Resuscitation of the critically ill patient. Use of branched-chain decision trees to improve outcome.
The algorithm approach provides criteria based on decision rules for expeditious monitoring, diagnostic and therapeutic decisions; algorithms are particularly useful in crisis situations, in which time is of great importance, for example, in the resuscitation of emergency patients. Because of its objectivity and usefulness as a teaching tool, this algorithmic approach is of practical benefit in the training of residents and students in teaching hospitals, as well as in the community hospital where less experienced physicians manage hypotensive emergency patients more infrequently. In a few instances there has been some reluctance to use the algorithm, but most often it was found to be useful in organizing the work-up and establishing therapeutic priorities. A second branched-chain decision tree was designed for use after the initial resuscitation in hemodynamically unstable patients who only had CVP catheters but were suspected of having hypovolemia. In this algorithm, MAP, HR, urine output, Hct, and CVP are used as criteria. The underlying premise under these conditions was to evaluate increments of volume therapy without exceeding safe CVP pressures (less than 18 mm Hg) in order to obviate fluid overloading. A third algorithm for ICU patients with pulmonary artery catheters was developed from decision rules based on objective physiologic, heuristic, survival data as the criteria for post-trauma and postoperative patients who were critically ill despite apparent success with the initial resuscitation and CVP algorithms. The improved mortality in prospective studies supports the hypothesis that compensatory responses of the survivors are the major determinants of outcome.


[Ultraviolet spectrophotometry of the cerebrospinal fluid in the differential diagnosis of strokes].
The authors describe the possibility of using the results of ultraviolet spectrophotometry of the cerebrospinal fluid (CSF) for the differential diagnosis of ischemic, hemorrhagic and mixed cerebral strokes. It was found that repeated examinations may be of a definite prognostic value. A correlational analysis showed that the greatest effect on the optical density of the CSF in cases of hemorrhagic strokes is exercised by protein and bilirubin and in cases of ischemic strokes by protein and aspartate aminotransferase (AST).


DNA repair deficiency in neurodegeneration.
Deficiency in repair of nuclear and mitochondrial DNA damage has been linked to several neurodegenerative disorders. Many recent experimental results indicate that the post-mitotic neurons are particularly prone to accumulation of unrepaired DNA lesions potentially leading to progressive neurodegeneration. Nucleotide excision repair is the cellular pathway responsible for removing helix-distorting DNA damage and deficiency in such repair is found in a number of diseases with neurodegenerative phenotypes, including Xeroderma Pigmentosum and Cockayne syndrome. The main pathway for repairing oxidative base lesions is base excision repair, and such repair is crucial for neurons given their high rates of oxygen metabolism. Mismatch repair corrects base mispairs generated during replication and evidence indicates that oxidative DNA damage can cause this pathway to expand trinucleotide repeats, thereby causing Huntington's disease. Single-strand breaks are common DNA lesions and are associated with the neurodegenerative diseases, ataxia-oculomotor apraxia-1 and spinocerebellar ataxia with axonal neuropathy-1. DNA double-strand breaks are toxic lesions and two main pathways exist for their repair: homologous recombination and non-homologous end-joining. Ataxia telangiectasia and related disorders with defects in these pathways illustrate that such defects can lead to early childhood neurodegeneration. Aging is a risk factor for neurodegeneration and accumulation of oxidative mitochondrial DNA damage may be linked with the age-associated neurodegenerative disorders Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis. Mutation in the WRN protein leads to the premature aging disease Werner syndrome, a disorder that features neurodegeneration. In this article we review the evidence linking deficiencies in the DNA repair pathways with neurodegeneration.


A water-stable EuIII-based MOF as a dual-emission luminescent sensor for discriminative detection of nitroaromatic pollutants.
A water-stable EuIII-based metal-organic framework (MOF) with dual-emission luminescence behavior, namely {[Eu4(INO)5(μ3-OH)2Cl4(H2O)]·(NO3)·(H2O)5}n (Eu-MOF; HINO = isonicotinic acid N-oxide), was successfully constructed by the solvothermal reaction of Eu3+ ions with the organic ligand HINO. The cationic 3D framework contains microporous channels with accessible Lewis-base sites and NO3- ions as balanced anions, which all contribute to the selective detection of multifarious analytes. This MOF shows ratiometric detection of acetone, Cr2O72- ions, and nitroaromatic compounds (NACs). In particular, it shows great recognition of four NACs in water, representing the first LnIII-MOF which can display distinguishing fluorescence phenomena on NACs rather than relying on the quenching effect. Furthermore, this is also the first example of a MOF-sensor for detecting these explosives discriminatively by ratiometric methods. Additionally, the mechanisms for luminescent responses towards different analytes have been discussed in detail.


CYCLIC STRUCTURE OF ADENOVIRUS DNA.
The deoxyribonucleic acids of adenovirus types 2, 7, and 12 were extracted and examined in the electron microscope. Cyclic forms of the molecules were found to exist in each viral type. The lengths of cyclic strands extracted from the most dense fraction of purified particles of adenovirus type 2 were highly variable, averaging about 2.5 microns. This corresponds to a molecular weight of about 5 million.


Development of an Inosine Hyperproducer from Bacillus licheniformis by Systems Metabolic Engineering.
Inosine is widely used in food, chemical, and medicine. This study developed Bacillus licheniformis into an inosine hyperproducer through systems metabolic engineering. First, purine metabolism was activated by deleting inhibitors PurR and YabJ and overexpressing the pur operon. Then, the 5-phosphoribosyl-1-pyrophosphate (PRPP) supply was increased by optimizing the glucose transport system and pentose phosphate pathway, increasing the inosine titer by 97% and decreasing the titers of byproducts by 36%. Next, to prevent the degradation of inosine, genes deoD and pupG coding purine nucleoside phosphorylase were deleted, accumulating 0.91 g/L inosine in the culture medium. Additionally, the downregulation of adenosine 5'-monophosphate (AMP) synthesis pathway increased the inosine titer by 409%. Importantly, enhancing the glycine and aspartate supply increased the inosine titer by 298%. Finally, the guanosine synthesis pathway was blocked, leading to strain IR-8-2 producing 27.41 g/L inosine with a 0.46 g inosine/g glucose yield and a 0.38 g/(L·h) productivity in a shake flask.


Structure of the T109S mutant of Escherichia coli dihydroorotase complexed with the inhibitor 5-fluoroorotate: catalytic activity is reflected by the crystal form.
Crystals of a single-point mutant (T109S) of Escherichia coli dihydroorotase (DHOase) with diminished activity grown in the presence of L-dihydroorotate (L-DHO) are tetragonal, with a monomer in the asymmetric unit. These crystals are extremely unstable and disintegrate shortly after formation, which is followed by the growth of orthorhombic crystals from the remnants of the tetragonal crystals or at new nucleation sites. Orthorhombic crystals, for which a structure has previously been reported [Thoden et al. (2001), Biochemistry, 40, 6989-6997; Lee et al. (2005), J. Mol. Biol. 348, 523-533], contain a dimer of DHOase in the asymmetric unit; the active site of one monomer contains the substrate N-carbamyl-L-aspartate (L-CA-asp) and the active site of the other monomer contains the product of the reaction, L-DHO. In the subunit with L-DHO in the active site, a surface loop (residues 105-115) is 'open'. In the other subunit, with L-CA-asp in the active site, the loop folds inwards, forming specific hydrogen bonds from the loop to the L-CA-asp. The tetragonal crystal form can be stabilized by crystallization in the presence of the inhibitor 5-fluoroorotate (FOA), a product (L-DHO) mimic. Crystals of the complex of T109S DHOase with FOA are tetragonal, space group P4(1)2(1)2, with unit-cell parameters a = b = 72.6, c = 176.1 A. The structure has been refined to R and R(free) values of 0.218 and 0.257, despite severe anisotropy of the diffraction. In this structure, the flexible loops are both in the 'open' conformation, which is consistent with FOA, like L-DHO, binding at both sites. The behaviour of the T109S mutant crystals of DHOase in the presence of L-DHO is explained by initial binding of L-DHO to both subunits, followed by slow conversion to L-CA-asp, with consequent movement of the flexible loop and dissolution of the crystals. Orthorhombic crystals are then able to grow in the presence of L-DHO and L-CA-asp.


Prevalence of active and latent TB among inmates in a prison hospital in Bahia, Brazil.
To determine the prevalence of latent and active TB among detainees in a prison hospital in Bahia, Brazil. A cross-sectional study with prospective data collection was carried out comprising 237 inmates in the Bahia State Prison Hospital between July 2003 and April 2004. A standardized questionnaire was applied and completed by medical students. The detainees were systematically submitted to the following tests: tuberculin skin test, chest X-ray (anteroposterior), sputum smear microscopy and culture for mycobacteria. The events of interest were active TB and latent TB. The mean age of the participants was 36.6 years, and 89.9% were male. Smoking and alcohol consumption were reported by 70.0% and 43.9% of the inmates, respectively. A history of treatment for TB was reported by 11.3% of the inmates. Of the inmates evaluated, 36.3% reported cough and 31.4% reported expectoration. Other less common symptoms were asthenia (in 26.2%), weight loss (in 23.1%), loss of appetite (in 17.7%), fever (in 11.3%) and hemoptysis (in 6.7%). Of the 86 inmates tested, none presented positive HIV serology. The prevalence of latent TB was 61.5% (96 of the 156 inmates submitted to tuberculin skin tests), whereas that of active TB was 2.5% (6 of the 237 inmates evaluated). The presence of cough was a determinant of active TB (prevalence ratio = 8.8; 95% CI: 1.04-73.9; p = 0.025). Active and latent TB are highly prevalent among inmates hospitalized in the Bahia State Prison Hospital. Our findings justify the need to implement public policies specifically directed towards the control of TB in this population.


