Molecular and functional characterization of ApPGRP from Anatolica polita in the immune response to Escherichia coli.
Peptidoglycan recognition proteins (PGRPs) widely distributed in invertebrates and vertebrates are pattern-recognition molecules in innate immunity. In the present study, a novel short PGRP gene, designated as ApPGRP was identified from Anatolica polita. The deduced amino acid sequence of ApPGRP is composed of 196 residues and contains a conserved PGRP domain at the C-terminus. The phylogenetic tree showed that ApPGRP shared high homology of amino acid sequence with TcPGRP2 from Tribolium castaneum. The recombinant protein of ApPGRP exhibited binding activity toward Escherichia coli and DAP-type PGN from E. coli. Quantitative real time PCR (RT-qPCR) analysis indicated that the relative expression level of ApPGRP was up-regulated significantly after E. coli and DAP-type PGN challenge in the fifth instar larvae of A. polita. Moreover, the expressions of three antimicrobial peptides (AMPs) (ApAttacin 1, ApAttacin 2 and ApColeoptericin) were significantly increased after E. coli and DAP-type PGN challenge. RNA interference (RNAi) experiments showed that the expressions of three AMPs in the larvae of A. polita were significantly decreased after injection of ApPGRP dsRNA. Furthermore, the expressions of three AMPs in the larvae injected were still significantly decreased after E. coli challenge compared to the control samples without dsRNA injection. The predicted 3D structure showed that the ApPGRP could form the protein core of five β-sheet and three ɑ-helices, which would be involved in specific PGN recognition. These results suggested that ApPGRP may play an important role in the immune response to E. coli infection and function as a receptor for antimicrobial peptide gene induction in Anatolica polita.


Elevated expression and cell cycle deregulation of a mitosis-associated target polypeptide of a carcinogen in hyperplastic and malignant rat hepatocytes.
A cytoplasmic polypeptide with a molecular weight of 14,000 (p14) was previously found to be the principal covalent target protein of the carcinogen, N-2-fluorenylacetamide (2-acetylaminofluorene), early during carcinogenesis in rat liver. The level of immunodetected p14 was markedly increased specifically during all stages of mitosis of hepatocytes in normal and regenerating partially hepatectomized livers. In addition, the polypeptide appeared to be immunologically and behaviorally related to a polypeptide with a molecular weight of 17,500 that is tightly bound to nucleosomes of chromatin in hepatocytes. This report describes the actions of the two polypeptides during hepatocarcinogenesis induced by ingestion of 3'-methyl-4-dimethylaminoazobenzene or N-2-fluorenylacetamide. Both carcinogens acted similarly in bringing about characteristic responses of the two polypeptides, detected immunohistochemically with specific rabbit antiserum and peroxidase-antiperoxidase complex. Four types of discrete hepatocytic lesions were observed. The earliest and least aberrant were hyperplastic foci of proliferating hepatocytes, which generally displayed markedly higher levels of immunostained p14 in cytoplasm, compared to levels in normal diploid hepatocytes. Furthermore, the very high concentrations of p14 were continuously present during cell interphase, in contrast to those in normal and regenerating hepatocytes, in which the elevation is restricted to the period of mitosis. Later-arising lesions were acidophilic adenomas of hepatocytes, which were characterized by quiescent morphology, fairly uniform and bulky eosinophilic cytoplasm, high levels of glycogen, and small delicate nuclei. These cells usually displayed little cytoplasmic p14. Coexistent hepatocytic lesions, i.e., mixed basophilic adenomas, exhibited considerable morphological heterogeneity, often slightly basophilic cytoplasm, and variable immunostain of cytoplasmic p14 during interphase. The fourth lesions were hepatocellular carcinomas, which continuously demonstrated much higher than normal levels of cytoplasmic p14 during cell interphase. During mitosis in the four types of hepatocytic lesions, the levels of immunostained p14 were usually further elevated above those in interphase, regardless of whether they were already very high during interphase in hyperplasia and malignancy, or low in acidophilic adenomas. All four kinds of carcinogen-altered lesions usually displayed little of the detectable Mr 17,500 polypeptide in nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)


Machine learning-based multimodal prediction of language outcomes in chronic aphasia.
Recent studies have combined multiple neuroimaging modalities to gain further understanding of the neurobiological substrates of aphasia. Following this line of work, the current study uses machine learning approaches to predict aphasia severity and specific language measures based on a multimodal neuroimaging dataset. A total of 116 individuals with chronic left-hemisphere stroke were included in the study. Neuroimaging data included task-based functional magnetic resonance imaging (fMRI), diffusion-based fractional anisotropy (FA)-values, cerebral blood flow (CBF), and lesion-load data. The Western Aphasia Battery was used to measure aphasia severity and specific language functions. As a primary analysis, we constructed support vector regression (SVR) models predicting language measures based on (i) each neuroimaging modality separately, (ii) lesion volume alone, and (iii) a combination of all modalities. Prediction accuracy across models was subsequently statistically compared. Prediction accuracy across modalities and language measures varied substantially (predicted vs. empirical correlation range: r = .00-.67). The multimodal prediction model yielded the most accurate prediction in all cases (r = .53-.67). Statistical superiority in favor of the multimodal model was achieved in 28/30 model comparisons (p-value range: <.001-.046). Our results indicate that different neuroimaging modalities carry complementary information that can be integrated to more accurately depict how brain damage and remaining functionality of intact brain tissue translate into language function in aphasia.


Hand-sewn esophagojejunostomy in transthoracic single-port assisted laparoscopic esophagogastrectomy for Siewert type II adenocarcinoma of the esophagogastric junction with esophageal invasion > 3 cm.
An optimal method for digestive tract reconstruction (DTR) in laparoscopic radical resection of Siewert type II adenocarcinoma of esophagogastric junction (AEG) has not yet been standardized. The aim of this study was to evaluate the safety and feasibility of a hand-sewn esophagojejunostomy (EJ) technique during transthoracic single-port assisted laparoscopic esophagogastrectomy (TSLE) for Siewert type II AEG with esophageal invasion > 3 cm. The perioperative clinical data and short-term outcomes for patients who underwent TSLE using hand-sewn EJ for Siewert type II AEG with esophageal invasion > 3 cm between March 2019 and April 2022 have been retrospectively reviewed. A total of 25 patients were eligible. All 25 patients were successfully operated. None was converted to open surgery or mortality. 84.00% of patients were male and 16.00% were female. The mean age, body mass index (BMI), and the American Society of Anesthesiologists (ASA) score were 67.88 ± 8.10 years, 21.30 ± 2.80 kg/m2, and 2.08, respectively. The average incorporated operative and hand-sewn EJ procedural times were 274.92 ± 57.46 and 23.36 ± 3.00 min, respectively. The length of extracorporeal esophageal involvement and proximal margin was 3.31 ± 0.26 cm and 3.12 ± 0.12 cm, respectively. The average time for the first oral feeding and hospital stay were 6 (3-14) and 7 (3-18) days, respectively. Two patients (8.00%) developed postoperative grade IIIa complications according to the Clavien-Dindo classification, including 1 case of pleural effusion and 1 case of anastomotic leakage, both of whom were cured by puncture drainage. Hand-sewn EJ in TSLE is safe and feasible for Siewert type II AEG. This method can ensure safe proximal margins and could be a good option with an advanced endoscopic suture technique for type II tumor with esophageal invasion > 3 cm.


The variety of clinical and histopathologic presentations of glomerulonephritis associated with latent syphilis.
Glomerulonephritis is a well established but rather uncommon complication of latent secondary syphilis. We present three cases of glomerulopathies associated with luetic infection, observed and managed in our institutions in the past three years. They illustrate a variety of clinicopathologic presentations of this nephropathy, from acute nephrotic syndrome through membranous glomerulopathy up to rapidly progressive glomerulonephritis. Regardless of the clinical course and histologic type, they were all characterized by strongly positive results of serologic tests for syphilis. Our observations suggest the necessity of eliminating luetic infection in aetiologic considerations of each newly diagnosed case of nephrotic syndrome.


Resource allocation for different types of vaccines against COVID-19: Tradeoffs and synergies between efficacy and reach.
Vaccine shortage and supply-chain challenges have caused limited access by many resource-limited countries during the COVID-19 pandemic. One of the primary decisions for a vaccine-ordering decision-maker is how to allocate the limited resources between different types of vaccines effectively. We studied the tradeoff between efficacy and reach of the two vaccine types that become available at different times. We extended a Susceptible-Infected-Recovered-Deceased (SIR-D) model with vaccination, ran extensive simulations with different settings, and compared the level of infection attack rate (IAR) under different reach ratios between two vaccine types under different resource allocation decisions. We found that when there were limited resources, allocating resources to a vaccine with high efficacy that became available earlier than a vaccine with lower efficacy did not always lead to a lower IAR, particularly if the former could vaccinate less than 42.5% of the population (with the selected study parameters) who could have received the latter. Sensitivity analyses showed that this result stayed robust under different study parameters. Our results showed that a vaccine with lower resource requirements (wider reach) can significantly contribute to reducing IAR, even if it becomes available later in the pandemic, compared to a higher efficacy vaccine that becomes available earlier but requires more resources. Limited resource in vaccine distribution is significant challenge in many parts of the world that needs to be addressed to improve the global access to life-saving vaccines. Understanding the tradeoffs between efficacy and reach is critical for resource allocation decisions between different vaccine types for improving health outcomes.


Cloning and characterization of keratin D, a murine endodermal cytoskeletal protein induced during in vitro differentiation of F9 teratocarcinoma cells.
We have identified a cDNA coding for the murine keratin D from a collection of clones representing F9 teratocarcinoma stem cell mRNA sequences. These sequences are synthesized specifically after the addition of retinoic acid and cAMP to the culture medium. The clone is 1,382 nucleotides long and contains the entire information for the active polypeptide, the complete 3' end and most, if not all, of the 5' non-coding region. The mRNA is found in hepatocytes, in PYS-2 cells (an endodermal cell line) and in differentiated (retinoic-acid-treated) F9 cells, but not in untreated F9 cells. The length of the mRNA is 1.4 kb, as estimated by Northern blot hybridization. Southern hybridization performed under very stringent conditions detects a single fragment hybridizing strongly with the cloned cDNA, suggesting that the mouse genome contains only one or very few copies of this gene. We present the first complete sequence of a keratin expressed in simple epithelia, i.e. keratin D, and discuss its structural features.


