANCE or its receptor, receptor activator of nuclear factor kappaB (RANK), results in severely osteopetrotic mice with no osteoclasts in their bones. TNF receptor-associated factor (TRAF) 6 is a key signaling adaptor for RANK, and its deficiency leads to similar osteopetrosis. Hence, the current paradigm holds that TRANCE-RANK interaction and subsequent signaling via TRAF6 are essential for the generation of functional osteoclasts. Surprisingly, we show that hematopoietic precursors from TRANCE-, RANK-, or TRAF6-null mice can become osteoclasts in vitro when they are stimulated with TNF-alpha in the presence of cofactors such as TGF-beta. We provide direct evidence against the current paradigm that the TRANCE-RANK-TRAF6 pathway is essential for osteoclast differentiation and suggest the potential existence of alternative routes for osteoclast differentiation.


Changes in reproductive behavior associated with the perception and individual experiences of the COVID-19 pandemic.
This study evaluates the impacts of the COVID-19 pandemic on the reproductive behavior of men and women during the most restrictive period of the pandemic in Czechia. At the end of this period, data was collected for the Czech GGS COVID Pilot-Follow-up Study (April 2021), which included additional questions on reproductive plans and perceptions of the pandemic related to fertility. The study focuses in detail on the evaluation of the favorability of fertility during this period by considering the impact of the pandemic on the lives of individuals in various life areas. It subsequently attempts to determine to what extent this evaluation and personal experiences of the pandemic affected reproductive behavior (the intended number of children, current attempts to conceive and short-term fertility intentions). In summary, in most cases the respondents considered the most severe period of the pandemic to be unfavorable in terms of childbirth. Women provided an overall negative assessment of the favorability of childbirth in this period, which was reflected in a reduction in the planned number of children, while the men who considered this period favorable declared a higher chance of short-term fertility intentions.


Sexual Identity Disorder and Psychosis in Klinefelter Syndrome: A Synthesis of Literature and a Case Report.
Klinefelter syndrome (KS) 47, XXY is the most frequent chromosomal abnormality causing hypogonadism in humans. This chromosomal abnormality of number in its classical form called homogeneous (supernumerary X) is generally the result of a meiosis accident. Several studies have suggested that individuals with KS are at greater risk of developing various psychiatric disorders, including depression and schizophrenia. The diagnosis is made based on subnormal testosterone with high pituitary gonadotropins and confirmed by determining the karyotype on a blood simple. We did a literature review using an electronic search in three databases: Pubmed/MEDLINE, Google Scholar, and PsychInfo. We found that since 1989, seven case reports with KS and mental disorders with similar and different characteristics of our case illustration of a patient with KS and psychosis were published.


"Living Between Two Different Worlds": Experiences of Leaving a High-Cost Religious Group.
The aim of this interdisciplinary study is to gain a comprehensive understanding of individuals' subjective experiences after leaving a high-cost religious group and how these experiences have affected their lives. In-depth interviews were done with 18 participants who had left different religious communities in Finland. The interviews were analysed through a thematic analysis. The results show that religious disaffiliation is a life change that may affect an individual's life in profound ways. Life after being a member of a high-cost group may involve experiences of fear, guilt, sorrow, pain, loss and even suffering on an existential level. These experiences can have serious implications for one's well-being and health. However, life after religious disaffiliation also includes many positive aspects, such as experiences of joy, freedom, relief, gratitude and empowerment.


Influence of the poly-3-hydroxybutyrate (PHB) granule-associated proteins (PhaP1 and PhaP2) on PHB accumulation and symbiotic nitrogen fixation in Sinorhizobium meliloti Rm1021.
Sinorhizobium meliloti cells store excess carbon as intracellular poly-3-hydroxybutyrate (PHB) granules that assist survival under fluctuating nutritional conditions. PHB granule-associated proteins (phasins) are proposed to regulate PHB synthesis and granule formation. Although the enzymology and genetics of PHB metabolism in S. meliloti have been well characterized, phasins have not yet been described for this organism. Comparison of the protein profiles of the wild type and a PHB synthesis mutant revealed two major proteins absent from the mutant. These were identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) as being encoded by the SMc00777 (phaP1) and SMc02111 (phaP2) genes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins associated with PHB granules followed by MALDI-TOF confirmed that PhaP1 and PhaP2 were the two major phasins. Double mutants were defective in PHB production, while single mutants still produced PHB, and unlike PHB synthesis mutants that have reduced exopolysaccharide, the double mutants had higher exopolysaccharide levels. Medicago truncatula plants inoculated with the double mutant exhibited reduced shoot dry weight (SDW), although there was no corresponding reduction in nitrogen fixation activity. Whether the phasins are involved in a metabolic regulatory response or whether the reduced SDW is due to a reduction in assimilation of fixed nitrogen rather than a reduction in nitrogen fixation activity remains to be established.


