Mapping the morbidity and mortality of Chagas disease in an endemic area in Brazil.
Chagas disease is among the 21 neglected diseases according to the World Health Organization. This study aimed to investigate the morbidity and mortality distribution of Chagas disease for identifying areas with greater prevalences and deaths of the disease in Northeast Brazil. A population-based ecological study was performed from 2016 to 2018 using data on acute Chagas disease patients from the Disease Notification Information System, chronic cases from the Chagas Disease and the referral Heart Failure Outpatient Clinic in Pernambuco, and Chagas disease-related mortality from the Mortality Information System. The unit of analysis were Pernambuco State mesoregions. The indicators were spatialized into thematic maps on the occurrence and mortality of the disease per 100,000 inhabitants. No cases of acute disease were reported in the period analyzed. Data on 801 chronic Chagas disease patients were analyzed. The population showed an average age of 62 years, with female predominance. The most prevalent comorbidity was systemic arterial hypertension and cardiologic involvement without ventricular dysfunction. The average chronic disease occurrence rate was 3.2/ 100,000 people/ year. As for deaths in the mortality system; in total, 350 deaths were recorded, showing male predominance, age ≥ 60 years, and chronic disease with cardiac involvement as the main mortality cause. The annual average mortality proportion was 1.6/100,000 people. The chronic case distribution showed spatial heterogeneity, with the highest rates of chronic disease and deaths observed in two mesoregions, with the main cause of death being heart-related. This highlights the need for more specialized services in areas with higher burden of the disease to avoid delay in the patients' care.


Dual-energy index variation when evaluating the potential ferromagnetism of ex vivo bullets.
An MRI is potentially hazardous for patients with retained ferromagnetic bullets. Recent studies have aimed to develop dual-energy computed tomography (DECT) as a screening tool for recognising highly ferromagnetic bullets. Inconsistent findings have been ascribed to inherent CT technology differences. Previous research demonstrated significant Hounsfield unit (HU) measurement variation among single-source CT machines. This study investigated the theoretical dual-energy index (DEI) variation between DECT machines when evaluating the potential ferromagnetic properties within the same sample of ex vivo bullets and metal phantoms. An experimental ex vivo study was conducted on eight metal phantoms and 10 unused bullets individually positioned in the same Perspex head phantom and scanned on two DECT machines. Two senior radiology registrars independently recorded the HU readings, and DEI values were calculated. Statistical analysis was performed using non-parametric methods for paired data, namely the Signed Rank Test. The DEI values based on mean HU readings between the DECT machines were compared. Inter- and intra-reader agreement was not statistically significant. The metal phantoms had poor interscanner agreement, with an overlap of the ferromagnetic and non-ferromagnetic ranges. The bullets had good interscanner agreement, with a similar ferromagnetic to non-ferromagnetic relationship. The use of DEI values negates the previous assumption that significant interscanner variability exists among different DECT technologies while assessing highly attenuative ex vivo bullets. This investigation demonstrated that even though HU readings may be variable, the implementation of the DEI equation translates this into comparable values with good interscanner agreement.


Adult circumcision and male sexual health: a retrospective analysis.
We aimed to evaluate possible associations of circumcision with several sexual dysfunctions and to identify predictors for the development of these outcomes post-operatively. Telephone surveys about sexual habits and dysfunctions before and after intervention were conducted post-operatively to patients that underwent circumcision in Centro Hospitalar Vila Nova de Gaia/Espinho during 2011. McNemar test was used for a matched-pairs analysis of pre- and post-operative data. Odds ratios, adjusted in a multivariate analysis, explored predictors of de novo sexual dysfunctions after circumcision. With intervention, there was an increase in frequency of erectile dysfunction (9.7% versus 25.8%, P = 0.002) and delayed orgasm (11.3% versus 48.4%, P < 0.001), and a significant symptomatic improvement in patients with pain with intercourse (50.0% versus 6.5%, P < 0.001). Significant predictors for de novo erectile dysfunction were diabetes mellitus (OR 9.81, P = 0.048) and lack of sexual desire (OR 8.76, P = 0.028). Less than three sex partners (OR 7.04, P = 0.007) and low sexual desire (OR 7.49, P = 0.029) were significant predictors for de novo delayed orgasm.


