Detection of coronary artery disease in patients with chest pain: A machine learning model based on magnetocardiography parameters.
Patients with chest pain and suspected of coronary artery disease(CAD) need further test to confirm the diagnosis. Magnetocardiography (MCG) is a non-invasive and emission-free technology which can detect and measure the weak magnetic fields created by the electrical activity of the heart. This study aimed to investigate the usefulness of the 10 MCG parameters to detect CAD in patients with chest pain by means of a machine learning method of multilayer perceptron(MLP) neural network. 209 patients who were suffering from chest pain and suspected of CAD were enrolled in this cross-sectional study. In all patients, 12-lead electrocardiography(ECG) and MCG test were performed before percutaneous coronary angiography(PCA). 10 MCG parameters were analyzed by MLP neural networks. 11 diagnostic models(M1 to M11) were established after MLP analysis. The accuracies ranged from 71.2% to 90.5%. Two models(M10 and M11) were further analyzed. The accuracy, sensitivity, specificity, PPV, NPV, PLR and NLR were 89.5%, 89.8%, 88.9%, 92.7%, 84.7%, 11.10 and 0.11, of M10, and were 90.0%, 91.4%, 87.7%, 92.1%, 86.6%, 7.43 and 0.10, of M11. By a method of MLP neural network, MCG is applicable in identifying CAD in patients with chest pain, which seems beneficial for detection of CAD.


Nephrology Provider Surprise Question Response and Hospitalizations in Older Adults with Advanced CKD.
Older adults with advanced non-dialysis-dependent chronic kidney disease (NDD-CKD) face a high risk of hospitalization and related adverse events. This prospective cohort study followed nephrology clinic patients ≥60 years old with NDD-CKD stages 4-5. After an eligible patient's office visit, study staff asked the patient's provider to rate the patient's risk of death within the next year using the surprise question ("Would you be surprised if this patient died in the next 12 months?") with a 5-point Likert scale response (1, "definitely not surprised" to 5, "very surprised"). We used a statewide database to ascertain hospitalization during follow-up. There were 488 patients (median age 72 years, 51% female, 17% black) with median estimated glomerular filtration rate 22 mL/min/1.73 m2. Over a median follow-up of 2.1 years, the rates of hospitalization per 100 person-years in the respective response groups were 41 (95% confidence interval [CI]: 34-50), "very surprised"; 65 (95% CI: 55-76), "surprised"; 98 (95% CI: 85-113), "neutral"; 125 (95% CI: 107-144), "not surprised"; and 120 (95% CI: 94-151), "definitely not surprised." In a fully adjusted cumulative probability ordinal regression model for proportion of follow-up time spent hospitalized, patients whose providers indicated that they would be "definitely not surprised" if they died spent a greater proportion of follow-up time hospitalized compared with those whose providers indicated that they would be "very surprised" (odds ratio 2.4, 95% CI: 1.0-5.7). There was a similar association for time to first hospitalization. Nephrology providers' responses to the surprise question for older patients with advanced NDD-CKD were independently associated with proportion of future time spent hospitalized and time to first hospitalization. Additional studies should examine how to use this information to provide patients with anticipatory guidance on their possible clinical trajectory and to target potentially preventable hospitalizations.


Inorganic carbon limitation during nitrogen conversions in sponge-bed trickling filters for mainstream treatment of anaerobic effluent.
Anaerobic sewage treatment is a proven technology in warm climate regions, and sponge-bed trickling filters (SBTFs) are an important post-treatment technology to remove residual organic carbon and nitrogen. Even though SBTFs can achieve a reasonably good effluent quality, further process optimization is hampered by a lack of mechanistic understanding of the factors influencing nitrogen removal, notably when it comes to mainstream anaerobically treated sewage. In this study, the factors that control the performance of SBTFs following anaerobic (i.e., UASB) reactors for sewage treatment were investigated. A demo-scale SBTF fed with anaerobically pre-treated sewage was monitored for 300 days, showing a median nitrification efficiency of 79% and a median total nitrogen removal efficiency of 26%. Heterotrophic denitrification was limited by the low organic carbon content of the anaerobic effluent. It was demonstrated that nitrification was impaired by a lack of inorganic carbon rather than by alkalinity limitation. To properly describe inorganic carbon limitation in models, bicarbonate was added as a state variable and sigmoidal kinetics were applied. The resulting model was able to capture the overall long-term experimental behaviour. There was no nitrite accumulation, which indicated that nitrite oxidizing bacteria were little or less affected by the inorganic carbon limitation. Overall, this study indicated the vital role of influent characteristics and operating conditions concerning nitrogen conversions in SBTFs treating anaerobic effluent, thus facilitating further process optimization.


NG-nitro-L-arginine (N5-[imino(nitroamino)methyl]-L-ornithine) impairs endothelium-dependent dilations by inhibiting cytosolic nitric oxide synthesis from L-arginine.
We studied the effects of the L-arginine analogue NG-nitro-arginine (L-NNA), in comparison with its D-isomer [D-NNA), on endothelium-dependent dilations of rabbit femoral arteries (RFA) and on the release of endothelium-derived relaxant factor (EDRF) from native and cultured endothelial cells. In addition, we examined the effects of L- and D-NNA on the L-arginine- and NADPH-dependent synthesis of nitric oxide (NO) in the cytosol of porcine aortic endothelial cells. L-NNA enhanced the noradrenaline-induced contraction of endothelium-intact, but not of endothelium-denuded segments of RFA, indicating an inhibition of basal EDRF release. L-NNA also inhibited significantly the endothelium-dependent dilations to acetylcholine (ACh). Both effects of L-NNA were attenuated by L-arginine. L-NNA rapidly inhibited the release of EDRF from cultured and native endothelial cells stimulated with thimerosal or ACh. L-NNA concentration-dependently and reversibly antagonized the L-arginine- and NADPH-dependent activation of a purified soluble guanylate cyclase (GC) by cytosol from freshly harvested porcine aortic endothelial cells, suggesting a direct competition between L-NNA and L-arginine at the level of endothelial NO-synthesis. D-NNA was ineffective in all instances. These results prove L-NNA to be a stereospecific inhibitor of the cytosolic NO formation from L-arginine in endothelial cells. Therefore, L-NNA will be a useful tool to elucidate the molecular mechanism of mammalian NO synthesis.


Climate and edaphic factors drive soil enzyme activity dynamics and tolerance to Cd toxicity after rewetting of dry soil.
The intense drying-rewetting cycle due to climate change can affect soil microbial community composition and function, resulting in long-term consequences for belowground carbon and nutrient dynamics. However, how climatic and edaphic factors influence the responses of enzymes to rewetting and their responses to additional perturbation (e.g., heavy metal pollution) after the drying-rewetting history are not well understood. In this study, we collected 18 surface soils from farmlands across various climate zones in China. We chose dehydrogenase (DHA) and alkaline phosphomonoesterase (ALP) as representative intracellular and extracellular enzymes, respectively, and investigated their tolerance to additional perturbation by adding metal ions (i.e., Cd2+) upon rewetting. In all soils, rewetting increased DHA activities but did not affect ALP activities compared to air-dried soils. Rewetting increased the tolerances of DHA and ALP to Cd stress, suggesting that the drying-rewetting history may reduce the susceptibility of soil enzymes to additional disturbance. The results demonstrate that differentiating enzymes based on their location in the soil will improve our ability to assess the stress response of microbial communities to drastic fluctuations in soil moisture, thereby better predicting the legacy of climate change on microbial function in soils contaminated with heavy metals.


Geospatial Variations and Neighborhood Deprivation in Drug-Related Admissions and Overdoses.
Drug overdoses are a national and global epidemic. However, while overdoses are inextricably linked to social, demographic, and geographical determinants, geospatial patterns of drug-related admissions and overdoses at the neighborhood level remain poorly studied. The objective of this paper is to investigate spatial distributions of patients admitted for drug-related admissions and overdoses from a large, urban, tertiary care center using electronic health record data. Additionally, these spatial distributions were adjusted for a validated socioeconomic index called the Area Deprivation Index (ADI). We showed spatial heterogeneity in patients admitted for opioid, amphetamine, and psychostimulant-related diagnoses and overdoses. While ADI was associated with drug-related admissions, it did not correct for spatial variations and could not account alone for this spatial heterogeneity.


Maternal and fetal hemodynamic effects of autologous blood donation during pregnancy.
Our objectives were to describe the maternal and fetal hemodynamic impact of a 1 U acute blood loss during pregnancy and to compare the hemodynamic responses of pregnant women with those of nonpregnant controls. With Doppler techniques cardiac output and total peripheral vascular resistance were determined in 16 pregnant women in the third trimester and in 16 nonpregnant volunteers before and during orthostatic stress, before and after donation of 450 ml of whole blood. In pregnant women the fetal umbilical artery systolic/diastolic ratio was also determined. Data were subjected to multiple logistic regression analysis. In both groups orthostasis provoked significant decreases in cardiac output and increases in total peripheral vascular resistance. The hemodynamic effects of blood loss were less pronounced and did not significantly change the hemodynamic response to orthostasis in either group. Neither orthostasis nor blood loss caused significant changes in the umbilical systolic/diastolic ratio. These data support the hemodynamic safety of uncomplicated autologous blood donation during late pregnancy.


Increased Neutrophil-to-Lymphocyte Ratio is Associated with Unfavorable Functional Outcome in Acute Ischemic Stroke.
Inflammatory response is the hallmark of secondary brain injury in stroke patients. Neutrophil-to-lymphocyte ratio (NLR) emerged as a marker for functional outcome in several diseases. To investigate the association between NLR on admission and during hospital stay and functional outcome in acute ischemic stroke (AIS). This observational study included all consecutive AIS patients admitted at a German stroke center covering 2011-2013. Patient characteristics and clinical data were retrieved from institutional databases. Multivariate analysis was conducted to investigate parameters associated with functional outcome. Receiver operating characteristic (ROC) analysis was performed to identify the best cutoff for NLR to discriminate between favorable and unfavorable functional outcome. To account for imbalances in baseline characteristics, propensity score matching was carried out to assess the influence of NLR on functional outcome. A total of 807 patients with AIS were included for analysis. Patients with worse functional outcome at 3 months were older and had worse clinical status on admission, higher rates of infectious complications, and an increased NLR. ROC analysis identified a NLR of 3.3 as best cutoff value to discriminate between favorable and unfavorable functional outcomes (area under the curve 0.693, p < 0.001, Youden's index = 0.318; p < 0.001; sensitivity 68.5%, specificity 63.9%). Propensity-matched analysis still demonstrated a higher rate of unfavorable functional outcome at 3 months in patients with NLR ≥ 3.3 [modified Rankin scale 3-6 at 3 months: NLR ≥ 3.3 51.5% vs. NLR < 3.3 36.4%; p = 0.002]. In AIS patients we identified NLR as an important predictor for unfavorable functional outcome.


