Reactions of the hydroperoxide anion with dimethyl methylphosphonate in an ion trap mass spectrometer: evidence for a gas phase alpha-effect.
The gas phase degradation reactions of the chemical warfare agent (CWA) simulant, dimethyl methylphosphonate (DMMP), with the hydroperoxide anion (HOO(-)) were investigated using a modified quadrupole ion trap mass spectrometer. The HOO(-) anion reacts readily with neutral DMMP forming two significant product ions at m/z 109 and m/z 123. The major reaction pathways correspond to (i) the nucleophilic substitution at carbon to form [CH(3)P(O)(OCH(3))O](-) (m/z 109) in a highly exothermic process and (ii) exothermic proton transfer. The branching ratios of the two reaction pathways, 89% and 11% respectively, indicate that the former reaction is significantly faster than the latter. This is in contrast to the trend for the methoxide anion with DMMP, where proton transfer dominates. The difference in the observed reactivities of the HOO(-) and CH(3)O(-) anions can be considered as evidence for an alpha-effect in the gas phase and is supported by electronic structure calculations at the B3LYP/aug-cc-pVTZ//B3LYP/6-31+G(d) level of theory that indicate the S(N)2(carbon) process has an activation energy 7.8 kJ mol(-1) lower for HOO(-) as compared to CH(3)O(-). A similar alpha-effect was calculated for nucleophilic addition-elimination at phosphorus, but this process--an important step in the perhydrolysis degradation of CWAs in solution--was not observed to occur with DMMP in the gas phase. A theoretical investigation revealed that all processes are energetically accessible with negative activation energies. However, comparison of the relative Arrhenius pre-exponential factors indicate that substitution at phosphorus is not kinetically competitive with respect to the S(N)2(carbon) and deprotonation processes.


Genome-wide association analysis on normal hearing function identifies PCDH20 and SLC28A3 as candidates for hearing function and loss.
Hearing loss and individual differences in normal hearing both have a substantial genetic basis. Although many new genes contributing to deafness have been identified, very little is known about genes/variants modulating the normal range of hearing ability. To fill this gap, we performed a two-stage meta-analysis on hearing thresholds (tested at 0.25, 0.5, 1, 2, 4, 8 kHz) and on pure-tone averages (low-, medium- and high-frequency thresholds grouped) in several isolated populations from Italy and Central Asia (total N = 2636). Here, we detected two genome-wide significant loci close to PCDH20 and SLC28A3 (top hits: rs78043697, P = 4.71E-10 and rs7032430, P = 2.39E-09, respectively). For both loci, we sought replication in two independent cohorts: B58C from the UK (N = 5892) and FITSA from Finland (N = 270). Both loci were successfully replicated at a nominal level of significance (P < 0.05). In order to confirm our quantitative findings, we carried out RT-PCR and reported RNA-Seq data, which showed that both genes are expressed in mouse inner ear, especially in hair cells, further suggesting them as good candidates for modulatory genes in the auditory system. Sequencing data revealed no functional variants in the coding region of PCDH20 or SLC28A3, suggesting that variation in regulatory sequences may affect expression. Overall, these results contribute to a better understanding of the complex mechanisms underlying human hearing function.


A possible mechanism for the anti-ketogenic action of alanine in the rat.
1. The anti-ketogenic effect of alanine has been studied in normal starved and diabetic rats by infusing l-alanine for 90min in the presence of somatostatin (10mug/kg body wt. per h) to suppress endogenous insulin and glucagon secretion. 2. Infusion of alanine at 3mmol/kg body wt. per h caused a 70+/-11% decrease in [3-hydroxybutyrate] and a 58+/-9% decrease in [acetoacetate] in 48h-starved rats. [Glucose] and [lactate] increased, but [non-esterified fatty acid], [glycerol] and [3-hydroxybutyrate]/[acetoacetate] were unchanged. 3. Infusion of alanine at 1mmol/kg body wt. per h caused similar decreases in [ketone body] (3-hydroxybutyrate plus acetoacetate) in 24h-starved normal and diabetic rats, but no change in other blood metabolites. 4. Alanine [3mmol/kg body wt. per h] caused a 72+/-9% decrease in the rate of production of ketone bodies and a 57+/-8% decrease in disappearance rate as assessed by [3-(14)C]acetoacetate infusion. Metabolic clearance was unchanged, indicating that the primary effect of alanine was inhibition of hepatic ketogenesis. 5. Aspartate infusion at 6mmol/kg body wt. per h had similar effects on blood ketone-body concentrations in 48h-starved rats. 6. Alanine (3mmol/kg body wt. per h) caused marked increases in hepatic glutamate, aspartate, malate, lactate and citrate, phosphoenolpyruvate, 2-phosphoglycerate and glucose concentrations and highly significant decreases in [3-hydroxybutyrate] and [acetoacetate]. Calculated [oxaloacetate] was increased 75%. 7. Similar changes in hepatic [malate], [aspartate] and [ketone bodies] were found after infusion of 6mmol of aspartate/kg body wt. per h. 8. It is suggested that the anti-ketogenic effect of alanine is secondary to an increase in hepatic oxaloacetate and hence citrate formation with decreased availability of acetyl-CoA for ketogenesis. The reciprocal negative-feedback cycle of alanine and ketone bodies forms an important non-hormonal regulatory system.


Finishing occlusion, degree of stability and the PAR index.
The occlusions of sixty-five patients, treated by one private specialist orthodontist with consistent philosophy and goals, were assessed according to the PAR Index. For each patient, pre-treatment, post-treatment and follow-up study models were assessed. Follow-up models were taken at least 6.5 years following the removal of all retention appliances. Mean-weighted PAR scores were calculated for the total sample and various sub-groups at each stage. Mean percentage changes in weighted PAR scores were also calculated. The relationships between the occlusal standards at the end of active treatment and at the end of the follow-up period, and specific diagnostic and treatment factors were then investigated to search for any factors that might be predictive of long-term post-treatment occlusal stability or instability. The overall mean-weighted pre-treatment, post-treatment and follow-up PAR scores were 25.5, 3.0 and 7.0, respectively. There was an 85.6 per cent decrease with treatment in the overall mean-weighted PAR score. This was followed by a 15.2 per cent increase in that overall mean during the follow-up period. In this introductory study, neither the PAR score at the end of active treatment, nor the amounts of occlusal change occurring during or after active treatment, were found to be predictive of the amount of post-treatment occlusal change.


Mathematics.
Current research in mathematics involves a wide variety of interlocking ideas, old and new. For example, results about the curves and surfaces defined by polynomial equations, as in algebraic geometry, appear in the study of solitary waves and also in the gauge theories in physics. Centuries-old problems in number theory have been solved, while others have been revealed as insoluble. The classification of all finite simple groups is nearly achieved (and the full treatment will be voluminous); the representation of groups aids in their application to the study of symmetry. These developments, and many others, attest to the vitality of mathematics.


Colorectal cancer screening: the old and the new.
As the second leading cancer killer of Americans, colorectal cancer represents a serious health threat to all Americans. All health care providers and patients should demand colorectal cancer screening, which has been shown to decrease the incidence and mortality of the disease through the detection and removal of the cancer's precursor lesion, the adenomatous polyp. Colonoscopy is the most effective screening tool because it identifies and treats more polyps than any other screening tool available today. However, cost issues have led to considerable controversy regarding its universal application as a screening tool.


Poly(amidoamine) Dendrimers with Carbonic Anhydrase Inhibitory Activity and Antiglaucoma Action.
Four generations of poly(amidoamine) (PAMAM) dendrimers decorated with benzenesulfonamide moieties were prepared by derivatizing the amino groups of the dendrimer with 4-carboxy-benzenesulfonamide functionalities. Compounds incorporating 4, 8, 16, and 32 sulfonamide moieties were thus obtained, which showed an increasing carbonic anhydrase (CA, EC 184.108.40.206) inhibitory action with the increase of the number of sulfamoyl groups in the dendrimer. Best inhibitory activity (in the low nanomolar-subnanomolar range) was observed for isoforms CA II and XII, involved among others in glaucoma. In an animal model of this disease, the chronic administration of such dendrimers for 5 days led to a much more efficient drop of intraocular pressure compared to the standard drug dorzolamide.


vanA-targeted oligonucleotide DNA probe designed to monitor vancomycin- and teicoplanin-resistant bacteria in surface waters.
The glycopeptide antibiotics teicoplanin and vancomycin are common to treat severe Gram-positive bacterial infections. The gene vanA confers high-level resistance to these antibiotics, and these phenomena have been shown to be transferable. Release of vancomycin- and teicoplanin-resistant bacteria to surface waters is, therefore, of particular concern since they might proliferate and spread in different environments. Monitoring of the fate of vanA gene in these waters provides information on the exposure and potential threats of those bacteria for the environment and public health. Therefore, this study aimed at preparing a 25-mer-oligonucleotide DNA probe based on the 909 bp BamHI-ClaI fragment from Enterococcus faecium plasmids pVEF1 and pVEF2 through the use of Vector NTI Express Software. The newly designed vanA probe displayed highly specific hybridization with vanA-positive Enterococcus faecalis tested at 46 °C, 55 % formamide, and 0.020 M NaCl stringency conditions. In situ fluorescein hybridizations under the same stringency conditions were also used to monitor river water samples by using fluorescein microscopy. The results showed that the vanA-targeted oligonucleotide DNA probe prepared was not only highly specific but also quantitative tool for monitoring vancomycin- and teicoplanin-resistant bacteria in surface waters.


Genetic variations in the xenobiotic-metabolizing enzymes CYP1A1, CYP1A2, CYP2C9, CYP2C19 and susceptibility to colorectal cancer among Turkish people.
Cytochrome P450 (CYP) enzymes are genetically polymorphic and play key roles in the metabolism of xenobiotics. Colorectal cancer (CRC) is one of the most common malignant tumors in Turkey as well as in the world. In this study, it was aimed both to evaluate the effects of CYP variants on the susceptibility to CRC and to predict the individual response of the Turkish people to xenobiotics metabolized by CYP enzymes. For that, we assessed the association of CYP1A1, CYP1A2, CYP2C9, and CYP2C19 polymorphisms in patients with CRC in the Turkish population through a case-control study. Distributions of the variants were determined in 104 patients with CRC and 183 healthy volunteers. As results, CYP1A1 6235T/C was significantly associated with CRC risk (odds ratio [OR]=2.53; 95% confidence interval [CI]=0.99-6.45; p=0.046). In a haplotype-based analysis, CYP1A1 haplotype C6235-A2455 might be associated with the development of CRC (OR=2.70; 95% CI=0.58-5.90; p=0.046). We believe that the findings are the first results of CYP allele distributions in the Turkish population and provide an understanding of the epidemiological studies that correlate therapeutic approaches and etiology of CRC especially in Turkish patients.