Cofactor engineering to regulate NAD+/NADH ratio with its application to phytosterols biotransformation.
Cofactor engineering is involved in the modification of enzymes related to nicotinamide adenine dinucleotides (NADH and NAD+) metabolism, which results in a significantly altered spectrum of metabolic products. Cofactor engineering plays an important role in metabolic engineering but is rarely reported in the sterols biotransformation process owing to its use of multi-catabolic enzymes, which promote multiple consecutive reactions. Androst-4-ene-3, 17-dione (AD) and androst-1, 4-diene-3, 17-dione (ADD) are important steroid medicine intermediates that are obtained via the nucleus oxidation and the side chain degradation of phytosterols by Mycobacterium. Given that the biotransformation from phytosterols to AD (D) is supposed to be a NAD+-dependent process, this work utilized cofactor engineering in Mycobacterium neoaurum and investigated the effect on cofactor and phytosterols metabolism. Through the addition of the coenzyme precursor of nicotinic acid in the phytosterols fermentation system, the intracellular NAD+/NADH ratio and the AD (D) production of M. neoaurum TCCC 11978 (MNR M3) were higher than in the control. Moreover, the NADH: flavin oxidoreductase was identified and was supposed to exert a positive effect on cofactor regulation and phytosterols metabolism pathways via comparative proteomic profiling of MNR cultured with and without phytosterols. In addition, the NADH: flavin oxidoreductase and a water-forming NADH oxidase from Lactobacillus brevis, were successfully overexpressed and heterologously expressed in MNR M3 to improve the intracellular ratio of NAD+/NADH. After 96 h of cultivation, the expression of these two enzymes in MNR M3 resulted in the decrease in intracellular NADH level (by 51 and 67%, respectively) and the increase in NAD+/NADH ratio (by 113 and 192%, respectively). Phytosterols bioconversion revealed that the conversion ratio of engineered stains was ultimately improved by 58 and 147%, respectively. The highest AD (D) conversion ratio by MNR M3N2 was 94% in the conversion system with soybean oil as reaction media to promote the solubility of phytosterols. The ratio of NAD+/NADH is an important factor for the transformation of phytosterols. Expression of NADH: flavin oxidoreductase and water-forming NADH oxidase in MNR improved AD (D) production. Besides the manipulation of key enzyme activities, which included in phytosterols degradation pathways, maintenance the balance of redox also played an important role in promoting steroid biotransformation. The recombinant MNR strain may be useful in industrial production.


Chemical pneumonitis following household exposure to hydrofluoric acid.
A previously healthy 26-year-old woman developed hemorrhagic alveolitis and adult respiratory distress syndrome (ARDS) following use of a hydrofluoric acid-containing household cleaning agent. Exposure was inhalational in nature. The circumstances and possible mechanism of chemical pneumonitis from low-dose inhalational exposure to hydrofluoric acid are discussed.


[The B cell in the pathogenesis of rheumatoid arthritis].
Classically, B-cells have been considered to play a secondary role in the pathogenesis of rheumatoid artritis, restricted to the production of auto-antibodies. Nevertheless, the unexpected good clinical response that the systemic depletion of B-cells has demonstrated in a well-controlled clinical trial in patients with rheumatoid artritis has revitalized the interest in this cell type in the pathogenesis of this autoimmune disease. Several evidences suggest that B-cells can regulate the course of the immune response through antibody productionindependent mechanisms. These mechanisms include antigen presentation and the release of soluble factors such as proinflammatory cytokines, metalloproteinases, and chemokines. This article reviews experimental data supporting that the participation of B-cells in the pathogenesis of rheumatoid arthritis occurs through multiple mechanisms.


Urgent call to action: Child rights and the state of child morbidity and mortality in Yemen.
More than 2.3 million children under the age of five in Yemen suffer from acute malnutrition. Approximately 450,000 are expected to suffer from severe acute malnutrition and may die if they do not receive urgent treatment. In this context, without security, stability, and better access for farmers to have the means to resume growing food, children and their families continue to sink deeper and deeper into hunger and malnutrition.


Enhanced perfluorooctane acid mineralization by electrochemical oxidation using Ti3+ self-doping TiO2 nanotube arrays anode.
Perfluorooctanoic acid (PFOA) is of increasing concern due to its worldwide application and extremely environmental persistence. Herein, we demonstrated the electrochemical degradation of PFOA with high efficiency using the Ti3+ self-doping TiO2 nanotube arrays (Ti3+/TiO2-NTA) anode. The fabricated Ti3+/TiO2-NTA anode exhibited vertically aligned uniform nanotubes structure, and was demonstrated good performance on the electrochemical degradation of PFOA in water. The degradation rate, total organic carbon (TOC) removal rate and defluorination rate of PFOA reached 98.1 %, 93.3 % and 74.8 %, respectively, after electrolysis for 90 min at low current density of 2 mA cm-2. The energy consumption (7.6 Wh L-1) of this electrochemical oxidation system using Ti3+/TiO2-NTA anode for PFOA degradation was about 1 order of magnitude lower than using traditional PbO2 anodes. Cathodic polarization could effectively prolong the electrocatalytic activity of the anode by regenerating Ti3+ sites. PFOA molecular was underwent a rapidly mineralization to CO2 and F-, with only low concentration of short-chain perflfluorocarboxylic acids (PFCAs) intermediates identified. A possible electrochemical degradation mechanism of PFOA was proposed, in which the initial direct electron transfer (DET) on the anode to yield PFOA free radicals (C7F15COO•) and hydroxyl radicals (•OH) oxidation were greatly enhanced. This presented study provides a novel approach for the purification of the recalcitrant PFOA from wastewaters.


Practice patterns in flexor tendon repair.
There is a considerable volume of literature describing new and supposedly superior methods of flexor tendon repair. The purpose of this study was to assess the flexor tendon techniques currently used in the Republic of Ireland. A postal survey was conducted of all consultant plastic surgeons and consultant orthopaedic surgeons who were members of the Irish Hand Surgery Society. The response rate was 90% (27/30). A simple running peripheral suture was used by 73% (P = 0.03) and the Kessler was the core suture of choice for 68% (P = 0.06). A significant number of respondents use non-absorbable suture materials for core (P = 0.0028) and peripheral suture (P < 0.0001). Seventy-seven percent sutured the flexor sheath where possible (P = 0.009). Notwithstanding the proposed advantages of newer techniques, it is evident from this study that the two-stranded Kessler core and simple running peripheral suture remains the most popular flexor tendon repair, with sheath closure preferred by the majority of respondents.


Comparison between multimodality imaging approaches for measurement of the tricuspid annulus in severe tricuspid regurgitation.
Tricuspid valve (TV) sizing is crucial for surgical or interventional procedures planning. Imaging TV is frequently challenging and often requires multimodal imaging techniques. Computed tomography (CT) is the gold standard for sizing. The authors compared measurements of the tricuspid annulus (TA) acquired using echocardiography and CT. Thirty-six patients with severe symptomatic tricuspid regurgitation were included in this retrospective analysis. During mid-diastole, the maximal two-dimensional (2D) TA diameter was directly measured in multiple views using transthoracic echocardiography (TTE) and transesophageal echocardiography (TEE). Three-dimensional (3D) TA size was assessed using cross-sectional long-axis and short-axis diameters, areas, and perimeters measured in the projected plane. The TA diameter was quantified by the perimeter measured on the CT images (CT imaging_indirect) and compared with echocardiography measurements. Tenting height and tenting area were also measured using TTE at mid systole. The long-axis dimensions measured using 3DTEE (3DTEE_direct) best correlated with the TA diameter (CT imaging_indirect) (R = 0.851, P = 0.0001) and the least discrepancies (difference 1.2 ± 2.4 mm, P = 0.012). The TA diameters quantified by the perimeters measured using 3DTEE (3DTEE_indirect) were smaller than the CT values (difference 2.5 ± 2.5 mm, P = 0.0001). The maximal dimensions directly measured by 2DTEE (2DTEE_direct) were modestly correlated with the CT values. Overall, the maximal dimensions by TTE_direct were less reliable than those by CT. TA eccentricity index correlated with the maximal tenting height and area. The patients with severe tricuspid regurgitation had a dilated and circular annulus. The long-axis TA dimensions (3DTEE_direct) were similar to the diameters (CT imaging_indirect).


Determination of methenamine, methenamine mandelate and methenamine hippurate in pharmaceutical preparations using ion-exchange HPLC.
An ion-exchange column high-performance liquid chromatography (HPLC) method has been developed for the determination of methenamine in methenamine and methenamine hippurate pharmaceutical preparations. The HPLC method uses a Zorbax SCX-300 column with acetonitrile-0.1M sodium perchlorate monohydrate (pH 5.8) (70:30, v/v) as the mobile phase at the flow rate of 1 mL/min. UV-detection was at 212 nm. The linear concentration plots for methenamine were linear over the concentration range of 0.25-50mM for methenamine and methenamine mandelate standards. The intra-day RSD precision was <1.25%, and for inter-day, <1.85%. The peaks for mandelic acid, hippuric acid and the other ingredients from placebo tablets do not interfere with the analysis for methenamine. The accuracy of this method was shown to be 99-101% by measuring the recovery of methenamine from spiked placebo tablets. The assay of methenamine from methenamine hippurate tablets and from a urinary antiseptic tablet containing methenamine were in the range of 98-102%. This HPLC method is a fast, simple and straightforward method for the analysis of methenamine in pharmaceutical preparations.


Initial experience with a radiology imaging network to newborn and intensive care units.
A digital image network has been installed in the James Whitcomb Riley Hospital for Children on the Indiana University Medical Center to create a limited all digital imaging system. The system is composed of commercial components, Philips/AT&T CommView system, (Philips Medical Systems, Shelton, CT; AT&T Bell Laboratories, West Long Beach, NJ) and connects an existing Philips Computed Radiology (PCR) system to two remote workstations that reside in the intensive care unit and the newborn nursery. The purpose of the system is to display images obtained from the PCR system on the remote workstations for direct viewing by referring clinicians, and to reduce many of their visits to the radiology reading room three floors away. The design criteria includes the ability to centrally control all image management functions on the remote workstations to relieve the clinicians from any image management tasks except for recalling patient images. The principal components of the system are the Philips PCR system, the acquisition module (AM), and the PCR interface to the Data Management Module (DMM). Connected to the DMM are an Enhanced Graphics Display Workstation (EGDW), an optical disk drive, and a network gateway to an ethernet link. The ethernet network is the connection to the two Results Viewing Stations (RVS) and both RVSs are approximately 100 m from the gateway. The DMM acts as an image file server and an image archive device. The DMM manages the image data base and can load images to the EGDW and the two RVSs. The system has met the initial design specifications and can successfully capture images from the PCR and direct them to the RVSs.(ABSTRACT TRUNCATED AT 250 WORDS)


The relationship between fluoride accumulation in tea plant and changes in leaf cell wall structure and composition under different fluoride conditions.
Tea plant is capable of hyper-accumulating fluoride (F) in leaves, suggesting drinking tea may cause excessive F intake in our body and threaten the health. This study investigated the changes in the structure, composition, and F content in the leaf cell wall of the tea (Camellia sinensis) under different F conditions to demonstrate the role of cell wall in F enrichment in tea plants. The cell wall was shown as the main part for F accumulation (67%-92%), with most of F distributed in the pectin fraction (56%-71%). With increasing F concentration, a significant increase (p < 0.05) was observed in the F content of cell wall and its components, the level of cell wall metal ions (i.e. Cu, Mg, Zn, Al, Ca, Ba, Mn), as well as the content of total cell wall materials, cellulose, and pectin. Meanwhile, the level of Cu, Mg, Zn, pectin, and cellulose was significantly positively correlated with the F content in the leaf cell wall. F addition was shown to increase the fluorescence intensity of LM19 and 2F4 antibody-labeled low-methylesterified homogalacturonans (HGs), while decrease LM20-labeled high-methylesterified HGs, coupled with an increase in the activity and gene expression of pectin methyl esterases (PMEs) in tea leaves. All these results suggest that F addition can increase pectin content and demethylesterification, leading to increased absorption of metal cations and chelation of F in the cell wall through the action of metal ions.