The Personal Social Networks of Resettled Bhutanese Refugees During Pregnancy in the United States: A Social Network Analysis.
Women comprise 50% of the refugee population, 25% of whom are of reproductive age. Female refugees are at risk for experiencing significant hardships associated with the refugee experience, including after resettlement. For refugee women, the strength of their personal social networks can play an important role in mitigating the stress of resettlement and can be an influential source of support during specific health events, such as pregnancy. A personal social network analysis was conducted among 45 resettled Bhutanese refugee women who had given birth within the past 2 years in the Akron Metropolitan Area of Northeast Ohio. Data were collected using in-depth interviews conducted in Nepali over a 6-month period in 2016. Size, demographic characteristics of ties, frequency of communication, length of relationship, and strength of connection were the social network measures used to describe the personal networks of participants. A qualitative analysis was also conducted to assess what matters were commonly discussed within networks and how supportive participants perceived their networks to be. Overall, participants reported an average of 3 close personal connections during their pregnancy. The networks were comprised primarily of female family members whom the participant knew prior to resettlement in the U.S. Participants reported their networks as "very close" and perceived their connections to be supportive of them during their pregnancies. These results may be used to guide future research, as well as public health programming, that seeks to improve the pregnancy experiences of resettled refugee women.


A New Recombinant Fibronectin/Cadherin Protein-Hydrophobically Modified Glycol Chitosan/Paclitaxel Layer-By-Layer Self-Assembly Strategy for the Postoperative Therapy of Osteosarcoma and Correlated Bone Injury.
Osteosarcoma is one of the most aggressive cancers which greatly threatens the health of adolescents and surgery is difficult to resect the whole piece of tumor tissue. The residual tumor cells might proliferate at the tumor site and invade into the blood circulation, leading to tumor recurrence and metastasis. Besides, the invasion of tumor cells could also lead to bone injury. We designed a recombinant fibronectin-cadherin fusion protein/hydrophobically modified glycol chitosan-PTX nanoparticles (rFN-CDH/HGC-PTX) layer-by-layer self-assembly polymer based on biphasic calcium phosphate ceramic (BCP) (BCP-PEI-(rFN/CDH-PTX/HGC)n-rFN/CDH). The SEM, FTIR, XPS and contact angle experiments proved the successful synthesis of the polymer. The chemotherapy drug PTX and bone-repairing-related rFN/CDH fusion protein could be stably released within one week and the in vitro experiments exhibited the efficacy of the polymer to kill residual tumor cells and promote the proliferation of osteoblast, confirming that our polymer was a superior material for postoperative osteosarcoma therapy.


Putative hybrids between two Anisakis cryptic species: molecular genotyping using High Resolution Melting.
The genus Anisakis includes nine recognized species and the complex of cryptic species Anisakis simplex s. l. is often associated with the human disease known as anisakiasis. During the last decades the use of nuclear ribosomal ITS allowed the identification and description of numerous anisakid nematodes and the discovery of recombinant genotypes or putative hybrids even in other parasitic helminths, such as those between A. simplex sensu stricto and A. pegreffii. The existence of pure hybrids of the two sibling species has been long debated due to the large recovery of larval forms from sympatric areas and the rare observation of adult hybrids. The aims of the present report were to identify anisakid nematodes collected from Stenella coeruleoalba using PCR-RFLP of ITS and to focus the interest on hybrid forms using a High Resolution Melting (HRM) and direct sequencing analyses, since the new record of putative hybrid at adult stage. The PCR-RFLP analysis enabled to identify A. simplex s.s., A. pegreffii, the heterozygous genotype of the two species and A. physeteris. The use of the genotyping approach based on HRM confirmed the profiles of the two species A. simplex s.s. and A. pegreffii, and of the hybrid individuals. The new record of adult hybrids in definitive hosts rekindles the long debate about their existence and their evolutionary meaning. Since the reproductive isolation between A. simplex s.s. and A. pegreffii is the assumption for their existence as separated species, the use of alternative molecular markers and population genetic studies on adult anisakids are recommended.


Brain maturation and its relevance to understanding criminal culpability of juveniles.
The purpose of this review is to summarize current understanding of the process of maturation in human brains during the juvenile period to young adulthood. We describe the methods used in such investigations, and outline the main findings regarding the course of brain development. There is much that we do not know about brain maturation, but there is congruence of evidence indicating that brain maturation is not complete until young adulthood (approximately age 21 years). Furthermore, the main index of maturation, which is rate of myelination, points to large variability in the rate of maturation among brain regions. Generally, maturation of association cortex is not complete by late adolescence, and within this cortex the prefrontal regions are last to mature. The review concludes by discussing the behavioral implications of these findings. The role of myelination is to focus and refine the operation of neural networks regulating behavior, and the frontal lobes specifically modulate and inhibit impulses, shaping behavior within the context of the environment in relation to planned action and long-term goals. Because the brain anatomy data indicate that people are not biologically prepared to exercise mature executive control required for full criminal intent until they reach adulthood, juveniles have diminished capacity in this regard and should not be eligible for the most severe punishment available for their crime.


Management of suspected pulmonary embolism (PE) by D-dimer and multi-slice computed tomography in outpatients: an outcome study.
A prospective outcome study designed to evaluate a simple strategy for the management of outpatients with suspected pulmonary embolism (PE), based on clinical probability, D-dimer, and multi-slice computed tomography (MSCT). A cohort of 432 consecutive patients admitted to the emergency department with suspected PE was managed by sequential non-invasive testing. Patients in whom PE was ruled out were not given anticoagulants, but were followed-up for 3 months. Normal D-dimer and low-intermediate clinical probability ruled out PE in 103 patients [24% (95% CI 20-28)]. Seventeen patients had normal D-dimer, but high clinical probability and proceeded to MSCT. All patients proved negative for PE. A total of 329 (76%) patients underwent MSCT examination. Pulmonary embolism was diagnosed in 93 patients [21.5% (95% CI 18-26)] and was ruled out by negative MSCT in 221 patients [51% (95% CI 46-56)]. MSCT scans were determined as inconclusive in 15 (4.5%) patients. No patient developed objectively verified venous thromboembolism (VTE) during the 3-month follow-up period. However, the cause of death was adjudicated as possibly related to PE in two patients, resulting in an overall 3-month VTE risk of 0.6% (95% CI 0-2.2%). The diagnostic algorithm yielded a definite diagnosis in 96.5% of the patients. This simple and non-invasive strategy combining clinical probability, D-dimer, and MSCT for the management of outpatients with suspected PE appears to be safe and effective.


Methylenetetrahydrofolate reductase gene A1298C polymorphism in pediatric stroke--case-control and family-based study.
Moderate hyperhomocysteinemia is one of the risk factors of pediatric stroke. Methylenetetrahydrofolate reductase (MTHFR) is an important enzyme, which regulates homocysteine metabolism, and some polymorphisms of gene encoding this enzyme are associated with a decreased activity of the enzyme. The aim of the study was to assess an association between the A1298C polymorphism and pediatric stroke. We also evaluated a possible synergistic effect of A1298C and C677T polymorphisms of this gene. The study group consisted of 88 children after ischemic stroke, 142 of their parents and 111 controls. The A1298C polymorphism was genotyped using the restriction fragment length polymorphism method. We used 2 study designs: a case-control model and a family-based association test. The Statistica 7.1 and EpiInfo 6 softwares were used in all analyses. We did not observe any statistically significant differences either in the transmission of the A allele in the family-based test or in the frequency of the A allele in the patients group compared with the controls. We also did not notice any significant additive or synergistic effects between the A1298C and C677T polymorphisms. An analysis of the results obtained in this study and a critical review of previously published studies indicate that the A1298C polymorphism of the MTHFR gene is not related to ischemic stroke in children.


Anterior horn cell degeneration and Bunina-type inclusions associated with dementia.
The ultrastructural features of Bunina type inclusions in the anterior horn cells of a patient dying of amyotrophic lateral sclerosis with dementia appear unique. The Bunina-type inclusions are electron dense aggregates containing, and surrounded by organelle-like membranes. These inclusions appear to be a special type of autophagic vacuole, possibly arising from altered mitochondria.


The molecular pathophysiology of concussive brain injury.
Concussion or mild traumatic brain injury (mTBI) is a condition that affects hundreds of thousands of patients worldwide. Understanding the pathophysiology of this disorder can help manage its acute and chronic repercussions. Immediately following mTBI, there are several metabolic, hemodynamic, structural, and electric changes that alter normal cerebral function. These alterations can increase the brain's vulnerability to repeat injury and long-term disability. This review evaluates current studies from the bench to the bedside of mTBI. Acute and chronic effects of concussion are measured in both animal and clinical studies. Also, the effect of repeat concussions is analyzed. Concussion-induced pathophysiology with regards to glucose metabolism changes, mitochondrial dysfunction, axonal injury, and structural damage are evaluated. Translational studies such as functional magnetic resonance imaging, magnetic resonance spectroscopy and diffusion tensor imaging prove to be effective clinical tools for both prognostic and treatment parameters. Understanding the neurobiology of concussion will lead to development and validation of physiological biomarkers of this common injury. These biomarkers (eg, laboratory tests, imaging, electrophysiology) will then allow for improved detection, better functional assessment and evidence-based return to play recommendations.


808-nm-Light-Excited Lanthanide-Doped Nanoparticles: Rational Design, Luminescence Control and Theranostic Applications.
808 nm-light-excited lanthanide (Ln3+ )-doped nanoparticles (LnNPs) hold great promise for a wide range of applications, including bioimaging diagnosis and anticancer therapy. This is due to their unique properties, including their minimized overheating effect, improved penetration depth, relatively high quantum yields, and other common features of LnNPs. In this review, the progress of 808 nm-excited LnNPs is reported, including their i) luminescence mechanism, ii) luminescence enhancement, iii) color tuning, iv) diagnostic and v) therapeutic applications. Finally, the future outlook and challenges of 808 nm-excited LnNPs are presented.