Online verification and management scheme of gateway meter flow in the power system by machine learning.
Currently, the calibration of electric energy meters often involves manual meter reading, dismantling inspection, or regular sampling inspection conducted by professionals. To improve work efficiency and verification accuracy, this research integrates machine learning into the scheme of online verification and management of gateway meter flow in the power system. The approach begins by applying the Faster Region Convolutional Neural Network (Faster-RCNN) model and the Single Shot MultiBox Detector (SSD) model to the recognition system for dial readings. Then, the collected measurement data is pre-processed, excluding data collected under light load conditions. Next, an estimation error model and a solution equation for the electricity meter are established based on the pre-processed data. The operation error of the electricity meter is estimated, and the estimation accuracy is verified using the limited memory recursive least squares algorithm (LMRLSA). Furthermore, business assistant decision-making is carried out by combining the remote verification results with the estimation outcomes. The proposed dial reading recognition system is tested using 528 images of meter readings, achieving an accuracy of 98.49%. In addition, the influence of various parameters on the error results of the electricity meter is also explored. The results demonstrate that a memory length ranging from 600 to 1,200 and a line loss error of less than 5% yield the most suitable accuracy for estimating the electricity meter error. Meanwhile, it is advisable to remove measurement data collected under light load to avoid unnecessary checks. The experiments manifest that the proposed algorithm can properly eliminate the influence of old measurement data on the error parameter estimation, thereby enhancing the accuracy of the estimation. The adjustment of the memory length ensures real-time performance in estimating meter errors and enables online monitoring. This research has certain reference significance for achieving the online verification and management of gateway meter flow in the power system.


Spontaneous occurrence of XYY primary spermatocytes in the Sitka deer mouse.
Analysis of microspread, silver-stained primary spermatocytes from chromosomally and phenotypically normal Peromyscus sitkensis revealed the occurrence of XYY zygotene and pachytene nuclei at low frequency in three of eight individuals examined. Observed pairing configurations of sex chromosomes included a trivalent and a Y bivalent-X univalent. The data suggest that premeiotic nondisjunction may be involved in the origination of XYY chromosomal conditions.


Working memory capacity predicts the beneficial effect of selective memory retrieval.
Selective retrieval of some studied items can both impair and improve recall of the other items. This study examined the role of working memory capacity (WMC) for the two effects of memory retrieval. Participants studied an item list consisting of predefined target and nontarget items. After study of the list, half of the participants performed an imagination task supposed to induce a change in mental context, whereas the other half performed a counting task which does not induce such context change. Following presentation of a second list, memory for the original list's target items was tested, either with or without preceding retrieval of the list's nontarget items. Consistent with previous work, preceding nontarget retrieval impaired target recall in the absence of the context change, but improved target recall in its presence. In particular, there was a positive relationship between WMC and the beneficial, but not the detrimental effect of memory retrieval. On the basis of the view that the beneficial effect of memory retrieval reflects context-reactivation processes, the results indicate that individuals with higher WMC are better able to capitalise on retrieval-induced context reactivation than individuals with lower WMC.


Development of an Interactive Dashboard to Analyse Physiological Signals in the Neonatal Intensive Care Unit.
Premature babies and those born with a medical condition are cared for within the neonatal intensive care unit (NICU) in hospitals. Monitoring physiological signals and subsequent analysis and interpretation can reveal acute and chronic conditions for these neonates. Several advanced algorithms using physiological signals have been built into existing monitoring systems to allow clinicians to analyse signals in real time and anticipate patient deterioration. However, limited enhancements have been made to interactively visualise and adapt them to neonatal monitoring systems. To bridge this gap, we describe the development of a user-friendly and interactive dashboard for neonatal vital signs analysis written in the Python programming language where the analysis can be performed without prior computing knowledge. To ensure practicality, the dashboard was designed in consultation with a neonatologist to visualise electrocardiogram, heart rate, respiratory rate and oxygen saturation data in a time-series format. The resulting dashboard included interactive visualisations, advanced electrocardiogram analysis and statistical analysis which can be used to extract important information on patients' conditions.Clinical Relevance- This will support the care of preterm infants by allowing clinicians to visualise and interpret physiological data in greater granularity, aiding in patient monitoring and detection of adverse conditions. The detection of adverse conditions could allow timely and potentially life-saving interventions for conditions such as sepsis and brain injury.


New horizons in the treatment of autoimmune diseases: immunoablation and stem cell transplantation.
The prevalence of autoimmune diseases (ADs) in Western countries is estimated to be from 3-7%, and the treatment of severe, relapsing/refractory cases is still not satisfactory. The concept of utilizing intense immunosuppression followed by allogeneic or even autologous hemolymphopoietic stem cells (HSCs) to treat AD is based on encouraging results in experimental animals and from serendipitous cases of patients with both ADs and malignancies who were allotransplanted for the latter. However, rare unexpected relapses despite donor immune engraftment have been reported following HSC transplantation for AD. Autologous transplantation is a more feasible procedure with lower toxicity than allogeneic transplantation. This article analyzes the experimental basis for stem cell transplantation in AD and discusses the most important clinical results of both allogeneic and autologous HSC transplants.


VEGF-A/VEGFR Inhibition Restores Hematopoietic Homeostasis in the Bone Marrow and Attenuates Tumor Growth.
Antiangiogenesis-based cancer therapies, specifically those targeting the VEGF-A/VEGFR2 pathway, have been approved for subsets of solid tumors. However, these therapies result in an increase in hematologic adverse events. We surmised that both the bone marrow vasculature and VEGF receptor-positive hematopoietic cells could be impacted by VEGF pathway-targeted therapies. We used a mouse model of spontaneous breast cancer to decipher the mechanism by which VEGF pathway inhibition alters hematopoiesis. Tumor-bearing animals, while exhibiting increased angiogenesis at the primary tumor site, showed signs of shrinkage in the sinusoidal bone marrow vasculature accompanied by an increase in the hematopoietic stem cell-containing Lin-cKit(+)Sca1(+) (LKS) progenitor population. Therapeutic intervention by targeting VEGF-A, VEGFR2, and VEGFR3 inhibited tumor growth, consistent with observed alterations in the primary tumor vascular bed. These treatments also displayed systemic effects, including reversal of the tumor-induced shrinkage of sinusoidal vessels and altered population balance of hematopoietic stem cells in the bone marrow, manifested by the restoration of sinusoidal vessel morphology and hematopoietic homeostasis. These data indicate that tumor cells exert an aberrant systemic effect on the bone marrow microenvironment and VEGF-A/VEGFR targeting restores bone marrow function.


Effects of dehydroepiandrosterone (DHEA) on cardiovascular risk factors in older women with frailty characteristics.
this analysis was to investigate the effects of dehydroepiandrosterone (DHEA) on cardiovascular risk factors in older women with frailty characteristics. the study was a double-blind, randomised, placebo-controlled trial of 99 women (mean 76.6 +/- 6.0 year) with the low DHEA-S level and frailty. participants received 50 mg/day DHEA or placebo for 6 months; all received calcium (1,000-1,200 mg/day diet) and supplement (combined) and cholecalciferol (1,000 IU/day). Women participated in 90-min twice weekly exercise regimens, either chair aerobics or yoga. assessment of outcome variables included hormone levels (DHEA-S, oestradiol, oestrone, testosterone and sex hormone-binding globulin (SHBG)), lipid profiles (total cholesterol, high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol and triglycerides), body composition measured by dual energy absorptiometry, glucose levels and blood pressure (BP). eighty-seven women (88%) completed 6 months of study; 88% were pre-frail demonstrating 1-2 frailty characteristics and 12% were frail with > or =3 characteristics. There were significant changes in all hormone levels including DHEA-S, oestradiol, oestrone and testosterone and a decline in SHBG levels in those taking DHEA supplements. In spite of changes in hormone levels, there were no significant changes in cardiovascular risk factors including lipid profiles, body or abdominal fat, fasting glucose or BP. research to date has not shown consistent effects of DHEA on cardiovascular risk, and this study adds to the literature that short-term therapy with DHEA is safe for older women in relation to cardiovascular risk factors. This study is novel in that we recruited women with evidence of physical frailty.


A pharmacoeconomic evaluation of the myocardial ischemia reduction with aggressive cholesterol lowering (MIRACL) study in Canada.
To determine a 16-week total healthcare cost and the cost-effectiveness of short-term, lipid-lowering therapy with atorvastatin 80 mg following acute coronary syndrome (ACS) in Canada. The expected costs per patient on atorvastatin 80 mg per day and placebo were compared using clinical outcome data from the MIRACL study and cost data from the Ontario Case Costing Project and the Ontario Schedule of Benefits. The cost per event avoided was also assessed. The clinical outcomes measured included: death, cardiac arrest, non-fatal myocardial infarction (MI), fatal MI, angina pectoris, stroke, congestive heart failure, and surgical or percutaneous coronary revascularizations. All direct medical costs from the perspective of the Canadian health care system were taken into account. The total expected cost per patient was 2,590 dollars in the placebo group and 2,639 dollars in the atorvastatin group. The incremental cost of atorvastatin treatment (49.26 dollars per patient) corresponded to a cost of 1,285 dollars per event avoided. The cost savings obtained through the reduction in events offset 86% of the cost of atorvastatin treatment. Budget impact analysis revealed that increased rates of atorvastatin usage following ACS were associated with large numbers of events avoided at a small additional cost when projected to the Canadian population. In Canada, the clinical benefits of intensive short-term atorvastatin treatment administered within 96 hours after ACS were associated with a favorable cost-effectiveness ratio. The incremental cost of atorvastatin is mostly offset by savings due to the reduction in events in patients treated with atorvastatin.