The molecular genetics of retinal photoreceptor proteins involved in cGMP metabolism.
Metabolism of cGMP is critically important for the functioning of phototransduction in the mammalian retina. In rod and cone photoreceptors, two types of antagonistic enzymes, guanylate cyclases and cGMP phosphodiesterases, carefully balance the available amount of the intracellular messenger. Guanylate cyclase produces cGMP and phosphodiesterase rapidly hydrolyzes cGMP upon bleaching of the photopigment. Regulation of their activity in light and dark, influence of Ca++, and feed-back mechanisms are currently under intense investigation. A molecular analysis on both the gene and protein levels will contribute significantly to our understanding of their respective roles in phototransduction. The two types of enzymes have been characterized molecularly to a very different extent. Rod phosphodiesterase was purified to homogeneity almost fifteen years ago, but photoreceptor guanylate cyclase has evaded all attempts for molecular characterization. Characterization of retinal guanylate cyclase cDNA(s), however, will most likely be achieved in the near future. Cone PDE was shown to be a distinct enzyme, different from, but related to, the rod enzyme. Molecular cloning has provided sequence information of two of the three subunits of rod PDE; the small inhibitory subunit has been expressed in bacterial expression vectors, giving us an elegant tool for exploring mechanisms of activation and inhibition. The gene encoding the alpha subunit was shown to be a member of a large gene family of cyclic nucleotide phosphodiesterases, present in many eucaryotes ranging from unicellular organisms (yeast) to mammals. While much has been achieved, many questions remain to be answered. The beta subunit of rod phosphodiesterase has evaded complete molecular characterization, and its origin (one gene and posttranslational modification of the gene product generating alpha and beta, alternative splicing, or two separate genes with distinct gene products) has not been elucidated. Mechanisms of interaction of subunits, activation and inhibition, the active site(s) of the enzyme are undefined. Virtually nothing is known about the molecular organization of the photoreceptor guanylate cyclase(s). Recent cloning of two apparently unrelated mammalian guanylate cyclases, however, containing a common homologous domain signals increasingly rapid progress in this field.


Far-ultraviolet astronomy on the astro-1 space shuttle mission.
The Astro-1 mission obtained observations related to a wide variety of current problems in astronomy during a 9-day flight of the space shuttle Columbia. Early results from one of the instruments, the Hopkins Ultraviolet Telescope, are reviewed here. Among these are new insights concerning the origin of the ultraviolet light from the old stellar population in elliptical galaxies, new evidence for a hot, gaseous corona surrounding the Milky Way, improved views of the physical conditions in active galactic nuclei, and a measurement of the ionization state of the local interstellar medium.


Polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive DNA sequences: strain variation in Cryptococcus neoformans.
Minisatellites and simple repetitive DNA sequence motifs are used as conventional oligonucleotide probes in DNA-hybridization-based fingerprinting. The same oligonucleotides can be used as single primers in the polymerase chain reaction (PCR) to generate individual PCR fingerprints. In this study, the simple repetitive sequences, (CA)8, (CT)8, (CAC)5, (GTG)5, (GACA)4 and (GATA)4, and a minisatellite core sequence derived from the wild-type phage M13 (5' GAGGGTGGCGGTTCT 3') were used as specific, single primers to amplify hypervariable repetitive DNA sequences during PCR analysis. The potential applications of this techniques are demonstrated with clinical isolates of the human pathogenic yeast, Cryptococcus neoformans. PCR fingerprint patterns have remained stable after long-term in vitro passage ( > 2 1/2 years to date). Hybridization of the primers to blots of electrophorectically separated chromosomes demonstrated that the target sequences recognized by most of the primers are dispersed through the entire yeast genome. Sequence analysis of the cloned bands obtained by PCR fingerprinting indicated that if the same or extremely similar, inversely oriented tandem repeats are located close to each other, when only one repeat-specific primer is used in the PCR, the region between these repeats is amplified. PCR fingerprinting has a wide range of current and potential applications to fungi, such as clarifying taxonomic questions, facilitating epidemiological studies and improving the diagnosis of mycotic diseases.


Computed tomography scans with intravenous contrast: low incidence of contrast-induced nephropathy in blunt trauma patients.
Computed tomography (CT) with intravenous (IV) contrast is an important step in the evaluation of the blunt trauma patient; however, the risk for contrast-induced nephropathy (CIN) in these patients still remains unclear. The goal of this study was to describe the rate of CIN in blunt trauma patients at a Level 1 trauma center and identify the risk factors of developing CIN. After internal review board approval, we reviewed our Level 1 trauma registry to identify blunt trauma patients admitted during a 1-year period. Chart review was used to identify patient demographics, creatinine levels, and vital signs. CIN was defined as an increase in creatinine by 0.5 mg/dL from admission after undergoing CT with IV contrast. Four percent of patients developed CIN during their admission following receipt of IV contrast for CT; 1% had continued renal impairment on discharge. No patients required dialysis during their admission. Diabetic patients had an increased rate of CIN, with 10% rate of CIN during admission and 4% at discharge. In multivariate analysis, only preexisting diabetes and Injury Severity Score (ISS) of greater than 25 were independently associated with risk for CIN. The rate of CIN in trauma patients following CT scan with IV contrast is low. Diabetes and ISS were independent risk factors of development of CIN in trauma patients. Epidemiologic/prognostic study, level III.


Evaluation of atrial fibrillation (AF) management and cardiovascular risk profile in AF patients: data from Turkish patients in the international observational cross-sectional REALISE AF trial.
To assess control of atrial fibrillation (AF) and cardiovascular (CV) risk profile of AF patients with previously established AF therapies. A total of 510 patients (mean (SD) age, 67.1 (12.3) years, 55.1% females) enrolled from 40 centers across Turkey were evaluated on a single-visit basis in terms of patient demographics, characteristics of underlying AF, the frequency and scoring of symptoms according to European Heart Rhythm Association AF cardiac symptoms classification, control of AF, cardiovascular (CV) risk profile, AF treatment and the consistency of current therapeutic practice with evidence-based guidelines. AF was controlled in 39.4% of patients based on sinus rhythm on the day of visit (10.2%) and AF with HR ≤80 bpm (29.2%). Permanent AF was the most commonly identified type of AF (56.0%). Symptoms were evident in 89.2% of patients either before V0 (78.8%) or at V0 (56.5%). Age (72.4%) and hypertension (70.0%) were the leading CV risk factors. Rate-control and rhythm-control strategies were chosen in 76.5 and 19.2% of patients at the enrollment visit. Mean (SD) of EQ-5D scores for VAS and for single index utility were 63.1 (19.8) and 0.62 (0.4), respectively. In this real life survey of AF patients from Turkey participating in the global contemporary, international, observational, cross-sectional REALISE AF survey, AF was determined to be not optimally controlled, leading patients to experience frequent symptoms, functional impairment and altered QoL,as well as frequent hospital admissions for cardiovascular events and a high requirement for procedures.


A leukotriene B4 receptor-2 is associated with paclitaxel resistance in MCF-7/DOX breast cancer cells.
Breast cancer is the most common malignancy in women. Although chemotherapeutic agents, such as paclitaxel, are effective treatments for the majority of breast cancer patients, recurrence is frequent and often leads to death. Thus, there is an urgent need to identify novel therapeutic targets that sensitise tumour cells to existing chemotherapy agents. The levels of leukotriene B4 receptor-2 (BLT2) in multidrug-resistant MCF-7/DOX cells were determined using quantitative PCR and FACS analysis. The potential role of BLT2 in the paclitaxel resistance of MCF-7/DOX cells was assessed using a pharmacological inhibitor and small interfering RNA knockdown, and the BLT2-associated resistance mechanism was assessed. The expression levels of BLT2 were markedly upregulated in MCF-7/DOX cells. The inhibition of BLT2 by pre-treatment with LY255283 or siBLT2 knockdown significantly sensitised MCF-7/DOX cells to paclitaxel and induced significant levels of apoptotic death, suggesting that BLT2 mediates paclitaxel resistance. We also demonstrated that BLT2-induced paclitaxel resistance was associated with the upregulation of P-glycoprotein. Finally, co-treatment with a BLT2 inhibitor and paclitaxel markedly reduced tumour growth in an MCF-7/DOX in vivo model. Together, our results demonstrate that BLT2 is a novel therapeutic target that sensitises drug-resistant breast cancer cells to paclitaxel.


Diffusion tensor imaging tractography in patients with intramedullary tumors: comparison with intraoperative findings and value for prediction of tumor resectability.
The aim of this retrospective study was to evaluate the predictive value of diffusion tensor (DT) imaging with respect to resectability of intramedullary spinal cord tumors and to determine the concordance of this method with intraoperative surgical findings. Diffusion tensor imaging was performed in 14 patients with intramedullary lesions of the spinal cord at different levels using a 3-T magnet. Routine MR imaging scans were also obtained, including unenhanced and enhanced T1-weighted images and T2-weighted images. Patients were classified according to the fiber course with respect to the lesion and their lesions were rated as resectable or nonresectable. These results were compared with the surgical findings (existence vs absence of cleavage plane). The interrater reliability was calculated using the kappa coefficient of Cohen. Of the 14 patients (7 male, 7 female; mean age 49.2 +/- 15.5 years), 13 had tumors (8 ependymomas, 2 lymphomas, and 3 astrocytoma). One lesion was proven to be a multiple sclerosis plaque during further diagnostic workup. The lesions could be classified into 3 types according to the fiber course. In Type 1 (5 cases) fibers did not pass through the solid lesion. In Type 2 (3 cases) some fibers crossed the lesion, but most of the lesion volume did not contain fibers. In Type 3 (6 cases) the fibers were completely encased by tumor. Based on these results, 6 tumors were considered resectable, 7 were not. During surgery, 7 tumors showed a good cleavage plane, 6 did not. The interrater reliability (Cohen kappa) was calculated as 0.83 (p < 0.003), which is considered to represent substantial agreement. The mean duration of follow-up was 12.0 +/- 2.9. The median McCormick grade at the end of follow-up was II. These preliminary data suggest that DT imaging in patients with spinal cord tumors is capable of predicting the resectability of the lesion. A further prospective study is needed to confirm these results and any effect on patient outcome.


[Informative significance of immunological studies in endocrine diseases in children].
The authors measured the content of circulating immune complexes (CIC) and serum immunoglobulins in children with diabetes mellitus and thyroid pathology. Diabetes mellitus was marked by the increase of the content of CIC and IgA and IgM. The steady rise of the blood content of CIC in patients with diabetes mellitus is a prognostically unfavourable sign of the development of microangiopathies. Children with thyroid diseases manifested dysimmunoglobulinemia and the rise of CIC content in blood. The IgG/IgM ratio can be used as the differential-diagnostic test of thyroid diseases.