A flow fluorimetric analysis of the cell cycle during growth and differentiation inDictyostelium discoideum.
We report a flow fluorimetric analysis of the DNA content of cells and nuclei from vegetative populations and various developmental stages of the cellular slime mouldDictyostelium discoideum using the dyes Hoechst 33258 and mithramycin. Nuclei from all of these populations showed an identical single DNA-content peak, indicating that most vegetative cells and most cells in all developmental stages are in one phase of the cell cycle. Our own data and findings in the literature indicate that this phase is G2. On the other hand, we also found that various stages, subpopulations of cells at early stages and the different differentiated cell types in the slug stage differ in DNA content per cell. Any particular population typically has one major peak of DNA content, with a modal value that is characteristic for the cell type and for the developmental stage. These differences presumably reflect differences in mitochondrial DNA content per cell.


Outcomes for abdominoperineal resections are not worse than those of anterior resections.
Local recurrence and cure rates following abdominoperineal resections have been reported to be much worse than sphincter-preserving anterior resections. We compared the oncologic outcomes of patients treated by abdominoperineal resections with those following sphincter-preserving anterior resections. The medical records of patients who underwent radical rectal resection for rectal carcinoma at the Colorectal Surgery Department, Singapore General Hospital, during the period from April 1989 to April 2002 were reviewed. A total of 791 cases were studied. Operative procedures were classified as either abdominoperineal resections or anterior resections with either straight or pouch anastomosis. Total mesorectal excision was routinely performed for carcinomas of the lower middle and lower third of the rectum. Sentinel events, including local and systemic recurrences or morbidity and mortality, were tracked prospectively. There were a total of 93 abdominoperineal resections (12.1 percent), 547 anterior resections with straight anastomoses (71 percent), and 130 anterior resections with pouch anastomoses (16.9 percent). Postoperative mortality was 2.6 percent and postoperative morbidity was 13.6 percent with an overall anastomotic leakage rate of 2.5 percent. The cumulative five-year local recurrence rate was 5.4 percent for abdominoperineal resections, 3.6 percent for anterior resections with straight anastomoses, and 3.8 percent for anterior resections with pouch anastomoses (P = 0.73 by log-rank test). The median time to local recurrence also did not differ significantly between the different procedures (abdominoperineal resections, 17 months, anterior resections with straight anastomoses, 18 months, anterior resections with pouch anastomoses, 13 months). Independent predictors for local recurrence included advanced tumor stage, tumor depth, and poorly differentiated tumors. The five-year cancer-specific survival was 70 percent. The type of anastomosis did not influence disease-free survival with median disease-free survival for patients who underwent abdominoperineal resections being 100 months, survival of anterior resections with straight anastomoses being 135 months, and survival of anterior resections with pouch anastomoses being 121 months (P = 0.33 by log-rank test). The independent factors for poor survival were age greater than 65 years, advanced tumor stage, tumor depth, and poorly differentiated tumors. Both abdominoperineal resections and sphincter-preserving anterior resections can be performed safely with low morbidity and mortality in a specialized high-volume hospital unit without compromising oncologic outcomes. With appreciation of the anatomic relations in total mesorectal excision and standardized consistent surgical technique, the oncologic outcomes of patients treated by abdominoperineal resections are not worse than those treated by sphincter-preserving anterior resections.


The isolation of adenoviruses from goats affected with peste des petits ruminants in Nigeria.
Mucosal scrapings from the large intestine of two goats that had died from peste des petits ruminants (PPR) in separate outbreaks in Nigeria were examined for viruses. A mixed viral infection of PPR virus (morbillivirus) and adenovirus was confirmed in both goats. The adenoviruses did not conform to any of the ovine and bovine serotypes recognised; the two isolates were considered different serotypes. It is concluded that, although the role of adenoviruses in the epizootiology of PPR in Nigeria is difficult to appraise, they are probably commensals. This is believed to be the first report of the isolation of adenoviruses from goats.


Reconstitution of the full transmembrane cadherin-catenin complex.
The dynamic regulation of epithelial adherens junctions relies on all components of the E-cadherin-catenin complex. Previously, the complexes have been partially reconstituted and composed only of α-catenin, β-catenin, and the E-cadherin cytoplasmic domain. However, p120-catenin and the full-length E-cadherin including the extracellular, transmembrane, and intra-cellular domains are vital to the understanding of the relationship between extracellular adhesion and intracellular signaling. Here, we reconstitute the complete and full-length cadherin-catenin complex, including full-length E-cadherin, α-catenin, β-catenin, and p120-catenin, into nanodiscs. We are able to observe the cadherin in nanodiscs by cryo-EM. We also reconstitute α-catenin, β-catenin, and p120-catenin with the E-cadherin cytoplasmic tail alone in order to analyze the affinities of their binding interactions. We find that p120-catenin does not associate strongly with α- or β-catenin and binds much more transiently to the cadherin cytoplasmic tail than does β-catenin. Overall, this work creates many new possibilities for biochemical studies understanding transmembrane signaling of cadherins and the role of p120-catenin in adhesion activation.


Imaging Paget's disease of bone--from head to toe.
Paget's disease of the bone is a common, non-inflammatory, metabolic, skeletal disorder of unknown aetiology characterized by an increase in osteoclast-mediated bone resorption and compensatory excessive osteoblast activation. Prevalence increases with age, and a pronounced geographical variation is well documented. The disease is often an incidental finding on a radiological examination requested for an unrelated indication. The osteolytic, mixed osteolytic/osteoblastic, and osteosclerotic phases may occur in the same patient and same bone in a synchronous or metachronous fashion. Radiological features in each phase mirror the histopathological appearances, and are distinctive enough to establish a diagnosis with confidence. Using multi-technique imaging, this review illustrates the most common and the not so common radiological patterns of involvement in Paget's disease of bone observed at our centre during the past 20 years.


Plasmids with temperature-dependent copy number for amplification of cloned genes and their products.
Miniplasmids (pKN402 and pKN410) were isolated from runaway-replication mutants of plasmid R1. At 30 degrees C these miniplasmids are present in 20--50 copies per cell of Escherichia coli, whereas at temperatures above 35 degrees C the plasmids replicate without copy number control during 2--3 h. At the end of this period plasmid DNA amounts to about 75% of the total DNA. During the gene amplification, growth and protein synthesis continue at normal rate leading to a drastic amplification of plasmid gene products. Plasmids pKN402 (4.6 Md) and pKN410 (10 Md) have single restriction sites for restriction endonucleases EcoRI and HindIII; in addition plamid pKN410 has a single BamHI site and carries ampicillin resistance. The plasmids can therefore be used as cloning vectors. Several genes were cloned into these vectors using the EcoRI sites; chromosomal as well as plasmid-coded beta-lactamase was found to be amplified up to 400-fold after thermal induction of the runaway replication. Vectors of this temperature-dependent class will be useful in the production of large quantities of genes and gene products. These plasmids have lost their mobilization capacity. Runaway replication is lethal to the host bacteria in rich media. These two properties contribute to the safe use of the plasmids as cloning vehicles.


Discovery and characterization of 2-aminobenzimidazole derivatives as selective NOD1 inhibitors.
NLR family proteins play important roles in innate immune response. NOD1 (NLRC1) activates various signaling pathways including NF-κB in response to bacterial ligands. Hereditary polymorphisms in the NOD1 gene are associated with asthma, inflammatory bowel disease, and other disorders. Using a high throughput screening (HTS) assay measuring NOD1-induced NF-κB reporter gene activity, followed by multiple downstream counter screens that eliminated compounds impacting other NF-κB effectors, 2-aminobenzimidazole compounds were identified that selectively inhibit NOD1. Mechanistic studies of a prototypical compound, Nodinitib-1 (ML130; CID-1088438), suggest that these small molecules cause conformational changes of NOD1 in vitro and alter NOD1 subcellular targeting in cells. Altogether, this inaugural class of inhibitors provides chemical probes for interrogating mechanisms regulating NOD1 activity and tools for exploring the roles of NOD1 in various infectious and inflammatory diseases.


Extra-articular subtalar arthrodesis with cancellous bone graft and internal fixation for children with myelomeningocele.
Extra-articular subtalar arthrodesis with cancellous bone graft and internal fixation was performed on 20 feet of 12 children with myelomeningocele, whose average age at operation was 7.4 years. Indication for surgery was progressive hindfoot valgus deformity causing difficulty in fitting orthoses. At average follow-up of 4.1 years, results were satisfactory in 18 of the 20 feet. Unsatisfactory results were a result of undercorrection at time of surgery. Extra-articular subtalar arthrodesis will correct hindfoot valgus deformity in children with myelomeningocele, and the use of internal fixation and iliac crest bone grafting contributed to the improved success rate in this series.


Effects of gellan gum and inulin on mixed-gel properties and molecular structure of gelatin.
Gellan gum (GG) is often added to gelatin (GL) to improve the gel property. GG-based or inulin (IL)-based hydrogels were developed. Rigid and brittle gels or smooth and delicate gels were prepared with GG and IL, respectively. This study aimed to explore the properties and interaction mechanisms of the mixed-gel system containing GL, GG, and IL, in which different ratios of GG-IL (0.4%) (10:0, 8:2, 6:4, 5:5, 4:6, 2:8, and 0:10) were added to GL (6%). Texture profiles, rheological properties, water mobility, intermolecular forces, circular dichroism (CD) spectra, and microstructures were analyzed. The results showed that addition of GG-IL could improve the hardness, chewiness, and cohesiveness of mixed-gel, besides maintaining appropriate springiness. Water mobility of the mixed-gel decreased while viscoelasticity increased upon the addition GG. At GG:IL = 2:8, the melting temperature of mixed-gel was far higher than that of GL gel itself. The GL-GG-IL gel showed decrease in nonspecific bonding and increase in hydrogen bonding compared with the GL gel. CD spectra indicated the promotion of GL unfolding by GG, hence suggesting the binding of GG to GL; binding ability was better at GG:IL >5:5. Cryo-SEM provided evidence for the formation of cross-linked network within GL-GG-IL. Overall, we concluded that addition of GG-IL to GL system would be most suitable for improving the properties of mixed-gel. This finding may be potentially applicable in the further development of gel food products, such as meat jellies and gummy jellies.


Association of Circulating Proteins with Death or Lung Transplant in Patients with Idiopathic Pulmonary Fibrosis in the IPF-PRO Registry Cohort.
Idiopathic pulmonary fibrosis (IPF) is a progressive and ultimately fatal disease with a variable clinical course. Biomarkers that predict patient outcomes are needed. We leveraged data from 300 patients in the multicenter IPF-PRO Registry to determine associations between circulating proteins and the composite outcome of respiratory death or lung transplant. Plasma collected at enrollment was analyzed using aptamer-based proteomics (1305 proteins). Over a median follow-up of 30.4 months, there were 76 respiratory deaths and 26 lung transplants. In unadjusted univariable analyses, 61 proteins were significantly associated with the outcome (hazard ratio > 2 or < 0.5, corrected p ≤ 0.05). In multivariable analyses, a set of 4 clinical measures and 47 unique proteins predicted the probability of respiratory death or lung transplant with an optimism-corrected C-index of 0.76. Our results suggest that select circulating proteins strongly associate with the risk of mortality in patients with IPF and confer information independent of clinical measures.