Effects of testosterone, estrogen, and dihydrotestosterone upon aggressive and sexual behavior of female rats.
Groups of female TMD rats were treated either with estradiol benzoate (EB), dihydrotestosterone propionate (DHTP), testosterone propionate (TP), EB + DHTP (EB/DHTP), or with oil. These groups of females were tested for social aggression and for masculine and feminine sexual behavior. In addition, patterns of masculine and feminine sexual responses during the aggressive encounters, were investigated. TP-treated females of the same strain were used as opponents in the tests for aggression. In accordance with previous results, EB did not activate aggression whereas TP treatment resulted in a significant increase in aggression in females. Aggressive responses were activated by adding DHTP to EB, up to levels equal to those activated by TP. Sexual responses were observed in the tests for aggression as well as in tests for sexual behavior. The results indicated that feminine and masculine sexual responses were affected significantly by hormonal treatment. Mounting behavior in the test for aggression was activated by TP and by EB/DHTP. Lordosis and proceptive responses were inhibited in these groups as compared to EB-treated females, both in tests for aggression and in tests for sexual behavior. The results are consistent with the idea that dihydrotestosterone inhibits feminine and activates masculine sexual activity. The results also indicate that EB and DHTP synergistically activate aggression.


Synthesis and characterization of a new and radiolabeled high-affinity substrate for H+/peptide cotransporters.
In this study we described the design, rational synthesis and functional characterization of a novel radiolabeled hydrolysis-resistant high-affinity substrate for H(+)/peptide cotransporters. L-4,4'-Biphenylalanyl-L-Proline (Bip-Pro) was synthesized according to standard procedures in peptide chemistry. The interaction of Bip-Pro with H(+)/peptide cotransporters was determined in intestinal Caco-2 cells constitutively expressing human H(+)/peptide cotransporter 1 (PEPT1) and in renal SKPT cells constitutively expressing rat H(+)/peptide cotransporter 2 (PEPT2). Bip-Pro inhibited the [(14)C]Gly-Sar uptake via PEPT1 and PEPT2 with exceptional high affinity (K(i) = 24 microm and 3.4 microm, respectively) in a competitive manner. By employing the two-electrode voltage clamp technique in Xenopus laevis oocytes expressing PEPT1 or PEPT2 it was found that Bip-Pro was transported by both peptide transporters although to a much lower extent than the reference substrate, Gly-Gln. Bip-Pro remained intact to > 98% for at least 8 h when incubated with intact cell monolayers. Bip-[(3)H]Pro uptake into SKPT cells was linear for up to 30 min and pH dependent with a maximum at extracellular pH 6.0. Uptake was strongly inhibited, not only by unlabeled Bip-Pro but also by known peptide transporter substrates such as dipeptides, cefadroxil, Ala-4-nitroanilide and delta-aminolevulinic acid, but not by glycine. Bip-Pro uptake in SKPT cells was saturable with a Michaelis-Menten constant (K(t)) of 7.6 microm and a maximal velocity (V(max)) of 1.1 nmol x 30 min(-1) x mg of protein(-1). Hence, the uptake of Bip-Pro by PEPT2 is a high-affinity, low-capacity process in comparison to the uptake of Gly-Sar. We conclude that Bip-[(3)H]Pro is a valuable substrate for both mechanistic and structural studies of H(+)/peptide transporter proteins.


ANTAGONISTIC PLEIOTROPY: AN INTERSPECIFIC DROSOPHILA COMPARISON.
Wild-type flies of 12 Drosophila species and semispecies were examined to determine whether correlation patterns between early- and late-life fitness characters predicted for individuals within a population by the antagonistic-pleiotropy hypothesis are reflected in comparisons of related species and semispecies that are known to differ in lifespan. Our goal was to determine whether the hypothesis is relevant to the evolution of life-history differences beyond the population level. Two fitness traits, egg production and percentage mating success, were observed at three ages: onset of reproductive age, one week later, and one month later. Age-dependent patterns of these traits do not consistently conform to predictions of the hypothesis. Species or semispecies that show reproductive vitality early in life need not be short-lived, and long lifespan need not be accompanied by a cost in early reproductive vitality, as measured by mating success and egg production. The two fitness traits can show different age-dependent patterns in the same species or semispecies. Potential explanations for the frequent inconsistency of the data with predictions of the hypothesis are discussed. Results support the idea that the hypothesis is only relevant to the evolution of life-history differences among individuals in the same breeding population confronted by the same environmental constraints.


Myocarditis in auto-immune or auto-inflammatory diseases.
Myocarditis is a major cause of heart disease in young patients and a common precursor of heart failure due to dilated cardiomyopathy. Some auto-immune and/or auto-inflammatory diseases may be accompanied by myocarditis, such as sarcoidosis, Behçet's disease, eosinophilic granulomatosis with polyangiitis, myositis, and systemic lupus erythematosus. However, data concerning myocarditis in such auto-immune and/or auto-inflammatory diseases are sparse. New therapeutic strategies should better target the modulation of the immune system, depending on the phase of the disease and the type of underlying auto-immune and/or auto-inflammatory disease.


Home Quarantine: A Double-Edged Sword During COVID-19 Pandemic for Hypertensive Disorders of Pregnancy and the Related Complications.
COVID-19 pandemic and worldwide quarantine seriously affected the physical and mental health of the general public. Our study aimed to investigate the effects of the COVID-19 quarantine on pregnancy outcomes among pregnant women with hypertensive disorders of pregnancy (HDP). This single-center retrospective cohort study collected complete clinical data of HDP patients with a history of home quarantine in The First Affiliated Hospital of Chongqing Medical University (Chongqing, China) in 2020 as well as the patients without home quarantine in 2018 and 2019. Then, the maternal and neonatal outcomes of two subtypes of HDP, gestational hypertension (GH) and preeclampsia/eclampsia (PE/E), were analyzed over the three years. The incidence of HDP increased from 0.84% in 2018 and 0.51% in 2019 to 2.30% in 2020. The data suggested that home quarantine was associated with higher gestational weight gain, obesity rates, blood pressure, and uric acid among the patients with HDP in 2020. Furthermore, HDP patients with a history of home quarantine may have worse neonatal outcomes, including lower newborn weight, shorter body length, lower Apgar score, and higher risk of fetal growth restriction. Our results suggested that COVID-19 quarantine may be a risk factor for poor pregnancy outcomes in HDP patients. Lifestyle guidance and antenatal care may be necessary for HDP patients with home quarantine in an epidemic outbreak.


The iPS technique provides hope for Parkinson's disease treatment.
More recently, reprogramming of somatic cells to an embryonic stem cell-like state presents a milestone in the realm of stem cells, making it possible to derive all cell types from any patients bearing specific genetic mutations. With the development of induced pluripotent stem (iPS) cells, we are now able to use the derivatives of iPS cells to study the mechanisms of disease and to perform drug screening and toxicology testing. In addition, differentiated iPS cells are now close to be used in clinical practice. Here we review the progress of iPS technique and the possible application in the area of Parkinson's disease treatment.


A prospective multi-centre, randomized study comparing the addition of tapering dexamethasone to other standard of care therapies for taxane-associated pain syndrome (TAPS) in breast cancer patients.
Taxane-associated pain syndrome (TAPS) is common with docetaxel and is characterised by myalgias and arthralgias starting 2-3 days after treatment and can last for up to 7 days. Anecdotal evidence suggests that corticosteroids can reduce TAPS. This multicentre, randomized trial evaluated the effect of additional tapering dexamethasone on TAPS. 130 breast cancer patients commencing docetaxel were randomized to dexamethasone premedication (8 mg/twice daily for 3 days) or dexamethasone premedication followed by tapering dexamethasone (4 mg/daily for 2 days followed by 2 mg/daily for 2 days). The primary endpoint was absolute change in FACT-Taxane questionnaire during the first chemotherapy cycle. Secondary endpoints: proportion of patients with clinically significant TAPS, QoL, pain and toxicity. 110/130 patients had complete data included in the primary analysis. The fall in FACT-Taxane scores was lower in the experimental group on day 5 (p = 0.05), but not on day 7 (p = 0.21). There was no difference in FACT-Taxane scores over the entire study duration (p = 0.59). Fewer patients in the experimental arm reported TAPS on day 5 (30 vs. 47%). There was a borderline significant attenuation of impairment of QoL with experimental treatment on day 5 (p = 0.06), but not day 7 (p = 0.53). Tapered schedule was associated with more dyspepsia and insomnia. A tapering schedule of dexamethasone was associated with a brief reduction in docetaxel-associated symptoms which was observed only during dexamethasone exposure and did not persist after discontinuation of the drug. ClinicalTrials.gov NCT03348696.


A comparative evaluation of intraoral and extraoral digital impressions: An in vivo study.
The accuracy of a dental impression is determined by two factors: "trueness" and "precision." The scanners used in dentistry are relatively new in market, and very few studies have compared the "precision" and "trueness" of intraoral scanner with the extraoral scanner. The aim of this study was to evaluate and compare accuracy of intraoral and extraoral digital impressions. Ten dentulous participants (male/female) aged 18-45 years with an asymptomatic endodontically treated mandibular first molars with adjacent teeth present were selected for this study. The prepared test tooth was measured using a digital Vernier caliper to obtain reference datasets. The tooth was then scanned using the intraoral scanner, and the extraoral scans were obtained using the casts made from the impressions. The datasets were divided into four groups and then statistically analyzed. The test tooth preparation was done, and dimples were made using a round diamond point on the bucco-occlusal, mesio-occlusal, disto-occlusal, and linguo-occlusal lines angles, and these were used to obtain reference datasets intraorally using a digital Vernier caliper. The test tooth was then scanned with the IO scanner (CS 3500, Carestream dental) thrice and also impressions were made using addition silicone impression material (3M™ ESPE) and dental casts were poured in Type IV dental stone (Kalrock-Kalabhai Karson India Pvt. Ltd., India) which were later scanned with the EO scanner (LAVA™ Scan ST Design system [3M™ ESPE]) thrice. The Datasets obtained from Intraoral and Extraoral scanner were exported to Dental Wings software and readings were obtained. Repeated measures ANOVA test was used to compare differences between the groups and independent t-test for comparison between the readings of intraoral and extraoral scanner. Least significant difference test was used for comparison between reference datasets with intraoral and extraoral scanner, respectively. A level of statistical significance of P < 0.05 was set. The precision values ranged from 20.7 to 33.35 μm for intraoral scanner and 19.5 to 37 μm for extraoral scanner. The mean deviations for intraoral scanner were 19.6 μm mesiodistally (MD) and 16.4 μm buccolingually (BL) and 24.0 μm MD and 22.5 μm BL for extraoral scanner. The mean values of the intraoral scanner (413 μm) for trueness were closest to the actual measurements (459 μm) than the extraoral scanner (396 μm). The intraoral scanner showed higher "precision" and "trueness" values when compared with the extraoral scanner.