Plasma Biomarkers of Neuropathogenesis in Hospitalized Patients With COVID-19 and Those With Postacute Sequelae of SARS-CoV-2 Infection.
Although patients hospitalized with COVID-19 frequently present with encephalopathy, those with mild initial COVID-19 disease who never required hospitalization also often develop neurologic symptoms as part of postacute sequelae of severe acute respiratory coronavirus type 2 (SARS-CoV-2) infection (neuro-PASC). The pathogenic mechanisms of COVID-19 encephalopathy and neuro-PASC are unknown. We sought to establish biochemical evidence of CNS injury in those patients and their association with neuropsychiatric manifestations and SARS-CoV-2 antigenemia. We recruited hospitalized, posthospitalized, and nonhospitalized patients with confirmed diagnosis of COVID-19 with neurologic symptoms in addition to healthy control (HC) subjects. Plasma neurofilament light chain (pNfL), plasma glial fibrillary acidic protein (pGFAP), and plasma SARS-CoV-2 Nucleocapsid antigen (pN Ag) were measured by HD-X Simoa analyzer (Quanterix) and compared with neuropsychiatric symptoms, patient-reported quality-of-life measures, and standardized cognitive assessments. Neuroglial scores (pGFAP/pNfL) were calculated to estimate the relative contribution of astroglial and neuronal involvement. We enrolled a total of 64 study participants, including 9 hospitalized patients with COVID-19 encephalopathy (CE), 9 posthospitalization neuro-PASC (PNP) patients, 38 nonhospitalized neuro-PASC (NNP) patients, and 8 HC subjects. Patients with CE were older, had higher pNfL and pGFAP concentrations, and more frequent pN Ag detection than all neuro-PASC groups. PNP and NNP patients exhibited similar PASC symptoms, decreased quality-of-life measures, and cognitive dysfunction, and 1 of the 38 (2.6%) NNP patients had pN Ag detectable 3 weeks postsymptoms onset. Patients with neuro-PASC presenting with anxiety/depression had higher neuroglial scores, which were correlated with increased anxiety on quality-of-life measures. pNfL, pGFAP, and pN Ag measurements indicate neuronal dysfunction and systemic involvement in hospitalized COVID-19 patients with encephalopathy. Detection of SARS-CoV-2 N Ag in blood 3 weeks after symptoms onset in a nonhospitalized patient suggests that prolonged antigenic stimulation, or possibly latent infection, may occur. Anxiety was associated with evidence of astroglial activation in patients with neuro-PASC. These data shed new light on SARS-Cov-2 neuropathogenesis and demonstrate the value of plasma biomarkers across the COVID-19 disease spectrum.


Symptoms of major depressive disorder subsequent to child maltreatment: Examining change across multiple levels of analysis to identify transdiagnostic risk pathways.
Major depressive disorder (MDD) is a prevalent psychiatric condition in the child maltreatment population. However, not all children who have been maltreated will develop MDD or MDD symptoms, suggesting the presence of unique risk pathways that explain how certain children develop MDD symptoms when others do not. The current study tested several candidate risk pathways to MDD symptoms following child maltreatment: neuroendocrine, autonomic, affective, and emotion regulation. Female adolescents (N = 110; age range = 14-19) were recruited into a substantiated child maltreatment or comparison condition and completed a laboratory stressor, saliva samples, and measures of emotion regulation, negative affect, and MDD symptoms. MDD symptoms were reassessed 18 months later. Mediational modeling revealed that emotion regulation was the only significant indirect effect of the relationship between child maltreatment and subsequent MDD symptoms, demonstrating that children exposed to maltreatment had greater difficulties managing affective states that in turn led to more severe MDD symptoms. These results highlight the importance of emotion dysregulation as a central risk pathway to MDD following child maltreatment. Areas of future research and implications for optimizing prevention and clinical intervention through the direct targeting of transdiagnostic risk pathways are discussed.


Perspectives for Monocyte/Macrophage-Based Diagnostics of Chronic Inflammation.
Low-grade chronic inflammation underlies the development of the most dangerous cardiometabolic disorders including type 2 diabetes and its vascular complications. In contrast to acute inflammation induced by bacteria and viruses, chronic inflammation can be driven by abnormal reaction to endogenous factors, including Th2 cytokines, metabolic factors like advanced glycation end products (AGEs), modified lipoproteins, or hyperglycemia. The key innate immune cells that recognize these factors in blood circulation are monocytes. Inflammatory programming of monocytes which migrate into tissues can, in turn, result into generation of tissue macrophages with pathological functions. Therefore, determination of the molecular and functional phenotype of circulating monocytes is a very promising diagnostic tool for the identification of hidden inflammation, which can precede the development of the pathology. Here we propose a new test system for the identification of inflammatory programming of monocytes: surface biomarkers and ex vivo functional system. We summarize the current knowledge about surface biomarkers for monocyte subsets, including CD16, CCR2, CX3CR1, CD64, stabilin-1 and CD36, and their association with inflammatory human disorders. Furthermore, we present the design of an ex vivo monocyte-based test system with minimal set of parameters as a potential diagnostic tool for the identification of personalized inflammatory responses.


Fluorogen-free aggregation induced NIR emission from gold nanoparticles.
Controlled diluted aqua regia addition leads to NIR (near infrared) luminescence from aggregated gold nanoparticles at 916 nm. This turn-on luminescence has been observed for gold nanoparticles, regardless of reductants used in their preparation. These aggregated nanoparticles are nontoxic and have been used for bioimaging in human liver carcinoma cells.


