[Study on Extraction Conditions of Water-Soluble Substances—Purity Test (4) for Polyvinylpolypyrrolidone Listed in Japan's Specifications and Standards for Food Additives].
The food additive polyvinylpolypyrrolidone is approved for use as a filter aid. The water-soluble substances test of polyvinylpolypyrrolidone often shows poor reproducibility. The instruction "boil gently while stirring using a stirrer" was considered critical, and so this issue was examined. The results showed that the use of a combination of both an oil bath and a stirrer provided good reproducibility without decomposition or other problems.


The independent effect of age of general practitioner on clinical practice.
To establish the extent to which general practitioner age alone explains variations in patient morbidity and treatment patterns. An on-going, national survey of general practice activity in Australia. A random sample of 5013 GPs with a minimum of 375 general practice Medicare items claimed in the previous 3 months. Each GP contributed details of 100 consecutive encounters, with about 1000 GPs sampled each year between 1998 and 2003. Effect of practitioner age on GP activity after removing the influence of measured confounding factors: doctor, patient and practice characteristics; number of problems; and morbidity managed at encounters. GP age played a significant role in practice style. In comparison with young GPs (< 35 years), older GPs provided more home visits (P < 0.001) and attendances at residential aged-care facilities (P = 0.044); were more likely to manage chronic problems (P < 0.001); had higher prescribing rates (P < 0.001), and lower rates of pathology ordering (P < 0.001) and non-pharmacological treatments (P < 0.001). Individual body system management rates also differed significantly between younger and older GPs. A GP's age plays a significant role in determining practice style. Our results have implications in terms of the ageing GP population and in the wider context of the ageing medical labour force.


Admission Lymphopenia is Associated With Discharge Disposition in Blunt Chest Wall Trauma Patients.
Lymphopenia contributes to the immune suppression observed in critical illness. However, its role in the immunologic response to trauma remains unclear. Herein, we assessed whether admission lymphopenia is associated with poor outcomes in patients with blunt chest wall trauma (BCWT). All adult patients with a Chest Abbreviated Injury Score (CAIS) ≥2 admitted to our Level I Trauma center between May 2009 and December 2018 were identified in our institution Trauma Registry. Patients with absolute lymphocyte counts (ALC) collected within 24 H of admission were included. Patients who died within 24 H of admission, had bowel perforation on admission, penetrating trauma, and burns were excluded. Demographics, injury characteristics, comorbidities, ALC, complications, and outcomes were collected. Lymphopenia was defined as an ALC ≤1000/µL. Association between lymphopenia and clinical outcomes of BCWT was assessed using multivariate analyses. P < 0.05 was considered significant. A total of 1394 patients were included; 69.7% were male; 44.3% were lymphopenic. On univariate analysis, lymphopenia was associated with longer in-hospital stay (11.6±10.2 versus 10.1±11.4, P = 0.009), in-hospital death (9.7% versus 5.8%, P = 0.006), and discharge to a healthcare facility (60.9% versus 46.4%, P < 0.001). Controlling for Injury Severity Score, age, gender, and comorbidities, the association between lymphopenia and discharge to another facility (SNF/rehabilitation facility/ACH) (OR = 1.380 [1.041-1.830], P = 0.025) remained significant. Lymphopenia on admission is associated with discharge requiring increased healthcare support. Routine lymphocyte count monitoring on admission may provide important prognostic information for BCWT patients.


CD44 and integrin matrix receptors participate in cartilage homeostasis.
Articular chondrocytes express the matrix receptors CD44 and integrins. Both of these receptors exhibit interactions with adjacent extracellular matrix macromolecules. In addition, both integrins and CD44 have the capacity for signal transduction as well as modulated interactions with the actin cytoskeleton. As such, both receptor families provide the chondrocytes a means to detect changes in matrix composition or to function as mechanotransducers. Disruption of CD44 or integrin-mediated cell-matrix interactions, either experimentally induced or when present in osteoarthritis, have profound effects on cartilage metabolism. Thus, CD44 and integrin receptors play a critical role in maintaining cartilage homeostasis.


On the biomechanics of human shoulder complex--I. Kinematics for determination of the shoulder complex sinus.
Effectiveness of the multi-segmented total-human-body models to predict realistically live human response depends heavily on the proper biomechanical description and simulation of the major articulating joints of the body. In these models, the most difficult and the least successful modelling of a joint has been the shoulder complex because of the lack of appropriate biomechanical data as well as the anatomical complexity of the region. This paper in Part I presents various aspects of a research program to collect three-dimensional kinematic data for the shoulder complex. A sonic digitizing technique which utilizes an overdeterminate number of sonic emitters on the moving body segment was used for the kinematics analysis. The numerical results are presented for three male subjects for their voluntary shoulder complex sinuses. The results are given in a locally-defined joint axis system as well as in the torso-fixed coordinate system in the form of globographic representation.


The ups and downs of general surgery resident experience in endocrine surgery: Analysis of 30 years of ACGME graduate case logs.
The aim of this study was to determine trends in the experience of general surgery residents with endocrine surgery cases. American Association of Endocrine Surgeons national general surgery case logs from 1989 through 2019 were reviewed. The numbers of individuals completing residency and the mean and median number of endocrine surgery cases by type and by level of operating resident surgeon were abstracted from annual data and analyzed. Descriptive statistics and linear regression analyses were performed modeling endocrine surgery cases over time and stratified by procedure type and resident level. The number of individuals completing general surgery residency each year increased from 981 to 1,219 (P < .001). The average total number of endocrine surgery cases performed increased from 17 to 33.2 (P < .001) but has declined since its peak at 36.9 in 2010 to 2011 (P = .014). Thyroid operations increased from 11.4 to 19.8 (P < .001) but peaked at 23.5 in 2010 to 2011 and have since declined (P < .001). Parathyroid operations more than doubled from 4.2 to 9.7 (P < .001). Adrenal operations increased from 1 to 2.2 (P < .001) and pancreatic endocrine operations increased from 0.2 to 1.5 (P < .001). Surgeon Chief endocrine surgery cases peaked at 14.4 in 2003 to 2004 but have since declined by 22.2% (P < .001). Surgeon Junior endocrine surgery cases increased overall (P < .001) but peaked at 22.8 in 2011 to 2012. There was increasing heterogeneity over time in trainee experience (P < .001). After having increased for 2 decades, the number of endocrine surgery cases performed by general surgery residents is currently in decline. Possible contributing factors include growth in the number of general surgery residents, variable and narrowed case mix, and encroachment by other learners.


Fitting Proportional Odds Model to Case-Control data with Incorporating Hardy-Weinberg Equilibrium.
Genetic association studies have been proved to be an efficient tool to reveal the aetiology of many human complex diseases and traits. When the phenotype is binary, the logistic regression model is commonly employed to evaluate the association strength of the genetic variants predispose to human diseases because the maximum likelihood estimator of the odds ratio based on case-control data is equivalent to that from the same model by taking the data as being arisen prospectively. This equivalence does not hold for the proportional odds model and using it to analyze the case-control data directly often results in a substantial bias. Through putting a parameter of the minor allele frequency in the modified likelihood function under the condition that the Hardy-Weinberg equilibrium law holds within controls, a consistent estimator is obtained. On the basis of it, we construct a score test statistic to test whether the genetic variant is associated with the diseases. Simulation studies show that the proposed estimator has smaller mean squared error than the existing methods when the genetic effect size is away from zero and the proposed test statistic has a good control of type I error rate and is more powerful than the existing procedures. Application to 45 single nucleotide polymorphisms located in the region of TRAF1-C5 genes for the association with four-level anticyclic citrullinated protein antibody from Genetic Analysis Workshop 16 further demonstrates its performance.


Clonal relatedness of carbapenem-resistant Acinetobacter baumannii: high prevalence of ST136pas in a burn center.
Carbapenem-resistant Acinetobacter baumannii (CRAB) is a global health crisis. This study aimed to determine the clonal relatedness of antibiotic-resistant A. baumannii isolates in hospitalized patients who suffered from burn wound infection. One hundred and six A. baumannii isolates from 562 patients with burn wound infections, were identified and examined for antimicrobial susceptibility. Detection and characterization of carbapenem-hydrolyzing class D OXA-type beta-lactamases (CHDLs) were performed by PCR assays. The clonal relatedness of A. baumannii isolates was determined by multilocus sequence typing (MLST) according to the Pasteur scheme, dual-sequence typing of blaOXA-51-like and ampC genes, and RAPD-PCR method. All isolates were carbapenem-resistant while susceptible to colistin, minocycline, doxycycline, and ampicillin-sulbactam. The intrinsic blaOXA-51-like was detected in all isolates, and blaOXA-23-like was identified in 92.5% of isolates. However, blaOXA-143-like and blaOXA-58-like genes were not detected among isolates. Four distinct blaOXA-51-like alleles were determined as follows: blaOXA-317 (67.0%), blaOXA-90 (9.4%), blaOXA-69 (17.0%), and blaOXA-64 (6.6%) and four ampC (blaADC) allele types including ampC-25 (6.6%), ampC-39 (9.4%), ampC-1 (17.0%), and blaADC-88 (67.0%) were identified. MLST (Pasteur scheme) analysis revealed four ST types including ST136 (singleton), ST1 (CC1), ST25 (CC25), and ST78 (singleton) in 71, 18, 7, and 10 of A. baumannii strains, respectively. Five RAPD clusters including A (1.9%), B (26.4%), C (57.5%), D (7.5%), and E (1.9%) were characterized and 5 (4.7%) strains were found to be singletons. The present study demonstrated that there was a high prevalence of blaOXA-23-like producing CRAB in the clinical setting. The majority of isolates belonged to ST136 (singleton). However, blaOXA-23-like producing multi-drug resistant international clones including ST1, and emerging lineages (e.g. ST25 and ST78) were also identified. Interestingly, in this study ST2 was not detected.


Transcutaneous ultrasound-guided pancreatic biopsy in cattle and its safety: a preliminary report.
This study describes a free-hand technique for percutaneous pancreatic biopsy in cattle with ultrasound-guidance using a 14G spinal biopsy needle. Its safety was evaluated based on 36 consecutive procedures. To assess the immediate effects of pancreatic biopsy, 31 cows were necropsied shortly after the procedure and examined. The remaining five cows were examined daily for eight days and then necropsied and examined. No life-threatening complications nor clinically detectable abnormalities were observed. Changes indicative of inflammation were not apparent in total and differential WBC counts or in total protein and fibrinogen concentrations. A small increase of amylase activity was detected in only one cow kept for the eight-day observation period. Serum lipase activity increased significantly on day four after biopsy. Urea nitrogen, creatinine, glucose and the activities of aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyltransferase remained within reference ranges. Changes in the peritoneum and pancreas observed at necropsy were negligible. We conclude that percutaneous ultrasound-guided pancreatic biopsy did not appear to influence the cow's condition adversely and the procedure provided an excellent method of obtaining a pancreatic specimen for histological examination. The procedure was considered safe, fast, cost-effective, and practical when performed properly. We believe that the technique can be used in cows with suspected pancreatic disease for making an ante mortem diagnosis.