[Grip strength changes in familial amyloidotic polyneuropathy].
Familial amiloidotic polyneuropathy (FAP) is an autossomical and dominant neurodegenerative disease related with systemic deposition of amyloid fibre mainly on peripheral nervous system. Clinically, is translated by an autonomous sensitive-motor polyneuropathy with beginning nearly always in foot, involving subsequently the hands. Until now, the unique available treatment for FAP disease is liver transplantation requiring medication that negatively affect muscle metabolism and force production mechanism. To our knowledge there are no quantitative characterizations of peak force in FAP patients or any comparison with healthy people. This knowledge will be extremely important to verify clinical and functional evolution of this disease and eventually prescribe an effective rehabilitation program. The purpose of this study was to analyse and compare levels of hand grip strength (peak force and endurance) in FAP patients with (FAPTx) or without (FAPNTx) a liver transplant with a group of healthy people (GC). The total sample of individuals where two hundred and six, assigned in 3 groups: 59 patients PAFNTx (23 males, 36 female; age 35 ± 8 years); 90 patients PAFTx (53 males, 37 females; age 34 ± 8 years) e 62 healthy persons (GC) (30 males, 32 females; age 33 ± 9 years). Grip strength was assessed by a portable grip dynamometer E-link (Biometrics Ltd, UK). All measurements were taken on standardized positions with standardized orders. The value noted to peak force was classified according to American College of Sports Medicine norms for grip strength. The 3 groups are differents (p < 0,05) for weigth, body mass index (BMI) and grip strength in both hands and endurance for left hand. Negative correlations between age and grip strength were found for PAFNTx and PAFTx but not for GC. According to our results FAP patients have lower values for grip strength in both hands than healthy subjects and consequently a worse classification in ACSM norms. Most patients present grip strength values lower than medium values or even precarious. These results could present predictable negative functional implications, showing also the necessity of a rehabilitation program with specificity at hand motor level.


DNA double-strand break repair defects in syndromes associated with acute radiation response: at least two different assays to predict intrinsic radiosensitivity?
Human diseases associated with acute radiation responses are rare genetic disorders with common clinical and biological features including radiosensitivity, genomic instability, chromosomal aberrations, and frequently immunodeficiency. To determine what molecular assays are predictive of cellular radiosensitivity whatever the genes mutations, the existence of a quantitative correlation between cellular radiosensitivity and unrepaired DNA double-strand breaks (DSB) repair defects was examined in a collection of 40 human fibroblasts representing 8 different syndromes. A number of techniques such as pulsed-field gel electrophoresis, plasmid assay and immunofluorescence with antibodies against MRE11, MDC1, 53BP1 and phosphorylated forms of H2AX, DNA-PK were applied systematically. Survival fraction at 2 Gy was found to be inversely proportional to the amount of unrepaired DSB, whatever the genes mutations and the assay applied. However, no single assay discriminates the full range of human radiosensitivity. Particularly, nuclear foci formed by the phosphorylation of H2AX do not predict well moderate radiosensitivities. Our findings suggest the existence of an ATM-dependent interplay between the activation of DNA-PK and MRE11. A classification of diseases according their cellular radiosensitivity, their molecular response to radiation and the functional assays permitting their evaluation is proposed.


The interferon inducible gene: Viperin.
The type I interferons (IFNs), IFN-α and -β, are key effector molecules of the immune response to viruses. The anti-viral action of IFNs on virus-infected cells and surrounding tissues is mediated by expression of hundreds of IFN-stimulated genes. Viperin (virus inhibitory protein, endoplasmic reticulum-associated, IFN-inducible) is an Interferon stimulated gene (ISG), which is induced by type I, II, and III IFNs or after infection with a broad range of DNA and RNA viruses. Recent evidence indicates that Viperin disrupts lipid rafts to block influenza virus budding and release and interferes with replication of hepatitis C virus by binding to lipid droplets, small organelles involved in lipid homeostasis that are essential for hepatitis C virus replication. Viperin is also induced by nonviral microbial products such as lipopolysaccharide (LPS) and by a wide range of bacteria, suggesting a broader role in innate antimicrobial defenses.


Chidamide (CS055/HBI-8000): a new histone deacetylase inhibitor of the benzamide class with antitumor activity and the ability to enhance immune cell-mediated tumor cell cytotoxicity.
Chidamide (CS055/HBI-8000) is a new histone deacetylase (HDAC) inhibitor of the benzamide class currently under clinical development in cancer indications. This study reports the in vitro and in vivo antitumor characteristics of the compound. Selectivity and potency of chidamide in inhibition of HDAC isotypes were analyzed by using a panel of human recombinant HDAC proteins. Tumor cell lines either in culture or inoculated in nude mice were used for the evaluation of the compound's antitumor activity. To investigate the immune cell-mediated antitumor effect, isolated peripheral blood mononuclear cells from healthy donors were treated with chidamide, and cytotoxicity and expression of relevant surface proteins were analyzed. Microarray gene expression studies were performed on peripheral white blood cells from two T-cell lymphoma patients treated with chidamide. Chidamide was found to be a low nanomolar inhibitor of HDAC1, 2, 3, and 10, the HDAC isotypes well documented to be associated with the malignant phenotype. Significant and broad spectrum in vitro and in vivo antitumor activity, including a wide therapeutic index, was observed. Chidamide was also shown to enhance the cytotoxic effect of human peripheral mononuclear cells ex vivo on K562 target cells, accompanied by the upregulation of proteins involved in NK cell functions. Furthermore, the expression of a number of genes involved in immune cell-mediated antitumor activity was observed to be upregulated in peripheral white blood cells from two T-cell lymphoma patients who responded to chidamide administration. The results presented in this study provide evidence that chidamide has potential applicability for the treatment of a variety of tumor types, either as a single agent or in combination therapies.


Negative symptoms and speech pauses in youths at clinical high risk for psychosis.
Aberrant pauses are characteristic of schizophrenia and are robustly associated with its negative symptoms. Here, we found that pause behavior was associated with negative symptoms in individuals at clinical high risk (CHR) for psychosis, and with measures of syntactic complexity-phrase length and usage of determiners that introduce clauses-that we previously showed in this same CHR cohort to help comprise a classifier that predicted psychosis. These findings suggest a common impairment in discourse planning and verbal self-monitoring that affects both speech and language, and which is detected in clinical ratings of negative symptoms.


Genetic mouse models of essential tremor: are they essential?
The causative genes for essential tremor (ET), one of the most common genetic neurological disorders, have eluded scientists despite intensive search. Two gene loci linked to ET, one on chromosome 3q13 and another on chromosome 2p24.1, have been identified, and a missense mutation in the HS1-BP3 gene on the 2p has been suggested as the cause of the disorder in about 10% of American ET patients. Therefore, the genetic basis for the vast majority of familial ET is still unknown. In this issue of the JCI, the gene coding for the gamma-aminobutyric acidA (GABA(A)) receptor alpha1 subunit is suggested as a potential candidate gene for ET, as mice lacking the gene express a phenotype that overlaps with some clinical characteristics of the human condition.


Abnormal rhythmic motor response in children with attention-deficit-hyperactivity disorder.
Children with attention-deficit-hyperactivity disorder (ADHD) have difficulties with motor control, inhibition of motor responses, motor flexibility, and motor preparedness. We proposed that motor abnormalities in ADHD might result, at least in part, from an abnormal neuronal oscillatory mechanism necessary for motor temporal regulation. The aim of this study was to assess pacing in children with ADHD, by testing for rhythmic abnormalities of motor activity using a tapping test. Twenty-seven children (21 males, six females; aged 6 to 14 years 6 months; mean age 11 years 4 months, SD 2 years 2 months) diagnosed with ADHD according to DSM-IV clinical criteria, and 33 controls (25 males, eight females; aged 6 to 14 years 6 months; mean 11 years 1 month, SD 2 years 2 months), underwent a finger-tapping test requiring rhythmic responses to frequencies from 1 to 6 Hz. All participants who were treated on a daily basis with methylphenidate (n = 22) were medication-free on the day of the test. Most of the children with ADHD responded at a constant rate regardless of stimulus frequency, a phenomenon only seen in a small number of the controls. This specific error pattern, also seen in Parkinson's disease, has been attributed to an abnormal oscillatory mechanism mediated by dopaminergic fronto-striatal circuitry, which might also be pathophysiologically relevant for ADHD.


I sell seashells by the seashore and my name is Jack: comment on Pelham, Mirenberg, and Jones (2002).
According to a new hypothesis based on implicit egotism, people gravitate toward cities, states, and careers with names similar to their own names. To support this hypothesis, B. W. Pelham, M. C. Mirenberg, and J. T. Jones (2002) reported a series of results regarding distributions of names in different cities, states, and jobs. In the present article, new analyses of the original data are reported, showing that the hypothesis is not supported for the large majority of names considered by the authors, and for some names even the opposite result is found. In addition, a meta-analysis reveals that either the data are unreliable, or the hypothesis cannot be supported in the whole population of names. Overall, the original data give no support of the idea that implicit egotism influences major life decisions.


Finite element analysis of stress caused by palatal orthodontic anchor screws.
This study used finite element (FE) analysis to investigate the stability of miniscrews (screws) placed at the median palate. FE models with variable suture maturity and screw-suture distances were used to examine the relationship with screw stability. Four groups were classified by extent of maturation of the midpalatal suture (0%, 60%, 75%, and 100%). The placement position was set at the center of the suture (0.0 mm), or 0.5, 1.0, and 1.5 mm to the side of the suture, and von Mises stress values in bone and screw displacement were compared among models. The stress value for the unsutured model, in which the screw was placed at the center of the suture, was greater than 30 MPa. Stress values for models in which screws were placed to the side (0.5-1.5 mm) were less than 28 MPa. Maximum screw displacement was greater in the 0.0-mm incomplete suture model than at other placement positions. Because bone conditions vary among patients, placement position and suture maturation should be examined on cone beam-computed tomography images, to ensure screw stability.


Burden of illness of progressive familial intrahepatic cholestasis in the US, UK, France, and Germany: study rationale and protocol of the PICTURE study.
Background: Progressive familial intrahepatic cholestasis (PFIC) is an ultra-rare disease with a considerable burden on pediatric patients and their caregivers, impacting quality of life (QoL). The mortality rates highlight a significant need for efficacious treatments. Real-world data on associated costs and QoL are needed to gauge the potential impact of new pharmacological treatments.Methods: Clinical and socio-economic burden of PFIC on patients/caregivers, health systems, and society will be assessed. Patient/caregiver- and physician-level retrospective cross-sectional data will be collected from the US, UK, France, and Germany, for PFIC types 1, 2, 3.A representative sample of physicians will provide clinical and resource utilization information using an electronic Case Report Form (eCRF). Patient/caregiver surveys will collect socio-economic and QoL data, enabling assessment of PFIC impact on QoL. Mean costs (direct medical/non-medical, indirect) will be calculated.The study materials were reviewed by medical professionals and patient representatives and received ethical approval from the University of Chester.Discussion: The study aims to reveal the unmet medical need, disease burden, resource utilization, and costs of PFIC, to raise awareness with policymakers and healthcare professionals, and provide support for the patient/caregiver community. As novel PFIC therapies recently emerged, this study will yield quantifiable data for health technology assessments.