Aggregation-independent modulation of proteoglycan binding by neutralization of C-terminal acidic residues in the chemokine macrophage inflammatory protein 1alpha.
Members of the chemokine family of proteins mediate their biological effects through interaction with a family of seven-transmembrane G-protein-coupled receptors. This interaction is complicated by the biochemical properties of chemokines, such as their ability to form self aggregates and their ability to bind to proteoglycans. With some chemokines there is a clear interrelationship between these interactions; the chemokine platelet factor 4 binds preferentially to proteoglycans in its aggregated form. Little is known about the role of aggregation in the proteoglycan binding of other chemokines. Here we demonstrate that the aggregation status of the chemokine macrophage inflammatory protein 1alpha (MIP-1alpha) has no detectable effect on its affinity for proteoglycans. Furthermore, we demonstrate that the alteration of acidic amino acid residues in MIP-1alpha influences the affinity of its interactions with heparin as these residues are progressively neutralized, leading to an enhanced binding affinity for heparin. Thus, with MIP-1alpha, aggregation is not a determinant of proteoglycan binding; however, overall charge does seem to have a major role in the interaction. These results therefore add to our understanding of the nature of the interaction between MIP-1alpha and proteoglycans and suggests that the basic amino acids might not be the sole regulators of proteoglycan binding.


Different estimation methods of spontaneous baroreflex sensitivity have different predictive value in heart failure patients.
Several methods have been developed so far to estimate cardiac baroreflex sensitivity (BRS) through the analysis of spontaneous fluctuations of systolic arterial pressure (SAP) and R-R interval. Their relative performance in predicting cardiac mortality in heart failure patients is not known. The aim of this study was to compare the prognostic value of a set of representative indexes of spontaneous BRS in these patients. We studied 228 stable, moderate-to-severe heart failure patients with reduced ejection fraction, in sinus rhythm, who underwent an 8-min supine recording of ECG, arterial blood pressure and respiration during paced breathing (15 breaths/min). BRS was estimated according to the following methods: sequence (BRSSeq); nonparametric transfer function in the low-frequency band (BRSTF_NP-LF); parametric spectral computed in the low-frequency and high-frequency bands (BRSPS-LF and BRSPS-HF); parametric transfer function computed in the low-frequency and high-frequency bands (BRSTF_P-LF and BRSTF_P-HF); model-based closed loop (BRSCL); and bivariate phase-rectified signal averaging (BRSPRSA). During a median follow-up of 36 months, 45 patients experienced a cardiac event. Only BRSTF_NP-LF, BRSPS-LF, BRSTF_P-LF and BRSPRSA were significantly associated with the outcome (P < 0.01), and statistical significance remained (P ≤ 0.03) after adjusting for clinical covariates. BRSTF_NP-LF and BRSPRSA also significantly improved the risk classification. This study shows that different spontaneous BRS indexes have different predictive value in patients with heart failure. It also shows that the prognostic information of BRS estimates is linked to SAP and RR oscillations in the low-frequency band.


Does autotitrating positive airway pressure therapy improve postoperative outcome in patients at risk for obstructive sleep apnea syndrome? A randomized controlled clinical trial.
Obstructive sleep apnea has been associated with postoperative complications. We hypothesized that postoperative autotitrating positive airway pressure (APAP) applied to patients at high risk for obstructive sleep apnea would shorten hospital stay and reduce postoperative complications. Included were patients aged 18 to 100 years scheduled for elective total knee or hip arthroplasty who were able to give informed consent. Patients without contraindication to positive airway pressure therapy were divided into a high- or low-risk group on the basis of the Flemons sleep apnea clinical score. Low-risk patients received standard care. High-risk patients were randomized to receive standard care or standard care plus postoperative APAP. All patients were administered a predismissal cardiorespiratory sleep study. The primary end point was length of stay, and secondary end points were a range of postoperative complications. One hundred thirty-eight patients were enrolled in the study (52 in the low-risk group, 86 in the high-risk group). Within the high-risk group, 43 were randomized to standard care and 43 to standard care plus postoperative APAP. There were no significant differences in the length of stay (P=.65) or any of the secondary end points between the randomized groups. On subgroup analysis of patients with an apnea-hypopnea index of ≥15, patients randomized to APAP had a longer postoperative stay (median, 5 vs 4 days; P=.02). The role for empirical postoperative APAP requires further study, but the findings did not show benefit for APAP applied postoperatively to positive airway pressure-naive patients at high risk for sleep apnea.