A dietary-driven redox modulation of gut microbiome-host interactions: the rescue of epithelial barrier and mucus production during dysbiosis by dietary nitrate.
Dietary nitrate, from green-leafy vegetables, is reduced to nitrite in the oral cavity and to nitric oxide ((•)NO) in the stomach. Gut microbiota is raising a great deal of interest since it has been recognized to be crucial to maintain gastrointestinal (GI) and systemic welfare. Dysbiosis (altered gut flora) is associated with increased epithelial permeability, deficient mucus production and with the activation of inflammatory pathways. Here we investigate whether dietary nitrate rescues GI physiology during dysbiosis. Four groups of Wistar rats were maintained in the animal facilities for 7 days during which, in addition to food, they had access to 1) water, 2) an antibiotic cocktail (neomycin, bacitracin, imipenem), 3) antibiotic cocktail sodium nitrate 10mM, 4) sodium nitrate. Animals were weighted daily. At the end of 7(th) day they were anesthetized (isoflurane) and euthanized. Ceca were collected and weighted. The stomach was isolated and occludin, claudin-5, ZO-1, iNOS and myeloperoxidase (MPO) were analyzed by western blot and immunofluorescence (n=4, triplicate). Values are mean±SEM. Antibiotic exposure prevented weight gain in all animals when compared to controls. Nitrate prevented body weight loss under dysbiosis. Ceca from animals with altered gut flora contained more fecal material than the ones from animals treated with nitrate under the same circumstances (18.47±0.99g vs 14.7±71.12g, p<0.05). Dysbiosis reduced occludin expression but supplementation with nitrate recovered this protein to control levels (p<0.05). No significant differences were observed for ZO-1 or claudin-5. Nitrate prevented MPO (115.70±5.36 vs 61.00±19.00, p<0.05; % of control) and iNOS (279.8±88.1 vs 57.3±18.4, p<0.05, % of control) overexpression under dysbiosis suggesting the prevention of an over stimulated inflammatory status in the gut. This data suggests that dietary nitrate may ensure gastric epithelial integrity and mucus production during dysbiosis. Supported by PTDC/AGR-ALI/115744/2009.


The impact of intravenous iron supplementation in elderly patients undergoing major surgery.
Age and preoperative anaemia are risk factors for poor surgical outcome and blood transfusion. The aim of this study was to examine the effect of iron supplementation in iron-deficient (ID) elderly patients undergoing major surgery. In this single-centre observational study, patients ≥ 65 years undergoing major surgery were screened for anaemia and ID. Patients were assigned to the following groups: A- (no anaemia); A-,ID+,T+ (no anaemia, iron-deficient, intravenous iron supplementation); A+ (anaemia); and A+,ID+,T+ (anaemia, iron-deficient, intravenous iron supplementation). Of 4,381 patients screened at the anaemia walk-in clinic, 2,381 (54%) patients were ≥ 65 years old and 2,191 cases were included in analysis. The ID prevalence was 63% in patients with haemoglobin (Hb) < 8 g/dl, 47.2% in patients with Hb from 8.0 to 8.9 g/dl, and 44.3% in patients with Hb from 9 to 9.9 g/dl. In severely anaemic patients, an Hb increase of 0.6 (0.4; 1.2) and 1.2 (0.7; 1.6) g/dl was detected with iron supplementation 6-10 and > 10 days before surgery, respectively. Hb increased by 0 (-0.1; 0) g/dl with iron supplementation 1-5 days before surgery, 0.2 (-0.1; 0.5) g/dl with iron supplementation 6-10 days before surgery, and 0.2 (-0.2; 1.1) g/dl with supplementation > 10 days before surgery (p < 0.001 for 1-5 vs. 6-10 days). Overall, 58% of A+,ID+,T+ patients showed an Hb increase of > 0.5 g/dl. The number of transfused red blood cell units was significantly lower in patients supplemented with iron (0 (0; 3)) compared to non-treated anaemic patients (1 (0; 4)) (p = 0.03). Patients with iron supplementation > 6 days before surgery achieved mobility 2 days earlier than patients with iron supplementation < 6 days. Intravenous iron supplementation increases Hb level and thereby reduces blood transfusion rate in elderly surgical patients with ID anaemia.


Death in the paddy field: Carcass decomposition and associated arthropods in subunits of a rice field landscape.
Although rice production landscapes are often the scene of homicides, vertebrate decomposition and associated arthropods have never been described from rice paddies. Tropical rice landscapes are typically composed of irrigated/non-irrigated rice fields, fallow land (i.e., uncultivated fields), and low earthen levees (bunds) used as access pathways. The specific microclimatic and environmental conditions associated with each subunit of rice field landscapes are likely to impact carcass decomposition as well as the arthropod species associated with carrion. Here, we tested the hypothesis that the combined effects of constraints on arthropod colonization and survival, scarcity of necromass, limited habitat architecture, and recurrent disturbance limit the ability of carrion-related arthropods to colonize corpses and large carcasses in tropical rice-fields. Our results from monitoring pig carcasses in Philippine rice fields indicated that vertebrate decomposition in irrigated fields was slower and incomplete when compared to non-irrigated fields and bunds. Carcasses were colonized by a small complex of carrion-related arthropods that differed in composition and relative species abundance between dry bunds and relatively humid rice paddies. Fire ants (Solenopsis germinata) were observed frequently on carcasses exposed on bunds, rarely in non-irrigated fields, and almost never in irrigated fields. The presence of fire ants was associated with reduced blow fly (Chrysomya megacephala) abundance. Taken together, this indicates that the arthropod fauna associated with carcasses in tropical rice fields is relatively simple in contrast to the generally high arthropod diversity reported for other ecosystems at tropical latitudes. The limited richness of the community also means that an understanding of the development of one abundant calliphorid blow fly, C. megacephala, may be sufficient to investigate deaths and homicides under conditions similar to the ones described in this study.


Co-sintering MSWI fly ash with electrolytic manganese residue and coal fly ash for lightweight ceramisite.
The MSWI fly ash (FA) is classified as hazardous waste and electrolytic manganese residue (EMR) as the harmful industrial waste. FA, water-washed FA (WFA), EMR and coal fly ash (CFA) were co-recycled to form lightweight MFCE ceramisites. The effects of FA, WFA and mixed MSWI fly ash on ceramisites were discussed. The approach to mixing FA and WFA increased the recycling amount of MSWI fly ash. The optimal mixture of 34.5% EMR, 24.1% CFA, 20.7% FA and 20.7% WFA sintered at 1160 °C for 12 min with a procedural heating rate (10 °C/min) and belonged to Class 800 artificial lightweight aggregate (GB/T 17431.1-2010); the quantity of MSWI fly ash in ceramisite was as high as 41.4%. Volatilization rates of Cd, Pb, Cu, Zn, Mn and Cr for ceramisite were higher 75.0, 24.2, 62.7, 133, 343 and 764% than FA respectively, attributed to the co-existence of chlorides and sulfates. The remained Zn, Cu, Pb, Mn and Cr were exchanged with Mg2+/Ca2+/Al3+ of diopside and wollastonite to form residual fractions. Our findings provided a feasibility method of co-recycling MSWI fly ash and electrolytic manganese residue to produce green lightweight aggregates.


Apnea at discharge and gastro-esophageal reflux in the preterm infant.
To determine the relationship between respiratory patterns and acid gastro-esophageal reflux (g-e reflux) prior to discharge of the formerly preterm infant. Forty-five infants of <32 weeks' gestation were studied at an average postmenstrual age of 37.2 weeks (SD 3.5). Following informed parental consent, a 12-hour multichannel recording including esophageal pH was obtained. Apneas of greater than 10 seconds were recorded, as well as the occurrence of bradycardia or desaturation. Acid g-e reflux (pH <4.0) occurred at least once in all of the infants; prevalence was between <1% and 41% of the 12-hour record (median 4.6%, interquartile range 0.5% to 9%). The number of reflux episodes ranged from 1 to 143 (median 23). The number of apneas (>10 seconds duration) ranged from 0 to 71, median 6. There was no correlation between apnea frequency or severity and reflux frequency or duration. There was no difference in apnea frequencies between the 5 minutes after the start of a reflux episode and the 5 minutes prior to each episode. Acid g-e reflux in the formerly preterm infant at discharge is frequent and may be prolonged; there is no association between reflux and apnea.


Coupling and dynamics of subunits in the hexameric AAA+ chaperone ClpB.
The bacterial AAA+ protein ClpB and its eukaryotic homologue Hsp104 ensure thermotolerance of their respective organisms by reactivating aggregated proteins in cooperation with the Hsp70/Hsp40 chaperone system. Like many members of the AAA+ superfamily, the ClpB protomers form ringlike homohexameric complexes. The mechanical energy necessary to disentangle protein aggregates is provided by ATP hydrolysis at the two nucleotide-binding domains of each monomer. Previous studies on ClpB and Hsp104 show a complex interplay of domains and subunits resulting in homotypic and heterotypic cooperativity. Using mutations in the Walker A and Walker B nucleotide-binding motifs in combination with mixing experiments we investigated the degree of inter-subunit coupling with respect to different aspects of the ClpB working cycle. We find that subunits are tightly coupled with regard to ATPase and chaperone activity, but no coupling can be observed for ADP binding. Comparison of the data with statistical calculations suggests that for double Walker mutants, approximately two in six subunits are sufficient to abolish chaperone and ATPase activity completely. In further experiments, we determined the dynamics of subunit reshuffling. Our results show that ClpB forms a very dynamic complex, reshuffling subunits on a timescale comparable to steady-state ATP hydrolysis. We propose that this could be a protection mechanism to prevent very stable aggregates from becoming suicide inhibitors for ClpB.


Discordant uptake of technetium-99m DISIDA and technetium-99m sulfur colloid in an amebic abscess.
Hepatobiliary imaging in a patient with an amebic abscess showed an early cold defect that later showed rim enhancement. A Tc-99m SC scan did not show prominent flow (making hepatoma unlikely) and showed the previously noted defect to appear larger and without rim enhancement. The differential damage to Kupffer cells and hepatocytes or edema may account for these findings. Amebic abscess should be included in the differential diagnosis of lesions that give increased Tc-99m IDA but cold Tc-99m SC images.