Th17 cell dynamics in HIV infection.
Th17 cells are a newly identified subtype of CD4 T cells that respond to bacterial and fungal antigens and are important in mucosal immunology. Because HIV infection results in loss of CD4 T cells as well as disruption to the gastrointestinal tract that causes microbial translocation and immune activation, Th17 cells potentially play an important role in HIV pathogenesis. Here we examine the relationship between Th17 cells and HIV disease pathogenesis. Th17 cells are preferentially lost from the gastrointestinal tract of HIV-infected individuals, which is not entirely due to direct infection, as Th17 cells can be infected in vivo, but are not preferentially infected. Long-term highly active antiretroviral therapy (HAART) can result in restoration of Th17 cells in the gastrointestinal, which may be associated with better disease prognosis. Furthermore, other cells, such as Vdelta1 T cells, can make IL-17 in vivo during HIV infection and may contribute to antibacterial immunity after loss of Th17 cells. Recent studies have improved our understanding of the role for Th17 cells during HIV infection; however, more studies are needed to discern better the detrimental consequences of loss of Th17 cells during HIV infection.


[Latissimus dorsi transfer for the treatment of irreparable craniodorsal tears of the rotator cuff].
Latissimus dorsi transfer is a relatively complicated procedure subsequently requiring an active, long-term physical therapy. The surgeon's performance and a patient's effort are therefore of worth consideration. However, sutures of massive rotator cuff tears do not always produce good results. The aim of the study is to evaluate the role of latissimus dorsi transfer in the treatment of these shoulder pathologies. A total of 21 patients were evaluated. Their age at the time of surgery ranged from 48 to 63 years, with an average of 54.8 years. A minimum follow-up was three years, the average was 47 months (70-36 months). The indication for surgery included a massive rotator cuff tear, without symptoms of arthropathy and with normal subscapularis muscle function, which had been resistant to conservative treatment for 6 months at least. In 13 patients, transfer was performed after previous surgery on the rotator cuff. The procedure was carried out in a lateral position. The latissimus dorsi tendon was cut off just at its attachment to the humerus, the muscle was mobilised and the musculo-tendinous flap was released to reach the proximal operative wound and to cover the greater tuberosity of the proximal humerus. A shoulder abduction splint was used to immobilise the upper extremity for 6 weeks. Active and supervised individual physical therapy followed. Subjective evaluation was excellent in 10, good in 9, and poor in 2 patients. The average Constant-Murley score improved by 32.25 points as compared before surgery (from 38.50 to 70.75). The evaluation of active arm elevation in the frontal plane showed improvement in 16 patients (77 %) by 40 degrees on average, no change in five patients (23%), and no deterioration in any of the patients. Active external rotation with the arm held at the body side improved by 14 degrees on average. Progression of radiographic changes indicating arthropathy was recorded in five patients.Post-operative haematoma, developed three patients it was treated by needle aspiration in one, and drainage in two patients. The results presented here are in agreement with those reported in the international literature.The method of latissimus dorsi transfer shows poorer outcomes after previous attempts to suture of massive rotator cuff tears. In patients with a restricted range of motion before surgery, the post-operative shoulder motion achieved at the final evaluation is also lower. When based on a correct indication, latissimus dorsi transfer with an uncomplicated post-operative therapy will result in improvement of shoulder function and pain relief, and it is therefore justified. One of the prerequisites for good results is the initial patient's consent with a supervised, active and long-term post-operative physical therapy. The transfer itself is a demanding surgical procedure associated with several risks and it should therefore be performed by an experienced and competent surgeon with a deep knowledge of shoulder girdle anatomy.


Repeated oronasal exposure to lipopolysaccharide induced mucosal IgA responses in periparturient dairy cows.
This study investigated the effects of repeated oronasal treatment with lipopolysaccharide (LPS) on the humoral immune responses in saliva, vaginal mucus, and the plasma markers of the acute phase response in periparturient dairy cows. One hundred pregnant Holstein cows were administered either 3 increasing doses of LPS (n = 50) as follows: 1) 0.01 µg/kg body weight (BW) on d -28, 2) 0.05 µg/kg BW on d -25, and -21, and 3) 0.1 µg/kg BW on d -18, and -14, or sterile saline solution (controls; n = 50) oronasally for 3 consecutive wk starting at 28 d before parturition. Intensive sampling was conducted on thirty cows (n = 15/group). Multiple saliva, vaginal mucus and blood samples were collected around parturition and analyzed for total immunoglobulin-(Ig)A, plasma serum amyloid A (SAA), lipopolysaccharide-binding protein (LBP), anti-LPS IgA, IgG, IgM, tumour necrosis factor(TNF)-α, and interleukin(IL)-1. Results regarding total secretory IgA (sIgA) antibodies showed greater concentrations in the saliva and an overall tendency for higher total sIgA in the vaginal mucus of the LPS-treated cows. Treatment had no effect on plasma sIgA, IgG, IgM anti-LPS antibodies, haptoglobin, SAA, LBP, TNF-α, and IL-1. Treatments by time interactions were observed for SAA and IL-1 with lowered concentrations of both variables in the plasma of LPS-treated cows after parturition. Overall, repeated oronasal LPS treatment clearly enhanced total sIgA antibodies in the saliva, stimulated their production in vaginal mucus shortly before calving, and lowered plasma IL-1 around parturition, but showed limited effects on markers of the acute phase response in the plasma in dairy cows around parturition.


Methionine metabolism by rat muscle and other tissues. Occurrence of a new carnitine intermediate.
Perfused rat hindquarter preparations were shown to incorporate radioactivity from [U-14C]methionine into citrate-cycle intermediates, lactate, alanine, glutamate, glutamine and CO2. During perfusion, large amounts of methionine were also oxidized to methionine sulphoxide. The capacity for transamination of methionine or its oxo analogue, 4-methylthio-2-oxobutyrate, by muscle extracts was demonstrated. Rat skeletal muscle, heart, liver and kidney mitochondria, when incubated with the latter plus radiolabelled carnitine, formed a newly identified carnitine derivative, 3-methylthiopropionylcarnitine. It is concluded that the capacity for oxidation of methionine by a trans-sulphuration-independent pathway occurs in several mammalian tissues. The extent of inter-organ handling of intermediates in this pathway(s) is discussed.


Family Medicine at the University of the West Indies: a model for the developing world.
Context and background: To describe the Family Medicine (FM) postgraduate training programme at the University of the West Indies (UWI). Actions and activities: This paper was created through a review of documents, and discussions with past and present coordinators and key stakeholders at four campuses in the English-speaking Caribbean (ESC). Despite intermittent setbacks the FM programme in the ESC has grown due to: (1) The presence of an umbrella institution in the UWI. (2) The role of the Caribbean College of Family Physicians providing a unifying force of advocacy and cooperation. (3) Collaboration of staff across four sites despite large distances, differing departmental and campus structures and financial models; and varying levels of local medical and public health support. (4) The use of a modular design for academic content, which means that students have comparable learning experiences. (5) Streamlining of exit examinations, thus sharing resources in the assessment process. (6) A strong presence of FM in the undergraduate curriculum. Despite a variety of timelines in programme development and funding mechanisms, over 150 physicians have graduated in FM in the past five years. We identify the unifying strategies and institutions which made this possible and present this model as an option for new programmes in the developing world.


P53 protein overexpression in de novo acute myeloid leukemia patients with normal diploid karyotype correlates with FLT3 internal tandem duplication and worse relapse-free survival.
Although ~50% of acute myeloid leukemia (AML) patients have a normal diploid karyotype by conventional cytogenetics at diagnosis, this patient subset has a variable disease course and outcome. Aberrant overexpression of the p53 protein is usually associated with TP53 alterations and a complex karyotype, but the prevalence and impact of p53 overexpression in AML with diploid cytogenetics is unknown. We examined 100 newly diagnosed AML patients to evaluate the impact of p53 expression status quantified in bone marrow core biopsy samples using immunohistochemistry and computer-assisted image analysis. A total of 24 patients had p53 overexpression defined as 3+ staining intensity in ≥5% of cells; this finding correlated with lower platelet counts (P = .002), absence of CD34 expression in blasts (P = .009), higher bone marrow blast counts (P = .04), and a higher frequency of FLT3 internal tandem duplication (P = .007). Overexpression of p53 independently predicted for shorter leukemia-free survival in patients who underwent allogeneic stem cell transplantation by univariate (P = .021) and multivariate analyses (P = .004). There was no correlation between MDM2 and p53 protein expression in this cohort. We conclude that p53 expression evaluated by immunohistochemistry in bone marrow biopsy specimens at the time of AML diagnosis may indicate distinct clinical characteristics in patients with normal diploid cytogenetics and is a potentially valuable tool that can enhance risk-stratification.


Water-mediated electrochemical nano-writing on thin ceria films.
Bias dependent mechanisms of irreversible cathodic and anodic processes on a pure CeO2 film are studied using modified atomic force microscopy (AFM). For a moderate positive bias applied to the AFM tip an irreversible electrochemical reduction reaction is found, associated with significant local volume expansion. By changing the experimental conditions we are able to deduce the possible role of water in this process. Simultaneous detection of tip height and current allows the onset of conductivity and the electrochemical charge transfer process to be separated, further elucidating the reaction mechanism. The standard anodic/cathodic behavior is recovered in the high bias regime, where a sizable transport current flows between the tip and the film. These studies give insight into the mechanisms of the tip-induced electrochemical reactions as mediated by electronic currents, and into the role of water in these processes, as well as providing a different approach for electrochemical nano-writing.


Symptom-based scoring technique by machine learning to predict COVID-19: a validation study.
Coronavirus disease 2019 (COVID-19) surges, such as that which occurred when omicron variants emerged, may overwhelm healthcare systems. To function properly, such systems should balance detection and workloads by improving referrals using simple yet precise and sensitive diagnostic predictions. A symptom-based scoring system was developed using machine learning for the general population, but no validation has occurred in healthcare settings. We aimed to validate a COVID-19 scoring system using self-reported symptoms, including loss of smell and taste as major indicators. A cross-sectional study was conducted to evaluate medical records of patients admitted to Dr. Sardjito Hospital, Yogyakarta, Indonesia, from March 2020 to December 2021. Outcomes were defined by a reverse-transcription polymerase chain reaction (RT-PCR). We compared the symptom-based scoring system, as the index test, with antigen tests, antibody tests, and clinical judgements by primary care physicians. To validate use of the index test to improve referral, we evaluated positive predictive value (PPV) and sensitivity. After clinical judgement with a PPV of 61% (n = 327/530, 95% confidence interval [CI]: 60% to 62%), confirmation with the index test resulted in the highest PPV of 85% (n = 30/35, 95% CI: 83% to 87%) but the lowest sensitivity (n = 30/180, 17%, 95% CI: 15% to 19%). If this confirmation was defined by either positive predictive scoring or antigen tests, the PPV was 92% (n = 55/60, 95% CI: 90% to 94%). Meanwhile, the sensitivity was 88% (n = 55/62, 95% CI: 87% to 89%), which was higher than that when using only antigen tests (n = 29/41, 71%, 95% CI: 69% to 73%). The symptom-based COVID-19 predictive score was validated in healthcare settings for its precision and sensitivity. However, an impact study is needed to confirm if this can balance detection and workload for the next COVID-19 surge.