Generalized estimating equation model and long-term exposure effect of antipsychotics on SH-SY5Y cells against oxidative stressors.
A comparison of the neuroprotective effect of different antipsychotics (APDs) over time on naïve SH-SY5Y against oxidative stressor insults using the generalized estimating equation (GEE). The hydrogen peroxide (H2O2), N-methyl-4-phenylpyridinium ion (MPP+), and β-amyloid peptide were used to treat cells with or without APDs (paliperidone, risperidone, olanzapine and haloperidol); cell survival and oxidative stress markers were measured and analyzed. Only haloperidol had higher baseline cytotoxicity than paliperidone. GEE showed the proper exposure time for evaluating the neuroprotection of APDs was 24 h, rather than 48 or 72 h. Paliperidone was superior to other APDs in protecting naïve SH-SY5Y, had the best effect against H2O2-, MPP+-induced cell death, and caused a significantly higher GST, lower HNE and protein carbonyl productions of naïve SH-SY5Y after stressor insults, which may implicate a molecular mechanism underlying its neuroprotective action. Repeated GEE measurements can correct for the correlation among the clusters to obtain a more accurate result for evaluating drug outcome. The interaction between drugs and stressors should be taken into account when determining the neuroprotective effect of APDs against different stressors. Paliperidone might be useful in alleviating oxidative stress induced by Aβ25-35 and MPP+, and provide neuroprotection against hydrogen peroxide in naïve SH-SY5Y.


An unusual nicotinamide derivative, 4-pyridone-3-carboxamide ribonucleoside (4PYR), is a novel endothelial toxin and oncometabolite.
Our recent studies identified a novel pathway of nicotinamide metabolism that involves 4-pyridone-3-carboxamide-1-β-D-ribonucleoside (4PYR) and demonstrated its endothelial cytotoxic effect. This study tested the effects of 4PYR and its metabolites in experimental models of breast cancer. Mice were divided into groups: 4T1 (injected with mammary 4T1 cancer cells), 4T1 + 4PYR (4PYR-treated 4T1 mice), and control, maintained for 2 or 21 days. Lung metastasis and endothelial function were analyzed together with blood nucleotides (including 4PYR), plasma amino acids, nicotinamide metabolites, and vascular ectoenzymes of nucleotide catabolism. 4PYR metabolism was also evaluated in cultured 4T1, MDA-MB-231, MCF-7, and T47D cells. An increase in blood 4PYR in 4T1 mice was observed at 2 days. 4PYR and its metabolites were noticed after 21 days in 4T1 only. Higher blood 4PYR was linked with more lung metastases in 4T1 + 4PYR vs. 4T1. Decreased L-arginine, higher asymmetric dimethyl-L-arginine, and higher vascular ecto-adenosine deaminase were observed in 4T1 + 4PYR vs. 4T1 and control. Vascular relaxation caused by flow-dependent endothelial activation in 4PYR-treated mice was significantly lower than in control. The permeability of 4PYR-treated endothelial cells was increased. Decreased nicotinamide but enhanced nicotinamide metabolites were noticed in 4T1 vs. control. Reduced N-methylnicotinamide and a further increase in Met2PY were observed in 4T1 + 4PYR vs. 4T1 and control. In cultured breast cancer cells, estrogen and progesterone receptor antagonists inhibited the production of 4PYR metabolites. 4PYR formation is accelerated in cancer and induces metabolic disturbances that may affect cancer progression and, especially, metastasis, probably through impaired endothelial homeostasis. 4PYR may be considered a new oncometabolite.


Laparoscopic Nissen fundoplication in childhood.
An obese, 10-year-old boy presented with symptoms of pain and apnea due to gastroesophageal reflux that was unresponsive to medication. A laparoscopic Nissen fundoplication was performed, and the child was discharged from the hospital within 36 hours of the operation. The rationale and technique of a laparoscopic Nissen fundoplication for symptomatic gastroesophageal reflux in childhood is described.


Titanium-manganese compound with a chiral Mn3Ti center.
We have shown here for the first time a facile route to the molecular compound [Mn3Ti(mu3-OCH2CH2OCH3)2(mu-OCH2CH2OCH3)3(mu-Cl)Cl2(OiPr)2] with a Mn3Ti motif, where the Ti atom is in the chiral position and the Mn atoms occupy nonchiral sites.


Endoscopic MRI: preliminary results of a new technique for visualization and staging of gastrointestinal tumors.
The principle of endoscopic ultrasonography--introducing the transducer of an external imaging method, such as ultrasonography, in combination with an endoscope into the gastrointestinal tract for higher-resolution imaging--has also been applied to magnetic resonance imaging (MRI). We report here on our preliminary experience with a new method of endoscopic MRI in the upper gastrointestinal tract. Endoscopic MRI was performed in 32 patients with esophageal (n = 8) or gastric diseases (n = 24), mostly tumors (n = 26). Most cancers were at an advanced stage (T3/T4). A prototype MRI endoscope connected to a 1.5 tesla MRI scanner was used. The accuracy of endoscopic MRI in visualizing and staging gastrointestinal lesions was assessed. The normal gastrointestinal wall consisted of three layers, and tumors were visualized as having a low signal intensity on both T1- and T2-weighted sequences. Destruction of the wall layers was found to be characteristic of malignancy. Sufficient images were obtained in seven of eight esophageal cases (88%), but in only 14 of 24 gastric cases (58%). In patients in whom adequate visualization was achieved, the endoscopic MRI results of local and regional staging were consistent with surgical, histopathological, CT and/or EUS results in all six esophageal cancer cases and in 89% (T stage) and 56% (N stage) of the nine patients with gastric cancer. These preliminary results of endoscopic MRI are the first to be reported in the English literature. They show the potential of the method for local and regional staging, three-dimensional visualization of lesions being a potential advantage. Further technical improvements are expected.


Canine intestinal contents vs. simulated media for the assessment of solubility of two weak bases in the human small intestinal contents.
This study was conducted to assess the relative usefulness of canine intestinal contents and simulated media in the prediction of solubility of two weak bases (dipyridamole and ketoconazole) in fasted and fed human intestinal aspirates that were collected under conditions simulating those in bioavailability/bioequivalence studies. After administration of 250 mL of water or 500 mL of Ensure plus [both containing 10 mg/mL polyethylene glycol (PEG) 4000 as nonabsorbable marker], intestinal aspirates were collected from the fourth part of the duodenum of 12 healthy adults and from the mid-jejunum of four Labradors. Pooled samples were analyzed for PEG, pH, buffer capacity, osmolality, surface tension, pepsin, total carbohydrates, total protein content, bile salts, phospholipids, and neutral lipids. The shake-flask method was used to measure the solubility of dipyridamole and ketoconazole in pooled human and canine intestinal contents and in fasted-state-simulating intestinal fluid (FaSSIF) and fed-state-simulating intestinal fluid (FeSSIF) containing various bile salts and pH-buffering agents. For both compounds, solubility in canine contents may be predictive of human intralumenal solubility in the fasting state but not in the fed state. The poor agreement of results in canine and human aspirates can be attributed to the higher bile salt content in canine bile. Solubility in FaSSIF containing a mixture of bile salts from crude bile predicted satisfactorily the intralumenal solubility of both drugs in the fasted state in humans. Solubility in FeSSIF, regardless of the identity of bile salts or of the buffering species, deviated from intralumenal values in the fed human aspirates by up to 40%. This was attributed to the lack of lipolytic products in FeSSIF, the higher bile salt content of FeSSIF, and the lower pH of FeSSIF. FaSSIF containing a mixture of bile salts from crude bile, and FeSSIF containing lipolytic products and, perhaps, having lower bile salt content but slightly higher pH, should be more useful than canine intestinal aspirates for predicting intralumenal solubilities in humans.


Bio-concentration of chromium--an in situ phytoremediation study at South Kaliapani chromite mining area of Orissa, India.
Mine waste water at South Kaliapani usually contains toxic levels of hexavalent Cr(VI). The present in situ study was conducted at South Kaliapani chromite mine area in Orissa state, India, to assess the phytoremediation ability of three plants, namely, rice (Oryza sativa L.), paragrass (Brachiaria mutica), and an aquatic weed (Eichhornia crassipes), in attenuating Cr(VI) from mine waste water and to correlate the bio-concentration factors (BCF) of Cr. Water hyacinth (E. crassipes) showed 24% to 54% reduction whereas paragrass (B. mutica) was able to reduce 18% to 33% of Cr(VI) from mine water. This reduction was studied over a period of 100 days of plant growth. The reduction was observed through a passage of a sum total of 2,000 sq. ft. cultivated plots and ponds separately. Reduction in Cr(VI) content in mine water varies with plant age as well as with the distance of passage. Cr accumulation and BCF values increased with high soil Cr levels as well as the age of plants. High BCF and transportation index (Ti) values, i.e., 10,924 and 32.09, respectively, were noted for water hyacinth. The Ti values indicated that the root-to-shoot translocation of Cr was very high after 100 days of growth. The total accumulation rate was maximum (8.29 mg Cr kg dry biomass(-1) day (-1)) in paragrass. The BCF values for roots were noted to be higher than those of leaves, stems, and grains of the 125-day-old plants. Hence, paragrass and water hyacinth may be used as tools of phytoremediation to combat the problem of in situ Cr contamination.