Evaluation of the adequacy of Hadlock's reference chart for identification of fetuses with growth restriction.
This study aimed to verify whether Hadlock's reference values for fetal weight identify fetuses below the 10th percentile in our population correctly. The fitness of the Hadlock reference range to our study population was tested by assessment of Z scores. We evaluated differences between the reference weight ranges proposed in our study and those recommended by Hadlock. Z scores for Hadlock reference values were non-normally distributed. The difference between the 50th percentile fetal weight proposed by our study model and that proposed by Hadlock was ≤1% at GAs ≥22 weeks and 2-3% at 19-21 weeks. For the 90th percentile level, the maximum difference at GAs ≥17 weeks was 1.5%. For the 10th percentile level, the differences were 2-4% in the third trimester, reaching 8% in week 20 and 13% at a GA of 14 weeks. The weight reference ranges of this study virtually overlap with the Hadlock ranges. We believe that only at lower gestational ages in the second trimester might some FGR diagnoses be missed in the population study with Hadlock's reference.


Incidental Findings on Pediatric Abdominal Computed Tomography At A Pediatric Trauma Center.
The increasing availability and use of computed tomography (CT) in pediatric abdominal trauma has increased the detection of incidental findings. While some of these findings are benign, others may require further evaluation for possible clinical importance. This study aimed to identify the frequency and type of incidental findings and their need for follow-up on abdominal CT in patients at a pediatric trauma center. This was a retrospective, observational study on trauma patients ≤21 years of age who presented to the emergency department between January 1, 2004 and July 31, 2016 and underwent CT scans of the abdomen and pelvis. Findings were classified as benign anatomic variants, benign pathologic lesions, and pathologic lesions requiring additional work-up. There were 1073 patients included in the study population, with a mean age of 15.5 years; 707 (66%) were males. A total of 418 incidental findings were identified in 345 patients. Of these, 290 (69%) were benign and 60 (14%) were likely benign pathologic that required possible outpatient monitoring. Of those requiring additional evaluation, 5 (1%) patients warranted further evaluation before discharge. Nearly one-third of patients had at least one radiographic finding unrelated to their injury. Of these, more than two-thirds did not require additional evaluation, but nearly one-third of patients required some form of further work-up.


Early impact of COVID-19 pandemic on paediatric surgical practice in Nigeria: a national survey of paediatric surgeons.
The novel coronavirus disease has had significant impact on healthcare globally. Knowledge of this virus is evolving, definitive care is not yet known and mortality is increasing. We assessed its initial impact on paediatric surgical practice in Nigeria, creating a benchmark for recommendations and future reference. Survey of 120 paediatric surgeons from 50 centres to assess sociodemographics and specific domains of impact of COVID-19 on their services and training in Nigeria. Valid responses were represented as categorical data and presented in percentages. Duplicate submissions for centres were excluded by combining and taking the mean of responses from centres with multiple respondents. Response rate was 74 (61%). Forty-six (92%) centres had suspended elective surgeries. All centres continued emergency surgeries but volume reduced in March by 31%. Eleven (22%) centres reported 13 suspended elective cases presenting as emergencies in March, accounting for 3% of total emergency surgeries. Twelve (24%) centres adopted new modalities for managing selected surgical conditions: non-operative reduction of intussusception in 1 (2%), antibiotic management of uncomplicated acute appendicitis in 5 (10%) and more conservative management of trauma and replacement of laparoscopic appendectomy with open surgery in 3 (6%), respectively. Low perception of adequacy of personal protective equipment (PPE) was reported in 35 (70%) centres. Forty (80%) centres did not offer telemedicine for patients' follow-up. Twenty-nine (58%) centres had suspended academic training. Perception of safety to operate was low in 37 (50%) respondents, indifferent in 24% and high in 26%. Majority of paediatric surgical centres reported cessation of elective surgeries while continuing emergencies. There was, however, an acute decline in the volume of emergency surgeries. Adequate PPE needs to be provided and preparations towards handling backlog of elective surgeries once the pandemic recedes. Further study is planned to more conclusively understand the full impact of this pandemic on children's surgery.


Controlled Switching of the Number of Skyrmions in a Magnetic Nanodot by Electric Fields.
Magnetic skyrmions are topological swirling spin configurations that hold promise for building future magnetic memories and logic circuits. Skyrmionic devices typically rely on the electrical manipulation of a single skyrmion, but controllably manipulating a group of skyrmions can lead to more compact and memory-efficient devices. Here, an electric-field-driven cascading transition of skyrmion clusters in a nanostructured ferromagnetic/ferroelectric multiferroic heterostructure is reported, which allows a continuous multilevel transition of the number of skyrmions in a one-by-one manner. Most notably, the transition is non-volatile and reversible, which is crucial for multi-bit memory applications. Combined experiments and theoretical simulations reveal that the switching of skyrmion clusters is induced by the strain-mediated modification of both the interfacial Dzyaloshinskii-Moriya interaction and effective uniaxial anisotropy. The results not only open up a new direction for constructing low-power-consuming, non-volatile, and multi-bit skyrmionic devices, but also offer valuable insights into the fundamental physics underlying the voltage manipulation of skyrmion clusters.


The role of cavitation in acoustically activated drug delivery.
Pluronic P105 micelles are potential candidates as chemotherapy drug delivery vehicles using ultrasonic stimulation as a release trigger. Acoustic power has been previously shown to release two anthracycline agents from these polymeric carriers. In this study, an ultrasonic exposure chamber with fluorescence detection was used to examine the mechanism of doxorubicin release from P105 micelles. Acoustic spectra were collected and analyzed, at the same spatial position as fluorescence data, to probe the role of cavitation in drug release. Our study showed a strong correlation between percent drug release and subharmonic acoustic emissions, and we attribute the drug release to collapse cavitation that perturbs the structure of the micelle and releases drug.


Closure of the internal opening for treatment of complex fistula-in-ano.
The surgical management of complex fistula-in-ano can be difficult, and often requires a seton suture or a colostomy. An alternative procedure is presented, based on principles that have been used successfully in the treatment of rectovaginal fistulas. The essential components of the technique are closure of the internal opening (when possible by an endorectal advancement flap), wide external drainage, and curettage of the fistulous tract. Seven patients with complex fistulas-in-ano were treated by this method. Six were cured and had excellent functional results up to 32 +/- 36 months postoperatively. The time to complete healing was 2.1 +/- 0.75 months. One patient developed a recurrent abscess. The procedure described is an alternative to conventional methods used in the treatment of complex fistula-in-ano.


Training in oral and maxillofacial surgery: a medicine-first perspective.
Specialist registration in oral and maxillofacial surgery (OMFS) requires dual medical and dental qualification involving at least eight years of undergraduate study. Training has continued to evolve since dual qualification was introduced and has often resulted in unwarranted repetition. If a time-based curriculum is necessary, second degree trainees should be allowed to pursue research and audit, and gain relevant clinical experience in lieu of repeating previously covered material. Junior surgical training could be integrated into the second degree. A programme that records competencies during the second degree may demonstrate equivalent to other aspects of junior training. One barrier is timetabling, which often restricts the integration of second degree trainees with OMFS units. Junior training in OMFS could be streamlined if the content was agreed nationally. This would also offer the opportunity for those key institutions that implement these changes to take on a prominent role in OMFS training.


Are anticholinergic effects responsible for the heterogeneous quinidine-induced modifications of heart muscle refractoriness?
Vagal tone is responsible for the heterogeneous reactivity of atrial and ventricular contractile tissues to quinidine. Acetylcholine may make atrial cells more sensitive to the effects of quinidine.


Association between anemia and bronchopulmonary dysplasia in preterm infants.
Anemia is commonly seen in preterm infants. It may reduce the capacity of hemoglobin to transport oxygen throughout the body and may result in tissue and organ dysfunction. This study aimed to investigate the effect of anemia on the development of bronchopulmonary dysplasia (BPD) in preterm infants. 243 infants who were admitted to BaYi Children's Hospital Affiliated to Clinical Medical College in Beijing Military General Hospital with gestational age (GA) less than 32 weeks from February, 2014 to February, 2015 were included in the study. Maternal and infant data were recorded. Multivarariate logistic regression analysis was performed to determine the association between anemia and BPD. Of 243 preterm infants, the incidence of anemia was higher in BPD patients than non-BPD patients (p < 0.001). Mean Hct in BPD patients was lower than non-BPD patients at different time points in 1d, 7d, 14d, and 21d. Controlling for other confounding factors, early anemia was associated with an increased risk of BPD. Number of transfusions is also a significant risk factor for BPD (p = 0.001). Therefore, prevention and treatment of early anemia is necessary and reducing number of transfusions may reduce the incidence of BPD in preterm infants.


Novel pathogenetic mechanisms and structural adaptations in ischemic mitral regurgitation.
Ischemic mitral regurgitation (MR) is a common complication of myocardial infarction thought to result from leaflet tethering caused by displacement of the papillary muscles that occurs as the left ventricle remodels. The author explores the possibility that left atrial remodeling may also play a role in the pathogenesis of ischemic MR, through a novel mechanism: atriogenic leaflet tethering. When ischemic MR is hemodynamically significant, the left ventricle compensates by dilating to preserve forward output using the Starling mechanism. Left ventricular dilatation, however, worsens MR by increasing the mitral valve regurgitant orifice, leading to a vicious cycle in which MR begets more MR. The author proposes that several structural adaptations play a role in reducing ischemic MR. In contrast to the compensatory effects of left ventricular enlargement, these may reduce, rather than increase, its severity. The suggested adaptations involve the mitral valve leaflets, the papillary muscles, the mitral annulus, and the left ventricular false tendons. This review describes the potential role each may play in reducing ischemic MR. Therapies that exploit these adaptations are also discussed.


SAPrIm, a semi-automated protocol for mid-throughput immunopeptidomics.
Human leukocyte antigen (HLA) molecules play a crucial role in directing adaptive immune responses based on the nature of their peptide ligands, collectively coined the immunopeptidome. As such, the study of HLA molecules has been of major interest in the development of cancer immunotherapies such as vaccines and T-cell therapies. Hence, a comprehensive understanding and profiling of the immunopeptidome is required to foster the growth of these personalised solutions. We herein describe SAPrIm, an Immunopeptidomics tool for the Mid-Throughput era. This is a semi-automated workflow involving the KingFisher platform to isolate immunopeptidomes using anti-HLA antibodies coupled to a hyper-porous magnetic protein A microbead, a variable window data independent acquisition (DIA) method and the ability to run up to 12 samples in parallel. Using this workflow, we were able to concordantly identify and quantify ~400 - 13000 unique peptides from 5e5 - 5e7 cells, respectively. Overall, we propose that the application of this workflow will be crucial for the future of immunopeptidome profiling, especially for mid-size cohorts and comparative immunopeptidomics studies.