Development of a sampling-based global sensitivity analysis workflow for multiscale computational cancer models.
There are two challenges that researchers face when performing global sensitivity analysis (GSA) on multiscale 'in silico' cancer models. The first is increased computational intensity, since a multiscale cancer model generally takes longer to run than does a scale-specific model. The second problem is the lack of a best GSA method that fits all types of models, which implies that multiple methods and their sequence need to be taken into account. In this study, the authors therefore propose a sampling-based GSA workflow consisting of three phases - pre-analysis, analysis and post-analysis - by integrating Monte Carlo and resampling methods with the repeated use of analysis of variance; they then exemplify this workflow using a two-dimensional multiscale lung cancer model. By accounting for all parameter rankings produced by multiple GSA methods, a summarised ranking is created at the end of the workflow based on the weighted mean of the rankings for each input parameter. For the cancer model investigated here, this analysis reveals that extracellular signal-regulated kinase, a downstream molecule of the epidermal growth factor receptor signalling pathway, has the most important impact on regulating both the tumour volume and expansion rate in the algorithm used.


Projecting ESRD Incidence and Prevalence in the United States through 2030.
Population rates of obesity, hypertension, diabetes, age, and race can be used in simulation models to develop projections of ESRD incidence and prevalence. Such projections can inform long-range planning for ESRD resources needs. We used an open compartmental simulation model to estimate the incidence and prevalence of ESRD in the United States through 2030 on the basis of wide-ranging projections of population obesity and ESRD death rates. Population trends in age, race, hypertension, and diabetes were on the basis of data from the Centers for Disease Control and Prevention's National Health and Nutrition Examination Survey and the US Census. The increase in ESRD incidence rates within age and race groups has leveled off and/or declined in recent years, but our model indicates that population changes in age and race distribution, obesity and diabetes prevalence, and ESRD survival will result in a 11%-18% increase in the crude incidence rate from 2015 to 2030. This incidence trend along with reductions in ESRD mortality will increase the number of patients with ESRD by 29%-68% during the same period to between 971,000 and 1,259,000 in 2030. The burden of ESRD will increase in the United States population through 2030 due to demographic, clinical, and lifestyle shifts in the population and improvements in RRT. Planning for ESRD resource allocation should allow for substantial continued growth in the population of patients with ESRD. Future interventions should be directed to preventing the progression of CKD to kidney failure.


Albumin in critically ill patients: controversies and recommendations.
Human albumin has been used as a therapeutic agent in intensive care units for more than 50 years. However, clinical studies from the late 1990s described possible harmful effects in critically ill patients. These studies' controversial results followed other randomized controlled studies and meta-analyses that showed no harmful effects of this colloid solution. In Brazil, several public and private hospitals comply with the Agência Nacional de Vigilância Sanitária (the Brazilian Health Surveillance Agency) recommendations for appropriate administration of intravenous albumin. This review discusses indications for albumin administration in critically ill patients and analyzes the evidence for metabolic and immunomodulatory effects of this colloid solution. We also describe the most significant studies from 1998 to the present time; these reveal an absence of incremental mortality from intravenous albumin administration as compared to crystalloid solutions. The National Health Surveillance Agency indications are discussed relative to the current body of evidence for albumin use in critically ill patients.


NFAT primes the human RORC locus for RORγt expression in CD4+ T cells.
T helper 17 (Th17) cells have crucial functions in mucosal immunity and the pathogenesis of several chronic inflammatory diseases. The lineage-specific transcription factor, RORγt, encoded by the RORC gene modulates Th17 polarization and function, as well as thymocyte development. Here we define several regulatory elements at the human RORC locus in thymocytes and peripheral CD4+ T lymphocytes, with CRISPR/Cas9-guided deletion of these genomic segments supporting their role in RORγt expression. Mechanistically, T cell receptor stimulation induces cyclosporine A-sensitive histone modifications and P300/CBP acetylase recruitment at these elements in activated CD4+ T cells. Meanwhile, NFAT proteins bind to these regulatory elements and activate RORγt transcription in cooperation with NF-kB. Our data thus demonstrate that NFAT specifically regulate RORγt expression by binding to the RORC locus and promoting its permissive conformation.


Methotrexate in combination with infliximab is no more effective than infliximab alone in patients with Crohn's disease.
Methotrexate and infliximab are effective therapies for Crohn's disease (CD). In the combination of maintenance methotrexate-infliximab trial, we evaluated the potential superiority of combination therapy over infliximab alone. In a 50-week, double-blind, placebo-controlled trial, we compared methotrexate and infliximab with infliximab alone in 126 patients with CD who had initiated prednisone induction therapy (15-40 mg/day) within the preceding 6 weeks. Patients were assigned randomly to groups given methotrexate at an initial weekly dose of 10 mg, escalating to 25 mg/week (n = 63), or placebo (n = 63). Both groups received infliximab (5 mg/kg of body weight) at weeks 1, 3, 7, and 14, and every 8 weeks thereafter. Prednisone was tapered, beginning at week 1, and discontinued no later than week 14. The primary outcome was time to treatment failure, defined as a lack of prednisone-free remission (CD Activity Index, <150) at week 14 or failure to maintain remission through week 50. Patients' baseline characteristics were similar between groups. By week 50, the actuarial rate of treatment failure was 30.6% in the combination therapy group compared with 29.8% in the infliximab monotherapy group (P = .63; hazard ratio, 1.16; 95% confidence interval, 0.62-2.17). Prespecified subgroup analyses failed to show a benefit in patients with short disease duration or an increased level of C-reactive protein. No clinically meaningful differences were observed in secondary outcomes. Combination therapy was well tolerated. The combination of infliximab and methotrexate, although safe, was no more effective than infliximab alone in patients with CD receiving treatment with prednisone. ClincialTrials.gov number, NCT00132899.


Sunitinib in metastatic renal cell carcinoma: recommendations for management of noncardiovascular toxicities.
The multitargeted tyrosine-kinase inhibitor sunitinib has emerged as one of the standards of care for good- and intermediate-risk metastatic renal cell carcinoma. Although generally associated with acceptable toxicity, sunitinib exhibits a novel and distinct toxicity profile that requires monitoring and management. Fatigue, diarrhea, anorexia, oral changes, hand-foot syndrome and other skin toxicity, thyroid dysfunction, myelotoxicity, and hypertension seem to be the most common and clinically relevant toxicities of sunitinib. Drug dosing and treatment duration are correlated with response to treatment and survival. Treatment recommendations for hypertension have been published but, currently, no standard guidelines exist for the management of noncardiovascular side effects. To discuss the optimal management of noncardiovascular side effects, an international, interdisciplinary panel of experts gathered in November 2009. Existing literature on incidence, severity, and underlying mechanisms of side effects as well as on potential treatment options were carefully reviewed and discussed. On the basis of these proceedings and the thorough review of the existing literature, recommendations were made for the monitoring, prevention, and treatment of the most common noncardiovascular side effects and are summarized in this review. The proactive assessment and consistent and timely management of sunitinib-related side effects are critical to ensure optimal treatment benefit by allowing appropriate drug dosing and prolonged treatment periods.


Use and Cost-Effectiveness of a Telehealth Service at a Centralized COVID-19 Quarantine Center in Taiwan: Cohort Study.
Telehealth is a recommended method for monitoring the progression of nonsevere infections in patients with COVID-19. However, telehealth has not been widely implemented to monitor SARS-CoV-2 infection in quarantined individuals. Moreover, studies on the cost-effectiveness of quarantine measures during the COVID-19 pandemic are scarce. In this cohort study, we aimed to use telehealth to monitor COVID-19 infections in 217 quarantined Taiwanese travelers and to analyze the cost-effectiveness of the quarantine program. Travelers were quarantined for 14 days at the Taiwan Yangmingshan quarantine center and monitored until they were discharged. The travelers' clinical symptoms were evaluated twice daily. A multidisciplinary medical team used the telehealth system to provide timely assistance for ill travelers. The cost of the mandatory quarantine was calculated according to data from the Ministry of Health and Welfare of Taiwan. All 217 quarantined travelers tested negative for SARS-CoV-2 upon admission to the quarantine center. During the quarantine, 28/217 travelers (12.9%) became ill and were evaluated via telehealth. Three travelers with fever were hospitalized after telehealth assessment, and subsequent tests for COVID-19 were negative for all three patients. The total cost incurred during the quarantine was US $193,938, which equated to US $894 per individual. Telehealth is an effective instrument for monitoring COVID-19 infection in quarantined travelers and could help provide timely disease management for people who are ill. It is imperative to screen and quarantine international travelers for SARS-CoV-2 infection to reduce the nationwide spread of COVID-19.


Real life experiences in HCV management in 2018.
Hepatitis C virus (HCV) infection is a major cause of chronic liver disease, with approximately 71 million chronically infected individuals worldwide. Treatment of chronic hepatitis C has considerably improved in the last few years thanks to the introduction of direct-acting antivirals able to achieve sustained virological response in more than 95% of patients. Successful anti-HCV treatment can halt liver disease progression and solve the HCV-related extra-hepatic manifestations, eventually reducing liver-related and overall mortality. Areas covered: With the aim to respond to unmet needs in patient's identification, universal access to antiviral therapy and treatment optimization in specific setting of HCV-infected patients, a group of Italian experts met in Stresa in May 2018. The summary of the considerations arising from this meeting and the final statements are reported in this paper. Expert commentary: All the advances on HCV cure may have a real clinical impact not only in individual patients but also at the social health level if they are applied to all infected patients, independently from the stage of liver disease. Further improvements are needed in order to attain HCV elimination, such as the development of an enhanced screening program working in parallel to the present treatment options.


[Development and comparison of real-time and conventional RT-PCR assay for detection of human coronavirus NL63 and HKU1].
We designed specific primers and fluorescence-labeled probes to develop real-time and conventional RT-PCR assays for detection of human coronavirus NL63 or HKU1. Subsequently, experiments were undertaken to assess diagnostic criteria such as specificity, sensitivity and reproducibility. The detection limit of the real-time RT-PCR assays was 10 RNA copies per reaction mixture. No cross-reactivity was observed between RNA samples derived from designed HCoV and other HCoV or human metapneumovirus. A total of 158 nasopharyngeal swab specimens collected from adult patients with acute respiratory tract infection in Beijing were screened for the presence of human coronavirus NL63 and HKU1 by using real-time RT-PCR and conventional RT-PCR method. The fluorescence quantitative RT-PCR method detected six specimens positive for human coronavirus NL63, five specimens positive for human coronavirus HKU1; and conventional RT-PCR method detected three HCoV-NL63 positive and three HCoV-HKU1 positive, respectively. The convention RT-PCR products of positive samples were obtained and sequence analysis confirmed the reliability of the above methods. In summary, the real-time RT-PCR assay for HCoV- NL63 or HKU1 was more sensitive than conventional RT-PCR and with less time (less than 4 hours) for completion. It may be suitable for molecular epidemiological surveillance and clinical diagnosis for human coronavirus NL63 and HKU1.