CYR61/CCN1 and WISP3/CCN6 are chemoattractive ligands for human multipotent mesenchymal stroma cells.
CCN-proteins are known to be involved in development, homeostasis and repair of mesenchymal tissues. Since these processes implicate recruitment of cells with the potential to be committed to various phenotypes, we studied the effect of CYR61/CCN1 and WISP3/CCN6 on migration of human bone marrow derived mesenchymal stroma cells (MSCs) in comparison to in vitro osteogenic differentiated MSCs using a modified Boyden chamber assay. CYR61 and WISP3 were purified as fusion proteins with a C-terminal Fc-tag from baculovirus infected SF21 cells using protein G sepharose columns. CYR61 and WISP3 stimulated cell migration of undifferentiated MSCs in a dose-dependent manner. CYR61 and WISP3 had similar effects on committed osteogenic precursor cells. Checkerboard analysis revealed that CYR61 and WISP3 stimulated true directed cell migration (chemotaxis) of MSCs and committed osteogenic precursors. In MSCs the chemotactic activity of WISP3 but not CYR61 was mediated through integrin alphanuss5. Our results indicate that CYR61 and WISP3 can function as soluble ligands transmitting chemotactic signals to human MSCs but differ in the involvement of integrin alphanuss5. This may be relevant for their possible role in connective tissue repair.


Efficiency of use of imported nitrogen, phosphorus, and potassium and potential for reducing phosphorus imports on Idaho dairy farms.
Eight commercial dairies from south central Idaho were surveyed to estimate the whole-farm surpluses of N, P, and K and to investigate the possibility of reducing P excretions through dietary manipulation. Nitrogen, P, and K imports and exports were monitored in a 12-mo period, and samples from the diets, feeds, feces, urine, and manure were collected at regular farm visits. Soils from manure-amended fields were sampled in the spring and fall. In all cases, the largest import of N, P, and K to the dairy was with purchased feeds. Major nutrient export items were milk and manure and forages, in the case of a dairy with a large land base (dairy F). Whole-farm N surplus varied from 90 to 599 t/yr (91 to 222 kg/yr per cow). The efficiency of use of imported N varied from 25 to 64%, with dairy F having the greatest efficiency of imported N use. Phosphorus and K surpluses were also significant (average of 29 and 182 t/yr and 12 and 76 kg per cow per year, respectively). During the study period, dairy F was a net exporter of K. The average efficiency of use of imported P and K was 66 and 58%, respectively. Soil P levels in the 30-cm layer were above state threshold standards, most likely from overapplication of manure. Soil nitrate-N concentrations were also high, but K concentrations were within the accepted range. Average P content of the lactating cow diets at the start of the study was 0.49% and was reduced to 0.38%. The estimated reduction in imported P due to the reduced dietary P levels was from 5.7 to 61.4 t/yr per farm, or on average 12 kg per cow per year. This study demonstrated that in addition to exports with milk and manure, export of nutrients with forages produced on the farm (dairy F) is a major factor in reducing whole-farm N, P, and K surpluses.


A fast, highly sensitive double-nested PCR-based method to screen fish immunobiomes.
Efficient methods for constructing 16S tag amplicon libraries for pyrosequencing are needed for the rapid and thorough screening of infectious bacterial diversity from host tissue samples. Here we have developed a double-nested PCR methodology that generates 16S tag amplicon libraries from very small amounts of bacteria/host samples. This methodology was tested for 133 kidney samples from the lake whitefish Coregonus clupeaformis (Salmonidae) sampled in five different lake populations. The double-nested PCR efficiency was compared with two other PCR strategies: single primer pair amplification and simple nested PCR. The double-nested PCR was the only amplification strategy to provide highly specific amplification of bacterial DNA. The resulting 16S amplicon libraries were synthesized and pyrosequenced using 454 FLX technology to analyse the variation of pathogenic bacteria abundance. The proportion of the community sequenced was very high (Good's coverage estimator; mean = 95.4%). Furthermore, there were no significant differences of sequence coverage among samples. Finally, the occurrence of chimeric amplicons was very low. Therefore, the double-nested PCR approach provides a rapid, informative and cost-effective method for screening fish immunobiomes and most likely applicable to other low-density microbiomes as well.


National Clearinghouse for Poison Control Centers.
The Clearinghouse was originally conceived to coordinate information flow among the then few Poison Control Centers. Services were to be provided through the state health departments where complete control over the individual state's poison centers was to reside. No authority to establish or regulate centers, or to set standards for staffing and operation of centers, or to designate regional centers was granted. However, growth in the numbers of centers, more sophistocation in the state of the art, more demands for product and human experience data, calls for research support, regionalization, standards setting, competency testing, and others has led to an increasing call for more and more Clearinghouse involvement in the entire poison control program. Though staffing and funding remains fairly constant, the Clearinghouse hopes to be as involved and to provide as great a service as possible to those involved in clinical toxicology and poison control.


Passage of Coxsackie virus (Connecticut-5 strain) in adult mice with production of pancreatic disease.
1. With Conn.-5 strain of Coxsackie virus, pancreatic disease can be regularly produced in adult mice. 2. The lesions consist of widespread necrosis, followed by repair; there occurs more or less complete loss of glandular acini, with fatty or fibrous replacement. The islands of Langerhans and pancreatic ducts persist. 3. Injection of virus suspensions by the intraperitoneal, subcutaneous, intramuscular, or intracerebral route is followed by selective necrosis of the pancreas. 4. The liver, in the earlier stages of the disease, is the seat of fat infiltration. There may be necrosis of individual hepatic cells, but the diffuse hepatitis described in suckling mice does not occur. In the later stages of the disease, the liver is not significantly altered. 5. Localized areas of fat necrosis, scattered through intra-abdominal adipose tissue, are usually present in the acute phase of the disease. These undergo fibrosis without calcification. 6. No lesions have been found in the skeletal muscle, even at the site of intramuscular injection. Central nervous system, heart, lungs, and peripheral fat lobules show no lesions comparable to those described in suckling mice. 7. Multiplication of virus takes place in the pancreas. Serial passage in adult mice, by injection of pancreas suspensions prepared from organs removed on the 4th day after infection, is readily accomplished. Five consecutive passages in adult mice have thus far been carried out. Pancreas suspension from 4th passage material produced typical disease in suckling mice when diluted 10(-6). No virus could be demonstrated in pancreas obtained 25 days after inoculation. 8. Complete protection against the pancreatic disease is obtained when the virus is neutralized, before injection, with Conn.-5 antiserum. Normal mouse serum and antiserum against the Ohio-R strain of Coxsackie virus have no protective effect. 9. Mice surviving the initial necrotizing effect of the virus, develop chronic pancreatic insufficiency. This is manifested by extreme weight loss-in some cases, 40 per cent or more of the body weight-and by hypoproteinemia, sometimes leading to anasarca. 10. The substitution of fox-chow which has been predigested with hog pancreas brings about a restoration of weight. 11. The possibility is considered that similar lesions of the pancreas in human beings may be due to virus infection.


[Postoperative intraabdominal bleedings].
Based on complex diagnosis and treatment of 58 patients with postoperative intraabdominal bleedings (PIB), clinical classification and adequate surgical policy are proposed. It is demonstrated that in PIB of degree II-III urgent relaparotomy is indicated. In PIB of degree I dynamic follow-up of drainages, hemodynamic and hematological indices are possible.


Single-domain antibodies targeting antithrombin reduce bleeding in hemophilic mice with or without inhibitors.
Novel therapies for hemophilia, including non-factor replacement and in vivo gene therapy, are showing promising results in the clinic, including for patients having a history of inhibitor development. Here, we propose a novel therapeutic approach for hemophilia based on llama-derived single-domain antibody fragments (sdAbs) able to restore hemostasis by inhibiting the antithrombin (AT) anticoagulant pathway. We demonstrated that sdAbs engineered in multivalent conformations were able to block efficiently AT activity in vitro, restoring the thrombin generation potential in FVIII-deficient plasma. When delivered as a protein to hemophilia A mice, a selected bi-paratopic sdAb significantly reduced the blood loss in a model of acute bleeding injury. We then packaged this sdAb in a hepatotropic AAV8 vector and tested its safety and efficacy profile in hemophilic mouse models. We show that the long-term expression of the bi-paratopic sdAb in the liver is safe and poorly immunogenic, and results in sustained correction of the bleeding phenotype in hemophilia A and B mice, even in the presence of inhibitory antibodies to the therapeutic clotting factor.


Bradyrhizobium namibiense sp. nov., a symbiotic nitrogen-fixing bacterium from root nodules of Lablab purpureus, hyacinth bean, in Namibia.
Four strains of symbiotic bacteria from root nodules of hyacinth bean (Lablab purpureus (L.) Sweet) from Namibia were previously identified as a novel group within the genus Bradyrhizobium. To confirm their taxonomic status, these strains were further characterized by taking a polyphasic approach. The type strain possessed 16S rRNA gene sequences identical to Bradyrhizobium paxllaeri LMTR 21T and Bradyrhizobiumicense LMTR 13T, the full-length sequences were identical to those retrieved from SAMN05230119 and SAMN05230120, respectively. However, the intergenic spacer sequences of the novel group showed identities of less than 93.1 % to described Bradyrhizobium species and were placed in a well-supported separate lineage in the phylogenetic tree. Phylogenetic analyses of six concatenated housekeeping genes, recA, glnII, gyrB, dnaK, atpD and rpoB, corroborated that the novel strains belonged to a lineage distinct from named species of the genus Bradyrhizobium, with highest sequence identities to Bradyrhizobiumjicamae and B. paxllaeri (below 93 %). The species status was validated by results of DNA-DNA hybridization and average nucleotide identity values of genome sequences. The combination of phenotypic characteristics from several tests, including carbon source utilization and antibiotic resistance, could be used to differentiate representative strains from recognized species of the genus Bradyrhizobium. Phylogenetic analysis of nodC and nifH genes placed the novel strains in a group with B. paxllaeri and B.lablabi. Novel strain 5-10T induces effective nodules on Lablab purpureus, Vigna subterranea, Vigna unguiculata and Arachis hypogaea. Based on our results, we conclude that our strains represent a novel species for which the name Bradyrhizobium namibiense sp. nov. is proposed, with type strain 5-10T[LMG 28789, DSM 100300, NTCCM0017 (Windhoek)].