A quantitative method for evaluating local perfusion using indocyanine green fluorescence imaging.
The purpose of this study is to introduce a new method, indocyanine green fluorescence imaging (ICG-FI), as an adjunct to distal pressure measurements in patients with peripheral arterial disease and symptomatic lower limb ischemia. A total of 34 patients with peripheral arterial disease, including 11 with claudication (Fontaine II), 7 with rest pain (FIII), and 16 with an ulcer or gangrene (FIV), were enrolled. After an intravenous injection of ICG (0.1 mg/kg), foot perfusion was recorded by an infrared light camera. Fluorescence intensity was plotted on a time-intensity curve using recorded images, allowing the calculation of new parameters. Severity of ischemia was assessed as the duration between the rising point and half value of maximum brightness (T½). The difference in the fluorescence intensity between 10 seconds after the rising point and baseline (PDE10) was compared with the transcutaneous oxygen pressure (tcPO2) at the same site (n=51). Median T½ was 23 seconds in FII, 41 seconds in FIII (P<0.05), and 17 seconds in FIV patients. PDE10 correlated moderately with tcpO2 (r2=0.5). A cut-off value (PDE10=28) predicted a critically ischemic limb (FIII and FIV), defined as tcpO2<30 mm Hg with a sensitivity of 100% and specificity of 86.6%. Local tissue perfusion can be quantitatively evaluated by using ICG fluorescence imaging. It is a safe, fast, noncontact method of imaging, which may be useful even at the ulcer itself and in the circumferential area.


Trypanosomatid infections in captive wild mammals and potential vectors at the Brasilia Zoo, Federal District, Brazil.
Conservation projects in zoos may involve translocation of captive animals, which may lead to pathogen spread. Neotropical mammals are important hosts of Trypanosoma cruzi and Leishmania spp. the etiological agents of Chagas disease and Leishmaniasis respectively. Studies of trypanosomatid-infected mammals and vectors (triatomines and sandflies) in zoos are important for the establishment of surveillance and control measures. We investigated trypanosomatid infections in captive wild mammals, triatomines and sandflies at the Brasília Zoo. We collected triatomines during active bimonthly surveys, sampled sandflies using light-traps and obtained blood samples from 74 mammals between 2016 and 2017. We used quantitative PCR to detect trypanosomatids in vectors and mammals. We found a colony of 19 Panstrongylus megistus in the porcupine unit and detected T. cruzi infections in five bugs. We captured 17 sandflies of four species including Nyssomyia whitmani and Lutzomyia longipalpis, but no Leishmania infection was detected. qPCR detected 50 T. cruzi-infected mammals belonging to 24 species and five groups of mammals (Carnivora, Cetartiodactyla, Perissodactyla, Pilosa and Primates); Leishmania DNA was detected in 23 mammals from 15 species, mainly carnivores. We detected trypanosomatid infections in 11 mammals born at the Brasília Zoo. Our results suggest vector-borne transmission of T. cruzi among maned wolves; measures to reduce the risk of new infections should therefore be taken. We also report sandfly presence and Leishmania-infected mammals at the Brasília Zoo. Translocation of wild mammals in and out of the Brasília Zoo should consider the risk of T. cruzi and Leishmania spread.


Ecotoxicology of mercury concentrations in arctic lamprey (Lethenteron camtschaticum).
Arctic lamprey (Lethenteron camtschaticum) is an important dietary resource for rural and indigenous communities in parts of Alaska, with some commercial use. As with many fish species harvested for human consumption, there are concerns regarding mercury concentrations ([Hg]) in Arctic lamprey that may impact human health. To date, information regarding the life cycle and diet of Arctic lamprey is scarce, with no published studies examining [Hg] in Arctic lamprey tissues. Our goals were to investigate the feeding ecology of Arctic lamprey from the Bering Sea, determine how diet and potential dietary shifts might influence [Hg] in muscle, and determine if current [Hg] may pose a human health risk. The mean total [Hg] in Arctic lamprey muscle (n = 98) was 19 ng/g wet-weight. Log transformed total [Hg] were not associated with any measured biological variables including length, mass, δ13C values, or δ15N values. A stable isotope mixing model estimated that capelin (Mallotus villosus) accounted for 40.0 ± 4.0% of the Arctic lamprey diet, while Pacific sand lance (Ammodytes hexapterus) and Pacific herring (Clupea pallasii) accounted for 37.8 ± 3.1% and 22.2 ± 3.5% respectively. Finally, diet percentage compositions shifted based on size class (i.e., medium versus large). These results indicated that feeding location, bioaccumulation, and biomagnification are not important drivers of [Hg] in Arctic lamprey and current [Hg] do not pose a human health risk. Taken together, this research further expands our knowledge of Arctic lamprey trophic ecology in the eastern Bering Sea.


Equally elevated concentrations of exhaled nitric oxide in nonatopic and low-sensitized atopic asthmatics.
Some studies show concentrations of exhaled nitric oxide (FENO) in nonatopic asthma and in healthy subjects to be similar, but include asthmatics on inhaled steroids, which is likely to interfere with the results. Comparison of FENO between nonatopic asthmatics, low-sensitized and high-sensitized atopic asthmatics, and healthy controls. We studied 85 non-smoking, steroid-naive young men with recently diagnosed symptomatic asthma and 10 healthy controls. FENO was measured according to European Respiratory Society Guidelines. In skin prick tests of 13 common aeroallergens, subjects with a total sum of prick wheals 3-10mm were regarded as low-sensitized and those with >10mm, as high-sensitized. Flow-volume spirometry, standardized histamine challenge, and an exercise test were also carried out. Prick tests revealed 14 subjects to be nonatopic and 71 atopic. In high-sensitized subjects with atopic asthma, the FENO median (25-75 quartiles) was significantly higher, 34.9 (21.3-53.8) parts per billion (ppb), than in subjects with nonatopic asthma, 15.2 (9.7-24.7)ppb (p<0.001), both being significantly higher than in healthy controls, 6.6 (5.2-8.5)ppb (p<0.001). FENO levels were similar in nonatopic and in low-sensitized atopic asthmatics, with no difference between them in bronchial responsiveness to histamine and exercise. Among steroid-naive young male asthmatics, FENO was equally elevated in nonatopic asthma and in low-sensitized atopic asthma but lower than in those with high-sensitized atopic asthma. These differences in FENO between asthma groups parallel the differences in airway function disturbance in terms of responsiveness to histamine or exercise.


Differential binding properties of B7-H1 and B7-DC to programmed death-1.
Programmed death-1 (PD-1) is a negative regulatory receptor expressed on activated T and B cells. Two ligands for PD-1, B7-H1 (PD-L1) and B7-DC (PD-L2), have been identified, but their binding properties have not been characterized yet. In this study, we generated soluble Ig fusion proteins of these molecules and examined the kinetics and relative affinities of the interactions between B7-H1 or B7-DC and PD-1 by flow cytometry and surface plasmon resonance. The interaction of B7-DC/PD-1 exhibited a 2-6-fold higher affinity and had different association/dissociation kinetics compared with the interaction of B7-H1/PD-1. Our results suggest that the differential binding properties of B7-H1 and B7-DC may be responsible for differential contributions of these two PD-1 ligands to immune responses.


Combination of two fiber-mutant adenovirus vectors, one encoding the chemokine FKN and another encoding cytokine interleukin 12, elicits notably enhanced anti-tumor responses.
For achieving optimal cancer immunotherapy, it is anticipated that both the activation and infiltration of immune cells into tumor are indispensable. In the present study, fiber-mutant adenovirus vectors (Ad) encoding chemokine FKN, (AdRGD-FKN), and cytokine interleukin 12, (AdRGD-IL-12), were constructed. The in vivo gene expression of AdRGD was confirmed and the combination of both FKN and IL-12 encoding Ad elicited synergistic anti-tumor activity in ovarian carcinoma, which induced tumor regression in all tumor-bearing mice, while using FKN alone did not show notable tumor-suppressive effect. The treatment with both IL-12 and FKN induced long-term specific immunity against OV-HM tumors in tumor-rejected mice. The results of immunohistochemical staining for CD3+ and perforin-positive cells suggested that the failure of using FKN alone was because of the inactivation of infiltrated immune cells. In contrast, cotransduction with IL-12 and FKN could induce more activated tumor-infiltrating immune cells than that transducted with FKN or IL-12 alone. The results indicated that using both chemokine and cytokine might be a powerful tool and a promising way for effective cancer immunotherapy.


Beta emitters rhenium-188 and lutetium-177 are equally effective in radioimmunotherapy of HPV-positive experimental cervical cancer.
Cervical cancer caused by the infection with the human papillomavirus (HPV) remains the fourth leading killer of women worldwide. Therefore, more efficacious treatments are needed. We are developing radioimmunotherapy (RIT) of HPV-positive cervical cancers by targeting E6 and E7 viral oncoproteins expressed by the cancer cells with the radiolabeled monoclonal antibodies (mAbs). To investigate the influence of different radionuclides on the RIT efficacy-we performed RIT of experimental cervical cancer with Rhenium-188 ((188) Re) and Lutetium-177 ((177) Lu)-labeled mAb C1P5 to E6. The biodistribution of (188) Re- and (177) Lu-labeled C1P5 was performed in nude female mice bearing CasKi cervical cancer xenografts and the radiation dosimetry calculations for the tumors and organs were carried out. For RIT the mice were treated with 7.4 MBq of either (188) Re-C1P5 or (177) Lu-C1P5 or left untreated, and observed for their tumor size for 28 days. The levels of (188) Re- and (177) Lu-C1P5 mAbs-induced double-strand breaks in CasKi tumors were compared on days 5 and 10 post treatment by staining with anti-gamma H2AX antibody. The radiation doses to the heart and lungs were similar for both (177) Lu-C1P5 and (188) Re-C1P5. The dose to the liver was five times higher for (177) Lu-C1P5. The doses to the tumor were 259 and 181 cGy for (177) Lu-C1P5 and (188) Re-C1P5, respectively. RIT with either (177) Lu-C1P5 or (188) Re-C1P5 was equally effective in inhibiting tumor growth when each was compared to the untreated controls (P = 0.001). On day 5 there was a pronounced staining for gamma H2AX foci in (177) Lu-C1P5 group only and on day 10 it was observed in both (177) Lu-C1P5 and (188) Re-C1P5 groups. (188) Re- and (177) Lu-labeled mAbs were equally effective in arresting the growth of CasKi cervical tumors. Thus, both of these radionuclides are candidates for the clinical trials of this approach in patients with advanced, recurrent or metastatic cervical cancer.


Modes of onset of epilepsy and differential diagnosis.
Epilepsy has protean manifestations and may be diagnosed when two unprovoked seizures have occurred. This determination is nearly always based on the available history because most seizures have stopped long before patients arrive at medical care. Great care must be taken, even by experts, to correctly interpret the history and there is strong evidence that incorrect diagnoses are frequent. An abnormal EEG cannot rule epilepsy in or out unless an actual seizure is recorded. When the diagnosis of epilepsy is based only on two seizures, the seizures are usually generalized tonic-clonic. Less "severe" seizure types usually occur multiple times before prompting a medical visit. Some patients present with what seems to be a first generalized tonic-clonic seizure but have a history of less severe attacks that have not brought them to medical attention - epilepsy can then be diagnosed. Others present with staring spells, episodes of confusion, body jerks, spasms, drops, loss of speech and social interactions and/or cognitive function, paroxysmal events during sleep, and febrile seizures. This chapter examines and considers the differential diagnoses for each of these modes of presentation. The consequences of a missed alternate diagnosis, such as cardiac arrhythmia, may be profound.