Gluconeogenesis from labeled carbon: estimating isotope dilution.
To estimate the rate of gluconeogenesis from steady-state incorporation of labeled 3-carbon precursors into glucose, isotope dilution must be considered so that the rate of labeling of glucose can be quantitatively converted to the rate of gluconeogenesis. An expression for the value of this isotope dilution can be derived using mathematical techniques and a model of the tricarboxylic acid (TCA) cycle. The present investigation employs a more complex model than that used in previous studies. This model includes the following pathways that may affect the correction for isotope dilution: 1) flux of 3-carbon precursor to the oxaloacetate pool via acetyl-CoA and the TCA cycle; 2) flux of 4- or 5-carbon compounds into the TCA cycle; 3) reversible flux between oxaloacetate (OAA) and pyruvate and between OAA and fumarate; 4) incomplete equilibrium between OAA pools; and 5) isotope dilution of 3-carbon tracers between the experimentally measured pool and the precursor for the TCA-cycle OAA pool. Experimental tests are outlined which investigators can use to determine whether these pathways are significant in a specific steady-state system. The study indicated that flux through these five pathways can significantly affect the correction for isotope dilution. To correct for the effects of these pathways an alternative method for calculating isotope dilution is proposed using citrate to relate the specific activities of acetyl-CoA and OAA.


Management of ureteral stones with pneumatic lithotripsy: report of 500 patients.
To evaluate the results of pneumatic lithotripsy (PL) with ureteroscopy in the treatment of ureteral stones. We reviewed, retrospectively, the records of 500 patients (366 male, 134 female) with ureteral calculi treated by PL with the Swiss Lithoclast. Of these patients, 124 (24.8%) were treated primarily and 376 (75.2%) were treated secondarily after unsuccessful extracorporeal shockwave lithotripsy (SWL). The results were evaluated 3 months after treatment by excretory urography, ultrasonography, or both. The over-all stone-free and fragmentation rates were 94.6% and 96.8%, respectively. These values were 97.1% and 98.5% for stones <or=10 mm and 83.7% and 89.1% for stones >10 mm, respectively. The main complications were stone migration (2.0%), urosepsis (3.0%), and ureteral perforation (1.4%). While SWL is generally excepted as a first-line treatment option in ureteral stones because of its noninvasive nature, in situ lithotripsy, and especially PL, has higher success rates with minimal morbidity. Thus, PL seems to be a good alternative in patients in whom SWL was unsuccessful or not indicated and in patients who need early stone removal.


Machine learning does not outperform traditional statistical modelling for kidney allograft failure prediction.
Machine learning (ML) models have recently shown potential for predicting kidney allograft outcomes. However, their ability to outperform traditional approaches remains poorly investigated. Therefore, using large cohorts of kidney transplant recipients from 14 centers worldwide, we developed ML-based prediction models for kidney allograft survival and compared their prediction performances to those achieved by a validated Cox-Based Prognostication System (CBPS). In a French derivation cohort of 4000 patients, candidate determinants of allograft failure including donor, recipient and transplant-related parameters were used as predictors to develop tree-based models (RSF, RSF-ERT, CIF), Support Vector Machine models (LK-SVM, AK-SVM) and a gradient boosting model (XGBoost). Models were externally validated with cohorts of 2214 patients from Europe, 1537 from North America, and 671 from South America. Among these 8422 kidney transplant recipients, 1081 (12.84%) lost their grafts after a median post-transplant follow-up time of 6.25 years (Inter Quartile Range 4.33-8.73). At seven years post-risk evaluation, the ML models achieved a C-index of 0.788 (95% bootstrap percentile confidence interval 0.736-0.833), 0.779 (0.724-0.825), 0.786 (0.735-0.832), 0.527 (0.456-0.602), 0.704 (0.648-0.759) and 0.767 (0.711-0.815) for RSF, RSF-ERT, CIF, LK-SVM, AK-SVM and XGBoost respectively, compared with 0.808 (0.792-0.829) for the CBPS. In validation cohorts, ML models' discrimination performances were in a similar range of those of the CBPS. Calibrations of the ML models were similar or less accurate than those of the CBPS. Thus, when using a transparent methodological pipeline in validated international cohorts, ML models, despite overall good performances, do not outperform a traditional CBPS in predicting kidney allograft failure. Hence, our current study supports the continued use of traditional statistical approaches for kidney graft prognostication.


Attenuating posttraumatic distress with omega-3 polyunsaturated fatty acids among disaster medical assistance team members after the Great East Japan Earthquake: the APOP randomized controlled trial.
On March 11, 2011, a magnitude 9.0 earthquake, the most powerful ever recorded in Japan, and a massive tsunami struck off the coast of the Sanriku region. A Disaster Medical Assistance Team, a mobile medical team with specialized training that is deployed during the acute phase of a disaster, was dispatched to areas with large-scale destruction and multiple injured and sick casualties. Previous studies have reported critical incident stress (i.e. posttraumatic stress disorder symptoms and depressive symptoms) among rescue workers as well as the need for screening and prevention for posttraumatic stress disorder. So far we have shown in an open trial that posttraumatic stress disorder symptoms in critically injured patients can be reduced by taking omega-3 fatty acids intended to stimulate hippocampal neurogenesis. This study is designed to determine the effectiveness of attenuating posttraumatic distress with omega-3 polyunsaturated fatty acids among Disaster Medical Assistance Team members after the Great East Japan Earthquake, and is named the APOP randomized controlled trial which is currently ongoing. First, we will provide psycho-education on posttraumatic distress, which is common in responders to the Disaster Medical Assistance Team members deployed to the disaster area. Second, observational research will be conducted to evaluate critical incident stress following the completion of medical activities. Third, team members who provide consent to participate in the intervention research will be randomly divided into a group given an omega-3 fatty acid supplement and a group not given the supplements. Outcome will be evaluated at 12 weeks after the supplements are shipped to the team members. Measures that address critical incident stress in disaster responders are important, but there is no substantial evidence that links such measures with prevention of posttraumatic stress disorder. Thus, any confirmation through this study that the intake of omega-3 fatty acid supplements serves as a simple preventative measure for critical incident stress will be of great significance. UMIN Clinical Trials Registry, UMIN000005367.


Macrophages Shed Excess Cholesterol in Unique Extracellular Structures Containing Cholesterol Microdomains.
Cells use various mechanisms to maintain cellular cholesterol homeostasis including efflux of cholesterol from the cellular plasma membrane to cholesterol acceptors such as HDLs (high-density lipoproteins). Little is known about the transfer of cholesterol from cells into the extracellular matrix. Using a unique monoclonal antibody that detects ordered cholesterol arrays (ie, cholesterol micro[or nano]-domains), we previously identified that particles containing these cholesterol domains accumulate in the extracellular matrix during cholesterol enrichment of human monocyte-derived macrophages and are found in atherosclerotic lesions. In this study, we further investigate these deposited particles containing cholesterol microdomains and discover their unexpected morphology. Although appearing spherical at the resolution of the conventional fluorescence microscope, super-resolution immunofluorescence and atomic force microscopy of in situ cholesterol microdomains, and immunoelectron microscopy of isolated cholesterol microdomains revealed that the microdomains are not vesicles or 3-dimensional crystals but rather appear as branching irregularly shaped deposits of varying size. These cholesterol microdomain-containing deposits are shed from the plasma membrane into the extracellular matrix. To date, research on cellular excretion of excess cholesterol has demonstrated cellular cholesterol efflux in the form of membranous vesicles and discoidal HDL particles released into the fluid-phase medium. Shedding of plasma membrane cholesterol microdomains provides an additional mechanism for cells such as macrophages to maintain plasma membrane cholesterol homeostasis. Furthermore, recognition that macrophages shed cholesterol microdomains into the extracellular matrix is important to our understanding of extracellular buildup of cholesterol in atherosclerosis.


Chlorhexidine preserves the hybrid layer in vitro after 10-years aging.
The present study investigated the ability of a chlorhexidine (CHX)-containing primer (0.2% aqueous solution) to inhibit dentinal enzymes, preserve the hybrid layer (HL) and remain within the HL, after 10 years of aging in artificial saliva at 37°C. Non-carious extracted molars were assigned to two groups, cut into slabs exposing middle/deep dentin, etched and bonded with Adper Scotchbond 1XT (SB1XT) with or without 0.2% CHX aqueous solution pretreatment. Composite build-ups were made, and the specimens were cut in 1-mm thick bonded sticks. In situ zymography was performed on freshly prepared specimens (T0) and specimens aged for 10 years (T10-yr) at 37°C in artificial saliva, to investigate endogenous gelatinolytic activity within the HL. At T10-yr, specimens were also decalcified and embedded in epoxy resin for TEM analysis. Micro-Raman spectroscopy was performed at T0 and T10-yr to evaluate the chemical profiles in intertubular dentin and the HL. In situ zymography showed less pronounced enzymatic activity in the CHX-pretreated group (p<0.05) regardless of aging, maintaining a similar level of fluorescence at T0 and T10-yr (p>0.05). TEM results showed that 98% of the HL had been degraded in the control group, while 95% of the HL was intact in the experimental group. Moreover, all the Raman spectra peaks assigned to CHX could be identified only in the CHX-pretreated group (T0 and T10-yr). In vitro, CHX remains in the HL after 10 years with its inhibitory effect preserved. This may be the underlying factor for HL preservation after this long aging period.