Exploring the endangerment mechanisms of Hipposideros pomona based on molecular phylogeographic methods.
The endangerment mechanisms of various species are a focus of studies on biodiversity and conservation biology. Hipposideros pomona is an endangered species, but the reasons behind its endangerment remain unclear. We investigated the endangerment mechanisms of H. pomona using mitochondrial DNA, nuclear DNA, and microsatellite loci markers. The results showed that the nucleotide diversity of mitochondria DNA and heterozygosity of microsatellite markers were high (π = 0.04615, H O = 0.7115), whereas the nucleotide diversity of the nuclear genes was low (THY: π = 0.00508, SORBS2: π = 0.00677, ACOX2: π = 0.00462, COPS7A: π = 0.00679). The phylogenetic tree and median-joining network based on mitochondrial DNA sequences clustered the species into three clades, namely North Vietnam-Fujian, Myanmar-West Yunnan, and Laos-Hainan clades. However, joint analysis of nuclear genes did not exhibit clustering. Analysis of molecular variance revealed a strong population genetic structure; IMa2 analysis did not reveal significant gene flow between all groups (p > .05), and isolation-by-distance analysis revealed a significant positive correlation between genetic and geographic distances (p < .05). The mismatch distribution analysis, neutral test, and Bayesian skyline plots revealed that the H. pomona population were relatively stable and exhibited a contraction trend. The results implied that H. pomona exhibits female philopatry and male-biased dispersal. The Hengduan Mountains could have acted as a geographical barrier for gene flow between the North Vietnam-Fujian clade and the Myanmar-West Yunnan clade, whereas the Qiongzhou Strait may have limited interaction between the Hainan populations and other clades. The warm climate during the second interglacial Quaternary period (c. 0.33 Mya) could have been responsible for species differentiation, whereas the cold climate during the late Quaternary last glacial maximum (c. 10 ka BP) might have caused the overall contraction of species. The lack of significant gene flow in nuclear microsatellite loci markers among the different populations investigated reflects recent habitat fragmentation due to anthropogenic activities; thus, on-site conservation of the species and restoration of gene flow corridors among populations need immediate implementation.


[Work-related MRSA CC398 carrier status had serious psychosocial consequences to a farmer].
We describe a case of work-related methicillin-resistant Staphylococcus aureus (MRSA) CC398 carrier status in a 33-year-old male working at a pig farm. The case was complicated since his pregnant girlfriend had also MRSA CC398 carrier status. Although she remained without contact to farm animals the MRSA could not be eradicated. Conflicting messages from healthcare workers complicated the case and the psychological consequences became traumatizing to the patient and caused sick leave and psychotherapy. This case shows the importance of better information of MRSA CC398-positive patients in the future, especially for MRSA-exposed occupations.


Relationship of frequent mantram repetition to emotional and spiritual well-being in healthcare workers.
Healthcare workers report high levels of stress in the workplace. To determine how to reduce stress, the authors examined the effectiveness of frequently repeating a mantram (a word with spiritual meaning) on emotional and spiritual well-being. A pretest-posttest design was used to measure stress, state/trait anxiety and anger, quality of life, and spiritual well-being in a convenience sample (N=42) of hospital workers completing a mantram intervention program. Significant improvements were found in stress (p < .001), trait-anxiety (p = .002), trait-anger (p = .02), quality of life (p = .001), and spiritual well-being (p = .003). When examining the effects of mantram practice, trait-anxiety and religious and spiritual well-being were significant (p < .05). Improvements in emotional and spiritual well-being may be mediated by frequent mantram repetition.