Morphological and functional types of tricuspid regurgitation: prognostic value in patients undergoing tricuspid annuloplasty during left-sided valvular surgery.
The nonuniform benefit of tricuspid annuloplasty may be explained by the proportionality of tricuspid regurgitation (TR) severity to right ventricular (RV) area. The purpose of this study was to delineate distinct morphological phenotypes of functional TR and investigate their prognostic implications in patients undergoing tricuspid annuloplasty during left-sided valvular surgery. The ratios of pre-procedural effective regurgitant orifice area (EROA) with right ventricular end-diastolic area (RVDA) were retrospectively assessed in 290 patients undergoing tricuspid annuloplasty. Based on optimal thresholds derived from penalized splines and maximally selected rank statistics, patients were stratified into proportionate (EROA/RVDA ratio ≤ 1.74) and disproportionate TR (EROA/RVDA ratio > 1.74). Overall, 59 (20%) and 231 (80%) patients had proportionate and disproportionate TR, respectively. Compared to those with proportionate TR, patients with disproportionate TR were older, had a higher prevalence of atrial fibrillation, lower pulmonary pressures, more impaired RV function, and larger tricuspid leaflet tenting area. Over a median follow-up of 4.1 years, 79 adverse events (47 heart failure hospitalizations and 32 deaths) occurred. Patients with disproportionate TR had higher rates of adverse events than those with proportionate TR (32% vs 10%; P = 0.001) and were independently associated with poor outcomes on multivariate analysis. TR proportionality outperformed guideline-based classification of TR severity in outcome prediction and provided incremental prognostic value to both the EuroSCORE II and STS score (incremental χ2 = 6.757 and 9.094 respectively; both P < 0.05). Disproportionate TR is strongly associated with adverse prognosis and may aid patient selection and risk stratification for tricuspid annuloplasty with left-sided valvular surgery.


A brush-like Cu2O-CoO core-shell nanoarray: an efficient bifunctional electrocatalyst for overall seawater splitting.
Herein, a brush-like Cu2O-CoO core-shell nanoarray on copper foam (Cu2O-CoO/CF) can achieve efficient oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) performance in alkaline seawater electrolyte. This Cu2O-CoO/CF shows overpotentials as low as 315 and 295 mV at 100 mA cm-2 for the OER and HER, respectively. Moreover, it could also be operated at 1.82 V with 100 mA cm-2 in a two-electrode electrolyzer and exhibits strong stability for at least 50 hours of electrolysis. The excellent performance and hierarchical structure advantages of Cu2O-CoO/CF provide new ideas for designing efficient seawater splitting electrocatalysts.


Fiber-integrated WGM optofluidic chip enhanced by microwave photonic analyzer for cardiac biomarker detection with ultra-high resolution.
Cardiac troponin I (cTnI) plays an important role in emergency diagnosis of cardiovascular diseases, which exists predominately in the form of cardiac troponin I-C (cTnI-C) complex. We proposed a fiber-integrated optofluidic chip immunosensor with time-delay-dispersion based microwave photonic analyzer (MPA) for cTnI-C detection. The whispering gallery mode (WGM) fiber probe was fabricated by embedding a polydopamine functionalized hollow glass microsphere (HGMS) into the etched capillary-fiber structure, and the WGMs could be excited through the efficient coupling between the thin-wall capillary and the HGMS. The reflective WGM optofluidic chip functioned as a wavelength tuner to construct fiber ring laser cavity, whose laser output wavelength was cTnI-C concentration-dependent. The tiny wavelength variation of sensing laser was converted into a radio-frequency (RF) response, which was retrieved by measuring the change of RF-domain free spectrum range (FSR) in time-delay-dispersion based MPA, and the quantitative detection of cTnI-C complex can be achieved with high resolution. Experimental results show that this immunosensor had a limit of detection (LOD) of 0.59 ng/mL, and a detection resolution of 1.2 fg/mL. The relative resolving power was 102-104-fold higher than that of others optical fiber cTnI biosensors. The proposed fiber-integrated optofluidic chip provides an innovative lab-on-chip diagnostic tool for myocardial damage.


Refraction-Assisted Solar Thermoelectric Generator based on Phase-Change Lens.
Solar thermoelectric generators (STEGs), which are used for various applications, (particularly small size electronic devices), have optical concentration systems for high energy conversion efficiency. In this study, a refraction-assisted STEG (R-STEG) is designed based on phase-change materials. As the phase-change material (PCM) changes phase from solid to liquid, its refractive index and transmittance also change, resulting in changes in the refraction of the sunlight transmitted through it, and concentration of solar energy in the phase-change lens. This innovative design facilitates double focusing the solar energy through the optical lens and a phase-change lens. This mechanism resulted in the peak energy conversion efficiencies of the R-STEG being 60% and 86% higher than those of the typical STEG at solar intensities of 1 kW m(-2) and 1.5 kW m(-2), respectively. In addition, the energy stored in PCM can help to generate steady electrical energy when the solar energy was removed. This work presents significant progress regarding the optical characteristic of PCM and optical concentration systems of STEGs.


Human papillomavirus 16L1-58L2 chimeric virus-like particles elicit durable neutralizing antibody responses against a broad-spectrum of human papillomavirus types.
The neutralizing antibodies elicited by human papillomavirus (HPV) major capsid protein L1 virus-like particle (VLP)-based vaccines are largely type-specific. An HPV vaccine inducing cross-neutralizing antibodies broadly will be cost-effective and of great value. To this end, we constructed HPV16L1-58L2 chimeric VLP (cVLP) by displaying HPV58 L2 aa.16-37 on the DE surface region of HPV16 L1. We found that vaccination with the HPV16L1-58L2 cVLP formulated with alum plus monophosphoryl lipid A (Alum-MPL) adjuvant elicited robust neutralizing antibodies in both mice and rabbits against all tested HPV types including HPV16/31/33/35/52/58 (genus α9), HPV18/39/45/59/68 (genus α7), HPV6/11 (genus α10), HPV2/27/57 (genus α4), and HPV5 (genus β1). Importantly, the cross-neutralizing antibody response was maintained at least 82 weeks in mice or 42 weeks in rabbits, and complete protection against HPV58 was observed at week 85 in mice. Our data demonstrate that HPV16L1-58L2 cVLP is an excellent pan-HPV vaccine candidate.


Structure and regulatory mechanism of Aquifex aeolicus NtrC4: variability and evolution in bacterial transcriptional regulation.
Genetic changes lead gradually to altered protein function, making deduction of the molecular basis for activity from a sequence difficult. Comparative studies provide insights into the functional consequences of specific changes. Here we present structural and biochemical studies of NtrC4, a sigma-54 activator from Aquifex aeolicus, and compare it with NtrC1 (a paralog) and NtrC (a homolog from Salmonella enterica) to provide insight into how a substantial change in regulatory mechanism may have occurred. Activity assays show that assembly of NtrC4's active oligomer is repressed by the N-terminal receiver domain, and that BeF3- addition (mimicking phosphorylation) removes this repression. Observation of assembly without activation for NtrC4 indicates that it is much less strongly repressed than NtrC1. The crystal structure of the unactivated receiver-ATPase domain combination shows a partially disrupted interface. NMR structures of the regulatory domain show that its activation mechanism is very similar to that of NtrC1. The crystal structure of the NtrC4 DNA-binding domain shows that it is dimeric and more similar in structure to NtrC than NtrC1. Electron microscope images of the ATPase-DNA-binding domain combination show formation of oligomeric rings. Sequence alignments provide insights into the distribution of activation mechanisms in this family of proteins.


A trifunctional Pt(II) complex alleviates the NHEJ/HR-related DSBs repairs to evade cisplatin-resistance in NSCLC.
Cisplatin, a representative of platinum-based drug, is clinically and widely used in the treatment of various types of malignant cancer. However, its non-selectivity to almost all the cell lines and resistance in long-term use severely limit its scope of use. As biotin-specific uptake systems are overexpressed in many types of tumors but rarely occur in normal tissues, making biotin a promising target for cancer treatment. In the study, we synthesized the Pt(II) complex C2 and determined its biological activities. The existence of biotin enhanced the ability of the complex to target tumors, while the introduction of a naphthalimide compound makes it possible to diagnose tumors and monitor their progress. We have also introduced a known Pt(II) complex DN604, which not only retains the excellent cytotoxicity of platinum drugs, but also inhibits the expression of DNA double-strand breaks (DSBs) repair-related NHEJ protein Ku70 and HR protein Rad51. In summary, we report a novel trifunctional Pt(II) complex that could target tumor cells, monitor tumor progression, and reverse DSBs repair-induced cisplatin-resistance.


[Effectiveness of a preventive intervention strategy based on structured telephone interviews in a working population with a moderate to high cardiovascular risk].
This study assesses the effectiveness of a structured telephone survey on cardiovascular prevention, in modifying lifestyle, on cardiovascular risk parameters, percentage of smoking cessation and overall cardiovascular risk (CVR). Quasi-experimental study of preventive intervention. Ibermutuamur (Spanish Accident and Health Insurance Company). Centres established throughout Spain. A total of 4,792 workers with moderate/high cardiovascular risk who had agreed to be contacted by phone. Subjects with a previous diagnosis of cardiovascular disease and those receiving treatment for hypertension, hypercholesterolemia or diabetes were excluded. A final total of 3,085 workers were contacted and were followed up by telephone surveys on the first, fourth and eighth month after the initial check up (CU) in order to emphasise cardiovascular health advice (Group A); we failed to contact 1,707 workers, who only attended the baseline and one year CUs (Group B). PRINCIPAL OUTCOMES: CUs included medical records and physical examination, with two blood pressure measurements, Body Mass Index (BMI), and biochemical parameters. Cardiovascular risk was stratified following the European cardiovascular SCORE. Individuals with a relative risk higher than 4 were also considered as high-risk. All workers were informed about their cardiovascular risk profile (CVRF) and healthy cardiovascular lifestyle measures. They were also given a letter for their General Practitioner (GP) to inform them on the worker's cardiovascular risk level. A total of 71.5% of the workers were over 45 years, 95.0% males, 76.6% manual workers ("Blue Collar") and 69.7% smokers. Both groups showed improvement in lipid parameters, blood pressure, smoking cessation and overall cardiovascular risk in the second CU. There were significant differences in favour of Group A as regards blood pressure, lipids (except HDL cholesterol), BMI, glycaemia, smoking cessation (A: 23.5%/B: 19.44%, P=0.001) and CVR stratum improvement (A: 46.6%/B: 37.7%, P=0.0001). The large majority (85%) of workers read preventive recommendations; 33% knew their risk level and 73% knew their CVRF. 52.9% gave the letter to the GP, which led them to start therapies on diet (47%), hypertension (19.5%), dyslipidaemia (16.7%), diabetes (4.4%) and smoking (2.9%) and no changes were made in 36.5% of cases. The results of this study suggests that cardiovascular prevention strategy based on structured telephone surveys on high/moderate CVR subjects to promote lifestyle changes could be effective at reducing CVR. A clinical trial is required for confirmation. Sending information on CVRF following routine medial CUs and Primary Care involvement, could contribute to the positive changes observed.