Efficacy and safety of esmirtazapine in adult outpatients with chronic primary insomnia: a randomized, double-blind placebo-controlled study and open-label extension.
Esmirtazapine (1.5-4.5 mg) has demonstrated short-term sleep-promoting effects in nonelderly outpatients with chronic insomnia. This phase 3, randomized, double-blind study (NCT00631657) and its open-label extension (NCT00750919) investigated efficacy and safety of long-term esmirtazapine treatment in adult outpatients with chronic insomnia. Participants were randomized to receive esmirtazapine 4.5 mg or placebo for 6 months; those receiving esmirtazapine were then rerandomized to esmirtazapine or placebo for an additional 7 days. Participants could enter the 6-month open-label extension with esmirtazapine 4.5 mg. The primary objective of the double-blind study was to assess long-term efficacy of esmirtazapine vs placebo on self-reported total sleep time. Assessing long-term safety and tolerability were secondary and primary objectives of the double-blind and extension studies, respectively. Overall, 457 participants received treatment in the double-blind study (esmirtazapine, n = 342; placebo, n = 115) and 184 participants (prior esmirtazapine, n = 136; prior placebo, n = 48) received esmirtazapine in the extension. In the double-blind study, a 48.7-minute increase in average nightly total sleep time was observed for esmirtazapine vs placebo (95% confidence interval, 35.0-62.5; P < .0001) at months 4-6. There was no evidence of residual effects on next-day alertness or daytime functioning and no evidence of rebound insomnia or withdrawal symptoms upon treatment discontinuation. Esmirtazapine was generally well tolerated; somnolence and weight gain were the most common adverse events. Esmirtazapine improved sleep duration vs placebo over at least 6 months. There was no evidence of next-day residual effects or of withdrawal symptoms or rebound insomnia following abrupt treatment discontinuation. Registry: ClinicalTrials.gov; Name: A 6-Month Efficacy and Safety Study of Org 50081 in Adult Patients With Chronic Primary Insomnia (21106/P05701/MK-8265-002); URL: https://clinicaltrials.gov/ct2/show/NCT00631657; Identifier: NCT00631657; and Registry: ClinicalTrials.gov; Name: Twenty-Six Week Extension Trial of Org 50081 (Esmirtazapine) in Outpatients With Chronic Primary Insomnia (176003/P05721/MK-8265-007); URL: https://clinicaltrials.gov/ct2/show/NCT00750919); Identifier: NCT00750919.


Partial purification and characterization of NAD-dependent 7alpha-hydroxysteroid dehydrogenase from Bacteroides thetaiotaomicron.
A NAD-dependent 7alpha-hydroxysteroid dehydrogenase was purified 18-fold over the activity in crude cell extracts prepared from Bacteroides thetaiotaomicron NCTC 10852 using Bio-Gel A 1.5-M column chromatography. A molecular weight of 320 000 was estimated for the partially purified intact enzyme. Substrate saturation kinetics were performed using the 18-fold purified enzyme and the lowest Km values were obtained for 3alpha,7alpha-dihydroxy bile acid and bile salt substrates including chenodeoxycholic acid (Km 0.048 mM), glycochenodeoxycholic acid (Km 0.083 mM) and taurochenodeoxycholic acid (Km 0.059 mM). In contrast, 3alpha,7alpha,12alpha-trihydroxy bile acid and bile salts had higher Km values, i.e. cholic acid (Km 0.22 mM), glycoholic acid Km 0.32 mM) and taurocholic acid Km 0.26 mM). NAD had a Km value of 0.20 mM. The possible physiological significance of 7alpha-hydroxy bile acid oxidation to intestinal bacteroides strains was accessed by determining the rate of conversion of [14C]-cholic acid to 7-ketodeoxy[14C]cholic acid by whole cell suspensions under different incubation conditions. The rate of biotransformation of bile acid to keto-bile acid incubated anaerobically under N2 gas increased markedly when potential electron acceptors such as fumarate (10 mM) or menadione (4 mM) was added exogenously. These results suggest that bile acid oxidation reactions may be linked to energy-generating systems in this bacterium.


Impact of two- and three-dimensional computed tomography use on intraobserver and interobserver reliabilities of pilon fracture classification and treatment recommendation.
Two-dimensional (2D) and three-dimensional (3D) computed tomography (CT) have been increasingly used in various intra-articular fractures including pilon fracture. However, no study has investigated intraobserver and interobserver reliabilities of pilon fracture classification using 3D CT images. (1) Intraobserver and interobserver agreements of fracture classifications and treatment recommendations will improve by using 2D CT images compared to using plain radiographs only; (2) agreements will improve by adding 3D CT images compared to adding 2D CT images; and (3) agreements of orthopedic residents rather than specialists will be influenced more by imaging modality. Ten orthopedic specialists and 10 residents completed a survey to classify the fractures according to the Rüedi-Allgöwer and AO/OTA classifications and to select treatment options using 25 pilon fracture images. The survey was conducted using plain radiographs, with 2D and 3D CT images introduced 3 and 6weeks later, respectively. Kappa coefficients were calculated to determine reliabilities. Intraobserver reliabilities for fracture classifications in specialists significantly improved by using 2D images compared to using plain radiographs only. Addition of 3D CT did not significantly improve intraobserver reliabilities compared to those with 2D CT. Use of 2D CT images significantly improved overall interobserver agreement of both classifications, with the improvement being greater for residents. Use of 3D CT images did not improve the interobserver reliability of both classifications. Overall interobserver reliabilities for treatment recommendations did not significantly differ according to the imaging modality. However, interobserver agreement among residents significantly improved from slight agreement using radiographs only to fair agreement using 2D CT images. Intraobserver and interobserver reliabilities of pilon fracture classification and treatment recommendations did not improve between using 3D and 2D CT. Using 2D CT images improved the intraobserver and interobserver reliabilities of the fracture classifications in specialists and the interobserver reliabilities of the fracture classifications and the treatment recommendations in residents. IV, case control study.


A review on polyhydroxyalkanoate production from agricultural waste Biomass: Development, Advances, circular Approach, and challenges.
Polyhydroxyalkanoates are biopolymers produced by microbial fermentation. They have excellent biodegradability and biocompatibility, which are regarded as promising substitutes for traditional plastics in various production and application fields. This review details the research progress in PHA production from lignocellulosic crop residues, lipid-type agricultural wastes, and other agro-industrial wastes such as molasses and whey. The effective use of agricultural waste can further reduce the cost of PHA production while avoiding competition between industrial production and food. The latest information on fermentation parameter optimization, fermentation strategies, kinetic studies, and circular approach has also been discussed. This review aims to analyze the crucial process of the PHA production from agricultural wastes to provide support and reference for further scale-up and industrial production.


Spatiotemporal variations and source identification of atmospheric nitrated and oxygenated polycyclic aromatic hydrocarbons in the coastal cities of the Bohai and Yellow Seas in northern China.
Gaseous and particulate nitrated polycyclic aromatic hydrocarbons (NPAHs, 12 species) and oxygenated polycyclic aromatic hydrocarbons (OPAHs, 4 species) in seven coastal cities of the Bohai and Yellow Seas were determined throughout the year. The annual arithmetical mean concentrations of ΣNPAH12 and ΣOPAH4 were 737 ± 475 pg/m3 and 35.3 ± 26.8 ng/m3. NPAHs and OPAHs existed mainly in the gaseous phase, accounted for 88.5% and 95.2% of the total concentrations. Air concentrations of ΣNPAH12 and ΣOPAH4 in the coastal cities of the Yellow Sea were significantly lower (p < 0.05) than those of the Bohai Sea. Air concentrations of ΣNPAH12 and ΣOPAH4 were significantly higher (p < 0.01) in winter than in summer. Strong secondary formation of atmospheric NPAHs and OPAHs occurred in all of the studied cities. The sequence of annual contribution of the emission sources of airborne NPAHs determined by positive matrix factorization was traffic exhaust > combustion of solid fuels (coal and biomass) > secondary formation, while for OPAHs, it was combustion of solid fuels > secondary formation > traffic exhaust. The combustion of solid fuels served as the main source of NPAHs and OPAHs in winter, while secondary formation was the predominant source in summer. Interregional transport may exert an important effect on the local atmospheric NPAHs and OPAHs by potential source contribution function analysis. The estimated incremental lifetime cancer risk (ILCR) due to inhalation exposure to specific NPAHs ranged from 2.9 × 10-12 to 6.2 × 10-6 (median at 4.8 × 10-9) was mainly attributed to exposure before the age of 16.


Evaluation of ICUs and weight of quality control indicators: an exploratory study based on Chinese ICU quality data from 2015 to 2020.
This study aimed to explore key quality control factors that affected the prognosis of intensive care unit (ICU) patients in Chinese mainland over six years (2015-2020). The data for this study were from 31 provincial and municipal hospitals (3425 hospital ICUs) and included 2 110 685 ICU patients, for a total of 27 607 376 ICU hospitalization days. We found that 15 initially established quality control indicators were good predictors of patient prognosis, including percentage of ICU patients out of all inpatients (%), percentage of ICU bed occupancy of total inpatient bed occupancy (%), percentage of all ICU inpatients with an APACHE II score ⩾15 (%), three-hour (surviving sepsis campaign) SSC bundle compliance (%), six-hour SSC bundle compliance (%), rate of microbe detection before antibiotics (%), percentage of drug deep venous thrombosis (DVT) prophylaxis (%), percentage of unplanned endotracheal extubations (%), percentage of patients reintubated within 48 hours (%), unplanned transfers to the ICU (%), 48-h ICU readmission rate (%), ventilator associated pneumonia (VAP) (per 1000 ventilator days), catheter related blood stream infection (CRBSI) (per 1000 catheter days), catheter-associated urinary tract infections (CAUTI) (per 1000 catheter days), in-hospital mortality (%). When exploratory factor analysis was applied, the 15 indicators were divided into 6 core elements that varied in weight regarding quality evaluation: nosocomial infection management (21.35%), compliance with the Surviving Sepsis Campaign guidelines (17.97%), ICU resources (17.46%), airway management (15.53%), prevention of deep-vein thrombosis (14.07%), and severity of patient condition (13.61%). Based on the different weights of the core elements associated with the 15 indicators, we developed an integrated quality scoring system defined as F score=21.35%xnosocomial infection management + 17.97%xcompliance with SSC guidelines + 17.46%×ICU resources + 15.53%×airway management + 14.07%×DVT prevention + 13.61%×severity of patient condition. This evidence-based quality scoring system will help in assessing the key elements of quality management and establish a foundation for further optimization of the quality control indicator system.


ACE inhibition in hypertension: focus on perindopril.
Ongoing developments in our understanding of cardiovascular disease, together with the introduction of new drugs to treat these conditions, has led to much debate over the optimal management of hypertension. The ALLHAT study showed no major differences in cardiovascular outcome among three major classes of antihypertensive drugs. Indeed, large meta-analyses have substantiated this view, and most experts would agree that BP reduction matters more than the choice of antihypertensive agent. However, recently published data from the ASCOT-BPLA trial for hypertensive patients at moderate risk of cardiac events have caused some experts to re-evaluate this view. The recent Blood Pressure Lowering Treatment Trialists' Collaboration publication confirmed this change. In the ASCOT-BPLA trial, antihypertensive therapy based on amlodipine+perindopril significantly reduced total and cardiovascular mortality as well as other clinically relevant outcomes in comparison with a traditional strategy based on atenolol and a thiazide diuretic, despite both regimens producing nonsignificantly different reductions in brachial BP. These findings suggest that amlodipine/perindopril may exert a beneficial effect by acting on other parameters such as central BP or BP variability. ACE inhibitors have been shown to have antiatherosclerotic and antithrombogenic effects, to improve endothelial dysfunction, and to prevent cardiac remodeling in patients with coronary heart disease. In this regard, perindopril, which has relatively high affinity for ACE and true 24-hour duration of action, is one of the most extensively studied ACE inhibitors. More recent data suggest that ACE inhibitors reduce arterial stiffness, an independent risk factor for cardiovascular events, and have a beneficial effect on central aortic BP, thus providing a possible explanation for the findings of ASCOT-BPLA and confirming that ACE inhibitors are an appropriate first choice for patients with hypertension.