Flotation of Streptomyces pilosus after lead accumulation.
The flotation of Streptomyces pilosus was studied in the absence and presence of adsorbed Pb(II) ions on the cell walls and with and without using sodium dodecylsulfate (SDS) as collector. Adsorption of SDS onto the organism in the absence and presence of Pb(II) ions was also investigated. It was found that in the absence of adsorbed Pb(II), very little flotation took place with or without the presence of SDS although SDS adsorbed to some extent on the organism. The presence of Pb(II) on the bacterium's cell walls significantly increased flotation even in the absence of the collector. Addition of SDS, however, further increased both flotation rate and recovery.


The relationship between hydrocortisone acetate distribution and efficacy in inflamed eyes: continuous vs intermittent delivery.
Hydrocortisone acetate was administered to rabbit eyes intermittently in 250mug doses (given twice daily or once hourly) or continuously at 10mug/hr from an experimental ocular therapeutic system. Tissue drug distribution was measured over a 1 to 4 day period in experimental conjunctivitis. Anti-inflammatory efficacy was evaluated over a 4 day period in the same model. Statistically significant improvement in inflammation was observed 48 hours after initiation of continuous delivery treatment and after 4 days of the twice daily eyedrop regimen. Drug uptake by ocular tissues correlated with drug efficacy. Continuous delivery at 10mug/hr produced conjunctival steriod concentrations exceeding 1mug/g during 8 through 24 hours of treatment, and average levels of 1mug/g from days 2 through 4. Conjunctival steroid concentations generally did not exceed 1mug/g in eyedrop treated eyes until the fourth treatment day, when their clinical improvement matched that observed in the continuously treated eyes.


Effect of 13-valent pneumococcal conjugate vaccine on experimental carriage of Streptococcus pneumoniae serotype 6B in Blantyre, Malawi: a randomised controlled trial and controlled human infection study.
The effect of childhood pneumococcal conjugate vaccine implementation in Malawi is threatened by absence of herd effect. There is persistent vaccine-type pneumococcal carriage in both vaccinated children and the wider community. We aimed to use a human infection study to measure 13-valent pneumococcal conjugate vaccine (PCV13) efficacy against pneumococcal carriage. We did a double-blind, parallel-arm, randomised controlled trial investigating the efficacy of PCV13 or placebo against experimental pneumococcal carriage of Streptococcus pneumoniae serotype 6B (strain BHN418) among healthy adults (aged 18-40 years) from Blantyre, Malawi. We randomly assigned participants (1:1) to receive PCV13 or placebo. PCV13 and placebo doses were prepared by an unmasked pharmacist to maintain research team and participant masking with identification only by a randomisation identification number and barcode. 4 weeks after receiving either PCV13 or placebo, participants were challenged with 20 000 colony forming units (CFUs) per naris, 80 000 CFUs per naris, or 160 000 CFUs per naris by intranasal inoculation. The primary endpoint was experimental pneumococcal carriage, established by culture of nasal wash at 2, 7, and 14 days. Vaccine efficacy was estimated per protocol by means of a log-binomial model adjusting for inoculation dose. The trial is registered with the Pan African Clinical Trials Registry, PACTR202008503507113, and is now closed. Recruitment commenced on April 27, 2021 and the final visit was completed on Sept 12, 2022. 204 participants completed the study protocol (98 PCV13, 106 placebo). There were lower carriage rates in the vaccine group at all three inoculation doses (0 of 21 vs two [11%] of 19 at 20 000 CFUs per naris; six [18%] of 33 vs 12 [29%] of 41 at 80 000 CFUs per naris, and four [9%] of 44 vs 16 [35%] of 46 at 160 000 CFUs per naris). The overall carriage rate was lower in the vaccine group compared with the placebo group (ten [10%] of 98 vs 30 [28%] of 106; Fisher's p value=0·0013) and the vaccine efficacy against carriage was estimated at 62·4% (95% CI 27·7-80·4). There were no severe adverse events related to vaccination or inoculation of pneumococci. This is, to our knowledge, the first human challenge study to test the efficacy of a pneumococcal vaccine against pneumococcal carriage in Africa, which can now be used to establish vaccine-induced correlates of protection and compare alternative strategies to prevent pneumococcal carriage. This powerful tool could lead to new means to enhance reduction in pneumococcal carriage after vaccination. Wellcome Trust.