Laminin and collagen modulate expression of the small leucine-rich proteoglycan fibromodulin in rat anterior pituitary gland.
The anterior pituitary is a complex organ consisting of five types of hormone-producing cells, non–hormone-producing cells such as folliculostellate (FS) cells and vascular cells (endothelial cells and pericytes). We have previously shown that FS cells and pericytes produce fibromodulin, a small leucine-rich proteoglycan (SLRP). SLRPs are major proteoglycans of the extracellular matrix (ECM) and are important in regulating cell signaling pathways and ECM assembly. However, the mechanism regulating fibromodulin expression in the anterior pituitary has not been elucidated. Here, we investigate whether fibromodulin expression is modulated by major anterior pituitary ECM components such as laminin and type I collagen. Using transgenic rats expressing green fluorescent protein (GFP) specifically in FS cells, we examine fibromodulin expression in GFP-positive (FS cells) and GFP-negative cells (e.g., pericytes, endocrine cells and endothelial cells). Immunostaining and Western blot analysis were used to assess protein expression in the presence and absence of laminin or type I collagen. We confirmed fibromodulin expression in the pituitary and observed the up-regulation of fibromodulin in FS cells in the presence of ECM components. However, neither laminin nor type I collagen affected expression in GFP-negative cells. This suggests that laminin and type I collagen support the function of FS cells by increasing fibromodulin protein expression in the anterior pituitary.


Bio-hybrid inorganic microparticles derived from CO2 for highly efficient and selective removal of antibiotics.
Antibiotics, which are the most important medication in human history, have brought global concerns due to their potential risk to human health and environment by accelerating the development of drug-resistant bacteria, and accumulating in the food chain system. Among antibiotics, oxytetracycline (OTC) is widely used in aquaculture, and its potential risk of toxicity to human by bioaccumulation has been reported. Therefore, the effective removal of OTC is highly needed. In this study, we report bio-hybrid inorganic microparticles (apt-mag-SiCC) for efficient capturing and facile magnet-based separation of oxytetracycline (OTC). These bio-hybrid inorganic microparticles are composed of magnetic separable silica coated calcium carbonate microparticles (mag-SiCC) derived from CO2, conjugated with oxytetracycline binding aptamers (OBA). These bio-hybrid inorganic microparticles were successfully synthesized, based on the characterization data obtained by SEM, FT-IR, EDAX, BET, and CLSM. About 6 μm sized bio-hybrid inorganic microparticles showed low non-specific adsorption to OTC and other molecules, and the selective capturing towards to the OTC in both buffer and tap water. Moreover, these bio-hybrid mineral microparticles were found to be stable, even after the repeated usages, maintaining the initial capturing efficiency. Using the newly synthesized bio-hybrid inorganic microparticles, we could successfully capture OTC by facile magnet-based separation. With advantages of theses bio-hybrid inorganic microparticles such as easy fabrication, low-price, and environmental friendliness, this novel material could be utilized in the drinking water treatment, in vitro medicinal diagnostics, or in vitro removal of antibiotics lining out from the blood (blood purification).


Assessment of whole-body MRI including diffusion-weighted sequences in the initial staging of breast cancer patients at high risk of metastases in comparison with PET-CT: a prospective cohort study.
The aim of this study was to assess the diffusion-weighted whole-body-MRI (WBMRI) in the initial staging of breast cancer at high risk of metastases in comparison with positron emission tomography (PET)-CT. Forty-five women were prospectively enrolled. The inclusion criteria were female gender, age >18, invasive breast cancer, an initial PET-CT, and a performance status of 0-2. The exclusion criteria were contraindication to WB-MRI and breast cancer recurrence. The primary outcome was the concordance of WB-MRI and PET-CT in the diagnosis of distant metastases, whereas secondary outcomes included their concordance for the primary tumor and regional lymph nodes (LN), as well as the agreement of WB-MRI interpretation between two radiologists. The mean age was 51.2 years with a median size of the primary tumor of 30 mm. Concordance between the two modalities was almost perfect for metastases staging, all sites included (k = 0.862), with excellent interobserver agreement. The accuracy of WB-MRI for detecting regional LN, distant LN, lung, liver, or bone metastases ranged from 91 to 96%. In 2 patients, WB-MRI detected bone metastases that were overlooked by PET-CT. WB-MRI showed a substantial agreement with PET-CT for staging the primary tumor, regional LN status, and stage (k = 0.766, k = 0.756, and k = 0.785, respectively) with a high interobserver agreement. WB-MRI including DWI could be a reliable and reproducible examination in the initial staging of breast cancer patients at high risk of metastases, especially for bone metastases and therefore could be used as a surrogate to PET-CT. Whole-body-MRI including DWI is a promising technique for detecting metastases in the initial staging of breast cancer at high risk of metastases. Whole-body-MRI (WB-MRI) was effective for detecting metastases in the initial staging of 45 breast cancer patients at high risk of metastases in comparison with PET-CT. Concordance between WB-MRI and PET-CT was almost perfect for metastases staging, all sites included, with excellent interobserver agreement. The accuracy of WB-MRI for detecting bone metastases was 92%.


Gastroportal shunt for portal hypertension in children.
Extrahepatic portal venous obstruction is the most common cause of portal hypertension in children. The Rex shunt has been used successfully to treat patients with extrahepatic portal hypertension. In the conventional Rex shunt, the internal jugular vein is used as a venous graft. Inevitably, such a procedure requires neck exploration and sacrifice of internal jugular vein. The authors describe a novel adaptation of gastroportal shunt, successfully carried out in 8 children with extrahepatic portal hypertension. The mean age of the 8 patients (6 boys and 2 girls) was 66.6 months at the time of operation. All children had portal hypertension. Seven had a history of upper gastrointestinal bleeding, and 4 had splenomegaly and hypersplenism. Gastroportal shunt was performed in all patients. The left gastric vein was mobilized and anastomosed to left portal vein. In 1 patient, the left gastric vein was not of adequate length and required a venous graft (the inferior mesenteric vein). All patients were followed up for 3 to 20 months (median, 9 months). The gastroportal shunt was successfully performed in all patients. The median operative time was 265 minutes (range, 205-360 minutes). Operative blood loss was 21 ± 7.4 mL, and the length of hospital stay varied from 9 to 19 days (median, 15 days). Intraoperative portal venous angiography demonstrated the patency of the shunt in all patients. Postoperatively, the complete blood count normalized, and the biochemistry tests were within reference range. Postoperative ultrasound confirmed shunt patency and satisfactory flow in the gastroportal shunt in each patient. The size of spleen decreased. There was no recurrence of variceal bleeding. The gastroportal shunt is an effective treatment of extrahepatic portal hypertension.


A model of waveform reliability in facial nerve electroneurography.
This study identifies four factors that should be considered when evaluating the reliability of new testing methods in facial nerve electroneurography (ENOG): (1) waveform peak-to-peak amplitude; (2) response variability; (3) statistical significance of day-to-day test/retest differences; and (4) waveform morphology. These factors were applied to data gathered on seven normal subjects who had three trials of facial-nerve ENOG on three separate test days in order to determine an optimal recording electrode montage. Both bipolar and monopolar recording-electrode montages were used on the nasal alae and on the nasolabial crease. Resultant peak-to-peak amplitudes of combined motor action potentials were recorded and measured using a clinical evoked-potential instrument. A biphasic waveform shape with optimal peak-to-peak point rather than the nasolabial crease. The contribution to the assessment of response reliability of each of the four factors is discussed.


"I don't want to hear statistics, I want real life stories": Systematic review and thematic synthesis of patient and caregiver experiences of Proton Beam Therapy.
Proton Beam Therapy (PBT) is an advanced form of radiotherapy, yet little evidence exists on patient experience to inform decision making and improve future care. We thematically synthesized the qualitative evidence of patient and caregivers' perceptions and experiences of PBT. Five electronic databases were systematically searched, using Medical Subject Headings (MeSH) terms and keywords. Two reviewers independently screened search results for qualitative studies relating to patients' and caregivers' experiences of PBT. The search generated 4,020 records, of which nine were eligible. Study quality (assessed by CASP checklist) varied. Qualitative results were analyzed using thematic synthesis. Three main themes were generated: decision making and perceptions, living in the PBT "bubble," and coping with the cancer treatment journey. PBT is not yet widely accessible worldwide, which uniquely influences the patient experience. Our review uncovers areas PBT providers could target to improve patient-centered care; however, additional primary qualitative research is recommended.


Preparation, physicochemical characterization and application of acetylated lotus rhizome starches.
Acetylated lotus rhizome starches were prepared, physicochemically characterized and used as food additives in puddings. The percentage content of the acetyl groups and degree of substitution increased linearly with the amount of acetic anhydride used. The introduction of acetyl groups was confirmed via Fourier transform infrared (FT-IR) spectroscopy. The values of the pasting parameters were lower for acetylated starch than for native starch. Acetylation was found to increase the light transmittance (%), the freeze-thaw stability, the swelling power and the solubility of the starch. Sensorial scores for puddings prepared using native and acetylated lotus rhizome starches as food additives indicated that puddings produced from the modified starches with superior properties over those prepared from native starch.


Mefunidone ameliorates diabetic kidney disease in STZ and db/db mice.
Diabetic kidney disease (DKD) is a major cause of end stage renal diseases worldwide. Despite successive interventions for delaying the progression of DKD, current treatments cannot reverse the pathological progression. Mefunidone (MFD) is a new compound with potent antifibrotic properties, but the effect of MFD on DKD remains unknown. Therefore, we investigated the protective effects of MFD in both models of the db/db type 2 diabetes (T2D) and streptozotocin (STZ)-induced type 1 diabetes (T1D) models. Compared with the model group, MFD treatment significantly reduced pathological changes observed by PAS staining, PASM staining, and Masson staining in vivo. To further elucidate the potential mechanisms, we discovered MFD treatment notably restored podocyte function, alleviated inflammation, abated ROS generation, inhibited the TGF-β1/SAMD2/3 pathway, suppressed the phosphorylation levels of MAPKs (ERK1/2, JNK, and P38), and reduced epithelial-to-mesenchymal transition(EMT). In conclusion, these findings demonstrate the effectiveness of MFD in diabetic nephropathy and elucidate its possible mechanism.