A Tc-99m HMPAO SPECT study of regional cerebral blood flow in drug-free schizophrenic patients with deficit and non-deficit syndrome.
Twenty-nine patients with DSM-IV diagnoses of schizophrenia were categorized into deficit syndrome (n=14) and non-deficit syndrome (n=15) subgroups on the basis of the Schedule for the Deficit Syndrome. The patients, who had all been free of antipsychotic medication for at least 3 weeks, and 17 sex- and age-matched normal controls were studied with single-photon emission computed tomography with Tc-99m HMPAO. Age at onset, Brief Psychiatric Rating Scale (BPRS) total scores, BPRS positive symptom subscores and duration of illness were similar between the two schizophrenic subgroups. As expected, the deficit patients had more negative symptoms than the non-deficit patients. There were no statistically significant correlations between clinical parameters and regional cerebral blood flow (rCBF) values. The deficit syndrome subgroup showed diminished rCBF in the frontal regions bilaterally, right parietal regions and right superior temporal region compared with the control groups. Deficit patients showed significantly lower rCBF perfusion ratios in the right superior and inferior frontal cortex than did the non-deficit patients. No differences were detected between the controls and the non-deficit schizophrenic patients in terms of rCBF perfusion indices. The results of the present study confirm previous reports of different patterns of rCBF in deficit vs. non-deficit schizophrenic subgroups.


A surface plasmon resonance assay for measurement of neuraminidase inhibition, sensitivity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drugs zanamivir and oseltamivir.
Antiviral resistance is currently monitored by a labelled enzymatic assay, which can give inconsistent results because of the short half-life of the labelled product, and variations in assay conditions. In this paper, we describe a competitive surface plasmon resonance (SPR) inhibition assay for measuring the sensitivities of wild-type neuraminidase (WT NA) and the H274Y (histidine 274 tyrosine) NA mutant to antiviral drugs. The two NA isoforms were expressed in High-five™ (Trichoplusia ni) insect cells. A spacer molecule (1,6-hexanediamine (HDA)) was conjugated to the 7-hydroxyl group of zanamivir, and the construct (HDA-zanamivir) was immobilized onto a SPR sensor chip to obtain a final immobilization response of 431 response units. The immobilized HDA-zanamivir comprised a bio-specific ligand for the WT and mutant proteins. The effects of the natural substrate (sialic acid) and two inhibitors (zanamivir and oseltamivir) on NA binding to the immobilized ligand were studied. The processed SPR data was analysed to determine 50% inhibitory concentrations (IC50-spr ), using a log dose-response curve fit. Although both NA isoforms had almost identical IC50-spr values for sialic acid (WT = 5.5 nM; H274Y mutant = 3.25 nM) and zanamivir (WT = 2.16 nM; H274Y mutant = 2.42 nM), there were significant differences between the IC50-spr values obtained for the WT (7.7 nM) and H274Y mutant (256 nM) NA in the presence of oseltamivir, indicating that oseltamivir has a reduced affinity for the H274Y mutant. The SPR inhibition assay strategy presented in this work could be applied for the rapid screening of newly emerging variants of NA for their sensitivity to antiviral drugs.


GIGANTEA-like genes control seasonal growth cessation in Populus.
Survival of trees growing in temperate zones requires cycling between active growth and dormancy. This involves growth cessation in the autumn triggered by a photoperiod shorter than the critical day length. Variations in GIGANTEA (GI)-like genes have been associated with phenology in a range of different tree species, but characterization of the functions of these genes in the process is still lacking. We describe the identification of the Populus orthologs of GI and their critical role in short-day-induced growth cessation. Using ectopic expression and silencing, gene expression analysis, protein interaction and chromatin immunoprecipitation experiments, we show that PttGIs are likely to act in a complex with PttFKF1s (FLAVIN-BINDING, KELCH REPEAT, F-BOX 1) and PttCDFs (CYCLING DOF FACTOR) to control the expression of PttFT2, the key gene regulating short-day-induced growth cessation in Populus. In contrast to Arabidopsis, in which the GI-CONSTANS (CO)-FLOWERING LOCUS T (FT) regulon is a crucial day-length sensor for flowering time, our study suggests that, in Populus, PttCO-independent regulation of PttFT2 by PttGI is more important in the photoperiodic control of growth cessation and bud set.


Chest pain centers: diagnosis of acute coronary syndromes.
Chest pain centers in the emergency department have generally been accepted as a safe, cost-effective, and rapid approach to the evaluation, triage, and management of patients with potential acute coronary syndromes. These centers were initially designed to enhance patient care by decreasing time to treatment for acute myocardial infarction (AMI) and rapidly identifying patients with unstable angina. They also included community outreach and educational objectives designed to reduce time from the onset of chest pain to ED presentation. In the past decade, health care financial constraints have created additional impetus to the development of chest pain centers. Cost reduction efforts have occurred to reduce hospitalizations, lengths of stay, and unnecessary treatments and procedures. Practitioners and administrators try to balance these goals with the imperative to provide high-quality patient care. Protocol-driven approaches have been developed for specific disease processes in emergency settings. The chest pain center concept is such an approach for patients with chest pain. Chest pain is the second most common ED presenting complaint and is a symptom related to the leading cause of death in the United States, coronary artery disease (CAD). One third of ED patients with chest pain will eventually have a diagnosis of acute coronary syndrome. Many patients with acute coronary syndromes have atypical presentations that are not diagnosed in the ED with the traditional diagnostic evaluation of a history, physical examination, and 12-lead ECG. If they are not admitted to the hospital for further evaluation, the diagnosis may be missed. The 2% to 5% of AMI patients who are inadvertently released home often have poor outcomes and result in a leading cause of malpractice suits in emergency medicine. More than one half of ED patients with chest pain have clinical findings after their initial evaluation consistent with acute coronary syndromes and are admitted to the hospital. Approximately one half of these patients, after evaluation in the hospital, are found not to have acute coronary syndromes. The cost for these negative inpatient cardiac evaluations has been estimated to be $6 billion in the United States each year. Today, chest pain centers serve as an integral component of many EDs. Their success and safety is the result of a focused, protocol-driven approach directed at the acute coronary syndrome continuum from unstable angina to transmural Q-wave myocardial infarction. New therapies for acute coronary syndromes make ED triage and risk stratification increasingly important. Although different chest pain center protocols have proved effective, all address the diagnosis and rapid treatment of acute myocardial necrosis, rest ischemia, and exercise-induced ischemia. Identifying patients with coronary artery disease in one of these stages in the spectrum of myocardial ischemia is the foundation for a successful chest pain center in the ED.


Reconsolidation or extinction: transcription factor switch in the determination of memory course after retrieval.
In fear conditioning, aversive stimuli are readily associated with contextual features. A brief reexposure to the training context causes fear memory reconsolidation, whereas a prolonged reexposure induces memory extinction. The regulation of hippocampal gene expression plays a key role in contextual memory consolidation and reconsolidation. However, the mechanisms that determine whether memory will reconsolidate or extinguish are not known. Here, we demonstrate opposing roles for two evolutionarily related transcription factors in the mouse hippocampus. We found that nuclear factor-κB (NF-κB) is required for fear memory reconsolidation. Conversely, calcineurin phosphatase inhibited NF-κB and induced nuclear factor of activated T-cells (NFAT) nuclear translocation in the transition between reconsolidation and extinction. Accordingly, the hippocampal inhibition of both calcineurin and NFAT independently impaired memory extinction, whereas inhibition of NF-κB enhanced memory extinction. These findings represent the first insight into the molecular mechanisms that determine memory reprocessing after retrieval, supporting a transcriptional switch that directs memory toward reconsolidation or extinction. The precise molecular characterization of postretrieval processes has potential importance to the development of therapeutic strategies for fear memory disorders.


[Changes in dipeptidyl carboxypeptidase activity during hypertension and its complications].
Using the fluorimetric method, the activity of dipeptidyl-carboxypeptidase (DCP) was studied in 85 patients suffering from essential hypertension. It was ascertained that the DCP activity in the blood serum of such patients was significantly higher than in normal subjects. The highest activity of the enzyme was observed in cases where essential hypertension was aggravated by chronic heart failure or an acute impairment of the cerebral circulation. Inflammatory processes in hypertensive patients were associated with a considerable fall in the serum enzyme activity.


The use of a Foley balloon catheter to control junctional hemorrhage in a dog with severe vascular injury secondary to penetrating trauma.
Penetrating trauma is commonly seen in dogs. The severity depends on the site of injury and tissue involved. Junctional hemorrhage can be especially challenging to control given the inaccessibility of the damaged vasculature. Methods described to control life-threatening hemorrhage in dogs include direct pressure, hemostatic gauze, hemostatic powder or granules, wound packing, tourniquets, and direct clamping of the vasculature. Foley balloon catheters (FBC) are commonly used to tamponade deep vascular hemorrhage in people, but the technique has not been previously described in the veterinary literature. To present a case of penetrating trauma (bite wound) in a dog with a transected left femoral artery and vein in which the life-threatening hemorrhage was initially controlled with tamponade using an FBC. A 7-year-old neutered male Terrier mix presented in hemorrhagic shock with an Animal Trauma Triage (ATT) of 7 and modifed Glasgow coma scale (MGCS) of 17 forty-five minutes after being attacked by another dog. The dog had sustained a deep penetrating wound to the left groin. Direct pressure and gauze packing at the site of injury were not successful at slowing the hemorrhage. A 10-Fr, 55-cm Foley catheter with a 5-mL balloon was inserted into the wound tract, and the balloon was inflated with 7.5 mL of sterile saline. Hemorrhage was controlled after inflation of the Foley balloon. CBC, blood biochemistries, abdominal point-of-care ultrasound, radiographs, prothrombin time, partial thromboplastin time, and whole blood viscoelastic testing were performed. Stabilization included fluid resuscitation, analgesics, antimicrobials, and epsilon aminocaproic acid. The dog was then anesthetized to definitively identify and control the hemorrhage. Transection of the left femoral artery and vein where identified and ligated. The dog fully recovered and was discharged 32 hours later. FBCs may be useful as an alternative technique for temporary control of life-threatening hemorrhage secondary to penetrating injuries in both the emergency department and prehospital settings.