One-year survey of opportunistic premise plumbing pathogens and free-living amoebae in the tap-water of one northern city of China.
In this study, qPCR was used to quantify opportunistic premise plumbing pathogens (OPPPs) and free-living amoebae in 11 tap water samples collected over four seasons from a city in northern China. Results demonstrated that the average numbers of gene copies of Legionella spp. and Mycobacterium spp. were significantly higher than those of Aeromonas spp. (p < 0.05). Legionella spp. and Mycobacterium spp. were 100% (44/44) positively detected while P. aeruginosa and Aeromonas spp. were 79.54% (35/44) and 77.27% (34/44) positively detected. Legionella pneumophila was only detected in 4 samples (4/44), demonstrating its occasional occurrence. No Mycobacterium avium or Naegleria fowleri was detected in any of the samples. The average gene copy numbers of target OPPPs were the highest in summer, suggesting seasonal prevalence of OPPPs. Average gene copy numbers of OPPPs in the taps of low-use-frequency were higher than in taps of high-use-frequency, but the difference was not significant for some OPPPs (p > 0.05). Moderate negative correlations between the chlorine concentration and the gene copy numbers of OPPPs were observed by Spearman analysis (rs ranged from -0.311 to -0.710, p < 0.05). However, no significant correlations existed between OPPPs and AOC, BDOC, or turbidity. Moderate positive correlations were observed between the target microorganisms, especially for Acanthamoeba spp., through Spearman analysis (p < 0.05). Based on our studies, it is proposed that disinfectant concentration, season, taps with different-use frequency, OPPP species, and potential microbial correlations should be considered for control of OPPPs in tap water.


Mitochondrial functions and content of microsomal and mitochondrial cytochromes in human cirrhosis.
1. Mitochondria and microsomal fractions have been isolated from liver biopsies from patients with alcoholic cirrhosis, cryptogenic cirrhosis or chronic aggressive hepatitis. 2. Cirrhotic livers yieled fewer mitochondria than normal liver. 3. The most significant change was a decrease in mitochondrial respiratory control. Cirrhotic microsomal fractions had a 50% diminution in cytochrome b5 and cytochrome P-450 content. 4. The two patients with chronic aggressive hepatitis showed similar mitochondrial and microsomal changes.


Internet-based therapy for adolescents with chronic fatigue syndrome: long-term follow-up.
Cognitive behavioral therapy (CBT) is known to be an effective treatment of adolescents with chronic fatigue syndrome (CFS), but its availability is limited. Fatigue in Teenagers on the Internet (FITNET), an Internet-based CBT program for adolescents with CFS, has been developed as an alternative to face-to-face CBT. Recently, its short-term effectiveness has been proven in a randomized clinical trial. Here we aimed to assess the long-term outcome of CFS in adolescents after FITNET treatment and after usual care. In addition, factors related to recovery at long-term follow-up (LTFU) for adolescents treated with the FITNET program were investigated. The study was an LTFU of participants of the FITNET trial. Data were completed for 112 (88.2%) of 127 approached FITNET study participants. Primary outcomes were fatigue severity (Checklist Individual Strength-20), physical functioning (87-item Child Health Questionnaire), and school/work attendance. After a mean follow-up of 2.7 years, 66 (58.9%) adolescents had recovered from CFS. Most adolescents who recovered directly after treatment with FITNET were still recovered at LTFU. At LTFU there was no difference between the recovery rates for the different treatment strategies (original randomization: FITNET [64%] versus any form of usual care [52.8%]). Per additional month of "pretreatment disease duration," the odds for recovery were 4% lower (odds ratio: 0.96; 95% confidence interval: 0.93-0.99; P = .016), and per added point on "focus on bodily symptoms" (Body Consciousness Scale) of the mother (0-20 points) the odds for recovery were 11% lower (odds ratio: 0.89; 95% confidence interval: 0.80-0.99; P = .029). The short-term effectiveness of Internet-based CBT on adolescent CFS is maintained at LTFU. At LTFU, usual care led to similar recovery rates, although these rates were achieved at a slower pace.


Presynaptic BK channel localization is dependent on the hierarchical organization of alpha-catulin and dystrobrevin and fine-tuned by CaV2 calcium channels.
Large conductance, calcium-activated BK channels regulate many important physiological processes, including smooth muscle excitation, hormone release and synaptic transmission. The biological roles of these channels hinge on their unique ability to respond synergistically to both voltage and cytosolic calcium elevations. Because calcium influx is meticulously regulated both spatially and temporally, the localization of BK channels near calcium channels is critical for their proper function. However, the mechanism underlying BK channel localization near calcium channels is not fully understood. We show here that in C. elegans the localization of SLO-1/BK channels to presynaptic terminals, where UNC-2/CaV2 calcium channels regulate neurotransmitter release, is controlled by the hierarchical organization of CTN-1/α-catulin and DYB-1/dystrobrevin, two proteins that interact with cortical cytoskeletal proteins. CTN-1 organizes a macromolecular SLO-1 channel complex at presynaptic terminals by direct physical interaction. DYB-1 contributes to the maintenance or stabilization of the complex at presynaptic terminals by interacting with CTN-1. We also show that SLO-1 channels are functionally coupled with UNC-2 calcium channels, and that normal localization of SLO-1 to presynaptic terminals requires UNC-2. In the absence of UNC-2, SLO-1 clusters lose the localization specificity, thus accumulating inside and outside of presynaptic terminals. Moreover, CTN-1 is also similarly localized in unc-2 mutants, consistent with the direct interaction between CTN-1 and SLO-1. However, localization of UNC-2 at the presynaptic terminals is not dependent on either CTN-1 or SLO-1. Taken together, our data strongly suggest that the absence of UNC-2 indirectly influences SLO-1 localization via the reorganization of cytoskeletal proteins. CTN-1 and DYB-1, which interact with cortical cytoskeletal proteins, are required for the presynaptic punctate localization of SLO-1 in a hierarchical manner. In addition, UNC-2 calcium channels indirectly control the fidelity of SLO-1 puncta localization at presynaptic terminals. We suggest that the absence of UNC-2 leads to the reorganization of the cytoskeletal structure that includes CTN-1, which in turn influences SLO-1 puncta localization.