Compressive Myelopathy in Congenital Kyphosis of the Upper Thoracic Spine: A Retrospective Study of 6 Cases.
A retrospective study. The goal of this retrospective study was to describe the uncommon presentation of neurological deficits in patients with congenital kyphosis of the upper thoracic spine (T1-T4). Congenital kyphosis is an uncommon deformity but can potentially lead to spinal cord compression and paraplegia, particularly in type I (failure of formation) deformities. Few reports have described compressive myelopathy associated with congenital kyphosis of the upper thoracic spine. Six patients with congenital kyphosis of the upper thoracic spine, including 2 adults and 4 pediatric patients, developed progressive or sudden onset of paraplegia. Angles of kyphosis ranged from 75 to 120 degrees. Magnetic resonance imaging demonstrated spinal cord thinning and compression at the kyphotic apex in all patients. All patients underwent decompressive and correctional surgery by single-stage posterior vertebral column resection or 2-stage anterior corpectomy fusion and posterior fixation. Neurological status was evaluated using the ASIA impairment classification and the motor score. Postoperatively, all patients had 25%-80% correction of kyphosis. All patients improved neurologically between 0 and 2 ASIA scales after surgery. Among them, an adolescent patient presenting as acute ASIA A improved to ASIA E within 1 year after surgery. Another adolescent patient deteriorated from preoperative ASIA C to ASIA A in the immediate postoperative period but improved to ASIA D within 1 year after surgery. Congenital kyphosis of the upper thoracic spine has a high incidence of compressive myelopathy. Duration from onset of paraplegia to surgical intervention and severity of preoperative paraplegia are 2 key factors in determining neurological prognosis after surgery.


Suppression of serum thyrotropin with thyroxine in patients with Graves' disease: effects on recurrence of hyperthyroidism and thyroid volume.
Based on findings that thyroxine may have a beneficial effect on the recurrence of Graves' hyperthyroidism, we prospectively studied the effects of a TSH suppressive treatment with thyroxine on the course of Graves' disease in fifty patients with recent onset of hyperthyroidism. After the normalization of serum tri-iodothyronine (T3) and thyroxine (T4) concentrations, one group of patients was randomly assigned to a combined treatment with carbimazole and a TSH suppressive dose of T4 for 12 months, followed by another 12 months of TSH suppressive therapy alone. The other group of patients also received carbimazole for one year, but T4 was only added as indicated to normalize elevated TSH serum concentrations, and patients received no therapy during the second year. By the end of the second year, a relapse of hyperthyroidism had occurred in 43% of the patients with and in 45% of the patients without suppressive T4 treatment. In those patients without a relapse of hyperthyroidism, initial thyroid size significantly (P = 0.01) decreased with time in both treatment groups. However, patients on suppressive T4 treatment tended to have a greater reduction in thyroid volume than patients with normal TSH serum concentrations (P = 0.05). In conclusion, we were unable to detect a preventive effect of exogenous TSH suppression on the recurrence of hyperthyroidism. However, our data suggest that TSH suppressive treatment may have a beneficial effect on thyroid enlargement in Graves' disease.


Live single cell imaging assays in glass microwells produced by laser-induced deep etching.
Miniaturization of cell culture substrates enables controlled analysis of living cells in confined micro-scale environments. This is particularly suitable for imaging individual cells over time, as they can be monitored without escaping the imaging field-of-view (FoV). Glass materials are ideal for most microscopy applications. However, with current methods used in life sciences, glass microfabrication is limited in terms of either freedom of design, quality, or throughput. In this work, we introduce laser-induced deep etching (LIDE) as a method for producing glass microwell arrays for live single cell imaging assays. We demonstrate novel microwell arrays with deep, high-aspect ratio wells that have rounded, dimpled or flat bottom profiles in either single-layer or double-layer glass chips. The microwells are evaluated for microscopy-based analysis of long-term cell culture, clonal expansion, laterally organized cell seeding, subcellular mechanics during migration and immune cell cytotoxicity assays of both adherent and suspension cells. It is shown that all types of microwells can support viable cell cultures and imaging with single cell resolution, and we highlight specific benefits of each microwell design for different applications. We believe that high-quality glass microwell arrays enabled by LIDE provide a great option for high-content and high-resolution imaging-based live cell assays with a broad range of potential applications within life sciences.


Contributing Factors and Mental Health Outcomes of First Suicide Attempt During Childhood and Adolescence: Results From a Nationally Representative Study.
To investigate whether risk factors for suicide attempts differ in children and adolescents and to categorize adulthood mental health outcomes of child and adolescent suicide attempters in the general population. Using a large (N = 34,653), nationally representative US adult sample, the 2004-2005 National Epidemiologic Survey on Alcohol and Related Conditions, we examined whether individuals who first attempted suicide during childhood (under the age of 13 years) differ from those who first attempted suicide during adolescence (13 through 17 years) in (1) contributing factors for first suicide attempt, including mental disorders and traumatic experiences that occurred before the first suicide attempt, parental history of mental disorders, and family poverty and (2) adulthood mental health outcomes, including lifetime and current prevalence of DSM-IV psychiatric disorders and quality of life measures. Suicide attempts during childhood (n = 104) were more strongly related to childhood maltreatment, while suicide attempts during adolescence (n = 415) were more strongly associated with major depressive episode. Compared to first suicide attempts during adolescence, first attempts during childhood were associated with increased risk for multiple suicide attempts (61.3% vs 32.6%), several psychiatric disorders (mania, hypomania, and panic disorder), and poorer social functioning during adulthood (all P values < .05). Suicide attempts in children and adolescents substantially differ in contributing factors and adulthood mental health outcomes. Preventing childhood maltreatment and early intervention for psychiatric disorders may have broad benefits to reduce not only the suffering of these children and adolescents, but also the burden of suicide.


Gastrin-releasing peptide: a potential growth factor expressed in human neuroblastoma tumors(1).
PURPOSE:Gastrin-releasing peptide (GRP) is a 27-amino acid neuropeptide that has been identified in the cytoplasm of many neuroendocrine tumors. Gastrin releasing peptide has been labeled as an autocrine growth factor in small cell lung carcinomas. Recent work has also shown this to be true in the growth of neuroblastoma cells in vitro. The purpose of this study was to demonstrate GRP and its receptor (GRP-R) in resected human neuroblastomas and to correlate the presence or absence with other known predictors of poor prognosis.To demonstrate the presence of GRP and GRP-R mRNA, total RNA was extracted from human neuroblastoma cells. A reverse transcription-polymerase chain reaction (RT-PCR) was then performed using specific primers. The products of the RT-PCR were then confirmed to be GRP and GRP-R cDNA by Southern blot analysis. The RT-PCR products were then sequenced, and these sequences were compared with the know sequences of GRP and GRP-R DNA.N = 19. GRP and GRP-R mRNA were present in all neuroblastoma specimens. Although no correlation with other known predictors of poor prognosis existed, transcripts of four different sizes (400, 450, 500, and 950 bp) were seen in the GRP-R transcripts. The sequences of the 950 bp-sized transcript reverse transcription PCR products were identical to the known GRP-R.We conclude that gastrin releasing peptide and gastrin releasing peptide receptor mRNA are present in all human neuroblastomas. Although qualitatively it appears to lack prognostic significance, its ubiquitous nature in the tumor suggests it may be a useful target on which to base future treatment modalities.


Design and Evaluation Process of a Personal and Motive-Based Competencies Questionnaire in Spanish-Speaking Contexts.
Most questionnaires used for managerial purposes have been developed in Anglo-Saxon countries and then adapted for other cultures. However, this process is controversial. This paper fills the gap for more culturally sensitive assessment instruments in the specific field of human resources while also addressing the methodological issues that scientists and practitioners face in the development of questionnaires. First, we present the development process of a Personal and Motive-based competencies questionnaire targeted to Spanish-speaking countries. Second, we address the validation process by guiding the reader through testing the questionnaire construct validity. We performed two studies: a first study with 274 experts and practitioners of competency development and a definitive study with 482 members of the general public. Our results support a model of nineteen competencies grouped into four higher-order factors. To assure valid construct comparisons we have tested the factorial invariance of gender and work experience. Subsequent analysis have found that women self-rate themselves significantly higher than men on only two of the nineteen competencies, empathy (p < .001) and service orientation (p < .05). The effect of work experience was significant in twelve competencies (p < .001), in which less experienced workers self-rate higher than experienced workers. Finally, we derive theoretical and practical implications.


Technical training in surgery: the trainee's view.
A questionnaire designed to investigate the technical training of general surgical trainees was distributed throughout Scotland. A total of 222 questionnaires were sent out and 179 (81 per cent) were returned. Of the responders, 38 per cent felt that the overall operative workload was too small and the majority thought that there was too little emphasis on supervised operating. Two-thirds of trainees were generally satisfied with the amount of unsupervised operating, although a substantial proportion (21 per cent) sometimes felt out of their depth when performing delegated emergency operations. Few had attended craft workshops or travelled to other centres, but of those who had, most considered both activities to be very useful. Photographic atlases and videos were not generally felt to be of great value by those who had used them. The majority of trainees did not believe that research activity had interfered excessively with their technical training.


Proline provides a nitrogen source in the retinal pigment epithelium to synthesize and export amino acids for the neural retina.
It is known that metabolic defects in the retinal pigment epithelium (RPE) can cause degeneration of its neighboring photoreceptors in the retina, leading to retinal degenerative diseases such as age-related macular degeneration. However, how RPE metabolism supports the health of the neural retina remains unclear. The retina requires exogenous nitrogen sources for protein synthesis, neurotransmission, and energy metabolism. Using 15N tracing coupled with mass spectrometry, we found human RPE can utilize the nitrogen in proline to produce and export 13 amino acids, including glutamate, aspartate, glutamine, alanine, and serine. Similarly, we found this proline nitrogen utilization in the mouse RPE/choroid but not in the neural retina of explant cultures. Coculture of human RPE with the retina showed that the retina can take up the amino acids, especially glutamate, aspartate, and glutamine, generated from proline nitrogen in the RPE. Intravenous delivery of 15N proline in vivo demonstrated 15N-derived amino acids appear earlier in the RPE before the retina. We also found proline dehydrogenase, the key enzyme in proline catabolism is highly enriched in the RPE but not the retina. The deletion of proline dehydrogenase blocks proline nitrogen utilization in RPE and the import of proline nitrogen-derived amino acids in the retina. Our findings highlight the importance of RPE metabolism in supporting nitrogen sources for the retina, providing insight into understanding the mechanisms of the retinal metabolic ecosystem and RPE-initiated retinal degenerative diseases.


[Nycthemeral gastric secretion in children under parenteral feeding].
The nycthemeral basal gastric secretion was studied in 11 children under continuous or cyclic total parenteral feeding (TPF). The suppression of the prandial stimulation shows nycthemeral changes in the secretory flow with a nocturnal minimum and a diurnal morning maximum, independent of the TPF infusion-conditions. The role of the infused aminoacids and lipid emulsion, as well as the duration of TPF is suggested to explain the increase or decrease in the basal acid flow. The authors suggest that an H2 antihistaminic should be used if the morning secretory peak induces a significant functional impairment.