The sedoheptulose kinase CARKL directs macrophage polarization through control of glucose metabolism.
Immune cells are somewhat unique in that activation responses can alter quantitative phenotypes upwards of 100,000-fold. To date little is known about the metabolic adaptations necessary to mount such dramatic phenotypic shifts. Screening for novel regulators of macrophage activation, we found nonprotein kinases of glucose metabolism among the most enriched classes of candidate immune modulators. We find that one of these, the carbohydrate kinase-like protein CARKL, is rapidly downregulated in vitro and in vivo upon LPS stimulation in both mice and humans. Interestingly, CARKL catalyzes an orphan reaction in the pentose phosphate pathway, refocusing cellular metabolism to a high-redox state upon physiological or artificial downregulation. We find that CARKL-dependent metabolic reprogramming is required for proper M1- and M2-like macrophage polarization and uncover a rate-limiting requirement for appropriate glucose flux in macrophage polarization.


Clopidogrel-induced hepatotoxicity after percutaneous coronary stenting.
Clopidogrel, an adenosine diphosphate receptor blocker, is widely used as an adjunctive antiplatelet therapy in acute coronary syndrome and percutaneous coronary stenting. The occurrence of hepatotoxicity is rare. We describe the occurrence of symptomatic liver disease in a 74-year-old man 5 weeks following commencement of therapy with clopidogrel. The reported cases of clopidogrel-induced hepatotoxicity are reviewed and the clinical significance of this event are discussed.


MicroRNA-21 Down-regulates Rb1 Expression by Targeting PDCD4 in Retinoblastoma.
Retinoblastoma (RB) is a children's ocular cancer caused by mutated retinoblastoma 1 (Rb1) gene on both alleles. Rb1 and other related genes could be regulated by microRNAs (miRNA) via complementarily pairing with their target sites. MicroRNA-21 (miR-21) possesses the oncogenic potential to target several tumor suppressor genes, including PDCD4, and regulates tumor progression and metastasis. However, the mechanism of how miR-21 regulates PDCD4 is poorly understood in RB. We investigated the expression of miRNAs in RB cell lines and identified that miR-21 is one of the most deregulated miRNAs in RB. Using qRT-PCR, we verified the expression level of several miRNAs identified by independent microarray assays, and analyzed miRNA expression patterns in three RB cell lines, including Weri-Rb1, Y79 and RB355. We found that miR-19b, -21, -26a, -195 and -222 were highly expressed in all three cell lines, suggesting their potential role in RB tumorigenesis. Using the TargetScan program, we identified a list of potential target genes of these miRNAs, of which PDCD4 is one the targets of miR-21. In this study, we focused on the regulatory mechanism of miR-21 on PDCD4 in RB. We demonstrated an inverse correlation between miR-21 and PDCD4 expression in Weri-Rb1 and Y79 cells. These data suggest that miR-21 down-regulates Rb1 by targeting PDCD4 tumor suppressor. Therefore, miR-21 could serve as a therapeutic target for retinoblastoma.


PP2A-based triple-strike therapy overcomes mitochondrial apoptosis resistance in brain cancer cells.
Mitochondrial glycolysis and hyperactivity of the phosphatidylinositol 3-kinase-protein kinase B (AKT) pathway are hallmarks of malignant brain tumors. However, kinase inhibitors targeting AKT (AKTi) or the glycolysis master regulator pyruvate dehydrogenase kinase (PDKi) have failed to provide clinical benefits for brain tumor patients. Here, we demonstrate that heterogeneous glioblastoma (GB) and medulloblastoma (MB) cell lines display only cytostatic responses to combined AKT and PDK targeting. Biochemically, the combined AKT and PDK inhibition resulted in the shutdown of both target pathways and priming to mitochondrial apoptosis but failed to induce apoptosis. In contrast, all tested brain tumor cell models were sensitive to a triplet therapy, in which AKT and PDK inhibition was combined with the pharmacological reactivation of protein phosphatase 2A (PP2A) by NZ-8-061 (also known as DT-061), DBK-1154, and DBK-1160. We also provide proof-of-principle evidence for in vivo efficacy in the intracranial GB and MB models by the brain-penetrant triplet therapy (AKTi + PDKi + PP2A reactivator). Mechanistically, PP2A reactivation converted the cytostatic AKTi + PDKi response to cytotoxic apoptosis, through PP2A-elicited shutdown of compensatory mitochondrial oxidative phosphorylation and by increased proton leakage. These results encourage the development of triple-strike strategies targeting mitochondrial metabolism to overcome therapy tolerance in brain tumors.