Effect of reactor configuration on the kinetics and nitrogen byproduct selectivity of urea electrolysis using a boron doped diamond electrode.
Electrochemical systems have emerged as an advantageous approach for decentralized management of source-separated urine with the possibility of recovering or removing nutrients and generating energy. In this study, the kinetics and byproduct selectivity of the electrolytic removal of urea were investigated using a boron doped diamond working electrode under varied operational conditions with a primary focus on comparing undivided and divided reactors. The urea removal rate in the undivided and divided reactors was similar, but the divided reactor had an increased required cell voltage needed to maintain the equivalent current density. The current efficiency was similar for 0.1, 0.25, and 0.5 A (33.3, 83.3, 167 mA/cm2), suggesting no interference from competing reactions at higher potentials. In a divided reactor, increasing the anolyte pH reduced the urea removal rate presumably from hydroxyl radical scavenging by hydroxide. Further, for all divided reactor experiments, the final pH was less than 1, suggesting that the transport of protons across the ion exchange membrane to the cathode was slower than the oxidation reactions producing protons. The nitrogen byproduct selectivity was markedly different in the undivided and divided reactors. In both reactors, nitrate (NO3-) formed as the main byproduct at the anode, but in the undivided reactor it was reduced at the stainless steel cathode to ammonia. In the presence of 1 M chloride, the urea removal kinetics improved from the generation of reactive chlorine species, and the byproduct selectivity was shifted away from NO3- to presumably chloramines and N2. Overall, these results indicate that the electrochemical reactor configuration should be carefully considered depending on the desired outcome of treating source-separated urine (e.g., nitrogen recovery, H2 generation).


Arithmetic memory networks established in childhood are changed by experience in adulthood.
Adult bilinguals show stronger access to multiplication tables when using the language in which they learned arithmetic during childhood (LA+) than the other language (LA-), implying language-specific encoding of math facts. However, most bilinguals use LA+ throughout their life, confounding the impact of encoding and use. We tested if using arithmetic facts in LA- could reduce this LA- disadvantage. We measured event related brain potentials while bilingual teachers judged the correctness of multiplication problems in each of their languages. Critically, each teacher taught arithmetic in either LA+ or LA-. Earlier N400 peak latency was observed in both groups for the teaching than non-teaching language, showing more efficient access to these facts with use. LA+ teachers maintained an LA+ advantage, while LA- teachers showed equivalent N400 congruency effects (for incorrect versus correct solutions) in both languages. LA- teachers also showed a late positive component that may reflect conflict monitoring between their LA+ and a strong LA-. Thus, the LA- disadvantage for exact arithmetic established in early bilingual education can be mitigated by later use of LA-.


Is blockchain for Internet of Medical Things a panacea for COVID-19 pandemic?
The outbreak of the COVID-19 pandemic has deeply influenced the lifestyle of the general public and the healthcare system of the society. As a promising approach to address the emerging challenges caused by the epidemic of infectious diseases like COVID-19, Internet of Medical Things (IoMT) deployed in hospitals, clinics, and healthcare centers can save the diagnosis time and improve the efficiency of medical resources though privacy and security concerns of IoMT stall the wide adoption. In order to tackle the privacy, security, and interoperability issues of IoMT, we propose a framework of blockchain-enabled IoMT by introducing blockchain to incumbent IoMT systems. In this paper, we review the benefits of this architecture and illustrate the opportunities brought by blockchain-enabled IoMT. We also provide use cases of blockchain-enabled IoMT on fighting against the COVID-19 pandemic, including the prevention of infectious diseases, location sharing and contact tracing, and the supply chain of injectable medicines. We also outline future work in this area.


Lack of control enhances accurate and inaccurate identification responses to degraded visual objects.
Recent studies have shown that lack of control induces illusory pattern perception. In this study, we demonstrate that lacking control also affected identification responses to degraded pictures of visual objects. As compared with control participants, participants in whom lack of control was experimentally induced produced identification responses to visual objects at a significantly higher level of degradation of the stimulus. Lacking control did not influence accuracy of identification, since the effect was present for both correct and incorrect identification responses, and did not encourage pure random guessing. Our results indicate that lacking control makes individuals more prone to achieve visual closure by attempting earlier identifications of perceptually degraded visual objects. Possible mechanisms involved in this effect are discussed.


Prognostic stratification of clinical and molecular epithelioid hemangioendothelioma subsets.
Epithelioid hemangioendothelioma is a low-grade malignant vascular tumor with an intermediate clinical behavior between benign hemangiomas and high-grade angiosarcomas. Pathologic or molecular factors to predict this clinical heterogeneity are not well defined. A WWTR1-CAMTA1 fusion is present in most classic epithelioid hemangioendothelioma, regardless of their clinical behavior, suggesting that additional genetic abnormalities might be responsible in driving a more aggressive biology. A small subset of cases show distinct morphology and are characterized genetically by a YAP1-TFE3 fusion. Two histologic grades have been described in classic epithelioid hemangioendothelioma of the soft tissue. However, proposed criteria do not apply to other clinical presentations and have not been assessed in the YAP1-TFE3 positive tumors. Furthermore, no previous studies have compared the survival of these two molecular subsets. In this study we investigate the clinicopathologic and molecular findings of a large cohort of 93 translocation-positive epithelioid hemangioendothelioma managed at our institution. Patient characteristics, histologic features, treatment outcomes, and genetic abnormalities were investigated and these factors were correlated with overall survival. In 18 patients (15 with WWTR1-CAMTA1 and 3 with YAP1-TFE3) Memorial Sloan Kettering-IMPACT targeted DNA sequencing was performed to identify secondary genetic alterations showing more than half of tumors had a genetic alteration beyond the disease-defining gene fusion. Patients with conventional epithelioid hemangioendothelioma with WWTR1-CAMTA1 fusion had a less favorable outcome compared with the YAP1-TFE3 subset, the 5-year overall survival being 59% versus 86%, respectively. Soft tissue epithelioid hemangioendothelioma were frequently solitary, followed an uneventful clinical course being often managed with curative surgery. Multifocality, pleural involvement, lymph node or distant metastases had a significantly worse outcome. Patients with pleural disease or lymph node metastases had an aggressive clinical course akin to high-grade sarcomas, with 22% and 30%, respectively, alive at 5 years, compared with >70% survival rate in patients lacking these two adverse factors.


The stability of several compounds in formalin fixed tissues and formalin-blood solutions.
Buffered formalin solutions were added to spiked blood samples containing diazepam, phenytoin, carbon monoxide and cyanide to give formalin-whole blood solutions of 5 and 8%. Sections of liver positive for desipramine, phenobarbital and phenytoin were placed in separate 5 and 8% formalin-water solutions. The formalin-blood solutions were monitored daily for 30 days, while the fixed liver and formalin-water samples were analyzed once a week for 4 weeks. In the formalin-blood solutions losses were found for diazepam and phenytoin over the 30-day period of at least 41% and 33%, respectively. Cyanide detection was not possible immediately after the addition of formalin and the presence of carboxyhemoglobin was difficult to detect after 1 week. In the liver, losses of phenobarbital and desipramine were greater than 60% while phenytoin showed little change. This study has revealed that the drugs examined at toxic concentrations can be detected, with variable recoveries, for up to 30 days after fixation with formalin. However, quantitative analysis for cyanide and carboxyhemoglobin may be significantly impaired in the presence of formaldehyde.


Self-directed affiliation and autonomy across acute and continuation phase cognitive therapy for recurrent depression.
Using Benjamin's (2000) Structural Analysis of Social Behavior, we evaluated change in "self-directed" affiliation and autonomy and prediction of treatment response and relapse/recurrence among adult outpatients with recurrent major depressive disorder consenting to acute phase cognitive therapy (A-CT; Beck, Rush, Shaw, & Emery, 1979; N=156); A-CT responders randomized (N=84) to 8 months of continuation phase cognitive therapy (C-CT; Jarrett, 1989; Jarrett et al., 1998; Jarrett & Kraft, 1997) or assessment-only control; and C-CT and control patients entering a 16-month, assessment-only follow-up (N=74). Self-directed affiliation and autonomy increased after A-CT, and C-CT further increased affiliation and autonomy. Affiliation and autonomy did not predict A-CT response, but lower affiliation and higher autonomy pre-A-CT predicted relapse/recurrence post-A-CT. We discuss potential clinical implications of these results and present case examples to illustrate patterns of change.


Ability of the post-operative ALBI grade to predict the outcomes of hepatocellular carcinoma after curative surgery.
The albumin-bilirubin (ALBI) grade has been validated as a significant predictor for hepatocellular carcinoma (HCC). However, there is little information about the impact of postoperative ALBI grade in patients with HCC who are undergoing liver resection. We enrolled 525 HCC patients who received primary resection from April 2001 to March 2017. The impact of the pre- and post-operative ALBI grades on overall survival (OS) and recurrence-free survival (RFS) were analyzed by multivariate analysis. During the follow-up period (mean, 65 months), 253 (48.1%) patients experienced recurrence, and 85 (16.2%) patients died. Multivariate analysis revealed that diabetes mellitus (DM) (p = 0.011), alpha-fetoprotein levels (AFP) (p < 0.001), low platelet count (p = 0.008), liver cirrhosis (p < 0.001), and the first year of ALBI grade after resection (p < 0.001) were independent predictors for RFS. Additionally, old age (p = 0.006), DM (p = 0.002), AFP (p = 0.027), and ALBI grade at the first year after resection (p < 0.001) were independent risk factors for poor liver-related survival. Patients with post-operative ALBI grades II/III had older age (p = 0.019), hypoalbuminemia (p = 0.038), DM (p = 0.043), and high stages of pTNM (p = 0.021). The post-operative ALBI grade is better for predicting the outcomes in HCC patients after curative hepatectomy than the pre-operative ALBI grade.


Applying a new pomelo peel derived biochar in microbial fell cell for enhancing sulfonamide antibiotics removal in swine wastewater.
A sequential anode-cathode double-chamber microbial fuel cell (MFC) is a promising system for simultaneously removing contaminants, recovering nutrients and producing energy from swine wastewater. To improve sulfonamide antibiotics (SMs)'s removal in the continuous operating of MFC, one new pomelo peel-derived biochar was applied in the anode chamber in this study. Results demonstrated that SMs can be absorbed onto the heterogeneous surfaces of biochar through pore-filling and π-π EDA interaction. Adding biochar to a certain concentration (500 mg/L) could enhance the efficiency in removing sulfamethoxazole, sulfadiazine and sulfamethazine to 82.44-88.15%, 53.40-77.53% and 61.12-80.68%, respectively. Moreover, electricity production, COD and nutrients removal were improved by increasing the concentration of biochar. Hence, it is proved that adding biochar in MFC could effectively improve the performance of MFC in treating swine wastewater containing SMs.