Python-assisted detection and photothermal inactivation of Salmonella typhimurium and Staphylococcus aureus on a background-free SERS chip.
In this study, a background-free surface-enhanced Raman scattering (SERS) chip with a sandwich configuration was fabricated to enable reliable detection and photothermal inactivation of multiple bacteria. The SERS chip consists of a graphene-coated, phenylboronic-modified plasmonic gold substrate (pAu/G/PBA), and two aptamer-functionalized core (gold)-shell (Prussian blue/Poly-L-lysine and 4-mercaptobenzonitrile/polydopamine) SERS tags (Au@PB@PLL@Apt and Au@MB@PDA@Apt). The detection signals rely on the characteristic and nonoverlapping Raman bands of the SERS tags within the Raman-silent region (1800-2800 cm-1), where no background signals from the sample matrix are observed, leading to improved detection sensitivity and accuracy. Considering the relatively large size of bacteria (e.g., micron level), a rapid Raman mapping technique was chosen over conventional point-scan methods to achieve more reliable quantitative analysis of bacteria. This technique involves collecting and analyzing intensity signals of SERS tags from all the scattering points with an average ensemble effect, which is facilitated by the use of Python. As a proof-of-concept, model bacterium of Salmonella typhimurium and Staphylococcus aureus were successfully detected using the SERS chip with a dynamic range of 10-107 CFU/mL. Additionally, the SERS chip demonstrated successful detection of these bacteria in whole blood samples. Moreover, the photothermal effect of pAu/G led to efficient bacteria elimination, achieving approximately 100% eradication. This study integrated a background-free SERS chip with a Python-assisted rapid Raman mapping technique, resulting in a reliable, rapid and accurate method for detecting and eliminating multiple bacteria, which may provide a promising alternative for multiple screening of bacteria in real samples.


Surface EMG during the Push-up plus Exercise on a Stable Support or Swiss Ball: Scapular Stabilizer Muscle Exercise.
[Purpose] Scapular stabilizer strengthening exercise is crucial for shoulder rehabilitation. The purpose of this study was to compare two types of push-up plus exercises, on a stable and unstable bases of support, using surface electromyography (EMG), to suggest an effective shoulder rehabilitation program. [Subjects and Methods] Ten healthy men volunteered for this study. All volunteers performed two sets of push-up plus exercise (standard push up and knee push up) on stable and unstable bases of support. The muscle activities of five important scapular stabilizer muscles (upper trapezius, middle trapezius, lower trapezius, serratus anterior, latissimus dorsi) were recorded during the exercise. [Results] The upper trapezius showed greater mean electric activation amplitude in the scapular retraction posture than in the scapular protraction posture, and the serratus anterior showed greater mean electric activation amplitude in the scapular protraction posture than in the scapular retraction posture. The root-mean-square normalized EMG values of the muscles were greater during the exercise performed on the unstable support than those on the stable support. [Conclusion] The standard push-up plus exercise on an unstable base of support helps to increase muscle activity, especially those of the upper/middle trapezius and serratus anterior.


Inhaled furosemide in the treatment of acute exacerbations of asthma
Inhalation of the loop diuretic furosemide has been found to protect against challenges with bronchoconstrictive agents. OBJECTIVE: To determine if furosemide has any objective and/or subjective therapeutic effect on adult patients with acute exacerbations of asthma, above and beyond the effects of B2 agonists and oral glucocorticoids. METHODS: 35 adult patients with acute exacerbations of asthma were randomized and recruited into the study if they had a prior history of asthma, and <10 pack-year smoking history, had not received pre-hospital B2 agonists, were not pregnant, and had no sulfonamide allergy. A double blind study design was employed. One group received albuterol 5 mg with placebo and one group received albuterol 5 mg with furosemide 40 mg in their nebulizer. Both groups received prednisone 80 mg po. PEFRs were obtained before initiation of treatment and 30 and 60 minutes thereafter. A 10 point dyspnea scale was also employed at initiation of treatment and 30 and 60 minutes thereafter. Physicians were permitted to give rescue albuterol treatments as deemed necessary. RESULTS: There was no difference in the treatment arms with respect to PEFR, dyspnea score, or number of rescue albuterol treatments at 30 and 60 minutes. CONCLUSIONS: We found no statistical benefit to adding furosemide to a regimen consisting of prednisone and optimal doses of B2 agonist. We conclude that there is not a current role for furosemide in acute exacerbations of asthma.


Description and discrimination of planar shapes using shape matrices.
A shape descriptor has been developed which can describe a shape independent of its translation, rotation, and scaling. The description is in the form of a matrix and it is obtained by a polar quantization of a shape. The quantization process takes into consideration not only a shape's outer geometry but its inner geometry as well. The descriptor is information preserving and if the quantization parameters are selected properly, it is possible to reconstruct an original shape from its description. In this technique, shape discrimination is possible by a simple EXCLUSIVE-OR operation on shape descriptions.


Clinical and microscopic hair features of griscelli syndrome associated with asymmetric crying facies in an infant.
Griscelli syndrome (GS) is a rare autosomal recessive disorder characterized by immunodeficiency and partial albinism. Asymmetric crying facies (ACF) is caused by congenital hypoplasia or agenesis of the depressor anguli oris muscle. Asymmetric crying facies may be isolated or it may be associated with various anomalies. We report a 3-month-old Turkish boy who had clinical and microscopic hair features of GS associated with ACF. To the best of our knowledge, this is the 1st case of this association to be reported in the literature.


Trastuzumab Deruxtecan‒Related Interstitial Lung Disease/Pneumonitis: Computed Tomography Imaging Patterns to Guide Diagnosis and Management.
Trastuzumab deruxtecan (T-DXd) is an antibody-drug conjugate approved for the treatment of several advanced cancers; however, severe or fatal interstitial lung disease/pneumonitis can occur. We characterized the computed tomography (CT) patterns of T-DXd‒related pneumonitis as a marker for its clinical severity. Ninety patients with advanced cancers who developed T-DXd‒related pneumonitis in two completed single-arm clinical trials were included. Three radiologists independently characterized the CT patterns of pneumonitis at diagnosis, for analyses of those patterns' relationships with clinical severity and pneumonitis outcome. T-DXd‒related pneumonitis most commonly presented with cryptogenic organizing pneumonia (COP) pattern, observed in 65 patients (72%), followed by a newly identified COP/hypersensitivity pneumonitis (HP) pattern (13%), acute interstitial pneumonia (AIP)/acute respiratory distress syndrome (ARDS) pattern (11%), and HP pattern (3%). A subset of cases with COP pattern demonstrated an atypical distribution with upper and peripheral lung involvement (6/65; 9%). CT patterns were associated with Common Terminology Criteria for Adverse Events severity grades of pneumonitis, with the AIP/ARDS pattern having higher grades compared with others (P < .0001). Fatal pneumonitis was more common in the AIP/ARDS pattern than in others (P = .005). The onset of pneumonitis was earlier in the AIP/ARDS pattern compared with others (median time to onset: at 17.9 v 32.7 weeks of therapy; P = .019). Pneumonitis was treated by withholding T-DXd with or without corticosteroids in most patients (78/90; 87%). T-DXd‒related pneumonitis most commonly demonstrated a COP pattern, with a subset having an atypical distribution. The AIP/ARDS pattern was indicative of severe, potentially fatal pneumonitis, and requires immediate clinical attention to mitigate serious adverse events.


Bile pigment--protein interactions. Coupled oxidation of cytochrome c.
A new methodology is described for the chemical modification of the heme prosthetic group of horse heart cytochrome c. The selective modification of the heme moiety of cytochrome c is facilitated by utilizing coupling oxidation conditions. Comparison of the absorption spectra of this chemically modified cytochrome c species in two different solvents (aqueous pyridine and carbon monoxide saturated 6 M guanidinium chloride) with those of two model compounds [bis(pyridine)(2,3,7,8,12,13,17,18-octaethyl-5-oxaporphyrinato)iron(II) tetrafluoroborate salt and (pyridine)carbonyl-(2,3,7,8,12,13,17,18-octaethyl-5-oxaporphyrinato)iron(II) tetrafluoroborate salt] shows that coupled oxidation of cytochrome c affords a new protein with a covalently bound iron(II) oxaporphyrin prosthetic group. Amino acid analysis of this protein-bound iron(II) oxaporphyrin species reveals that only limited modification of the primary structure of the apoprotein occurs during coupled oxidation of cytochrome c. This protein-bound iron(II) oxaporphyrin species is also interconvertible to a protein-bound bilatriene species under hydrolytic conditions. The synthetic utility of the coupled oxidation of cytochrome c for the preparation of chromoproteins which possess covalently bound iron(II) oxaporphyrin and bilatriene prosthetic groups is considered.


Regulation of the class II MHC pathway in primary human monocytes by granulocyte-macrophage colony-stimulating factor.
GM-CSF stimulates the growth and differentiation of hematopoietic progenitors and also affects mature cell function. These effects have led to the use of GM-CSF as a vaccine adjuvant with promising results; however, the mechanisms underlying GM-CSF-mediated immune potentiation are incompletely understood. In this study, we investigated the hypothesis that the immune stimulatory role of GM-CSF is in part due to effects on class II MHC Ag presentation. We find that, in primary human monocytes treated for 24-48 h, GM-CSF increases surface class II MHC expression and decreases the relative level of the invariant chain-derived peptide, CLIP, bound to surface class II molecules. GM-CSF also increases expression of the costimulatory molecules CD86 and CD40, but not the differentiation marker CD1a or CD16. Furthermore, GM-CSF-treated monocytes are better stimulators in a mixed leukocyte reaction. Additional analyses of the class II pathway revealed that GM-CSF increases total protein and RNA levels of HLA-DR, DM, and DOalpha. Expression of class II transactivator (CIITA) types I and III, but not IV, transcripts increases in response to GM-CSF. Furthermore, GM-CSF increases the amount of CIITA associated with the DR promoter. Thus, our data argue that the proinflammatory role of GM-CSF is mediated in part through increased expression of key molecules involved in the class II MHC pathway via induction of CIITA.