Increased angiogenesis and migration of dermal microvascular endothelial cells from patients with psoriasis.
Psoriasis displays both increased angiogenesis and microvascular dilation in the skin, while human dermal microvascular endothelial cells (HDMECs) are involved in angiogenesis and microvascular dilation. Whether the functions of HDMECs are altered in psoriatic skin versus healthy skin remain unknown. Here, we isolated HDMECs from the skin of 10 patients with psoriasis and 10 healthy subjects and compared angiogenesis, proliferation, migration and cell metabolism between psoriatic HDMECs and normal HDMECs. We found that the morphology of primary HDMECs was comparable between psoriatic HDMECs and normal HDMECs. After passage, psoriatic HDMECs displayed larger cell size and wider intercellular space. In addition to DiI-Ac-LDL (DiI-labelled acetylated low-density lipoprotein) uptake, expression levels of CD31, vWF (von Willebrand factor) and LYVE-1 were comparable in psoriatic HDMECs versus normal HDMECs. However, psoriatic HDMECs exhibited increased tube formation (numbers of nodes and meshes, p < 0.05) and migration (numbers of migrated cells, p < 0.001) and reductions in proliferation (growth rates, p < 0.05) and energy metabolism (oxygen consumption rate and extracellular acidification rate, p < 0.05) compared with normal HDMECs. Therefore, psoriatic HDMECs display an increased angiogenesis and migration and decreased proliferation and metabolic activity, suggesting a pathogenic role of HDMECs in psoriasis.


An evaluation of the recognised systemic inflammatory biomarkers of chronic sub-optimal inflammation provides evidence for inflammageing (IFA) during multiple sclerosis (MS).
The pathogenesis of the human demyelinating disorder multiple sclerosis (MS) involves the loss of immune tolerance to self-neuroantigens. A deterioration in immune tolerance is linked to inherent immune ageing, or immunosenescence (ISC). Previous work by the author has confirmed the presence of ISC during MS. Moreover, evidence verified a prematurely aged immune system that may change the frequency and profile of MS through an altered decline in immune tolerance. Immune ageing is closely linked to a chronic systemic sub-optimal inflammation, termed inflammageing (IFA), which disrupts the efficiency of immune tolerance by varying the dynamics of ISC that includes accelerated changes to the immune system over time. Therefore, a shifting deterioration in immunological tolerance may evolve during MS through adversely-scheduled effects of IFA on ISC. However, there is, to date, no collective proof of ongoing IFA during MS. The Review addresses the constraint and provides a systematic critique of compelling evidence, through appraisal of IFA-related biomarker studies, to support the occurrence of a sub-optimal inflammation during MS. The findings justify further work to unequivocally demonstrate IFA in MS and provide additional insight into the complex pathology and developing epidemiology of the disease.


Epidemiology of Asthma-Chronic Obstructive Pulmonary Disease Overlap.
Asthma COPD Overlap has consistently reported to be associated with an increase burden of disease but the impact on lung function decline and mortality varies by study. The prevalence increases with age but the relationship with gender also varies with the study population. The variability in the prevalence and clinical characteristics of ACO is linked to differences in how chronic obstructive pulmonary disease (COPD) and asthma are defined, including diagnostic criteria (spirometry-based vs. clinical or symptom-based diagnoses vs. claims data), the population studied, the geographic region and environment and a consensus approach to the diagnosis of ACO is needed to allow meaningful and consistent epidemiologic information to be generated about this condition.


Ultrasensitive and on-site eDNA detection for the monitoring of crown-of-thorns starfish densities at the pre-outbreak stage using an electrochemical biosensor.
The coral reef crisis has significantly intensified over the last decades, mainly due to severe outbreaks of crown-of-thorns starfish (COTS). Current ecological monitoring has failed to detect COTS densities at the pre-outbreak stage, thus preventing early intervention. In this work, we developed an effective electrochemical biosensor modified by a MoO2/C nanomaterial, as well as a specific DNA probe that could detect trace COTS environmental DNA (eDNA) at a lower detection limit (LOD = 0.147 ng/μL) with excellent specificity. The reliability and accuracy of the biosensor were validated against the standard methods by an ultramicro spectrophotometer and droplet digital PCR (p > 0.05). The biosensor was then utilized for the on-site analysis of seawater samples from SYM-LD and SY sites in the South China Sea. For the SYM-LD site suffering an outbreak, the COTS eDNA concentrations were 0.33 ng/μL (1 m, depth) and 0.26 ng/μL (10 m, depth), respectively. According to the ecological survey, the COTS density was 500 ind/hm2 at the SYM-LD site, verifying the accuracy of our measurements. At the SY site, COTS eDNA was also detected at 0.19 ng/μL, but COTS was not found by the traditional survey. Hence, larvae were possibly present in this region. Therefore, this electrochemical biosensor could be used to monitor COTS populations at the pre-outbreak stages, and potentially serve as a revolutionary early warning method. We will continue to improve this method for picomolar or even femtomolar detection of COTS eDNA.


The microRNA miR-29 controls innate and adaptive immune responses to intracellular bacterial infection by targeting interferon-γ.
Interferon-γ (IFN-γ) has a critical role in immune responses to intracellular bacterial infection. MicroRNAs (miRNAs) are important in the regulation of innate and adaptive immunity. However, whether miRNAs can directly target IFN-γ and regulate IFN-γ production post-transcriptionally remains unknown. Here we show that infection of mice with Listeria monocytogenes or Mycobacterium bovis bacillus Calmette-Guérin (BCG) downregulated miR-29 expression in IFN-γ-producing natural killer cells, CD4(+) T cells and CD8(+) T cells. Moreover, miR-29 suppressed IFN-γ production by directly targeting IFN-γ mRNA. We developed mice with transgenic expression of a 'sponge' target to compete with endogenous miR-29 targets (GS29 mice). We found higher serum concentrations of IFN-γ and lower L. monocytogenes burdens in L. monocytogenes-infected GS29 mice than in their littermates. GS29 mice had enhanced T helper type 1 (T(H)1) responses and greater resistance to infection with BCG or Mycobacterium tuberculosis. Therefore, miR-29 suppresses immune responses to intracellular pathogens by targeting IFN-γ.


The omicron (B.1.1.529) SARS-CoV-2 variant of concern does not readily infect Syrian hamsters.
The emergence of SARS-CoV-2 variants of concern (VoCs) has exacerbated the COVID-19 pandemic. End of November 2021, a new SARS-CoV-2 variant namely the omicron (B.1.1.529) emerged. Since this omicron variant is heavily mutated in the spike protein, WHO classified this variant as the 5th variant of concern (VoC). We previously demonstrated that the ancestral strain and the other SARS-CoV-2 VoCs replicate efficiently in and cause a COVID19-like pathology in Syrian hamsters. We here wanted to explore the infectivity of the omicron variant in comparison to the ancestral D614G strain in the hamster model. Strikingly, in hamsters that had been infected with the omicron variant, a 3 log10 lower viral RNA load was detected in the lungs as compared to animals infected with D614G and no infectious virus was detectable in this organ. Moreover, histopathological examination of the lungs from omicron-infected hamsters revealed no signs of peri-bronchial inflammation or bronchopneumonia.


Mobility and persistence of the herbicide fomesafen in soils cultivated with bean plants using SLE/LTP and HPLC/DAD.
A method has been optimized and validated for the determination of fomesafen in soils using solid-liquid extraction with low-temperature partitioning (SLE/LTP) and analysis by liquid chromatography with a high-efficiency diode array detector (HPLC/DAD). The method was used to evaluate the persistence and mobility of this herbicide in different soils cultivated with bean plants. Recovery values were ≥98.9 %, with variations in the repeatability coefficients of ≤15 %, and a detection limit of 7.3 μg kg(-1). Half-life values of fomesafen were between 60 and 71 days in soil cultivated using a no-till system and 99 and 114 days in soil cultivated using a conventional tillage system. The mobility of fomesafen was moderate and mainly influenced by the organic matter content, pH, and soil type. In Red-Yellow Argisol, which has a higher content of organic matter, the leaching of fomesafen was less pronounced. In Red-Yellow Latosol, which has smaller amounts of organic matter and high pH, the leaching of fomesafen was more pronounced.


Nature of Medical Malpractice Claims Against Radiation Oncologists.
To examine characteristics of medical malpractice claims involving radiation oncologists closed during a 10-year period. Malpractice claims filed against radiation oncologists from 2003 to 2012 collected by a nationwide liability insurance trade association were analyzed. Outcomes included the nature of claims and indemnity payments, including associated presenting diagnoses, procedures, alleged medical errors, and injury severity. We compared the likelihood of a claim resulting in payment in relation to injury severity categories (death as referent) using binomial logistic regression. There were 362 closed claims involving radiation oncology, 102 (28%) of which were paid, resulting in $38 million in indemnity payments. The most common alleged errors included "improper performance" (38% of closed claims, 18% were paid; 29% [$11 million] of total indemnity), "errors in diagnosis" (25% of closed claims, 46% were paid; 44% [$17 million] of total indemnity), and "no medical misadventure" (14% of closed claims, 8% were paid; less than 1% [$148,000] of total indemnity). Another physician was named in 32% of claims, and consent issues/breach of contract were cited in 18%. Claims for injury resulting in death represented 39% of closed claims and 25% of total indemnity. "Improper performance" was the primary alleged error associated with injury resulting in death. Compared with claims involving death, major temporary injury (odds ratio [OR] 2.8, 95% confidence interval [CI] 1.29-5.85, P=.009), significant permanent injury (OR 3.1, 95% CI 1.48-6.46, P=.003), and major permanent injury (OR 5.5, 95% CI 1.89-16.15, P=.002) had a higher likelihood of a claim resulting in indemnity payment. Improper performance was the most common alleged malpractice error. Claims involving significant or major injury were more likely to be paid than those involving death. Insights into the nature of liability claims against radiation oncologists may help direct efforts to improve quality of care and minimize the risk of being sued.