Association of periodontal disease and tooth loss with rheumatoid arthritis in the US population.
To test for an association of periodontitis and tooth loss with rheumatoid arthritis (RA). The third National Health and Nutrition Examination Survey (NHANES III) is a nationally representative cross-sectional survey of noninstitutionalized civilians. We included participants aged > or = 60 years who had undergone both musculoskeletal and dental examinations. RA was defined based on American College of Rheumatology criteria. Dental examinations quantified decayed and filled surfaces, missing teeth, and periodontitis. Periodontitis was defined as at least 1 site exhibiting both attachment loss and a probing depth of > or = 4 mm. We classified dental health status as (1) no periodontitis, (2) periodontitis, or (3) edentulous (i.e., complete tooth loss). We performed multivariate multinomial logistic regression models with dental health status as the dependent and RA as the independent variables. The sample consisted of 4461 participants, of whom 103 were classified as having RA. Participants with RA had more missing teeth (20 vs 16 teeth; p < 0.001), but less decay (2% vs 4%; p < 0.001) than participants without RA. After adjusting for age, sex, race/ethnicity, and smoking, subjects with RA were more likely to be edentulous [odds ratio (OR) 2.27, 95% confidence interval (CI) 1.56 3.31] and have periodontitis (OR 1.82, 95% CI 1.04 3.20) compared with non-RA subjects. In participants with seropositive RA there was a stronger association with dental health status, in particular with edentulism (OR 4.5, 95% CI 1.2 17). RA may be associated with tooth loss and periodontitis.


Skin symptoms in the construction industry: occurrence and determinants.
In the construction industry, a relatively high hand eczema prevalence can be expected due to exposure to irritating and allergenic agents. As part of a regular program of voluntary medical examinations, a questionnaire including items on health symptoms and working circumstances is administered to construction industry personnel. We studied 152,200 male workers (response rate 52%). Associations between possible risk factors and self-reported skin symptoms and skin hypersensitivity were assessed using log-binomial regression analysis. Prevalence of skin symptoms on the hands was 25.4% among construction workers, 14.6% among office personnel. Nuisance due to dust exposure was the most important work-related determinant for skin symptoms [Prevalence Ratio (PR) 1.59, 95% confidence interval (CI): 1.55-1.63]. Cross-sectional findings were supported by longitudinal analyses in a study population subset. Skin symptoms are common among construction workers. Nuisance due to dust exposure was associated with higher prevalences of skin symptoms.


Gene expression profiling of HeLa cells in G1 or G2 phases.
The cell division cycle is regulated through both transcriptional and post-transcriptional mechanisms. The altered expression of a number of genes at the mRNA level is known to be essential for progression through the cell cycle, however, a comprehensive gene expression profile of human cells remains to be completed. Here we sought to monitor the differential gene expression of genes after the transition of G2 cells into G1 prior to the restriction point. GeneChip containing microarrays of oligonucleotides corresponding to over 12 000 human genes were employed to profile differential gene expression in G1 and G2. After three independent experiments the resultant data was filtered and a set of genes was compiled based on at least threefold-altered expression, no background noise in determining expression and observation in all experiments. Our analysis identified 154 genes that were elevated in G2 phase of cells as compared to early G1 phase including 15 novel genes. This number included mRNAs whose upregulation is known to occur in G2 phase. Surprisingly only 19 genes were upregulated in G1 phase, among these six genes were novel. Some of these genes are candidates for transition through early G1. This gene inventory for G1 and G2 phases of cell cycle will provide the basis for understanding of cell cycle delay as a result of DNA damage.


Homology and architecture of the caudal basket of Pachycephalosauria (Dinosauria: Ornithischia): the first occurrence of myorhabdoi in Tetrapoda.
Associated postcranial skeletons of pachycephalosaurids, most notably those of Stegoceras and Homalocephale, reveal enigmatic osseous structures not present in other tetrapod clades. The homology and functional significance of these structures have remained elusive as they were originally interpreted to be abdominal ribs or gastralia, and more recently have been interpreted as de novo structures in the tail. Analysis of these structures in nearly all pachycephalosaurid skeletons has facilitated a complete description of their architecture, and the establishment of patterns consistent with those of myorhabdoid ossifications--ossifications of the myoseptal tendons associated with myomeres. The presence and structure of myorhabdoid ossifications are well established for teleost fish, but this marks their first recognition within Tetrapoda. These elements are both structurally and histologically distinct from the deep, paraxial ossified tendon bundles of other ornithischian clades, although they may have performed a similar function in the stiffening of the tail. These myorhabdoi are not de novo structures, but are instead ossifications (and therefore more amenable to fossilization) of the normally unossified plesiomorphic caudal myosepta of vertebrates. The ubiquitous ossification of these structures in pachycephalosaurids (all specimens preserving the tail also exhibit myorhabdoid ossifications) suggests it is a likely synapomorphic condition for Pachycephalosauria.