Parallel repulsive logic regression with biological adjacency.
Logic regression, an extension of generalized linear models with Boolean combinations of binary variables as predictors, is a useful tool in exploring interactions among single-nucleotide polymorphisms (SNPs) in genome-wide association studies. However, since the search space defined by all possible combinations of SNPs, their complements, and logical operators in Boolean expressions can be exceedingly large in such studies, objective function optimization is slow and likely to be trapped in many local solutions, resulting in model over-fitting. We introduce a new search algorithm, parallel repulsive logic regression (PRLR), to efficiently estimate parameters of a logic regression to find a best model within the large space of SNP interactions by incorporating: (i) relevant biological adjacency matrix between SNPs to define similarity of estimation paths or trees, which are derived from physical SNP positions on chromosomes and/or memberships in biological gene pathways; and (ii) two repulsive forces to counter the similarity between and within estimation paths considered in parallel, which are introduced as penalty terms in the objective function. We compare our method's performance for identifying biologically-meaningful SNP interactions through simulations and with real genetic-epidemiological data. PRLR's detection-accuracy measures outperform existing approaches, especially in terms of positive predictive value and sensitivity for detecting SNP-SNP interactions.


Gastrostomy tube placement by radiological versus endoscopic methods in an acute care setting: a retrospective review of frequency, indications, complications and outcomes.
To describe the current practice of placing gastrostomy tubes (endoscopic and radiological), patient characteristics, indications for enteral support, complications and outcomes over a 13-month period, and explore factors that influenced complications and outcomes. Second, to provide Canadian data regarding feeding tube placement because no current literature reflecting these practices for Canadian hospitals is available. Retrospective chart reviews were conducted. Patients who had initial percutaneous endoscopic gastrostomy (PEG) or percutaneous radiological gastrostomy (PRG) tubes inserted for nutritional purposes were included in the study. A total of 136 charts which included 30 PEG and 44 PRG procedures were reviewed. The PRG group was older than the PEG group (mean [+/-SD+/-5D; age 68+/-19 years versus 55+/-21 years, respectively; P=0.008). Patients in PEG group had longer lengths of hospital stay and more intensive care unit admissions than the PRG group (P=0.029). The main reason for tube insertion was dysphagia/aspiration (PEG [60%] and PRG [77%]). Minor complications were comparable between the two groups (P=0.678). There were three cases of major complications overall. More subjects in the PRG group died (18%) while in hospital than in the PEG group (3%) (P=0.055). No procedure-related deaths occurred in either group. Both methods of tube insertion provided a safe route for nutrition delivery despite a significant cost differential with PEGs costing 44% more than PRGs. Characteristics such as age, presence of ascites and severity of disease influenced the method of insertion despite the lack of current guidelines. Overall, the present study provides new descriptive data in a Canadian context.


Risk Factors for Depression in Patients with Chronic Obstructive Pulmonary Disease.
BACKGROUND Depression is a major comorbidity in Chronic Obstructive Pulmonary Disease (COPD). The aim of this study was to explore cytokine levels, and socio-demographic and clinical factors that may affect COPD-related depression. MATERIAL AND METHODS This prospective study enrolled 53 consecutive COPD patients, without any other lower respiratory tract diseases, psychiatric disorders, family history or cognitive disorders, who were hospitalized in the Department of Respiratory Medicine of Hefei First People's Hospital, China between October 2015 and October 2016. All patients were investigated for depression, and the serum levels of C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-8 (IL-8) and Tumor Necrosis Factor-α (TNF-α) were measured. The factors that may affect COPD-related depression were analyzed. RESULTS There were 40 (75.47%) patients with depression. There were differences in gender, smoking, time of cough, education, forced expiratory volume in the first second (FEV1) and serum CRP levels between COPD patients with or without depression. In a univariate analysis, gender, smoking, education, FEV1, time of cough and serum CRP level were associated with depression. In a multivariate logistic regression model, serum CRP level and FEV1 were risk factors for depression. CONCLUSIONS Male patients, heavy smoking, higher academic qualifications, cough, and high serum CRP level were linked to higher incidence rate of depression. High serum CRP level and low FEV1 were the risk factors for COPD-related depression.


The structure- and metal-dependent activity of Escherichia coli PgaB provides insight into the partial de-N-acetylation of poly-β-1,6-N-acetyl-D-glucosamine.
Exopolysaccharides are required for the development and integrity of biofilms produced by a wide variety of bacteria. In Escherichia coli, partial de-N-acetylation of the exopolysaccharide poly-β-1,6-N-acetyl-D-glucosamine (PNAG) by the periplasmic protein PgaB is required for polysaccharide intercellular adhesin-dependent biofilm formation. To understand the molecular basis for PNAG de-N-acetylation, the structure of PgaB in complex with Ni(2+) and Fe(3+) have been determined to 1.9 and 2.1 Å resolution, respectively, and its activity on β-1,6-GlcNAc oligomers has been characterized. The structure of PgaB reveals two (β/α)(x) barrel domains: a metal-binding de-N-acetylase that is a member of the family 4 carbohydrate esterases (CE4s) and a domain structurally similar to glycoside hydrolases. PgaB displays de-N-acetylase activity on β-1,6-GlcNAc oligomers but not on the β-1,4-(GlcNAc)(4) oligomer chitotetraose and is the first CE4 member to exhibit this substrate specificity. De-N-acetylation occurs in a length-dependent manor, and specificity is observed for the position of de-N-acetylation. A key aspartic acid involved in de-N-acetylation, normally seen in other CE4s, is missing in PgaB, suggesting that the activity of PgaB is attenuated to maintain the low levels of de-N-acetylation of PNAG observed in vivo. The metal dependence of PgaB is different from most CE4s, because PgaB shows increased rates of de-N-acetylation with Co(2+) and Ni(2+) under aerobic conditions, and Co(2+), Ni(2+) and Fe(2+) under anaerobic conditions, but decreased activity with Zn(2+). The work presented herein will guide inhibitor design to combat biofilm formation by E. coli and potentially a wide range of medically relevant bacteria producing polysaccharide intercellular adhesin-dependent biofilms.


First-line Therapeutic Strategies for Myelodysplastic Syndromes.
The precise diagnostic tests and subsequent prognostic stratification for patients with myelodysplastic syndrome (MDS) are often cumbersome, yet they are the basis of successful therapy. Diverse treatment options are available for these patients; however, the decisions in real-life are often not grounded on the available evidence. Although the International Prognostic Scoring System and revised International Prognostic Scoring System are still driving the medical approach to MDS patients, additional variables must be considered when therapeutic intervention is needed. A rational scheme for first-line therapy is described that allows for the possibility of selecting the optimal individual therapy for MDS patients.


Intensive Glycemic Therapy in Type 2 Diabetes Patients With Cardiac Autonomic Dysfunction: The ACCORD Trial.
To assess whether the presence of cardiac autonomic dysfunction denoted by low heart rate variability (HRV) modifies the effect of intensive glycemic therapy on outcomes in patients with type 2 diabetes. This study included 7946 participants in the ACCORD (Action to Control Cardiovascular Risk in Diabetes) trial from January 2001 through June 2009. Heart rate variability measures included standard deviation of all normal-to-normal intervals (SDNN) and root mean square of successive differences between normal-to-normal intervals (rMSSD). Abnormal values were defined based on less than the 10th percentile for SDNN and rMSSD. Compared with standard therapy, intensive therapy was associated with improved primary outcome (composite of cardiovascular events) in the low-HRV group (SDNN: HR, 0.57; 95% CI, 0.39 to 0.84; rMSSD: HR, 0.57; 95% CI, 0.38 to 0.84), but not in the normal-HRV group (SDNN: HR, 0.90; 95% CI, 0.77 to 1.05; rMSSD: HR, 0.90; 95% CI, 0.77 to 1.05). A similar pattern was found for coronary heart disease. Conversely, intensive therapy had a neutral effect on all cause death in the low-HRV group (SDNN: HR, 0.88; 95% CI, 0.54 to 1.41; rMSSD: HR, 0.71; 95% CI, 0.43 to 1.17;), but increase risk of all-cause death in the normal-HRV group (SDNN: HR, 1.21; 95% CI, 1.00 to 1.46; rMSSD: HR, 1.25; 95% CI, 1.03 to 1.51). Intensive therapy induced a greater risk of hypoglycemia in the normal-HRV group than that in the low-HRV group. Cardiac autonomic dysfunction expressed as low HRV identified subpopulations in ACCORD with more benefits and less harms from intensive therapy.


Joint effects of inflammation and androgen metabolism on prostate cancer severity.
Multiple pathways of prostate carcinogenesis have been proposed, including those involving androgen metabolism and inflammation. These pathways are not independent, and may act together in prostate cancer etiology: androgens promote both inflammatory processes and serve as mitogens in prostate tumor growth. To explore the possible joint effects of these pathways in prostate cancer severity, we studied 1,090 Caucasian prostate cancer cases to evaluate whether tumor severity is influenced by a history of benign prostatic hyperplasia (BPH) interacting with genotypes involved in inflammation or androgen metabolism including MSR1, RNASEL, AR, CYP3A4, CYP3A43, CYP3A5 and SRD5A2. We observed a statistically significant interaction between a number of genotypes and BPH. After considering the potential for false positive associations, the only remaining significant associations involved CYP3A43 P340A genotypes and history of BPH on both Gleason grade (interaction p-value = 0.026) and tumor stage (interaction p-value = 0.017). These results suggest that androgen metabolism may act in concert with inflammatory phenotypes such as BPH in determining prostate cancer severity.


[Indices of humoral antimeasles postvaccinal immunity].
Inoculation to children of live measles L-16 vaccine was found to cause changes in the content of the main serum classes of immunoglobulins, A, M and G, and synthesis of specific antibody of various physico-chemical nature. Between these two values only a relative correlation was observed at 10--14 and 28--32 days after vaccination. Changes in the synthesis of serum immunoglobulins in the majority of the children were not great and statistically insignificant. Only in 2 children with hypergammaglobulinemia inoculation of measles L-16 vaccine induced intensive (statistically significant) increase in serum M and G globulins. The results indicate the necessity of further study of the effects of live measles vaccine on the immunological status of children which could probably reveal the causes of occurrence of unusual postvaccination reactions and complications.