Problems with radiolabelling egg albumin for the Farr ammonium sulphate technique for antibody estimation.
High concentrations of passive haemagglutinating antibodies were raised in sheep immunised with egg albumin (Ea) in Feund's adjuvant. Further attempts to study the antigen binding capacities and avidities of these antisera by the Farr ammonium sulphate technique were hindered by unsuccessful attempts to label Ea with 125Iodine by either the chloramine T or sodium nitrite-HCl methods. This problem was attributed to the unavailability of tyrosine in the native protein. This was solved by labelling Ea with the Bolton-Hunter reagent which does not require the presence of tyrosine. However labelled Ea binds non-specifically to normal serum to give high control values of circa 40% binding. Despite this it was possible to estimate antibody concentrations and avidities of lamb sera.


Attention and emotional responses to sexual stimuli and their relationship to sexual desire.
Little is known about why individuals vary in their levels of sexual desire. Information processing models, like Barlow's (Journal of Consulting and Clinical Psychology 54:140-148, 1986) model of sexual functioning, suggest that individuals with higher sexual desire attend more and respond with more pleasant emotions to sexual cues than individuals with lower levels of sexual desire. In this study, 69 participants (36 women, 33 men) completed a dot detection task measuring attention capture by sexual stimuli and a startle eyeblink modulation task indexing the valence of emotional response to affective stimuli. Participants with high levels of sexual desire were slower to detect targets in the dot detection task that replaced sexual images but did not differ in startle eyeblink responses to sexual stimuli. The results suggest that the amount of attention captured by sexual stimuli is a stronger predictor of a person's sexual desire level than the valence of the emotional responses elicited by such stimuli.


Methenyltetrahydrofolate synthease deficiency (MTHFS deficiency): Novel mutation and brain MRI findings: A case report and glance to other cases.
This is a case report of Methenyl Tetrahydrofolate synthetase deficiency (MTHFS deficiency) characterized by global developmental delay, cerebral hypomyelination, severe spastic tonicity in extremities, and microcephaly. Mutation in the MTHFS gene was reported in the Whole Exome Sequencing (WES) and confirmed with Sanger sequencing of parents. It is of great significance to report since it would be the first case of MTHFS mutation reported from Iran and the fourth throughout the world with novel mutation and brain imaging.


Appendix E: Rapid-iTM: Closed Vitrification Device by Vitrolife.
Cryopreservation of gametes and embryos is a growing technique in numerous reproductive fields including human-assisted reproduction. With improved understanding of embryo physiology and optimized culture conditions, there are now more embryos than ever to vitrify for potential use in subsequent cycles. Many gametes and embryos have been cryopreserved in open systems, but there are concerns with regard to contamination from the liquid nitrogen and also cross-contamination between patients' germplasm. The development of the Rapid-i™, a closed vitrification device that does not use direct contact with liquid nitrogen during vitrification or subsequent storage, will be discussed as well as clinical protocols for human oocytes and embryos.


Air pollutants and development of interstitial lung disease in patients with connective tissue disease: a population-based case-control study in Taiwan.
The aim of this study was to assess the association between air pollutant exposure and interstitial lung disease (ILD) in patients with connective tissue diseases (CTDs). A nationwide, population-based, matched case-control study in Taiwan. Using the 1997-2013 Taiwanese National Health Insurance Research Database, we identified patients with newly diagnosed CTD during 2001-2013, including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), dermatomyositis (DMtis)/polymyositis (PM) and primary Sjögren's syndrome (pSS). Patients with newly diagnosed ILD during 2012-2013 were identified as ILD cases, and selected patients with CTD without ILD matching (1:4) the CTD cases for CTD diagnosis, age, gender, disease duration and year of ILD diagnosis date were identified as non-ILD controls. Data of hourly level of air pollutants 1 year before the index date were obtained from the Taiwan Environmental Protection Agency. The association between ILD and air pollutant exposure was evaluated using logistic regression analysis shown as adjusted ORs (aORs) with 95% CIs after adjusting for potential confounders. We identified 505 newly diagnosed CTD-ILD patients, including 82 with SLE, 210 with RA, 47 with SSc, 44 with DMtis/PM and 122 with pSS. Ozone (O3) exposure (per 10 ppb) was associated with a decreased ILD risk in patients with CTD (aOR, 0.51; 95% CI, 0.33 to 0.79) after adjusting for potential confounders. A previously unrecognised inverse correlation was found between O3 exposure and ILD in patients with RA and SSc. Further studies are warranted to explore the underlying mechanisms.


Stage II and stage III periodontitis clinical burdens of HIV-1 undergoing antiretroviral therapy.
The aim of this retrospective cross-sectional study was to estimate the association of HIV-1 infection under highly active antiretroviral treatment (HAART) on the clinical parameters of periodontitis. A total of 205 patients were divided in two groups: 74 HIV + and 131 HIV - . Periodontal probing depth (PPD), clinical attachment loss (CAL), bleeding on probing (BOP), and visible supragingival biofilm (VSB) were recorded. The association of HIV-1 infection with the presence of at least 3 sites with PPD ≥ 5 mm and/or CAL ≥ 4 mm in non-adjacent teeth was estimated using binary logistic regression models. The variables HIV-1 infection (OR = 5.53, p < 0.0001, 95% CI: 2.45-13.64), age [range 35-50 years old (OR = 5.73, p < 0.0001, 95% CI: 2.49-13.20); > 50 years old (OR = 6.29, p = 0.002, 95% CI: 1.94-20.42)], and VSB (OR = 23.68, p < 0.0001, 95% CI: 8.07-69.53) showed a significant direct association with BOP outcome. HIV-1 infection under HAART did not have association with the presence of at least 3 sites with PPD ≥ 5 mm and/or CAL ≥ 4 mm. However, HIV patients on HAART had direct association of HIV-1 infection with BOP and an inverse association with PPD. These results support that monitoring gingival bleeding associated with oral prophylaxis would be beneficial in the prevention and management of periodontitis in HIV-1 patients on HAART.


Assessment of protozoa in Yunnan Yellow cattle rumen based on the 18S rRNA sequences.
Library of ruminal protozoal 18S rRNA of Yunnan Yellow Cattle has been constructed in the present study. Phylogenic analysis of sequences was meanwhile employed to reveal the diversity of protozoa in the rumen of Yunnan Yellow Cattle. One Yellow Cattle was fed malt meal (YCRPB) and the other was fed wheat straw (YCRPS). A protozoa-specific primer (P-SSU-342f) and a eukarya-specific primer (Medlin B) were used to amplify a 1,360-bp fragment of DNA encoding protozoal small subunit (SSU) ribosomal RNA from rumen fluid. The results showed as follows: A total of 121 clones were obtained and fell into four genera identified as Entodinium (66.9%), Dasytricha (5.8%), Isotricha (9.1%), and Diplodinium (18.2%). Within the genus Entidinium, 48 of the YCRPB sequences and 33 of the YCRPS sequences clustered with the Entodinium caudatum. 7 of the YCRPB sequences were phylogenetically placed within the genus Dasytricha. 11 of the YCRPB sequences were related with high confidence to Isotricha intestinalis. 22 of the YCRPS sequences were phylogenetically placed within the genus Diplodinium. The predominant protozoal genus identified in the rumen fluid belonged to the Entodinium group, and the divergences between two cattle may due to diet and individual differences.


Denitrifying bacterial community dominantly drove nitrogen removals in vertical flow constructed wetlands as impacted by macrophyte planting patterns.
The study aims to identify relations of denitrifying bacterial and fungal communities to nitrogen removals in vertical flow wetland microcosms (VFWMs) using four macrophyte species (Iris pseudacorus, Canna glauca, Scirpus validus and Cyperus alternifolius) and three species richness levels (unplanted, monocultured and 4-species mixture) as fixed factors. Results showed that among four macrophyte species, only Canna glauca planting significantly decreased nitrate removal by 87.7% in the VFWMs. The 4-species mixture improved TN and nitrate removals by 84.0% and 91.3%, but decreased ammonium removal by 94.5%. Heatmap and nonmetric multidimensional scaling analyses identified a significant difference in denitrifying bacterial community structure across macrophyte richness levels, but did not identify the difference in denitrifying fungal communities. The redundancy analysis revealed that denitrifying bacterial community individually explained 99.4% and 93.0% variance of nitrogen removals among four macrophyte species and across macrophyte richness levels, while the fungal community only explained 30.7% and 21.8% variance of nitrogen removals. Overall, the macrophyte richness and bacterial denitrifiers are the critical factors of nitrogen removals in the VFWMs, thus providing useful data to design a vertical flow constructed wetland at a full scale.


Pneumatic retinopexy for inferior retinal breaks.
To introduce a pneumatic retinopexy positioning for the management of retinal detachment from inferior retinal breaks after failed scleral buckle. Retrospective review of interventional consecutive case series. Seventeen eyes of 17 patients followed up from January 1993 to February 2005. Internal tamponade was achieved by 10 degrees Trendelenburg positioning, 10 degrees neck hyperextension, 10 degrees ocular supraduction and a large intravitreal gas bubble. Visual acuity, intraocular pressure, and retinal flattening were evaluated. All patients tolerated the procedure. Retinal reattachment was maintained in 15 of 17 patients (88.2%) followed up for a mean of 2.8 years (range, 0.1-11.5 years). Two patients experienced redetachment after 5 days and 4 months respectively. Six patients required antiglaucoma medications during the technique. Graduated body-neck-eye tilt allowed retinal reattachment in most eyes with inferior retinal detachment after failed scleral buckle.


Development of a cloud point extraction and spectrophotometry-based microplate method for the determination of nitrite in human urine and blood.
A novel and simple method for the sensitive determination of trace amounts of nitrite in human urine and blood has been developed by combination of cloud point extraction (CPE) and microplate assay. The method is based on the Griess reaction and the reaction product is extracted into nonionic surfactant Triton-X114 using CPE technique. In this study, decolorization treatment of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity of nitrite detection. Multi-sample can be simultaneously detected thanks to a 96-well microplate technique. The effects of different operating parameters such as type of decolorizing agent, concentration of surfactant (Triton X-114), addition of (NH4)2SO4, extraction temperature and time, interfering elements were studied and optimum conditions were obtained. Under the optimum conditions, a linear calibration graph was obtained in the range of 10-400 ng mL(-1) of nitrite with limit of detection (LOD) of 2.5 ng mL(-1). The relative standard deviation (RSD) for determination of 100 ng mL(-1) of nitrite was 2.80%. The proposed method was successfully applied for the determination of nitrite in the urine and blood samples with recoveries of 92.6-101.2%.