The implementation of liquid chromatography tandem mass spectrometry for the official control of lipophilic toxins in seafood: single-laboratory validation under four chromatographic conditions.
We performed a comprehensive study to assess the fit for purpose of four chromatographic conditions for the determination of six groups of marine lipophilic toxins (okadaic acid and dinophysistoxins, pectenotoxins, azaspiracids, yessotoxins, gymnodimine and spirolides) by LC-MS/MS to select the most suitable conditions as stated by the European Union Reference Laboratory for Marine Biotoxins (EURLMB). For every case, the elution gradient has been optimized to achieve a total run-time cycle of 12 min. We performed a single-laboratory validation for the analysis of three relevant matrices for the seafood aquaculture industry (mussels, pacific oysters and clams), and for sea urchins for which no data about lipophilic toxins have been reported before. Moreover, we have compared the method performance under alkaline conditions using two quantification strategies: the external standard calibration (EXS) and the matrix-matched standard calibration (MMS). Alkaline conditions were the only scenario that allowed detection windows with polarity switching in a 3200 QTrap mass spectrometer, thus the analysis of all toxins can be accomplished in a single run, increasing sample throughput. The limits of quantification under alkaline conditions met the validation requirements established by the EURLMB for all toxins and matrices, while the remaining conditions failed in some cases. The accuracy of the method and the matrix effects where generally dependent on the mobile phases and the seafood species. The MMS had a moderate positive impact on method accuracy for crude extracts, but it showed poor trueness for seafood species other than mussels when analyzing hydrolyzed extracts. Alkaline conditions with EXS and recovery correction for OA were selected as the most proper conditions in the context of our laboratory. This comparative study can help other laboratories to choose the best conditions for the implementation of LC-MS/MS according to their own necessities.


Whole-genome sequencing of Leptospira interrogans from southern Brazil: genetic features of a highly virulent strain.
BACKGROUND Leptospirosis is the most widespread zoonotic disease. It is caused by infection with pathogenic Leptospira species, of which over 300 serovars have been described. The accurate identification of the causative Leptospira spp. is required to ascertain the pathogenic status of the local isolates. OBJECTIVES This study aimed to obtain the complete genome sequence of a virulent Leptospira interrogans strain isolated from southern Brazil and to describe its genetic features. METHODS The whole genome was sequenced by next-generation sequencing (Ion Torrent). The genome was assembled, scaffolded, annotated, and manually reviewed. Mutations were identified based on a variant calling analysis using the genome of L. interrogans strain Fiocruz L1-130 as a reference. FINDINGS The entire genome had an average GC content of 35%. The variant calling analysis identified 119 single nucleotide polymorphisms (SNPs), from which 30 led to a missense mutation. The structural analyses identified potential evidence of genomic inversions, translocations, and deletions in both the chromosomes. MAIN CONCLUSIONS The genome properties provide comprehensive information about the local isolates of Leptospira spp., and thereby, could facilitate the identification of new targets for the development of diagnostic kits and vaccines.


Monitoring the extracorporeal photopheresis by immunophenotyping in a Sézary syndrome patient.
The extracorporeal photopheresis is indicated as immunomodulatory treatment in primary cutaneous T-cell lymphoma. There are no guidelines about the most suitable immunophenotypic panel to monitor and correlate clinical evolution and laboratory parameters. In this study we characterize a patient with Sézary syndrome stage III, who performed 98 sessions of photopheresis; in the last 30, the expression of T-cells in peripheral blood was evaluated. The patient had exfoliative dermatitis (mainly in the lower limbs) and several analytical disorders: anemia, neutropenia and lymphocytosis; abnormal number of effector T-cells and a CD4/CD8 ratio higher than 10; atypical cells (CD7, CD26) with aberrant phenotype (characteristic of malignant T-cell clones); and regulatory T-cells indicating an immunotolerance state. The clinical evolution verified can be related with the therapeutic scheme adopted. We propose a multiparameter flow cytometry approach to monitor patients with Sézary syndrome that realize photopheresis, including the aberrant cases.