Laparoscopic Radical Resection versus Routine Surgery for Colorectal Cancer.
For patients with colorectal cancer, minimally invasive surgical methods, particularly laparoscopic methods, are now the preferred course of therapy. This research is performed to investigate the effects of laparoscopic radical resection on patients with colorectal cancer. A total of 100 colorectal cancer patients treated in our hospital from January 2017 to January 2019 were enrolled. The subjects were divided into observation (n = 50) and control (n = 50) groups and treated with laparoscopic surgery and laparotomy, respectively. As well as postoperative complications and survival rates, the levels of inflammatory substances, stress response, immunological function, and perioperative markers were compared between the two groups. There was no significant difference in the postoperative exhaust time between the two groups (P > 0.05). Compared with the control group, the observation group showed longer operation time, faster recovery of intestinal function, shorter hospital stay, and less intraoperative bleeding amount (P < 0.05). The serum contents of hs-CRP, TNF-α, IL-6, norepinephrine, adrenaline, and cortisol at 1 d, 3 d, and 5 d after surgery were significantly higher than before in both groups (P < 0.05). Moreover, the serum contents of hs-CRP, TNF-α, IL-6, norepinephrine, adrenaline, and cortisol in the observation group were significantly lower than that in the control group (P < 0.05). At 10 days following surgery, immune index levels had dramatically increased in both groups, with noticeably higher immune index levels in the observation group than in the control group (P < 0.05). There were no appreciable differences in the two groups' 2-year survival rates (P > 0.05), but the complication rate was much greater in the control group (P < 0.05). To sum up, after laparoscopic surgery, patients had fewer complications, shorter hospital stay, lower inflammatory factor expression, less stress response, better immune function, less trauma, faster recovery, and improved quality of life.


Multiple pathways suppress telomere addition to DNA breaks in the Drosophila germline.
Telomeres protect chromosome ends from being repaired as double-strand breaks (DSBs). Just as DSB repair is suppressed at telomeres, de novo telomere addition is suppressed at the site of DSBs. To identify factors responsible for this suppression, we developed an assay to monitor de novo telomere formation in Drosophila, an organism in which telomeres can be established on chromosome ends with essentially any sequence. Germline expression of the I-SceI endonuclease resulted in precise telomere formation at its cut site with high efficiency. Using this assay, we quantified the frequency of telomere formation in different genetic backgrounds with known or possible defects in DNA damage repair. We showed that disruption of DSB repair factors (Rad51 or DNA ligase IV) or DSB sensing factors (ATRIP or MDC1) resulted in more efficient telomere formation. Interestingly, partial disruption of factors that normally regulate telomere protection (ATM or NBS) also led to higher frequencies of telomere formation, suggesting that these proteins have opposing roles in telomere maintenance vs. establishment. In the ku70 mutant background, telomere establishment was preceded by excessive degradation of DSB ends, which were stabilized upon telomere formation. Most strikingly, the removal of ATRIP caused a dramatic increase in telomeric retrotransposon attachment to broken ends. Our study identifies several pathways that suppress telomere addition at DSBs, paving the way for future mechanistic studies.


Deciphering the immune landscape of head and neck squamous cell carcinoma: A single-cell transcriptomic analysis of regulatory T cell responses to PD-1 blockade therapy.
Immunotherapy is changing the Head and Neck Squamous Cell Carcinoma (HNSCC) landscape and improving outcomes for patients with recurrent or metastatic HNSCC. A deeper understanding of the tumor microenvironment (TME) is required in light of the limitations of patients' responses to immunotherapy. Here, we aimed to examine how Nivolumab affects infiltrating Tregs in the HNSCC TME. We used single-cell RNA sequencing data from eight tissues isolated from four HNSCC donors before and after Nivolumab treatment. Interestingly, the study found that Treg counts and suppressive activity increased following Nivolumab therapy. We also discovered that changes in the CD44-SSP1 axis, NKG2C/D-HLA-E axis, and KRAS signaling may have contributed to the increase in Treg numbers. Furthermore, our study suggests that decreasing the activity of the KRAS and Notch signaling pathways, and increasing FOXP3, CTLA-4, LAG-3, and GZMA expression, may be mechanisms that enhance the killing and suppressive capacity of Tregs. Additionally, the result of pseudo-temporal analysis of the HNSCC TME indicated that after Nivolumab therapy, the expression of certain inhibitory immune checkpoints including TIGIT, ENTPD1, and CD276 and LY9, were decreased in Tregs, while LAG-3 showed an increased expression level. The study also found that Tregs had a dense communication network with cluster two, and that certain ligand-receptor pairs, including SPP1/CD44, HLA-E/KLRC2, HLA-E/KLRK1, ANXA1/FPR3, and CXCL9/FCGR2A, had notable changes after the therapy. These changes in gene expression and cell interactions may have implications for the role of Tregs in the TME and in response to Nivolumab therapy.


Synergistic Impacts of Plant-Growth-Promoting Bacteria and Selenium Nanoparticles on Improving the Nutritional Value and Biological Activities of Three Cultivars of Brassica Sprouts.
Due to the growing world population and increasing environmental stress, improving the production, nutritional quality, and pharmaceutical applications of plants have become an urgent need. Therefore, current research was designed to investigate the impact of seed priming using plant-growth-promoting bacteria (PGPB) along with selenium nanoparticles (SeNPs) treatment on chemical and biological properties of three Brassica oleracea cultivars [Southern star (VA1), Prominence (VA2), Monotop (VA3)]. With this aim, one out of five morphologically different strains of bacteria, namely, JM18, which was further identified via 16S rRNA gene sequencing as a Nocardiopsis species with strong plant-growth-promoting traits, isolated from soil, was used. To explore the growth-promoting potential of Nocardiopsis species, seeds of three varieties of B. oleracea were primed with JM18 individually or in combination with SeNP treatment. Seed treatments increased sprout growth (fresh and dry weights) and glucosinolate accumulation. The activity of myrosinase was significantly increased through brassica sprouts and consequently enhanced the amino-acid-derived glucosinolate induction. Notably, a reduction in effective sulforaphane nitrile was detected, being positively correlated with a decrease in epithiospecifier protein (EP). Consequently, the antioxidant activities of VA2 and VA3, determined by the ferric reducing antioxidant power (FRAP) assay, were increased by 74 and 79%, respectively. Additionally, the antibacterial activities of JM18-treated cultivars were improved. However, a decrease was observed in SeNP- and JM18 + SeNP-treated VA2 and VA3 against Serratia marcescens and Candida glabrata and VA1 against S. marcescens. In conclusion, seed priming with the JM18 extract is a promising method to enhance the health-promoting activities of B. oleracea sprouts.


Dry formulations of the biocontrol agent Candida sake CPA-1 using fluidised bed drying to control the main postharvest diseases on fruits.
The biocontrol agent Candida sake CPA-1 is effective against several diseases. Consequently, the optimisation of a dry formulation of C. sake to improve its shelf life and manipulability is essential for increasing its potential with respect to future commercial applications. The present study aimed to optimise the conditions for making a dry formulation of C. sake using a fluidised bed drying system and then to determine the shelf life of the optimised formulation and its efficacy against Penicillium expansum on apples. The optimal conditions for the drying process were found to be 40 °C for 45 min and the use of potato starch as the carrier significantly enhanced the viability. However, none of the protective compounds tested increased the viability of the dried cells. A temperature of 25 °C for 10 min in phosphate buffer was considered as the optimum condition to recover the dried formulations. The dried formulations should be stored at 4 °C and air-packaged; moreover, shelf life assays indicated good results after 12 months of storage. The formulated products maintained their biocontrol efficacy. A fluidised bed drying system is a suitable process for dehydrating C. sake cells; moreover, the C. sake formulation is easy to pack, store and transport, and is a cost-effective process. © 2017 Society of Chemical Industry.


The influence of substrate curvature on neurite outgrowth is cell type dependent.
Damage to axonal tracts of the central nervous system results in costly and permanent disability. The observations of aborted neurite outgrowth and disorganized scarring in injured central nervous system tissue have motivated the hypothesis that engineered bridging devices might facilitate regeneration. It is thought that both the shape and surface chemistry are important design parameters, however, their relative importance is poorly understood. Previously, we utilized smooth cylindrical surfaces to demonstrate that surfaces designed with directionally varying curvature bias in a stereotyped way postnatal dorsal root ganglion axonal regeneration in the direction of minimum curvature independent of surface chemistry. In the present study, we extend this analysis to include adult dorsal root ganglion neurons and cerebellar granule cells, cell types more representative of the challenge faced clinically. We found that axonal outgrowth of both the adult neuron and the central neuron was less sensitive to substrate curvature than the outgrowth of the postnatal neurons. These differences were quantified by constructing distributions describing the probability of outgrowth for a defined range of surface curvatures. Both the adult neuron and the central neuron exhibited a higher probability of extension in high-curvature directions compared to the postnatal neuron implying that surface geometry may not be as potent a cue in directing the regeneration of these neurons. A microtubule-stabilizing agent enhanced the sensitivity to curvature of the adult neuron, partially reversing the increased probability of growing in a high-curvature direction. The results suggest novel methods to enhance directed neuron regeneration using bridging substrates.


Incidence, impact and predictors of residual device patency after left atrial appendage closure with the LACbes device.
The residual device patency (RDP) after left atrial appendage closure (LAAC) with the LACbes device has not been specifically explored in atrial fibrillation (AF) patients. This study aims to explore the incidence, impact and predictors of RDP detected by cardiac computed tomography angiography (CCTA) post LAAC. AF patients implanted with the LACbes device were prospectively enrolled. CCTA device surveillance was performed at 3 months post-procedure. Major adverse events (MAEs), including stroke/transient ischemic attack, major bleeding and all-cause death, were evaluated. Among 141 patients with CCTA surveillance, 56 (39.7%) showed no visible leak and 85 (60.3%) showed RDP. During the median follow-up of 443 [232, 706] days, the presence of RDP was not associated with an increased risk of MAEs (adjusted hazard ratio [HR]: 4.07, 95% confidence interval [CI]: 0.49-34.24, p = 0.196), while peri-device leak (PDL) at the lobe was associated with heightened risks of MAEs (adjusted HR: 6.85, 95% CI: 1.62-28.89, p = 0.009). In patients with PDL at the lobe, antiplatelet after 6 months (HR: 0.20, 95% CI: 0.05-0.91, p = 0.038) was independent protective predictor of MAEs. Besides, current smoking (odds ratio [OR]: 7.52, 95% CI: 2.68-21.08, p < 0.001) and maximum diameter of LAA orifice (OR: 1.16, 95% CI: 1.00-1.34, p = 0.048) were independent predictors of PDL at the lobe. Presence of PDL at the device lobe detected by CCTA at 3-month post LAAC with LACbes is associated with unfavorable prognosis in AF patients. ClinicalTrials.gov Identifier: NCT03788941.