Isolation and chemical identification of inhibitors of plasma ligand binding.
The binding by serum albumin of many drugs and endogenous metabolites is impaired in humans and animals with renal failure. Unknown solute(s) retained in renal failure have been extracted from uremic fluids. When added to normal plasma they induce a similar binding defect. Similar activity can be extracted from normal urine. We have devised a series of extraction and purification techniques that yielded three binding inhibitory ligands from normal human urine in sufficient quantity and of a high degree of purity. Rigorous methods have been applied to determine chemical identity of the ligands. Purification steps consisted of: adsorption at pH 3.0 to polystyrene-divinylbenzene resin (XAD-2); elution from the resin with methanol followed by drying and solution in dilute formic acid; passage through SP-Sephadex to remove cations, especially yellow-brown pigments; adsorption to the anion exchanger QAE-Sephadex, and separation into three zones of inhibitory activity with a formic acid gradient; purification to homogeneity with C-8 or C-18 silica reversed-phase chromatography. Using this isolation procedure, followed by mass spectroscopy and nuclear magnetic resonance spectroscopy, we have shown that the binding inhibitory activity is due not to one ligand, but to a family of aromatic acids. To date hippurate, beta-(m-hydroxyphenyl)-hydracrylate and p-hydroxyphenylacetate have been identified as binding inhibitors. Other active ligands remain to be identified.


Groupthink: one peril of group cohesiveness.
A group's aim is to make well-conceived, well-understood, well-accepted and realistic decisions to reach their agreed-upon goals. This aim applies equally to their own goals and those occasionally imposed by outsiders such as hospital administration, accreditation committees and the federal government. Effective groupwork requires group cohesion with its components of trust, risk taking, mutual support, and group esteem. With constant vigilance the group can maintain its positive dynamics, so that the unhealthy state of groupthink does not undermine its existence.


Design and validation of a novel assessment tool for laparoscopic suturing of the vaginal cuff during hysterectomy.
The aim of this study was to design and validate a new assessment tool for laparoscopic vaginal cuff suturing in a box trainer. A primary grading tool was constructed including a novel checklist component and a previously validated global rating scale. A four-step validation process was then undertaken. Face validity was determined by a survey of a panel of experts in the field of minimally invasive gynecologic surgery. Content validity was assessed via analysis of consistency and variability of the experts' ratings, and items were removed or rephrased according to the experts' comments. Overall, five novices and five expert laparoscopic surgeons were videotaped performing suture closure of a latex vaginal cuff model in a box trainer. The videotapes were reviewed by two raters. Discriminate validity, along with interrater and intrarater reliabilities, was assessed by analysis of the video ratings. The Simulation, Training, Research, and Technology Utilization System center at Brigham and Women's Hospital, a tertiary medical center in Boston, MA. The final assessment tool is presented. We have validated an assessment tool for vaginal cuff suturing in a box trainer.


Monotherapy and combination therapy with acitretin for mycosis fungoides: results of a retrospective, multicentre study.
Retinoids have long been used in the treatment of cutaneous T-cell lymphomas. However, data on acitretin use for mycosis fungoides (MF) are very limited. To evaluate treatment outcomes of acitretin in patients with MF attending three academic referral centres in different regions of Greece. Data on effectiveness, safety and drug survival of acitretin as monotherapy or as adjuvant regimen were collected in a multicentre, register-based, retrospective study. Overall, 128 patients (64.8% male; mean age at MF diagnosis 59.7 years) were included. Folliculotropic MF was present in 24 (18.8%) cases. Most patients (n = 118; 92.2%) had early-stage disease (≤IIA) at acitretin initiation. In all, 28 (21.9%) patients received acitretin monotherapy, while 100 (78.1%) subjects on acitretin concomitantly received phototherapy (n = 65; 50.8%) or topical steroids (n = 27; 21.1%). Acitretin was given as a first-line agent in 73 (57%) cases. A 77.3% overall response rate was noted: 44.5% and 32.8% for complete and partial responses, respectively. Acitretin was more effective as first-line than as a subsequent agent (P = 0.008). A trend towards better response was observed in the combination arm compared to patients receiving acitretin alone (P = 0.056). Median time to best response was 6.9 months (IQR 4.4-9.4); median duration of response was 23.7 months (IQR 11.9-35.4). Overall, the mean length of all treatment patterns was 569 days (SD 718.8). Therapy was discontinued in 5 (3.9%) cases due to drug intolerance. Adverse effects were recorded in 62 (48.4%) cases with dyslipidaemia (n = 31; 24.2%), xerosis (n = 24; 18.6%) and hair loss (n = 10; 7.8%) being the most commonly recorded. Acitretin, either alone or as adjuvant, showed a stable long-term effectiveness in this cohort, especially when used in the first-line setting. This RAR-selective agonist may serve as an attractive option for treatment of MF and should be further evaluated.


Single dose epidural hydromorphone in labour pain: maternal pharmacokinetics and neonatal exposure.
Epidural hydromorphone could be useful in obstetric analgesia as there is a need for a more water-soluble opioid than sufentanil or fentanyl with prolonged analgesic effect. To our knowledge, the pharmacokinetics of epidural hydromorphone has not been evaluated in parturients. In this pilot study, seven healthy parturients were given a single epidural dose of hydromorphone for labour pain. One parturient received 1.5 mg, two 0.75 mg and four 0.5 mg of hydromorphone hydrochloride. Dose was decreased due to nausea and pruritus. Hydromorphone's effect, adverse effects and plasma concentrations were evaluated. Neonatal drug exposure was evaluated by umbilical vein and artery opioid concentration at birth. Neonatal outcomes were assessed using Apgar and the Neurologic Adaptive Capacity Score (NACS). All patients received additional levobupivacaine doses on parturients' requests. The first dose was requested at a median of 163 min (range 19-303 min) after hydromorphone administration. A total of 12 opioid related expected adverse events were reported by seven parturients. All newborn outcomes were uneventful. Hydromorphone's distribution and elimination after single epidural dose seem similar to that reported for non-pregnant subjects after intravenous hydromorphone administration, but further research is required to confirm this observation. The optimal dose of hydromorphone in labour pain warrants further evaluation.


Kidney development and perspectives for organ engineering.
Organ transplantation is currently the best strategy for treating end stage renal disease (ESRD) but the numbers of donor kidneys available are not sufficient to meet the needs of the ever-increasing ESRD population. Therefore, developments in the field of tissue engineering are necessary to provide alternative treatments. Decellularization and three-dimensional (3D) bioprinting strategies may serve as attractive novel options. Since successful tissue engineering requires an in -depth understanding of organ development and regulatory pathways, we discuss signaling in renal development and the composition of the renal extracellular matrix before presenting progress in the decellularization and 3D bioprinting fields.


Recombinant production of anti-HIV protein, griffithsin, by auto-induction in a fermentor culture.
Griffithsin (GRFT) is a novel anti-HIV protein isolated from the red alga Griffithia sp. The potent anti-viral activity of GRFT against both laboratory and primary isolates of HIV at picomolar concentrations makes this protein an attractive candidate microbicide to prevent sexual transmission of HIV. Here, we describe the recombinant production and purification of a biologically active hexa-histidine-tagged GRFT (His-GRFT) from Escherichia coli. To facilitate a large-scale production of recombinant His-GRFT, we tested different expression conditions to optimize the expression in the cytoplasm of E. coli to increase the overall production of soluble His-GRFT. Attempts to express His-GRFT in shake flask cultures resulted in a modest yield of soluble His-GRFT, with a large accumulation of the protein in inclusion bodies. The use of a fermenter and of a rich, auto-inducing medium allowed the total amount of His-GRFT per liter to be increased by about 45-fold, with approximately 70% of the protein expressed in the soluble fraction. N-terminal sequencing and MALDI-TOF analyses of the recombinant His-GRFT confirmed that the initial methionine residue was cleaved off. Recombinant His-GRFT showed equivalent activity with natural GRFT, both in respect to gp120-binding characteristics as well as anti-HIV activity. Size-exclusion chromatography analysis showed that both native GRFT and recombinant His-GRFT existed as homodimers in solution. The expression system described in this work provides a basis for the mass production of GRFT to allow further studies of the protein and investigation of therapeutic and preventive strategies against HIV.


A novel Abeta isoform pattern in CSF reflects gamma-secretase inhibition in Alzheimer disease.
LY450139 (semagacestat) inhibits gamma-secretase, a key enzyme for generation of amyloid beta (Abeta), the peptide deposited in plaques in Alzheimer disease (AD). Previous data have shown that LY450139 lowers plasma Abeta, but has no clear effect on Abeta1-40 or Abeta1-42 levels in cerebrospinal fluid (CSF). By using targeted proteomics techniques, we recently identified several shorter Abeta isoforms, such as Abeta1-16, that in experimental settings increase during gamma-secretase inhibitor treatment, and thus may serve as sensitive biochemical indices of the treatment effect. Here, we test the hypothesis that these shorter Abeta isoforms may be biomarkers of gamma-secretase inhibitor treatment in clinical trials. In a phase II clinical trial, 35 individuals with mild to moderate AD were randomized to placebo (n = 10) or LY450139 (100 mg (n = 15) or 140 mg (n = 10)) and underwent lumbar puncture at baseline and after 14 weeks of treatment. The CSF Abeta isoform pattern was analyzed with immunoprecipitation combined with MALDI-TOF mass spectrometry. The CSF levels of Abeta1-14, Abeta1-15, and Abeta1-16 showed a dose-dependent increase by 57% and 74%, 21% and 35%, and 30% and 67%, respectively in the 100-mg and 140-mg treatment groups. Abeta1-40 and Abeta1-42 were unaffected by treatment. CSF Abeta1-14, Abeta1-15, and Abeta1-16 increase during gamma-secretase inhibitor treatment in AD, even at doses that do not affect Abeta1-42 or Abeta1-40, probably because of increased substrate availability of the C99 APP stub (APP beta-CTF) induced by gamma-secretase inhibition. These Abeta isoforms may be novel sensitive biomarkers to monitor the biochemical effect in clinical trials. Clinical Trials.gov NCT00244322.


Induction of the SOS response by bacteriophage lytic development in Salmonella enterica.
Infection of Salmonella enterica with lytic mutants of either P22 or SE1 bacteriophages triggers the expression of its DNA damage-inducible SOS response through a lexA-dependent pathway. This induction of the SOS system strictly requires the presence of the bacteriophage kil gene. Accordingly, plasmid overexpression of the kil gene also promotes the S. enterica SOS network induction. Furthermore, S. enterica Gifsy prophages are induced following the infection with SE1 and P22 lytic derivatives. The observed data reveal a hitherto unknown SOS system-mediated fail-safe mechanism of resident prophages against infection with heteroimmune lytic bacteriophages and suggest a novel role for the kil family of proteins.