Examination of Cross-Sectional Associations of Neighborhood Deprivation and Alcohol Outlet Density With Hazardous Drinking Using a Twin Design.
The purpose of this study was to examine whether neighborhood socioeconomic deprivation and alcohol outlet density are associated with hazardous drinking using a co-twin design to control for confounding because of genetic and shared environmental factors. The study sample included cross-sectional data from 1,996 same-sex adult twin pairs (mean age = 36.6; 65.9% female) from the Washington State Twin Registry. The Singh Index was used to characterize neighborhood social deprivation for participants' census tract of residence. Geocoded alcohol outlet data were used to create a measure of census tract alcohol outlet density. The three-item Alcohol Use Disorders Identification Test-Consumption scale (AUDIT-C) was used to measure the level of hazardous drinking. Poisson mixed-effects models were used to examine associations between neighborhood factors and AUDIT-C score. Covariates included household income, level of education completed, non-White race, sex, and rurality of residence. Accounting for covariates, there was a statistically significant within-pair association between neighborhood socioeconomic deprivation and a higher level of hazardous drinking. There was no within-pair association between the density of alcohol outlets and hazardous drinking. Associations did not differ by zygosity. The socioeconomic conditions of the neighborhood may play an important role in the development of alcohol misuse even after accounting for genetic and shared environmental influences. Twin designs may be a promising complementary approach to investigating the role of neighborhood characteristics on alcohol and substance use. Further research is needed to better understand the ways through which and for whom neighborhood characteristics may influence hazardous drinking.


MACF1 Mutations Encoding Highly Conserved Zinc-Binding Residues of the GAR Domain Cause Defects in Neuronal Migration and Axon Guidance.
To date, mutations in 15 actin- or microtubule-associated genes have been associated with the cortical malformation lissencephaly and variable brainstem hypoplasia. During a multicenter review, we recognized a rare lissencephaly variant with a complex brainstem malformation in three unrelated children. We searched our large brain-malformation databases and found another five children with this malformation (as well as one with a less severe variant), analyzed available whole-exome or -genome sequencing data, and tested ciliogenesis in two affected individuals. The brain malformation comprised posterior predominant lissencephaly and midline crossing defects consisting of absent anterior commissure and a striking W-shaped brainstem malformation caused by small or absent pontine crossing fibers. We discovered heterozygous de novo missense variants or an in-frame deletion involving highly conserved zinc-binding residues within the GAR domain of MACF1 in the first eight subjects. We studied cilium formation and found a higher proportion of mutant cells with short cilia than of control cells with short cilia. A ninth child had similar lissencephaly but only subtle brainstem dysplasia associated with a heterozygous de novo missense variant in the spectrin repeat domain of MACF1. Thus, we report variants of the microtubule-binding GAR domain of MACF1 as the cause of a distinctive and most likely pathognomonic brain malformation. A gain-of-function or dominant-negative mechanism appears likely given that many heterozygous mutations leading to protein truncation are included in the ExAC Browser. However, three de novo variants in MACF1 have been observed in large schizophrenia cohorts.


Effects of Chinese herbal medicine Yiqi Huaju Qingli Formula in metabolic syndrome patients with microalbuminuria: a randomized placebo-controlled trial.
Microalbuminuria (MAU) is a key component of metabolic syndrome (MetS) and is an early sign of diabetic nephropathy as well. Although routine Western medicine treatments are given to MetS patients to control high blood pressure, hyperglycemia and dyslipidemia, some patients still experience progressive renal lesions and it is necessary to modify and improve the treatment strategy for MetS patients. To investigate the efficacy of Yiqi Huaju Qingli Herb Formula, a compound traditional Chinese herbal medicine, in MetS patients with MAU when it is combined with routine Western medicine treatment. Sixty patients with MetS were randomized into the Chinese herbal formula group (CHF, Yiqi Huaju Qingli formula treatment in combination with Western medicine) and control group (placebo in combination with Western medicine). All treatments were administered for 12 weeks. Urinary microalbumin (MA), urinary albumin-to-creatinine ratio (UACR), 24-hour total urine protein (24-hTP), body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (2-hPPG), glycosylated hemoglobin (HbA1c), homeostasis model assessment for insulin resistance (HOMA-IR), blood lipid profile and blood pressure were observed. Compared with the control group, CHF treatment significantly decreased BMI (P<0.05), WC (P<0.01) and WHR (P<0.01). Both groups had significant decreases in FPG, 2-hPPG, HbA1c, HOMA-IR, MA, and UACR, with CHF treatment showing better effects on these parameters compared with the control treatment (P<0.05). Both treatments significantly reduced the levels of total cholesterol, low-density lipoprotein cholesterol and triacylglycerol (TAG), and a greater reduction in TAG was observed with CHF treatment (P<0.05). The level of high-density lipoprotein cholesterol did not change in the control group after treatment (P>0.05), whereas it significantly increased with CHF treatment (P<0.01). Compared with before the treatment, significant decreases in systolic blood pressure, diastolic blood pressure and mean arterial blood pressure were observed in both groups (P<0.01). However, there was no significant difference between the two groups (P>0.05). Combined treatment of Yiqi Huaju Qingli Formula and Western medicine significantly alleviated MAU, which may correlate with the improvement of insulin sensitivity and glucose and lipid metabolism. TRIAL REGISTRATION IDENTIFIER: This trial was registered in the Chinese Clinical Trial Registry with the identifier ChiCTR-TRC-11001633.