Cysteine homeostasis under inhibition of protein synthesis in Escherichia coli cells.
Increased intracellular cysteine poses a potential danger to cells due to the high ability of cysteine to reduce free iron and promote the Fenton reaction. Here, we studied ways to maintain cysteine homeostasis in E. coli cells while inhibiting protein synthesis with valine or chloramphenicol. When growing wild-type bacteria on minimal medium with sulfate, an excess of cysteine resulting from the inhibition of protein synthesis is mainly incorporated into glutathione (up to 90%), which, therefore, can be considered as cysteine buffer. The share of hydrogen sulfide, which is the product of cysteine degradation by cysteine synthase B (CysM), does not exceed 1-3%, the rest falls on free cysteine, exported from cells. As a result, intracellular free cysteine is maintained at a low level (about 0.1 mM). The lack of glutathione in the gshA mutant increases H2S production and excretion of cysteine and leads to a threefold increase in the level of intracellular cysteine in response to valine and chloramphenicol. The relA mutants, exposed to valine, produce more H2S, dramatically accelerate the export of glutathione and accumulate more cysteine in the cytoplasm than their parent, which indicates that the regulatory nucleotide (p)ppGpp is involved in maintaining cysteine homeostasis. Disruption of cysteine homeostasis in gshA and relA mutants increases their sensitivity to peroxide stress.


Stereotactic radiosurgery optimization with hippocampal-sparing in patients treated for brain metastases.
Cranial irradiation is associated with significant neurocognitive sequelae, secondary to radiation-induced damage to hippocampal cells. It has been shown that hippocampal-sparing (HS) leads to modest benefit in neurocognitive function in patients with brain metastases, but further improvement is possible. We hypothesized that improved benefits could be seen using HS in patients treated with stereotactic radiation (HS-SRS). Our study evaluated whether the hippocampal dose could be significantly reduced in the treatment of brain metastases using SRS, while maintaining target coverage. Sixty SRS plans were re-planned to minimize dose to the hippocampus while maintaining target coverage. Patients with metastases within 5 mm of the hippocampus were excluded. Minimum, mean, maximum and dose to 40% (mean equivalent dose in 2 Gy per fraction, EQD2 to the hippocampus) were compared between SRS and HS-SRS plans. Median number of brain metastases was two. Compared to baseline SRS plans, hippocampal-sparing plans demonstrated Dmin was reduced by 35%, from 0.4 Gy to 0.3 Gy (p-value 0.02). Similarly, Dmax was reduced by 55%, from 8.2 Gy to 3.6 Gy, Dmean by 52%, from 1.6 Gy to 0.5 Gy, and D40 by 50%, from 1.8 Gy to 0.9 Gy (p-values <0.001). Our study demonstrated that further reduction of hippocampal doses of more than 50% is possible in the treatment of brain metastases with SRS using dose optimization. This could result in significantly improved neurocognitive outcomes for patients treated for brain metastases.


Rehabilitation of the geriatric patient. Stroke rehabilitation, the rehabilitation team, and the usefulness of functional assessment.
Salient features of rehabilitation in the elderly are illustrated through the case history of a stroke patient. The efforts of the multidisciplinary rehabilitation team and the consideration of alternative settings and levels of care are critical factors in the restoration and preservation of function in the elderly. Systematic functional assessment, using instruments such as the Barthel index, is recommended to document patient outcomes.


Extracellular vesicle- and particle-mediated communication shapes innate and adaptive immune responses.
Intercellular communication among immune cells is vital for the coordination of proper immune responses. Extracellular vesicles and particles (EVPs) act as messengers in intercellular communication, with important consequences for target cell and organ physiology in both health and disease. Under normal physiological conditions, immune cell-derived EVPs participate in immune responses by regulating innate and adaptive immune responses. EVPs play a major role in antigen presentation and immune activation. On the other hand, immune cell-derived EVPs exert immunosuppressive and regulatory effects. Consequently, EVPs may contribute to pathological conditions, such as autoimmune and inflammatory diseases, graft rejection, and cancer progression and metastasis. Here, we provide an overview of the role of EVPs in immune homeostasis and pathophysiology, with a particular focus on their contribution to innate and adaptive immunity and their potential use for immunotherapies.


IL4 receptor polymorphism is associated with increased risk of sudden deafness in Korean population.
The interleukin 4 receptor (IL4R) polymorphism Q576R (rs 180275) has been well known to be associated with atopy and other inflammatory diseases. A single nucleotide polymorphism (SNP) A > G transition potentiates the binding specificity of the adjacent tyrosine residue. In this study we investigated the possible relationship between sudden deafness (SD) and IL4R polymorphism Q576R in 97 Korean SD patients and 613 controls using pyrosequencing method. The odds ratio (OR) for SD associated with the G vs. A allele was 2.58 [P < 0.0001, 95% confidence interval (CI) = 1.84-3.60]. We then sub-grouped SD into Tinnitus positive (+) and Tinnitus negative (-). G allele in Tinnitus (+) is significantly associated with the development of Tinnitus (+) [X(2) = 32.02, P < 0.0001, OR (95% CI) = 2.74 (1.91-3.93)] but not with Tinnitus (-). Taken together these results suggest that G allele could be a risk factor for SD.


Financial Effect of Unbundling Moderate Sedation from Procedural Codes in Radiology.
To assess and quantify the financial effect of unbundling newly unbundled moderate sedation codes across major payors at an academic radiology practice. Billing and reimbursement data for 23 months of unbundled moderate sedation codes were analyzed for reimbursement rates and trends. This included 10,481 and 28,189 units billed and $443,257 and $226,444 total receipts for codes 99152 (initial 15 minutes of moderate sedation) and 99153 (each subsequent 15 minute increment of moderate sedation), respectively. Five index procedures-(i) central venous port placement, (ii) endovascular tumor embolization, (iii) tunneled central venous catheter placement, (iv) percutaneous gastrostomy placement, and (v) percutaneous nephrostomy placement-were identified, and moderate sedation reimbursements for Medicare and the dominant private payor were calculated and compared to pre-bundled reimbursements. Revenue variation models across different patient insurance mixes were then created using averages from 4 common practice settings among radiologists (independent practices, all hospitals, safety-net hospitals, and non-safety-net hospitals). Departmental reimbursement for unbundled moderate sedation in FY2018 and FY2019 totaled $669,701.34, with high per-unit variability across payors, especially for code 99153. Across the 5 index procedures, moderate sedation reimbursement decreased 1.3% after unbundling and accounted for 3.9% of procedural revenue from Medicare and increased 11.9% and accounted for 5.5% of procedural revenue from the dominant private payor. Between different patient insurance mix models, estimated reimbursement from moderate sedation varied by as much as 29.9%. Departmental reimbursement from billing the new unbundled moderate sedation codes was sizable and heterogeneous, highlighting the need for consistent and accurate reporting of moderate sedation. Total collections vary by case mix, patient insurance mix, and negotiated reimbursement rates.


Nausea and vomiting of pregnancy and association with pregnancy outcome.
The pattern of nausea and/or vomiting of pregnancy were investigated in a group of 414 predominantly white, upper middle-class women in Albany, New York; patterns were ascertained before their eighty-eighth day of gestation and followed up to the end of pregnancy. Of these, 89.4% reported at least some symptoms. This frequency is higher than reported previously, perhaps in part because extensive attempts were made in this study to ascertain symptoms. The incidence of vomiting was about 55%. Women with no symptoms of nausea and/or vomiting of pregnancy experienced a significantly greater proportion of nonviable pregnancy outcomes (fetal death). Increased intake of niacin during the first trimester was associated with decreased infant birth weight. This may be the result of quicker fetal maturation due to increased levels of protein intake.


Expression of EMSY gene in sporadic ovarian cancer.
The majority of familial breast and ovarian cancers arise from mutations in the BRCA1 and BRCA2 genes. Amplification at the 11q13.5 locus is commonly observed in breast and ovarian cancers. In 2003, Hughes-Davies et al. identified a novel gene (EMSY) at this locus which binds BRCA2 within a region deleted in some cancers. Although little is known about the cellular role of EMSY, indirect evidence suggests that this nuclear protein is capable of silencing the activation potential of BRCA2. In this study we aimed to investigate expression of the EMSY gene and its protein product in sporadic ovarian cancer. Real-time quantitative RT-PCR was performed in 50 ovarian cancer and 17 normal ovarian tissue samples. Overexpression of the EMSY gene was found in 6/50 cases (12%), but in none of the control samples. To determine the EMSY protein by Western blotting, semi-quantitative analysis of the EMSY protein was performed using the Scion Image Gel Analysis Program. Statistical analysis was performed using SPSS 11.5. All patients having EMSY overexpression also displayed increased levels of the EMSY protein. Sporadic ovarian cancer shows overexpression of EMSY at a prevalence similar to that found in breast cancer and the overexpression is correlated with the protein level.


Immunoscintigraphy of colorectal carcinoma with an anti-CEA monoclonal antibody: a critical review.
An anti-CEA monoclonal antibody (FO23C5), belonging to the IgG1a class, was used to perform several preclinical and clinical studies on radioimmunoscintigraphy (RIS) in patients with colorectal carcinoma. Preliminary screening on different tissues showed high specificity of this antibody for gastric and colorectal carcinomas. A pilot study on 51 patients with 64 localizations of colorectal carcinoma was realized, followed by a multicenter validation study in which, among 509 patients bearing CEA secreting tumors, 254 patients had primary or relapsed colorectal tumors. High sensitivity and specificity values were obtained by these studies and many "unsuspected" localizations were recorded. In order to better define the clinical utility of this approach, a prospective trial was run on 59 patients previously submitted to surgery for colorectal carcinoma and with suspected local relapses. A comparative evaluation of RIS, CT scan, US and MRI was performed. RIS and MRI have the highest value of accuracy (86%) followed by CT scan (68%) and US (54%). No adverse reactions were noticed in any of the patients examined.


Epidemiology and clinical characteristics of drowning patients presenting to a pediatric emergency department from 2017 to 2020.
Drowning is a common mechanism of injury in the pediatric population that often requires hospitalization. The primary objective of this study was to describe the epidemiology and clinical characteristics of pediatric drowning patients evaluated in a pediatric emergency department (PED), including the clinical interventions and outcomes of this patient population. A retrospective cohort study was conducted of pediatric patients evaluated in a mid-Atlantic urban pediatric emergency department from January 2017 to December 2020 after a drowning event. Eighty patients ages 0-18 were identified, representing 57 79 unintentional events and 1 intentional self-injury event. The majority of patients (50%) were 1-4 years of age. The majority (65%) of patients 4 years of age or younger were White, whereas racial/ethnic minority patients accounted for the majority (73%) of patients 5 years of age or older. Most drowning events (74%) occurred in a pool, on Friday through Saturday (66%) and during the summer (73%). Oxygen was used in 54% of admitted patients and only in 9% of discharged patients. Cardiopulmonary resuscitation (CPR) was performed in 74% of admitted patients and 33% of discharged patients. Drowning can be an intentional or unintentional source of injury in pediatric patients. Among the patients who presented to the emergency department for drowning, more than half received CPR and/or were admitted, suggesting high acuity and severity of these events. In this study population, outdoor pools, summer season and weekends are potential high yield targets for drowning prevention efforts.