The effects of a 2-year physical education program (SPARK) on physical activity and fitness in elementary school students. Sports, Play and Active Recreation for Kids.
This study evaluated a health-related physical education program for fourth- and fifth-grade students designed to increase physical activity during physical education classes and outside of school. Seven schools were assigned to three conditions in a quasi-experimental design. Health-related physical education was taught by physical education specialists or trained classroom teachers. Students from these classes were compared with those in control classes. Analyses were conducted on 955 students with complete data. Students spent more minutes per week being physically active in specialist-led (40 min) and teacher-led (33 min) physical education classes than in control classes (18 min; P < .001). After 2 years, girls in the specialist-led condition were superior to girls in the control condition on abdominal strength and endurance (P < .001) and cardiorespiratory endurance (P < .001). There were no effects on physical activity outside of school. A health-related physical education curriculum can provide students with substantially more physical activity during physical education classes. Improved physical education classes can potentially benefit 97% of elementary school students.


Metabolites of a tobacco-specific carcinogen in urine from newborns.
Cigarette smoking during pregnancy can result in fetal exposure to carcinogens that are transferred from the mother via the placenta, but little information is available on fetal uptake of such compounds. We analyzed samples of the first urine from newborns whose mothers did or did not smoke cigarettes for the presence of metabolites of the potent tobacco-specific transplacental carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). The urine was collected and analyzed for two metabolites of NNK, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronide (NNAL-Gluc). Gas chromatography and nitrosamine-selective detection, with confirmation by mass spectrometry, were used in the analyses, which were performed without knowledge of the origin of the urine samples. NNAL-Gluc was detected in 22 (71%) of 31 urine samples from newborns of mothers who smoked; NNAL was detected in four of these 31 urine samples. Neither compound was detected in the 17 urine samples from newborns of mothers who did not smoke. The arithmetic mean level of NNAL plus NNAL-Gluc in the 27 newborns of smokers for which both analytes were quantified was 0.14 (95% confidence interval [CI] = 0.083-0.200) pmol/mL. The levels of NNAL plus NNAL-Gluc in the urine from these babies were statistically significantly higher than those in the urine from newborns of nonsmoking mothers (geometric means = 0.062 [95% CI = 0.035-0.110] and 0.010 [considered as not detected; no confidence interval], respectively; two-sided P<.001). NNAL plus NNAL-Gluc levels in the 18 positive urine samples in which both analytes were quantified ranged from 0.045 to 0.400 pmol/mL, with an arithmetic mean level of 0.20 (95% CI = 0.14-0.26) pmol/mL, about 5%-10% of the levels of these compounds detected in the urine from adult smokers. Two metabolites of the tobacco-specific transplacental carcinogen NNK can be detected in the urine from newborns of mothers who smoked cigarettes during pregnancy.


Effects of Introducing Xpert MTB/RIF on Diagnosis and Treatment of Drug-Resistant Tuberculosis Patients in Indonesia: A Pre-Post Intervention Study.
In March 2012, the Xpert MTB/RIF assay (Xpert) was introduced in three provincial public hospitals in Indonesia as a novel diagnostic to detect tuberculosis and rifampicin resistance among high risk individuals. This study assessed the effects of using Xpert in place of conventional solid and liquid culture and drug-susceptibility testing on case detection rates, treatment initiation rates, and health system delays among drug-resistant tuberculosis (TB) patients. Cohort data on registration, test results and treatment initiation were collected from routine presumptive patient registers one year before and one year after Xpert was introduced. Proportions of case detection and treatment initiation were compared using the Pearson Chi square test and median time delays using the Mood's Median test. A total of 975 individuals at risk of drug-resistant TB were registered in the pre-intervention year and 1,442 in the post-intervention year. After Xpert introduction, TB positivity rate increased by 15%, while rifampicin resistance rate reduced by 23% among TB positive cases and by 9% among all tested. Second-line TB treatment initiation rate among rifampicin resistant patients increased by 19%. Time from client registration to diagnosis was reduced by 74 days to a median of a single day (IQR 0-4) and time from diagnosis to treatment start was reduced by 27 days to a median of 15 days (IQR 7-51). All findings were significant with p<0.001. Compared to solid and liquid culture and drug-susceptibility testing, Xpert detected more TB and less rifampicin resistance, increased second-line treatment initiation rates and shortened time to diagnosis and treatment. This test holds promise to improve rapid case finding and management of drug-resistant TB patients in Indonesia.


Smoking Patterns and Smoking Cessation Willingness-A Study among Beneficiaries of Government Welfare Assistance in Poland.
This study examines the prevalence and tobacco use patterns among adult social assistance beneficiaries and their interest in quitting. The results are based on data collected in a cross-sectional survey conducted among adults in the Piotrkowski district. A sample of 3636 social assistance beneficiaries produced a total of 1817 respondents who completed face-to-face questionnaires. Overall, 37.1% of the respondents, including 52.8% men and 29.6% women, were current smokers. Over one third of the smokers reported their willingness to quit. In the study population, several characteristics were significantly associated with the current daily smoking: male gender, low educational attainment, unemployment or temporary employment, lack of awareness of smoking-associated health risks, use of e-cigarettes, and exposure to environmental tobacco smoke (ETS). The intention to quit smoking among the daily smokers was positively correlated with their awareness of smoking-associated health risks, lack of previous quit attempts, and low exposure to ETS. Smoking prevalence among social assistance recipients tends to be higher than in the general population, but more than half of the smokers are willing to quit. There is an urgency to develop policies tailored to the needs of these disadvantaged population groups.


Reduction in energy for electrochemical disinfection of E. coli in urine simulant.
We report the development of novel modes of operation for electrochemical disinfection of E. coli in human urine simulant with an aim to minimize the energy required for disinfection. The system employs boron-doped diamond electrodes and will be part of an energy neutral, water and additive free outdoor toilet being developed for use in developing countries. Disinfection had been previously demonstrated with voltage being continuously applied to the electrode until disinfection was achieved. In the present study, a new pulsed mode of operation is investigated. This includes a continuous on mode, where oxidants are generated until disinfection is achieved, a single cycle mode, where oxidants are generated for a fixed time and the water is circulated so allow already generated oxidants to disinfect, and a pulsed mode with different duty cycles, which is like the single cycle mode but with multiple cycles. Disinfection was achieved with pulsed mode operation with a 68% energy reduction compared to the continuous on mode. Energy saving was most likely achieved by lengthening the contact time of the disinfectant with the bacteria and increased generation of non-chlorine disinfecting oxidants.


Reduction of blood culture contamination rate by an educational intervention.
The efficacy of an educational intervention to prevent blood culture contamination (BCC) in internal medicine was studied in two medical wards in a busy tertiary-care hospital in which blood cultures were obtained by physicians rather than dedicated phlebotomists. Baseline BCC rates were 5.7% and 7.1% in intervention and control wards, respectively (p 0.6), compared with 1.95% and 6.7%, respectively, post-intervention (p < 0.001). Following multivariate analysis, only an absence of intervention was an independent variable associated with BCC. Thus simple educational intervention reduced BCC in internal medicine and was considered to be cost-effective.


Effect of succinylation (3-carboxypropionylation) on the conformation and immunological activity of ovalbumin.
The epsilon-amino groups of ovalbumin were modified with succinic anhydride; as many as 16 lysine residues were succinylated (3-carboxypropionylated). The five succinylated derivatives thus prepared were homogeneous with respect to the extent of chemical modification as shown by electrophoretic and immunological data. Succinylation of the amino groups altered electrophoretic mobility and isoionic pH of ovalbumin in the expected direction. U.v.-absorption and fluorescence spectra suggested changes in the microenvironment of the chromophores in the modified proteins. The difference-spectral results showed greater exposure of tyrosine and tryptophan residues in the succinylated ovalbumin. Increase in susceptibility to tryptic digestion, Stokes radius and intrinsic viscosity of native ovalbumin, which was observed on successive increase in the chemical modification, demonstrated a conformational change that was proportional to the extent of modification. The loss of immunological reactivity caused by chemical modification also indicated a conformational change in succinylated ovalbumin. The fact that the intrinsic viscosity of maximally modified ovalbumin was less than one-third of that for the completely denatured protein in 6M-guanidinium chloride suggested that the modified protein contained significant residual native structure. The latter presumably accommodates some antigenic determinants accounting for 37% residual immunological activity observed with maximally succinylated ovalbumin.


Tracking of familial resemblance for resting blood pressure over time in the Québec Family Study.
The etiology of familial resemblance for systolic (SBP) and diastolic (DBP) blood pressure, both within a single time point as well as across time points, was assessed to determine how familial etiologies underlying a trait may change across time. SBP and DBP measurements were taken roughly 12 years apart in family members participating in the longitudinal Québec Family Study. A longitudinal (bivariate) familial correlation model yields 3 types of correlations: intraindividual cross-time (e.g., father's BP at time 1 with his own BP at time 2); interindividual within-time (e.g., father time 1 with child time 1); and interindividual cross-time (e.g., father time 1 with child time 2). In addition, the change in BP across time (i.e., time 1-time 2) is examined using a univariate family correlation model. This combined method is useful in assessing the degree to which the same familial factors are operating across time (interindividual cross-time correlations), as well as the degree to which different heritable components are involved across time (change score). Maximal heritabilities for SBP were about 70% at each time point, while for DBP the heritability was larger at time 1 (87%) than time 2 (39%). Both the change scores (48% for SBP and 54% for DBP) and the cross-time comparisons (58% to 72% for SBP and 63% to 65% for DBP) evidenced significant familial resemblance. These results illustrate how simple methodologies can be used to specify how familial etiologies underlying a trait may change across time. For BP, the model includes unique familial factors that are specific to each time measurement, and an additional familial factor which is common to both time points. The factors leading to differences in longitudinal familial resemblance for BP (i.e., the unique factors) may be primarily genetic in origin, while those leading to stability across time may include both genetic and familial environmental effects. Sex and/or age interactions with the genotypes are also suggested.