Up-regulation of human cervical cancer proto-oncogene contributes to hepatitis B virus-induced malignant transformation of hepatocyte by down-regulating E-cadherin.
Hepatocellular carcinoma (HCC) is one of the most fatal human malignancies, Human cervical cancer proto-oncogene (HCCR) aberrantly expressed in a number of malignant tumors, including HCC. HCC is associated with Hepatitis B virus (HBV) infection in a large percentage of cases. To explore the regulation and function of HCCR expression in the development of HCC, we detected HCCR expression in HBV expressing hepatocytes. Results showed that the expression of HCCR was higher in HBV-expressing hepatocytes than that in control cells. Examining different components of HBV revealed that the HBx promotes HCCR expression in hepatocytes via the T-cell factor (TCF)/β-catenin pathway. HCCR expression in HBx transgenic mice increased with as the mice aged and developed tumors. We also found that overexpression of HCCR in hepatocytes promoted cell proliferation, migration, and invasion and reduced cell adhesion. Suppressing HCCR expression abolished the effect of HBx-induced hepatocyte growth. In addition, HCCR represses the expression of E-cadherin by inhibition its promoter activity, which might correlate with the effects of HCCR in hepatocytes. Taken together, these results demonstrate that HBx-HCCR-E-cadherin regulation pathway might play an important role in HBV-induced hepatocarcinogenesis. They also imply that HCCR is a potential risk marker for HCC and/or a potential therapeutic target.


Ultrasound assessment of vocal fold paresis: a correlation case series with flexible fiberoptic laryngoscopy and adding the third dimension (3-D) to vocal fold mobility assessment.
Perioperative examination ofthe vocal folds with flexible fiberoptic laryngoscopy is not always feasible. Prior studies suggest vocal fold ultrasound may provide a useful screening tool, however, correlation to laryngoscopic findings is necessary. The purpose of the case series was to validate vocal fold ultrasound in the adult population and to correlate the ultrasound findings to the assessment provided by flexible fiberoptic laryngoscopy. This IRB approved study accrued sixteen patients. Vocal fold ultrasound performed by the anesthesiologist was correlated with the laryngoscopy performed by the otolaryngologist. Assessment of vocal fold motion was congruent in thirteen patients with normal vocal fold mobility; however, there was discordance between the findings in three patients. Vocal fold ultrasound may be useful to screen for vocal fold motion abnormalities in the adult population. Abnormal findings on vocal fold ultrasound should be confirmed with subsequent laryngoscopy.


Brief report: Exploring the benefits of a peer-tutored physical education programme among high school students with intellectual disability.
The purpose of this study was to explore possible benefits of a peer-tutored physical education programme (PTPE) in comparison with school physical education (SPE) in high school students with intellectual disability. Nineteen students with intellectual disabilities (15 boys, mean age 17.4 ± 1.7 years) were monitored during three PTPE and three SPE classes. A factorial RM-ANOVA was used to test differences on objective measured physical activity (PA), enjoyment and exertion during the two conditions, considering participants' weight condition as independent factor. During PTPE, participants reported higher light intensity PA, enjoyment and exertion than during SPE. Participants with overweight showed less inactive time and higher light intensity PA during PTPE than during SPE. The peer-tutored programme was beneficial for adolescents with intellectual disability, particularly for those in overweight condition. The higher enjoyment found during PTPE may encourage exercise participation of students with intellectual disability.