An economic analysis of patient controlled remifentanil and epidural analgesia as pain relief in labour (RAVEL trial); a randomised controlled trial.
To compare the costs of a strategy of patient controlled remifentanil versus epidural analgesia for pain relief in labour. We performed a multicentre randomised controlled trial in 15 hospitals in the Netherlands, the RAVEL trial. Costs were analysed from a health care perspective alongside the RAVEL trial. Pregnant women of intermediate to high risk beyond 32 weeks gestation who planned vaginal delivery. Women were randomised before the onset of labour, to receive either patient controlled remifentanil or epidural analgesia when pain relief was requested during labour. Primary outcome for effectiveness was satisfaction with pain relief, expressed as the area under the curve (AUC). A higher AUC represents higher satisfaction with pain relief. Here, we present an economic analysis from a health care perspective including costs from the start of labour to ten days postpartum. Health-care utilization was documented in the Case Report Forms and by administering an additional questionnaire. The costs in the patient controlled remifentanil group (n = 687) and in the epidural group (n = 671) were €2900 versus €3185 respectively (mean difference of -€282 (95% CI -€611 to €47)). The (non-significant) higher costs in the epidural analgesia group could be mainly attributed to higher costs of neonatal admission. From an economic perspective, there is no preferential pain treatment in labouring intermediate to high risk women. Since patient controlled remifentanil is not equivalent to epidural analgesia with respect to AUC for satisfaction with pain relief we recommend epidural analgesia as the method of choice. However, if appropriately counselled on effect and side effects there is, from an economic perspective, no reason to deny women patient controlled remifentanil.


Cystatin C and NT-proBNP as prognostic biomarkers in Fabry disease.
Fabry disease (FD) is a lysosomal storage disorder caused by mutations in the α-galactosidase A gene. It is characterized by the deposition of the incompletely metabolized substrate globotriaosylceramide in several cell types and multisystem involvement. Major morbidity results from renal, cardiac and cerebrovascular pathology, mediated by endothelial dysfunction. We examined the potential utility of Cystatin C and natriuretic peptides as biomarkers in FD, and evaluated serum levels in 89 FD patients with varying degrees of disease severity. The results revealed that as a prognostic marker, Cystatin C is a good and cost effective indicator of early renal dysfunction and/or heart failure in FD. It is also more useful than serum creatinine in detecting mild renal damage and small decreases in glomerular filtration. In addition, the natriuretic peptide NT-proBNP, was elevated in patients with FD and cardiac involvement, and found to be an adequate detection marker, not only of cardiac involvement, but also of diastolic dysfunction.


Sensorineural hearing loss alters recovery from short-term adaptation in the C57BL/6 mouse.
Several strains of laboratory mouse (Mus musculus) have a pattern of hearing loss which resembles that found in humans. The C57BL/6 strain of mouse has a genetic defect that results in degeneration of the organ of Corti, originating in the basal, high-frequency region and then proceeding apically over time. The end result is a severe-to-profound sensorineural hearing loss (SNHL) by 14 months of age. In contrast, auditory function of the CBA strain remains normal through its early life span then slowly declines later in life, much like that typified by human presbycusis. The purpose of the present study was to compare ABR (peak 5) forward masking recovery functions in young, normal-hearing CBA and C57BL/6 mice to hearing-impaired C57BL/6 mice. ABR audiograms were obtained prior to collecting the tone-on-tone forward masking data. Masking was defined as a 50% reduction in the P5 component of the ABR, elicited and masked by 12 kHz tone bursts, using masker/probe time delays from 0 to 100 ms. Time constants were computed from an exponential model fit to the recovery functions (masker level vs. time delay). In hearing-impaired animals there was a significant increase in recovery from short-term adaptation as measured by the time constants, as well as a significant latency shift in the P5 component. The effects of SNHL on the recovery of the P5 component from short-term adaptation was comparable to that reported behaviorally for human hearing-impaired listeners and physiologically from the inferior colliculus (IC) of chinchillas suffering permanent threshold shifts.


B-mode and M-mode Ultrasonography of Tongue Movements during Swallowing.
This study investigated the possibility of using B-mode and M-mode ultrasonography to differentiate between the visceral and somatic swallowing patterns. Tongue movements during empty swallowing by 11 volunteers with a visceral swallowing pattern were traced in B-mode and M-mode ultrasound imaging before and after successful myofunctional therapy. Thirteen subjects with a somatic swallowing pattern served as the control group. We examined and compared the B-mode sequences and M-mode images of at least six of each study subject's swallowing processes for reproducible characteristics and parameters. We observed wide intraindividual variability, making interindividual comparison difficult or meaningless. We identified no qualitative characteristic features in B-mode imaging that could be designated as solely visceral or physiological swallowing movements. There was no obvious intraindividual reproducibility in the M-mode sonogram of the swallowing act. Given the experimental set-up we used (without a submandibular cushioning pad), it was not possible to differentiate individual swallowing phases, that is, the initial, transporting, and returning phases, on the M-mode images based on characteristic trace points, as described in the literature. We determined the amplitude and velocity of vertical tongue movement as well as total swallowing duration on the M-mode images. These parameters revealed wide intra- and interindividual variability. No qualitative differences could be established in B-mode imaging with the ultrasound method selected for this study. The parameters that could be measured on the M-mode images are not suitable for differentiating between visceral and somatic swallowing.


Featured Article: Electroporation-mediated gene delivery of surfactant protein B (SP-B) restores expression and improves survival in mouse model of SP-B deficiency.
Surfactant Protein B Deficiency is a rare but lethal monogenetic, congenital lung disease of the neonate that is unresponsive to any treatment except lung transplantation. Based on the potential that gene therapy offers to treat such intractable diseases, our objective was to test whether an electroporation-based gene delivery approach could restore surfactant protein B expression and improve survival in a compound knockout mouse model of surfactant protein B deficiency. Surfactant protein B expression can be shut off in these mice upon withdrawl of doxycycline, resulting in decreased levels of surfactant protein B within four days and death due to lung dysfunction within four to seven days. Control or one of several different human surfactant protein B-expressing plasmids was delivered to the lung by aspiration and electroporation at the time of doxycycline removal or four days later. Plasmids expressing human surfactant protein B from either the UbC or CMV promoter expressed surfactant protein B in these transgenic mice at times when endogenous surfactant protein B expression was silenced. Mean survival was increased 2- to 5-fold following treatment with the UbC or CMV promoter-driven plasmids, respectively. Histology of all surfactant protein B treated groups exhibited fewer neutrophils and less alveolar wall thickening compared to the control groups, and electron microscopy revealed that gene transfer of surfactant protein B resulted in lamellar bodies that were similar in the presence of electron-dense, concentric material to those in surfactant protein B-expressing mice. Taken together, our results show that electroporation-mediated gene delivery of surfactant protein B-expressing plasmids improves survival, lung function, and lung histology in a mouse model of surfactant protein B deficiency and suggest that this may be a useful approach for the treatment of this otherwise deadly disease. Impact statement Surfactant protein B (SP-B) deficiency is a rare but lethal genetic disease of neonates that results in severe respiratory distress with no available treatments other than lung transplantation. The present study describes a novel treatment for this disease by transferring the SP-B gene to the lungs using electric fields in a mouse model. The procedure is safe and results in enough expression of exogenous SP-B to improve lung histology, lamellar body structure, and survival. If extended to humans, this approach could be used to bridge the time between diagnosis and lung transplantation and could greatly increase the likelihood of affected neonates surviving to transplantation and beyond.


Establishment of the model of white blood cell membrane chromatography and screening of antagonizing TLR4 receptor component from Atractylodes macrocephala Koidz.
A model of white blood cell membrane chromatography (WB-CMC) was established to screen active component from Atractylodes macrocephala Koidz. The component can antagonize Toll-like receptor 4 (TLR4) and inhibit inflammatory reaction. In the model of WB-CMC, cell membrane stationary phase (CMSP) was prepared by immobilizing the rabbit white blood cell membrane (WBCM) onto the surface of silica carrier and taxinol was used as a model molecule. The active component which can act on WBCM and its receptor (such as TLR4) as an effective target in A. macrocephala was determined by using a replacement experiment. The anti-inflammatory effects of the active component were tested by using pharmacological methods in vivo. The results indicated that the retention characteristics of atractylenolide I as active component was similar to that of taxinol in the model of WB-CMC. And so, atractylenolide I acted on the WBCM and TLR4 and its anti-inflammatory activity was related with antagonizing TLR4. Therefore, the interaction between the active component and WBCM and its receptor can be simulated by the model of WB-CMC in vitro. This model can be used to screen active components and to study effective characteristics for acting on definite targets.


The relationship between early-life conditions in the home country and adult outcomes among child immigrants in the United States.
We examine the impact of health and economic conditions at birth on the adult outcomes of child immigrants using the Children of Immigrants Longitudinal Study. Our sample consists of children from 39 countries who were brought to the United States before the age of 13. We estimate immigrant outcomes as a function of the infant mortality rate (IMR) and GDP per capita of their home country in the year of birth, controlling for birth-year, year-of-arrival and country-of-birth fixed effects, as well as demographic characteristics. IMR has a significant negative impact on English reading ability and GPA in middle school. IMR significantly decreases first job prestige, years of schooling, working hours and log earnings. Some of these effects appear to be working through the lower middle school GPA. IMR does not influence self-rated health or labor market participation in adulthood, and there is no statistically significant relationship between GDP per capita and adult outcomes. Detrimental effects of IMR are significantly lower for children who arrived younger and whose parents have high school degree or above. Our estimates are of economic significance: the impact of being born in 1975 versus 1976 in Nicaragua in terms of the impact of IMR on earnings is equal to the gender effect on earnings. Our results cannot be explained by selection on observables: the pre-migration characteristics of children and parents are not associated significantly with the health and economic conditions at birth. Also, several tests show that our results cannot be explained by potential selection on unobservables.