The type III secretion injectisome, a complex nanomachine for intracellular 'toxin' delivery.
The type III secretion injectisome is a nanomachine that delivers bacterial proteins into the cytosol of eukaryotic target cells. It consists of a cylindrical basal structure spanning the two bacterial membranes and the peptidoglycan, connected to a hollow needle, eventually followed by a filament (animal pathogens) or to a long pilus (plant pathogens). Export employs a type III pathway. During assembly, all the protein subunits of external elements are sequentially exported by the basal structure itself, implying that the export apparatus can switch its substrate specificity over time. The length of the needle is controlled by a protein that it also secreted during assembly and presumably acts as a molecular ruler.


Alternative rearing systems in pigs: consequences on stress indicators at slaughter and meat quality.
The objective of this study was to evaluate the effects of three alternative (ALT) rearing systems for growing pigs (outdoor: 150 m2/pig; straw bedding: 1.30 m2/pig; and hut with access to a courtyard: 1.30 m2/pig) compared with a conventional system (fully slatted floor: 0.65 m2/pig, considered as control), on pre-slaughter stress indicators in relation with meat quality. To that end, the number of skin lesions on whole carcasses, as well as blood creatine kinase (CK) activity and urine levels in cortisol and catecholamines (adrenaline and noradrenaline) were determined at slaughter. Glycolytic potential (GP) and ultimate pH of the semimembranosus muscle were also measured. The global correlation network calculated between all these parameters shows that the indicators of pre-slaughter muscle activity (plasma CK) and/or stress indicators (e.g. adrenaline) are negatively (r=-0.26, P<0.01; r=-0.29, P<0.05, respectively) correlated with muscle GP and positively (r=0.17, P<0.05; r=0.44, P<0.001, respectively) with meat ultimate pH. Although some traits measured were sensitive to the degree of pre-slaughter mixing, they differed across rearing systems. The differences were most pronounced for the comparison of outdoors v. slatted floor. The lower levels of plasma CK and urinary catecholamines, and the lower number of carcass skin lesions of pigs reared outdoors, were related to a lower meat ultimate pH. Thus, ALT rearing systems influence animal welfare and meat quality, by providing enriched environmental conditions to the animals.


Directing the Self-Assembly Behaviour of Porphyrin-Based Supramolecular Systems.
The self-assembly behaviour of a library of tetra-amidated porphyrin molecules decorated with a variety of solubilizing wedges is investigated as dilute solutions in methylcyclohexane. Small changes in the solubilising wedge of the porphyrins resulted in different aggregated states, as evidenced by CD and UV/Vis absorption spectroscopy. The porphyrins form co-facially stacked H-aggregates, slip-stacked J-aggregates or a mixture of both. Detailed thermodynamic and kinetic analysis showed that in all cases the formation of J-aggregates proceeds via an isodesmic mechanism whereas H-aggregates are formed via a cooperative mechanism. It is shown that these aggregates assemble in a parallel pathway, in which both compete for the monomer, compared to a sequential pathway, in which one of the aggregates interconverts into the other. Interestingly, kinetic analysis of porphyrins that only form H-aggregates in thermodynamic equilibrium revealed that the competing pathway towards J-aggregates is operational in these systems as well. Our findings show that the balance between H- and J-aggregates depends on remarkably small changes in the architecture of the solubilising wedges.


Perceived burdensomeness and suicide-related behaviors in clinical samples: current evidence and future directions.
The interpersonal-psychological theory of suicide identifies perceived burdensomeness as a primary component of suicidal desire and a possible point of intervention for suicide prevention. A growing literature has explored the relationship between perceived burdensomeness and suicide-related behaviors. The aim of this review is to integrate the evidence, identify critical gaps in the evidence-base, and explore implications for translation to prevention and intervention science. Papers published that reported on the association between perceived burdensomeness and suicide-related behaviors were included. The literature indicates (a) significant cross-sectional associations between perceived burdensomeness and suicidal ideation and suicide attempts and (b) that perceived burdensomeness acts as either a moderator or a mediator of the association between risk and protective factors and suicide-related behaviors. Research is needed to examine the longitudinal association between perceived burdensomeness and suicide-related behaviors, develop additional measurement approaches, generalize findings to other samples, and begin translating findings to prevention and intervention science.


EPGAT: Gene Essentiality Prediction With Graph Attention Networks.
Identifying essential genes and proteins is a critical step towards a better understanding of human biology and pathology. Computational approaches helped to mitigate experimental constraints by exploring machine learning (ML) methods and the correlation of essentiality with biological information, especially protein-protein interaction (PPI) networks, to predict essential genes. Nonetheless, their performance is still limited, as network-based centralities are not exclusive proxies of essentiality, and traditional ML methods are unable to learn from non-euclidean domains such as graphs. Given these limitations, we proposed EPGAT, an approach for Essentiality Prediction based on Graph Attention Networks (GATs), which are attention-based Graph Neural Networks (GNNs), operating on graph-structured data. Our model directly learns gene essentiality patterns from PPI networks, integrating additional evidence from multiomics data encoded as node attributes. We benchmarked EPGAT for four organisms, including humans, accurately predicting gene essentiality with ROC AUC score ranging from 0.78 to 0.97. Our model significantly outperformed network-based and shallow ML-based methods and achieved a very competitive performance against the state-of-the-art node2vec embedding method. Notably, EPGAT was the most robust approach in scenarios with limited and imbalanced training data. Thus, the proposed approach offers a powerful and effective way to identify essential genes and proteins.


Ezetimibe blocks the internalization of NPC1L1 and cholesterol in mouse small intestine.
The multiple transmembrane protein Niemann-Pick C1 like1 (NPC1L1) is essential for intestinal cholesterol absorption. Ezetimibe binds to NPC1L1 and is a clinically used cholesterol absorption inhibitor. Recent studies in cultured cells have shown that NPC1L1 mediates cholesterol uptake through vesicular endocytosis that can be blocked by ezetimibe. However, how NPC1L1 and ezetimibe work in the small intestine is unknown. In this study, we found that NPC1L1 distributed in enterocytes of villi and transit-amplifying cells of crypts. Acyl-CoA cholesterol acyltransferase 2 (ACAT2), another important protein for cholesterol absorption by providing cholesteryl esters to chylomicrons, was mainly presented in the apical cytoplasm of enterocytes. NPC1L1 and ACAT2 were highly expressed in jejunum and ileum. ACAT1 presented in the Paneth cells of crypts and mesenchymal cells of villi. In the absence of cholesterol, NPC1L1 was localized on the brush border of enterocytes. Dietary cholesterol induced the internalization of NPC1L1 to the subapical layer beneath the brush border and became partially colocalized with the endosome marker Rab11. Ezetimibe blocked the internalization of NPC1L1 and cholesterol and caused their retention in the plasma membrane. This study demonstrates that NPC1L1 mediates cholesterol entering enterocytes through vesicular endocytosis and that ezetimibe blocks this step in vivo.


Robust and Reproducible Protocol for Phage Genome "Rebooting" Using Transformation-Associated Recombination (TAR) Cloning into Yeast Centromeric Plasmid.
Production of infectious bacteriophage based on its genome is one of the necessary steps in the pipeline of editing phage genomes and creating synthetic bacteriophages. This process is called "rebooting" of the phage genome. In this chapter, we describe key steps required for successful genome "rebooting" using a native host or intermediate host. A detailed protocol is given for the "rebooting" of the genome of T7 bacteriophage specific to Escherichia coli and bacteriophage KP32_192 that infects Klebsiella pneumoniae.


The Benjamin H. Kean Travel Fellowship in Tropical Medicine: Assessment of Impact at 15 Years.
The Benjamin H. Kean Fellowship in Tropical Medicine is an American Society of Tropical Medicine and Hygiene initiative that provides medical students with funding for international clinical or research experiences lasting at least 1 month. Of the 175 Kean fellows from 1998 to 2013, 140 had current available e-mails, and 70 of the 140 (50%) responded to a survey about their fellowship experience. Alumni indicated that the Kean Fellowship had a high impact on their career plans with regard to preparation for (N = 65, 94.2%) and inspiration to pursue (N = 59, 88.1%) a career in tropical medicine and global health. Continued involvement in tropical medicine and global health was common: 52 alumni (74.3%) were currently working in tropical medicine or global health, 49 (71.0%) had done so in the interim between the Kean fellowship and their current position; and 17 of 19 Kean fellows (89.4%) who had completed all medical training and were now in professional practice continued to work in tropical medicine and global health. Alumni had been highly productive academically, publishing a total of 831 PubMed-indexed manuscripts, almost all on tropical medicine or global health topics, in the period between their fellowship year and 2013. Alumni reported strengths of the fellowship including funding, networking, and flexibility, and suggested that more networking and career mentoring would enhance the program. The Benjamin H. Kean fellowship program has been highly successful at inspiring and fostering ongoing work by trainees in tropical medicine and global health.


Comparative Outcome of Computed Tomography-guided Percutaneous Radiofrequency Ablation, Partial Nephrectomy or Radical Nephrectomy in the Treatment of Stage T1 Renal Cell Carcinoma.
Renal cell carcinoma (RCC) accounts for 3% of all cancers in adults. The indications for Radiofrequency Ablation (RFA) for renal carcinomas include T1a (tumor 4 cm or less, limited to the kidney), elderly patients, renal impairment, comorbidities, poor surgical candidate, and multiple bilateral renal masses. We retrospectively reviewed medical records, specifically investigating the indications, complications and outcomes of RFA and nephrectomy for treatment of RCC in a tertiary medical center with a predominantly Hispanic patient population. Forty-nine patients with RCC were evaluated. Nine patients had RFA, 9 had partial nephrectomy and 31 had radical nephrectomy. All patients among the 3 groups had stage T1N0M0 RCC at diagnosis. Tumor recurrence was observed in 2 (22%) patients that had RFA, one (11%) patient that had partial nephrectomy and no patients that had radical nephrectomy. One patient had recurrence of the tumor at the opposite kidney pole from the initial RFA site 4 years later. This particular patient did not have any tumor recurrence at the site of the initial RFA. A second RFA was performed on the recurrent tumor with no recurrence upon subsequent follow up visits. The second patient had recurrence of the RCC on 1 year follow that was discovered to be sarcomatoid RCC, which is an aggressive type with a poor prognosis. Our results support the clinical utility of RFA in patients with stage T1 RCC who are poor surgical candidates or those with reduced renal function. The clinical utility of RFA as an equally effective approach when compared to partial nephrectomy in patients with stage T1 RCC that meet strict indications for the procedure. The treatment choice should be individualized and based on the characteristics of the renal tumor such as size, location and histological type of RCC. We conclude that RFA presents a safe treatment choice for patients with RCC if long term follow up is maintained.