Arytenoid dislocation related to an uneventful endotracheal intubation: a case report.
Invasive methods currently applied to the respiratory tract may result in impaired movement of the cricoarytenoid joint with hoarseness and immobility of the vocal ligament. Hoarseness after tracheal intubation is reported as a high incidence in patients who receive general anaesthesia. In most cases, the symptoms are temporary and improve within several days. We report this case for emphasizing that early diagnosis of arytenoid cartilage dislocation is important even in nontraumatic cases. We present the case of a 19-year-old Caucasian male who developed arytenoid cartilage dislocation associated with uneventful endotracheal intubation and anesthesia. Arytenoid subluxation should be considered whenever any of the symptoms mentioned occur following endolaryngeal manipulation, and they become persistent, as recovery becomes difficult if appropriate treatment is not started immediately.


Reduced emotional and cardiovascular reactivity to emotionally evocative stimuli in major depressive disorder.
Major Depressive Disorder (MDD) is a highly debilitating mental health concern that affects a large number of adults in the United States. The emotional context insensitivity (ECI) hypothesis argues that individuals with MDD disengage from the environment to defend themselves from futile activity. In the current study, electrocardiogram and pupillometry were recorded from 50 participants (MDD n=25, never depressed control n=25) during the display of emotionally evocative images, sounds, and movie clips. Individuals with MDD reported reduced change in happiness to positively- and negatively-valenced images and sounds. Heart rate reactivity also was reduced in individuals with MDD when viewing images and watching movie clips. These results suggest that individuals with MDD may have some difficulty engaging with certain environmental stimuli.


Heparin-induced thromboembolism: a case report.
A case of heparin-induced thrombosis of the brachial artery with thrombocytopenia is described. With discontinuation of the heparin, the circulating platelet volumes returned to normal. The role of pre-existing arterial disease and sepsis in patients exhibiting this phenomenon is unresolved.


Impact of prior trauma exposure on the development of PTSD symptoms after suspected acute coronary syndrome.
To determine the association between PTSD symptoms due to prior trauma and prior trauma type with PTSD symptoms after suspected acute coronary syndrome (ACS). A consecutive sample of patients presenting to the emergency department (ED) for suspected ACS were surveyed. Logistic regression was used to estimate the odds of elevated ACS-related PTSD symptoms [PCL-S ≥ 33] at 1-month associated with PTSD symptoms due to prior trauma and prior trauma type at the time of suspected ACS, adjusting for demographics, comorbidities, depression, and etiology of ACS symptoms. Of 984 patients, 81.6% reported ≥1 prior trauma type and 22.5% reported PTSD symptoms due to prior trauma at the time of suspected ACS. One month later, 18.0% had ACS-related PTSD symptoms. Patients with versus without PTSD symptoms due to prior trauma at the time of the suspected ACS had increased odds of ACS-related PTSD symptoms one month later (42.1% vs 9.9%; aOR 4.49, 95% CI:3.05-6.60; p < .001). Prior life-threatening illness was the only trauma type significantly associated with ACS-related PTSD symptoms (aOR 1.57, 95% CI:1.03-2.39; p = .04). PTSD symptoms from prior trauma and history of life-threatening medical illness at the time of suspected ACS increased risk of ACS-related PTSD symptoms one month later.


Simultaneous dermatophytosis and keratomycosis caused by Trichophyton interdigitale infection: a case report and literature review.
Dermatophytosis is a fungal infectious disease caused by dermatophytes, which produce protease and keratinase to digest keratin, leading to the colonization, invasion, and infection of the stratum corneum of the skin, hair shafts, and nails. Trichophyton interdigitale belongs to Trichophyton mentagrophytes complex, which is the common pathogen causing dermatophytosis. Fungal keratitis, also called keratomycosis, is an infectious disease of cornea. Here, we report a case of simultaneous dermatophytosis and keratomycosis caused by Trichophyton interdigitale. A 67-year-old man presented with extensive erythema all over the body since 4 years ago, fungal infection of left eye for 2 years, and loss of vision in the eye. These symptoms had become aggravated in the last month. Dermatological examinations showed extensive erythematous plaques with clear borders and scales, scattered red papules with ulceration, and scabs throughout the body. Onychomycosis was observed on the nails of left hand, conjunctival infection with secretion and loss of vision were noted in left eye. Hyaline septate hyphae were observed under direct microscopic examination, fungal culture and internal transcribed spacer sequencing revealed T. interdigitale. Histopathological examination suggested infectious granuloma. A diagnosis of dermatophytosis and keratomycosis caused by T. interdigitale with loss of vision in left eye was made. The patient was treated with luliconazole cream (two applications per day) and itraconazole (100 mg, BID, PO). Complete clinical remission was achieved after 1 month. Subsequently, the patient underwent left eye enucleation in the ophthalmology department. In the present study, we reported a case of simultaneous dermatophytosis and keratomycosis caused by T. interdigitale, and reviewed the literature on corneal infection caused by Trichophyton. A total of 10 articles with 45 patients were published between 1973 and 2018. The pathogen of 27 patient were identified to species level. There were T. schoenleinii (17), T. mentagrophytes (4), T. verrucosum (3), T. rubrum (1), T. erinacei (1), and T. interdigitale (1). Five patients had corneal trauma, one had contact lens use history. Direct microscopic examination, fungal culture, and analysis of physiological characteristics were the main methods of identification. Early diagnosis and prompt treatment may help improve the management and outcomes.


Purification, characterization and anti-proliferation activities of polysaccharides extracted from Viscum coloratum (Kom.) Nakai.
Three polysaccharides, VCP1, VCP2 and VCP3 were isolated from Viscum coloratum (Kom.) Nakai using DEAE-cellulose chromatography. VCP1 (32KDa) was composed of glucose, galactose, arabinose, rhamnose and mannose, while VCP2 (280KDa) and VCP3 (21KDa) were consisted of glucose, galactose, arabinose, rhamnose, mannose, glucuronic acid and galacturonic acid. The optical rotation was measured at 20+1°C. The characteristic absorptive bands of purified fraction were determined by FT-IR. (13)C NMR spectroscopy analysis showed that VCP1 was a neutral polysaccharide, and VCP2 and VCP3 were RG-I type pectin. The linkage patterns of VCP2 were evaluated by methylation analysis: 1,5-linked Araf, 1,4-linked Galp, 1,2-linked Rhap, and 1,2,4-linked Rhap. The degree of esterification was 50%. The anti-proliferation ability against HepG2 cells and HepG2.2.15 cells of VCP2 was stronger than VCP1 and VCP3. So the polysaccharides from Viscum coloratum (Kom.) Nakai could be used as potential natural sources with inhibiting tumor cells proliferation.


[Instrument for the assessment of middle-aged and older adults' physical activity: design, eliability and application of the German-PAQ-50+].
Existing physical activity questionnaires have focused either on young and middle-aged adults or on the elderly. They have mainly assessed only a portion of possible physical activities or contained nation-specific sports. As there is no gold standard for a questionnaire-based assessment of physical activity in the over-50 population, recommendations for such a questionnaire relating to German-speaking countries were developed. This work included a systematic literature research, a survey of experts, and the design of a questionnaire based on validated measuring instruments. Finally, to test its reliability and application in the field, the complete questionnaire, including a retest, was applied by telephone interview (n = 57). The test-retest-correlation was r = 0.60 for the total time of physical activity and r = 0.52 for total energy expenditure. The researchers determined that the instrument is comprehensive in its coverage of all relevant domains of physical activity for the over-50 population; it is economically feasible and showed good acceptance.


Cryotechniques for the Long-Term Conservation of Embryogenic Cultures from Woody Plants.
Since its development in the 1960s, plant cryopreservation is considered an extraordinary method of safe long-term conservation of biological material, as it does not induce genetic alterations and preserve the regeneration potential of the stored material. It is based on the storage of explants at cryogenic temperatures, such as the one of liquid nitrogen (-196 °C), where the metabolism within the cells is suspended; thus, the time for these cells is theoretically "stopped". Cryopreservation is particularly important for embryogenic cultures, as they require periodic subculturing for their maintenance, and this, in turn, increases the risk of losing the material, as well as its embryogenic potential. Periodic re-initiation of embryogenic cultures is possible; however, it is labor intensive, expensive, and particularly difficult when working with species for which embryogenic explants are available only during a limited period of the year. Among various methods of cryopreservation available for embryogenic cultures, slow cooling is still the most common approach, especially in callus cultures from softwood species. This chapter briefly reviews the cryopreservation of embryogenic cultures in conifers and broadleaf trees, and describes as well a complete protocol of embryogenic callus cryopreservation from common ash tree (Fraxinus excelsior L.) by slow cooling.


Six-year changes in health-related quality of life in gastric bypass patients versus obese comparison groups.
Few studies have evaluated the long-term outcomes of bariatric surgery patients in relation to obese individuals not participating in weight loss interventions. Our objective was to evaluate the 6-year changes in health-related quality of life (HRQOL) in gastric bypass (GB) patients versus 2 obese groups not undergoing surgical weight loss. The study setting was a bariatric surgery practice. A total of 323 GB patients were compared with 257 individuals who sought but did not undergo gastric bypass and 272 population-based obese individuals using weight-specific (impact of weight on quality of life-lite) and general (medical outcomes study short-form 36 health survey) HRQOL questionnaires at baseline and 2 and 6 years later. At 6 years, compared with the controls, the GB group exhibited significant improvements in all domains of weight-specific and most domains of general HRQOL (i.e., all physical and some mental/psychosocial). The 6-year percentage of excess weight loss correlated significantly with improvements in both weight-specific and physical HRQOL. The HRQOL scores were fairly stable from 2 to 6 years for the GB group, with small decreases in HRQOL corresponding to some weight regain. GB patients demonstrated significant improvements in most aspects of HRQOL at 6 years compared with 2 nonsurgical obese groups. Despite some weight regain and small decreases in HRQOL from 2 to 6 years postoperatively, the HRQOL was relatively stable. These results support the effectiveness of weight loss achieved with gastric bypass surgery for improving and maintaining long-term HRQOL.


Hyaluronan is an abundant constituent of the extracellular matrix of Xenopus embryos.
The spatiotemporal distribution of hyaluronan (HA), a major constituent of the vertebrate extracellular matrix, was analyzed during early embryonic development of Xenopus laevis. This polysaccharide is abundantly present in ventricular structures such as the blastocoel, the archenteron as well as later on in the hepatic cavity, the brain ventricles and the developing heart. At the blastula stage, HA was detected in the extracellular matrix of the ecto- and mesodermal primordia. Shortly before gastrulation, it becomes enriched at the basal site of the superficial cell layer of the ectoderm. During gastrulation, enhanced synthesis of HA takes place in the involuting marginal zone, shortly before invagination starts, hence, resulting in a torus-like deposition in the deep layer of the equatorial mesodermal primordium. After gastrulation, HA appears to accumulate within the extracellular matrix demarcating the primary germ layers. During tailbud stages, it is found highly enriched in many mesodermal derivatives, e.g., in mesenchyme, the heart, precordal cartilage and the lung primordia. Furthermore, extracellular matrix of the ventral mesodermal cell layer in the trunk region and the immediate proximity of blood vessels contain high amounts of HA.