Nitrogen use aggravates bacterial diversity and network complexity responses to temperature.
Rising temperature affects microbial composition and function in agriculture field, especially under nitrogen fertilization. In this study, we investigated the bacterial community of paddy soil incubated at controlled temperatures (5 °C, 15 °C, 25 °C, and 35 °C). Results showed that the response of bacterial communities to temperature was not uniform. Temperature elevation from 15 to 25 °C abruptly shifted the soil bacterial community, whereas elevation from 5 to 15 °C and from 25 to 35 °C had a marginal effect. The bacterial α-diversity was higher at 5 °C and 15 °C, owing to the massively distributed taxa with low abundance. However, as the temperature increased to 25 °C and 35 °C, these taxa were diminished, whereas Firmicutes significantly increased, resulting in a strong decline in α-diversity. Simultaneously, bacterial network complexity significantly increased at 25 °C and 35 °C, indicating the bacteria had closer interactions. Nitrogen application aggravated the variation in bacterial diversity and network complexity among temperatures. Interestingly, most complex network was observed under higher temperatures in fertilized soils. Collectively, these results indicate that nitrogen exacerbates the response of the soil bacterial community to temperature, and association between diversity and network complexity may be present.


Smooth muscle antibodies and cryoglobulinemia are associated with advanced liver fibrosis in Brazilian hepatitis C virus carriers.
Cryoglobulinemia and non-organ-specific-autoantibody are biomarkers of autoimmunity of the chronic infection caused by hepatitis C virus (HCV). In this work, we report the association between the presence of smooth muscle antibodies (SMA) and cryoglobulinemia and chronic liver disease in HCV carriers. Sixty-five untreated HCV patients, 38 women and 27 men were included in this study. Cryoglobulinemia was tested by cryoprecipitation, SMA by indirect fluorescent antibody test, and liver fibrosis and hepatocellular inflammation activity was investigated by histology of liver biopsy using the METAVIR score. The prevalence of SMA in the patients was 33.8% and cryoglobulinemia was demonstrated in 36.9% patients. Cryoglobulinemia and SMA seropositivity was associated with advanced fibrosis (p < 0.05). The presence of SMA and cryoglobulinemia was not associated with hepatocellular inflammation activity, age, carrier gender or HCV genotype. We concluded that liver biopsy should be recommended for HCV carriers that are seropositive for SMA or cryoglobulinemia.


Contrast echocardiography: evidence-based recommendations by European Association of Echocardiography.
This paper examines the evidence for contrast echocardiography, both for improving assessment of left ventricular structure and function compared with unenhanced echocardiography and for the identification of myocardial perfusion. Based on the evidence, recommendations are proposed for the clinical use of contrast echocardiography.


Acid-induced change in ozone-reactive site in indole ring of tryptophan.
It is well established that ozone as well as oxygen activated by tryptophan 2,3-dioxygenase or indoleamine 2,3-dioxygenase cleave the 2,3-C=C bond of the indole ring of tryptophan to produce N-formylkynurenine. In the present study, however, we found that exposure of tryptophan to aqueous ozone at and below pH 4.5 generated a different compound. The compound was identified as kynurenine by high performance liquid chromatography and mass spectrometry. Exposure of N-formylkynurenine to acidic ozone did not generate a significant amount of kynurenine, indicating that the kynurenine was not produced via N-formylkynurenine. Acidic ozone thus appears to cleave the 1, 2-NAC bond in place of the 2,3-C=C bond of the indole ring, followed by liberation of the 2-C atom. The 1,2-NAC bond and 2,3-C=C bond are likely to undergo changes in their nature of bonding on acidification, enabling ozone to react with the former bond but not with the latter bond.


Organization of nosocomial infection control measures and local networks for infectious disease control in middle-scale hospitals in Japan.
The aims of this study were to assess nosocomial infection control measures at middle-scale hospitals throughout Japan. Of the 823 hospitals participating in this questionnaire-based survey, more than half of the middle-scale hospitals have implemented nosocomial infection control measures, including infection surveillance or infection control rounds, while acknowledging a shortage of infection control staff. These hospitals most frequently consulted public health centers to obtain information and advice. Improved nosocomial infection control in middle-scale hospitals requires sufficient staffing and a local network, with active participation by public health centers.


Heparin-induced priapism.
Heparin-induced priapism constitutes a special form of pharmaco-induced prolonged erection, but the pathophysiological principles are not yet definitely clear. Heparin-induced antiplatelet-antibodies may lead to the aggregation of thrombocytes and thus alter the penile blood flow leading to low-flow priapism. Alternatively, this condition may be explained by initial high-flow priapism that later turns into ischemic priapism. The question remains whether hemorrhage with subsequent organisation of the hematoma and late fibrosis constitutes a pathogenetic factor. Besides this pathogenetic discussion, this paper presents the differential diagnosis of priapism as well as diagnostic and therapeutic procedures.


RBE-weighted dose and its impact on the risk of acute coronary event for breast cancer patients treated with intensity modulated proton therapy.
To evaluate the relative biological effectiveness (RBE)-weighted dose to the heart and to estimate RBE uncertainties when assuming a constant RBE of 1.1, for breast cancer patients receiving intensity-modulated proton therapy (IMPT). Further, to study the impact of RBE uncertainties on the risk of an acute coronary event (ACE). We analyzed 20 patients who received IMPT to either the left breast (n = 10) or left chest wall (n = 10) and regional lymph nodes. The Monte Carlo simulation engine, MCsquare, was used to simulate the dose-averaged linear energy transfer (LETd) map. The RBE-weighted dose to the heart and its substructures was calculated using three different RBE models. The risk of ACE was estimated per its linear relationship with mean heart dose (MHD) as established by Darby et al. The median MHD increased from 1.33 GyRBE assuming an RBE of 1.1 to 1.64, 1.87, and 1.99 GyRBE when using the RBE-weighted dose models. The median values (and ranges) of the excess absolute risk of ACE were 0.4% (0.1%-0.8%) when assuming an RBE of 1.1, and 0.6% (0.2%-1.0%), 0.6% (0.2%-1.1%), and 0.7% (0.2%-1.1%) with the RBE-weighted models. For our patient cohort, the maximum excess absolute risk of ACE increased by 0.3% with the RBE-weighted doses compared to the constant RBE of 1.1, reaching an excess absolute ACE risk of 1.1%. The interpatient LETd variation was small for the relevant high-dose regions of the heart. All three RBE models predicted a higher biological dose compared to the clinical standard dose assuming a constant RBE of 1.1. An underestimation of the biological dose results in underestimation of the ACE risk. Analyzing the voxel-by-voxel biological dose and the LET map alongside clinical outcomes is warranted in the development of a more accurate normal-tissue complication probability model.


Tapped-inductor bi-directional Cuk converter with high step-up/down conversion ratio and its optimum design.
A bidirectional DC-DC converter is required for an energy storage system. High efficiency and a high step-up and step-down conversion ratio are the development trends. In this research, a series of bidirectional high-gain Cuk circuits was derived by combining tapped inductors and bidirectional Cuk. After analyzing and comparing the characteristics of each circuit, a bidirectional high-gain Cuk circuit with a tapped-inductor (reverse coupling) was proposed. The proposed converter has a simple structure and a high voltage gain in both the step-down (Buck) and step-up (Boost) operation modes. The voltage stress of S2 was low. The voltage stress of S1 was high, however, and this is a disadvantage of the proposed converter. The proposed circuit's characteristics were thoroughly examined, including the voltage gain characteristics and the design of the main parameters. We established a power loss model of the new topology, and the tapped-inductor turn ratio was optimized for high efficiency. Finally, a 400 W experimental implementation of the converter was shown to achieve efficiencies of 93.5% and 92.4% in the step-up and step-down modes, respectively. These findings verified the validity of the proposed circuit's theoretical analysis.


A Novel Ultrasound-Based Lower Extremity Motion Tracking System.
Tracking joint motion of the lower extremity is important for human motion analysis. In this study, we present a novel ultrasound-based motion tracking system for measuring three-dimensional (3D) position and orientation of the femur and tibia in 3D space and quantifying tibiofemoral kinematics under dynamic conditions. As ultrasound is capable of detecting underlying bone surface noninvasively through multiple layers of soft tissues, an integration of multiple A-mode ultrasound transducers with a conventional motion tracking system provides a new approach to track the motion of bone segments during dynamic conditions. To demonstrate the technical and clinical feasibilities of this concept, an in vivo experiment was conducted. For this purpose the kinematics of healthy individuals were determined in treadmill walking conditions and stair descending tasks. The results clearly demonstrated the potential of tracking skeletal motion of the lower extremity and measuring six-degrees-of-freedom (6-DOF) tibiofemoral kinematics and related kinematic alterations caused by a variety of gait parameters. It was concluded that this prototyping system has great potential to measure human kinematics in an ambulant, non-radiative, and noninvasive manner.


Maternal-fetal HLA-DR relationships and pregnancy-induced hypertension.
Some studies have found an increased prevalence of pregnancy-induced hypertension among women sharing HLA antigens with their spouses or fetuses, thus supporting the hypothesis that maternal sensitization to fetal HLA alloantigens reduces the risk for pregnancy-induced hypertension. However, not all studies have confirmed these findings. No investigators have examined the four different types of maternal-fetal HLA relationships in their studies of pregnancy-induced hypertension. Our goal was to examine such associations to test further the HLA-allosensitization hypothesis. We conducted a cohort study of pregnancy-induced hypertension among 683 nulliparous women. Women and their neonates were typed for HLA-A, -B, -DR, and -DQ antigens using serologic techniques to establish maternal-fetal relationships. We found an increased prevalence of pregnancy-induced hypertension when the fetus, but not the mother, was potentially exposed to HLA-DR alloantigens (maternal allogenicity) compared with the other three conditions combined (P < .003). Controlling for confounding factors, the increased prevalence of pregnancy-induced hypertension persisted in situations of maternal HLA-DR allogenicity (P < .007). Based upon our observations and other immunologic studies of pregnancy-induced hypertensive and uncomplicated pregnancies, we conclude that a maternal humoral response against fetal anti-HLA-DR immunoglobulin (IgG) antibody may influence the development of pregnancy-induced hypertension. This could occur when an immunocompetent fetus is exposed to maternal HLA-DR alloantigens, maternal exposure to fetal HLA-DR alloantigens alloantigens, maternal exposure to fetal HLA-DR alloantigens is not possible, and fetal IgG antibody bears paternally inherited markers allogeneic to the mother.