Evaluation of a test to detect circulating Aspergillus fumigatus antigen in a survey of immunocompromised patients with proven or suspected invasive disease.
An ELISA for the detection and measurement of Aspergillus antigenaemia has been developed and evaluated by examining sera submitted over a 12-month period from immunocompromised patients with a likelihood of invasive aspergillosis. Results from proven cases of invasive aspergillosis confirmed at post-mortem and specimens from individuals with suspected disease showed that tests on single serum samples were often negative. Multiple specimens from the same patient greatly increased the frequency of detection. Repeated monitoring of sera from a single patient showed wide fluctuations in antigen level, which was considered to be due partly to the medical regimen to which the patient was subject. Control sera from healthy laboratory personnel were consistently negative, but a number of 'at-risk' patients without other evidence of invasive aspergillosis sometimes had low amounts of antigen. Concentrations of Aspergillus antigen of 100 ng ml-1 or higher were considered to be strongly suggestive of fungal invasion.


The influence of lip function on the sagittal and transversal development of the maxilla in cleft patients.
Cleft patients who have undergone surgery often show considerable inhibition in the sagittal development of the mid-face after completed growth. This is partly attributed to scar contraction in the lip region following surgery. The present investigation deals with the etiological factors involved in the development disturbances caused by the cleft in the maxilla of 100 cleft patients. Particular attention was paid as to the unfavourable effect of the operated cleft lip. This was tested electromyographically and correlation between the electromyographic and the skeletal measurements was sought. Electromyographic changes in lip activity during orthodontic treatment was also investigated. The effect of these changes is discussed.


Selective accumulation and growth inhibition of hybrid liposomes to human hepatocellular carcinoma cells in relation to fluidity of plasma membranes.
Hybrid liposomes (HLs), composed of l-α-dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene(23) dodecyl ether, have selectively inhibited the growth of human hepatocellular carcinoma (HCC) cells without affecting normal hepatocytes to trigger apoptosis via caspase-3 activation. Furthermore, HLs distinguished between the HCC and normal cells which had higher and lower membrane fluidities respectively, then fused and accumulated preferentially into the membranes of HCC cells. It is noteworthy that the anti-cancer activity of HLs correlated well with the fluidity of cell membranes for HCC and other cancer cells. These results suggest that HLs could target cancer cell-membranes in relation to their lipid fluidity that provide the possibility of novel nanotherapy for intractable cancer.


Absence of detectable alterations in the putative tumor suppressor gene BTRC in cerebellar medulloblastomas and cutaneous basal cell carcinomas.
The sonic hedgehog signal transduction pathway is aberrantly activated in the majority of cutaneous basal cell carcinomas and a subset of cerebellar medulloblastomas. The latter tumors may also show activation of the wingless (Wnt) signaling pathway. In Drosophila, the F-box/WD40-repeat containing protein Slimb has been shown to function as an intracellular negative regulator of both pathways. The BTRC gene (beta-transducin repeat-containing protein) is a human homolog of Slimb that is located at 10q24.3, a chromosome region frequently deleted in medulloblastomas. Here, we report on the mutational analysis of BTRC in 91 human tumors, including 66 primitive neuroectodermal tumors (PNETs) of the central nervous system (62 medulloblastomas and 4 supratentorial PNETs) and 25 cutaneous basal cell carcinomas (BCCs). These analyses revealed no tumor-associated BTRC mutations. BTRC transcripts were expressed in non-neoplastic brain tissue, normal skin, as well as in all PNETs and BCCs analyzed by real-time reverse transcription-PCR. Two novel BTRC transcript variants were expressed at higher levels in non-neoplastic brain tissue than in normal skin and the investigated tumors. Taken together, our results indicate that PNETs and BCCs neither show mutations nor loss of mRNA expression of BTRC. Therefore, BTRC alteration does not appear to be involved in the pathogenesis of these neoplasms.


Development and Validation of Multiclass Chiral Pesticide Residues Analysis Method in Tea by QuEChERS Combined Liquid Chromatography Quadruple-Linear Ion Trap Mass Spectrometry.
No single pure enantiomeric pesticide residues was investigated and set regulations for tea quality safety and risk assessment. Due to lack of chiral pesticide analysis method and data, the Maximum residue limits (MRLs) about the chiral pesticides in tea was unknown. An analytical method for the determination of chiral pesticide residues by QuEChERS combined chiral liquid chromatography quadruple/linear ion trap mass spectrometry (LC-MS/MS-Qtrap) was developed and applied to the analysis of various teas. The mean recoveries for pesticides enantiomers ranged from 75.9% to 112.4%. Reproducibility represented by relative standard deviation percentage was 10% or less. Good linearity was achieved for all enantiomers with determination coefficients (r2) greater than 0.99. The detection of limit (CCα) and quantification of limit (CCβ) were 0.2 ∼1 µg/kg and 0.5∼5 µg/kg, respectively. The method was suitable for monitoring the enantiomeric pesticide residues in various teas. Enantioselective multiclass pesticide residues were determined in various teas by LC-MS/MS-Qtrap, additional Qtrap scan functions greatly enhance the performance of screening, confirmation, and identification of chiral pesticides.


Study on synergistic effect of ozone and monochloramine on the degradation of chloromethylisothiazolinone biocide.
In this study, it was found that monochloramine induced the formation of reactive species during ozonation of chloromethylisothiazolinone (CMIT). CMIT was found recalcitrant to chloramine. However, chloramine promoted the degradation of CMIT by ozonation significantly. Hydroxyl radicals contributed most to CMIT degradation (87%) during ozone/chloramine synergistic oxidation process (SOP). The hydroxyl radical exposure during ozone/chloramine SOP was around 7.9 times higher than that of ozonation process. The hydroxyl radical yield of ozone/chloramine SOP was estimated to be 32%. The reaction mechanisms between ozone and chloramine were postulated to include the oxygen transfer reaction to form singlet oxygen, and the formation of hydroxyl radical by the insertion pathway or electron transfer pathway. Chloramine dosage and pH are essential influencing factors. The degradation of CMIT increased from 41% to 74% with increasing chloramine dosage (0-20 μM), and then decreased to 65% when chloramine dosage continually increased to 40 μM. Ozone/chloramine SOP showed better performance at acidic or neutral conditions than basic condition. Based on the intermediates identified, the degradation pathway of CMIT during ozone/chloramine SOP included the oxidation of sulfur atom and the substitution of chlorine group by hydroxyl group. The oxidation of sulfur atom induced lower toxicities of transformation products. The toxicities of hydroxylation products were lower to fish and algae, but higher to daphnia. Based on the GC-ECD results, only trichloromethane (1.94 μg/L) was detected after ozone/chloramine SOP, accounting for 0.17% (μM/μM) of the CMIT removal.


Perforation Mechanisms, Risk Stratification, and Management in the Post-Coronary Artery Bypass Grafting Patient.
Coronary perforations during chronic total occlusion percutaneous coronary intervention (CTO PCI) is a most frequent major complication and the incidence is significantly higher compared with non-CTO PCI. Patients with prior history of coronary bypass have more major adverse events when perforation occurs compared with patients without prior bypass surgery. In this article, the authors discuss the unique challenges in identification and timely treatment of perforations in patients with prior bypass surgery.


Blue native polyacrylamide gel electrophoresis and the monitoring of malate- and oxaloacetate-producing enzymes.
We demonstrate a facile blue native polyacrylamide gel electrophoresis (BN-PAGE) technique to detect two malate-generating enzymes, namely fumarase (FUM), malate synthase (MS) and four oxaloacetate-forming enzymes, namely pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), citrate lyase (CL) and aspartate aminotransferase (AST). Malate dehydrogenase (MDH) was utilized as a coupling enzyme to detect either malate or oxaloacetate in the presence of their respective substrates and cofactors. The latter four oxaloacetate-forming enzymes were identified by 2,6-dichloroindophenol (DCIP) and p-iodonitrotetrazolium (INT) while the former two malate-producing enzymes were visualized by INT and phenazine methosulfate (PMS) in the reaction mixtures, respectively. The band formed at the site of enzymatic activity was easily quantified, while Coomassie staining provided information on the protein concentration. Hence, the expression and the activity of these enzymes can be readily evaluated. A two-dimensional (2D) BN-PAGE or SDS-PAGE enabled the rapid purification of the enzyme of interest. This technique also provides a quick and inexpensive means of quantifying these enzymatic activities in normal and stressed biological systems.


Action of some natural polypeptides on the longitudinal muscle of the guinea pig ileum.
The structure--activity relationship of some natural tachykinins was investigated in the longitudinal muscle of the guinea pig ileum. The order of potency was eledoisin greater than uperolein greater than substance P greater than phyllomedusin greater than physalaemin. This ratio of activity is different from that observed in other experimental conditions, and in different in vivo or in vitro preparations; it is suggested that the N-terminal part of the molecule of the tachykinins plays a role in determining the degree of potency of these compounds.


Bioactive hydrogel microcapsules for guiding stem cell fate decisions by release and reloading of growth factors.
Human pluripotent stem cells (hPSC) hold considerable promise as a source of adult cells for treatment of diseases ranging from diabetes to liver failure. Some of the challenges that limit the clinical/translational impact of hPSCs are high cost and difficulty in scaling-up of existing differentiation protocols. In this paper, we sought to address these challenges through the development of bioactive microcapsules. A co-axial flow focusing microfluidic device was used to encapsulate hPSCs in microcapsules comprised of an aqueous core and a hydrogel shell. Importantly, the shell contained heparin moieties for growth factor (GF) binding and release. The aqueous core enabled rapid aggregation of hPSCs into 3D spheroids while the bioactive hydrogel shell was used to load inductive cues driving pluripotency maintenance and endodermal differentiation. Specifically, we demonstrated that one-time, 1 h long loading of pluripotency signals, fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-β1, into bioactive microcapsules was sufficient to induce and maintain pluripotency of hPSCs over the course of 5 days at levels similar to or better than a standard protocol with soluble GFs. Furthermore, stem cell-carrying microcapsules that previously contained pluripotency signals could be reloaded with an endodermal cue, Nodal, resulting in higher levels of endodermal markers compared to stem cells differentiated in a standard protocol. Overall, bioactive heparin-containing core-shell microcapsules decreased GF usage five-fold while improving stem cell phenotype and are well suited for 3D cultivation of hPSCs.