Dysiscalarones A-E, scalarane sesterterpenoids with nitric oxide production inhibitory activity from marine sponge Dysidea granulosa.
Dysiscalarones A-E (1-5), five new scalarane-type bishomoscalarane sesterterpenoids, were isolated from marine sponge Dysidea granulosa collected from the South China Sea, together with two known ones, honulactone A (6) and phyllofolactone I (7). The new structures were determined by extensive spectroscopic analysis including HR-ESI-MS and 1D and 2D NMR data, and their absolute configurations were assigned by single crystal X-ray diffraction analyses. The inhibitory activity of all the seven isolates on the production of nitric oxide (NO) stimulated by lipopolysaccharide (LPS) in mouse RAW 264.7 macrophages was evaluated. Of these metabolites, dysiscalarones A-B (1-2), honulactone A (6), and phyllofolactone I (7) showed inhibitory activities with respective IC50 values of 16.4, 18.5, 2.6, and 3.7 μM, which suggested that the γ-methylated α,β-unsaturated γ-lactone might be the functional group. In addition, all the seven metabolites showed no significant cytotoxicity against lung cancer PC9 cell line at the concentration of 20 μM.


METTL3-mediated N6-methyladenosine modification is critical for epithelial-mesenchymal transition and metastasis of gastric cancer.
As one of the most frequent chemical modifications in eukaryotic mRNAs, N6-methyladenosine (m6A) modification exerts important effects on mRNA stability, splicing, and translation. Recently, the regulatory role of m6A in tumorigenesis has been increasingly recognized. However, dysregulation of m6A and its functions in tumor epithelial-mesenchymal transition (EMT) and metastasis remain obscure. qRT-PCR and immunohistochemistry were used to evaluate the expression of methyltransferase-like 3 (METTL3) in gastric cancer (GC). The effects of METTL3 on GC metastasis were investigated through in vitro and in vivo assays. The mechanism of METTL3 action was explored through transcriptome-sequencing, m6A-sequencing, m6A methylated RNA immunoprecipitation quantitative reverse transcription polymerase chain reaction (MeRIP qRT-PCR), confocal immunofluorescent assay, luciferase reporter assay, co-immunoprecipitation, RNA immunoprecipitation and chromatin immunoprecipitation assay. Here, we show that METTL3, a major RNA N6-adenosine methyltransferase, was upregulated in GC. Clinically, elevated METTL3 level was predictive of poor prognosis. Functionally, we found that METTL3 was required for the EMT process in vitro and for metastasis in vivo. Mechanistically, we unveiled the METTL3-mediated m6A modification profile in GC cells for the first time and identified zinc finger MYM-type containing 1 (ZMYM1) as a bona fide m6A target of METTL3. The m6A modification of ZMYM1 mRNA by METTL3 enhanced its stability relying on the "reader" protein HuR (also known as ELAVL1) dependent pathway. In addition, ZMYM1 bound to and mediated the repression of E-cadherin promoter by recruiting the CtBP/LSD1/CoREST complex, thus facilitating the EMT program and metastasis. Collectively, our findings indicate the critical role of m6A modification in GC and uncover METTL3/ZMYM1/E-cadherin signaling as a potential therapeutic target in anti-metastatic strategy against GC.


Genome-wide identification and expression analysis of the calmodulin-binding transcription activator (CAMTA) family genes in tea plant.
As a type of calmodulin binding protein, CAMTAs are widely involved in vegetative and reproductive processes as well as various hormonal and stress responses in plants. To study the functions of CAMTA genes in tea plants, we investigated bioinformatics analysis and performed qRT-PCR analysis of the CAMTA gene family by using the genomes of 'ShuChaZao' tea plant cultivar. In this study, 6 CsCAMTAs were identified from tea plant genome. Bioinformatics analysis results showed that all CsCAMTAs contained six highly conserved functional domains. Tissue-specific analysis results found that CsCAMTAs played great roles in mediating tea plant aging and flowering periods. Under hormone and abiotic stress conditions, most CsCAMTAs were upregulated at different time points under different treatment conditions. In addition, the expression levels of CsCAMTA1/3/4/6 were higher in cold-resistant cultivar 'LongJing43' than in the cold-susceptible cultivar 'DaMianBai' at cold acclimation stage, while CsCAMTA2/5 showed higher expression levels in 'DaMianBai' than in 'LongJing43' during entire cold acclimation periods. In brief, the present results revealed that CsCAMTAs played great roles in tea plant growth, development and stress responses, which laid the foundation for deeply exploring their molecular regulation mechanisms.


Association between transient pulmonary congestion during acute myocardial infarction and high incidence of death in six months.
In a prospective multicenter study of 866 patients after acute myocardial infarction (AMI), an increased or excessive mortality rate (13%) was confined to the first 6 months after AMI. In the subsequent 18 months of follow-up, the mortality rate (4%) was similar to that in coronary patients in chronic stable condition. Analysis of patients who died in the first 6 months revealed that 55% had had pulmonary congestion at the time of the index AMI. Neither these patients nor the others who died in the early period were found to have more severe ventricular dysfunction, more malignant arrhythmias or more severe ischemia than patients who died after 6 months. The reason for the high and early mortality in patients with pulmonary congestion is not clear, particularly because 30% had reasonable ventricular function, with an ejection fraction of more than 40%. However, given the poor prognosis of these patients, early and aggressive diagnostic efforts should be undertaken to exclude jeopardized regions remote from the initial AMI.


Scalable fabrication of bimetal modified polyacrylonitrile (PAN) nanofibrous membranes for photocatalytic degradation of dyes.
Fabricating nanostructured fibrous membranes as photocatalyst would facilitate the effective treatment of dyeing effluents. However, creating such photocatalytic nanofibrous membranes has proven tremendously challenging. In this work, we report a scalable strategy to prepare copper-iron bimetal modified polyacrylonitrile (PAN) nanofibrous membranes by electrospinning technology. The resultant PAN membranes exhibit an ultra-fine fiber diameter (600 nm), large surface area (5.34 m2 g-1) and excellent photocatalytic capacity for reactive blue 19, reactive red 195 and acid orange 7 (>99.99%) within 60 min. The successful synthesis of such intriguing materials could provide a versatile platform for further exploring nano-sized photocatalyst fibrous membranes for degradation of dyes.


Acetylcholinesterase activity at the synapses of presynaptic boutons with presumed alpha-motoneurons in chicken ventral horn. Light- and electron-microscopic studies.
Acetylcholinesterase (AChE) activity at the synapses of presynaptic boutons on presumed alpha-motoneurons in the chicken ventral horn was studied histochemically at the light- and electron-microscope levels. At the light-microscope level, many dot-like AChE-active sites were observed on the soma and dendrites of presumed alpha-motoneurons. On electron microscopy, reaction products for AChE activity were observed mainly in the synaptic clefts of the four kinds of presynaptic boutons: (1) S type boutons, (2) boutons containing small, spherical, dense cored vesicles (diameter range, 60-105 nm) and spherical, clear vesicles, (3) boutons containing medium-sized, spherical, dense cored vesicles (65-115 nm) and spherical, clear vesicles, and (4) boutons containing large, spherical, dense cored vesicles (80-130 nm) and spherical, clear vesicles. In the light of previous physiological and biochemical studies, the present results suggest the possibility that each of these presynaptic boutons which are AChE-active in their synaptic clefts may contain acetylcholine, substance P, or enkephalins which acts as a neurotransmitter or modulator.


Demonstration of the interaction between glycosaminoglycans and fibronectin in the basement membrane of the living chicken embryo.
A method utilizing microinjection of glycosaminoglycan-degrading enzymes in the chicken blastoderm prior to embryo culture and immunostaining for fibronectin have been applied to demonstrate an interaction between glycosaminoglycans and fibronectin in the basement membrane of the epiblast. Fixation of tissue in a mixture of formaldehyde and cetylpyridinium chloride allows detection of fibronectin only in those zones of the embryo that are not colonized by mesoblast cells. The epithelial-mesenchymal interface thus remains unstained. After degradation of glycosaminoglycans in the living organism, it is shown that this particular site, in fact, also contains fibronectin that is masked in vivo by, at least, hyaluronate. This interaction between both compounds is, during gastrulation, constantly correlated with mesoblast migration. Since previous studies have shown that the degradation of hyaluronate determines the behaviour of mesoblast cells, it is proposed that remodelling of the interaction between these compounds is necessary for mesoblast migration to occur.


Public involvement. Contributory factors.
A joint project involving a community health council, primary care trust and GPs identified more than 100 people keen to become involved in local services. The venture has led to a shadow patients' forum being set up. GPs and practice managers have proved important in involving the local population.


Ion flow in cochlear hair cells and the regulation of hearing sensitivity.
This paper discusses how ion transport proteins in the hair cells of the mammalian cochlea work to produce a sensitive but stable hearing organ. The transport proteins in the inner and outer hair cells are summarized (including their current voltage characteristics), and the roles of these proteins in determining intracellular Ca(2+), membrane potential, and ultimately cochlear sensitivity are discussed. The paper also discusses the role of the Ca(2+) sequestration sacs in outer hair cells in the autoregulation of hair cell membrane potential and cochlear gain, and how the underdamped control of Ca(2+) within these sacs may produce the observed slow oscillations in cochlear sensitivity and otoacoustic emissions after cochlear perturbations, including perilymphatic perfusions and prolonged low-frequency tones. The relative insensitivity of cochlear gain to short-term changes in the endocochlear potential is also discussed.


BAR-Seq clonal tracking of gene-edited cells.
Gene editing by engineered nucleases has revolutionized the field of gene therapy by enabling targeted and precise modification of the genome. However, the limited availability of methods for clonal tracking of edited cells has resulted in a paucity of information on the diversity, abundance and behavior of engineered clones. Here we detail the wet laboratory and bioinformatic BAR-Seq pipeline, a strategy for clonal tracking of cells harboring homology-directed targeted integration of a barcoding cassette. We present the BAR-Seq web application, an online, freely available and easy-to-use software that allows performing clonal tracking analyses on raw sequencing data without any computational resources or advanced bioinformatic skills. BAR-Seq can be applied to most editing strategies, and we describe its use to investigate the clonal dynamics of human edited hematopoietic stem/progenitor cells in xenotransplanted hosts. Notably, BAR-Seq may be applied in both basic and translational research contexts to investigate the biology of edited cells and stringently compare editing protocols at a clonal level. Our BAR-Seq pipeline allows library preparation and validation in a few days and clonal analyses of edited cell populations in 1 week.


Brodie abscess of the femoral capital epiphysis in a 2-year-old child caused by Kingella kingae.
We report the case of a Brodie abscess of the femoral capital epiphysis from which Kingella kingae was isolated. This is to the best of our knowledge the first report of a Brodie abscess of the femoral capital epiphysis from which K. kingae was isolated.