Mortality and incidence rates of stomach cancer in the JACC Study.
The objective of this study was to examine the mortality and incidence of stomach cancer in the Japan Collaborative Cohort Study (JACC Study), and compare them with those of the general population of Japan. The cohort study involved 127,477 people living in 45 municipalities in Japan. The deaths due to stomach cancer were ascertained by death certificates. The age-adjusted mortality rate per 100,000 was calculated based on 110,792 subjects aged between 40 and 79 years at the baseline in all study areas. The incident cases were defined as the subjects in whom stomach cancer had developed, or subjects who died of stomach cancer during the observed period of survey for cancer incidence. The age-adjusted incidence rate per 100,000 was ascertained and calculated in 64,820 people aged 40-79 years at the baseline living in 24 study municipalities with cancer registries. Then, the mortality rate was compared with the mortality data published in vital statistics of Japan of 1995, and the incidence rate compared with the incidence data published in The Research Group for Populationbased Cancer Registration in Japan of 1992. During more than 10 years of follow-up, there were 582 male deaths and 287 female deaths due to stomach cancer. The age-adjusted mortality rate was 93.4/100,000 person-year (95% confidence interval [CI]: 84.6-102.2) in males and 31.1 (95% CI: 27.0-35.3) in females. There were 646 incident cases in males and 370 in females. The age-adjusted incidence rate was 245.3 / 100,000 person-year (95% CI: 221.6-268.9) and 94.8 (95% CI: 83.0-106.6) in males and females, respectively. The mortality rate of stomach cancer in the JACC Study was lower than that in the vital statistics in Japan. Similar incidence rate of stomach cancer seems to be found between data of the JACC Study and that of the Research Group for Population-based Cancer Registration, but care is needed to interpret this similarity, because it might be due to different degree of completeness of incidence survey between the 2 studies.


The effect of oxytocin on the anthropomorphism of touch.
One of the leading hypotheses regarding the mechanism underlying the social effects of oxytocin (OT) is the "social salience hypothesis", which proposes that OT alters the attentional salience of social cues in a context-dependent manner. Recently, OT was implicated in the process of anthropomorphism; specifically, OT was found to increase the tendency to ascribe social meaning to inanimate stimuli. However, the precise component of social interaction that contributes to this effect remains unclear. Because OT plays a role in the response to touch, whether or not objects are touching in a social context may represent the prominent trigger. Given that OT plays a major role in both anthropomorphism and touch, it is reasonable to assume that OT enhances anthropomorphism specifically for non-human touch, further clarifying its role in altering the perceptual salience of social cues. Here, we examined whether intranasal delivery of OT influences anthropomorphism for touch in inanimate objects. To that end, we implicitly measured the emotional reactions of participants (N=51) to photos that depicted two humans or two inanimate objects either touching or not touching. We asked them to rate whether they will include each photo in an emotional album and found that OT treatment increased the likelihood of inclusion in an emotional album to photos that contain touch, particularly between inanimate objects. In a follow-up experiment we found that the more human the inanimate objects were perceived, the more included they were in the emotional album. Our findings demonstrate that OT can enhance the social meaning of touch between two inanimate objects and advance our understanding of the mechanisms underlying the ability of OT to anthropomorphize environmental cues.


Effect of back squat depth on lower-body postactivation potentiation.
Postactivation potentiation (PAP) refers to increased muscular force generation after previous muscular activity. Various studies have used different squat variations as a PAP stimulus; however, different squat depths can have different mechanical and physiological demands that could yield different PAP levels and subsequent performance. The study aimed to compare the effects of the parallel (PS) and quarter (QS) squat on PAP. Twenty-seven, semiprofessional, male rugby union players (mean ± SD, 18 ± 2 years, 87.2 ± 5.4 kg, 180.7 ± 5.1 cm) performed a countermovement jump (BL-CMJ) followed by a 10-minute rest. Subsequently, they performed 3 PS or QS, at each squat's respective 3-repetition maximum load, in a randomized counterbalanced order. After a 5-minute rest, another countermovement jump (CMJ) was performed (POST-CMJ). Countermovement jump height (JH), peak power (PP), impulse (I), and flight time (FT) were recorded using a contact mat. BL-CMJ and POST-CMJ pairwise comparisons for all variables were conducted for each squat type to examine performance changes. Delta values were compared to examine whether one squat produced better CMJ results. Both squats induced PAP for all the variables (p < 0.05), although PS produced better results than QS (p < 0.05; JH, 4.6 ± 2 vs. 3.5 ± 2 cm; I, 15 ± 6 vs. 12 ± 5 N·s; PP, 285 ± 109 vs. 215 ± 96 W; FT, 34 ± 23 vs. 26 ± 11 milliseconds for PS vs. QS). This is the first study to demonstrate that different squat types can induce PAP and that PS is more beneficial for subsequent CMJ performance compared with QS. It is suggested that the deeper depth of PS, which increases gluteus maximum activation and work produced, is responsible for the increased CMJ performance.


Coevolution of costly mate choice and condition-dependent display of good genes.
Females often choose their mates, instead of mating at random, even when a father contributes nothing but genes to his offspring. Costly female preferences for males with exaggerated traits that reduce viability, such as the peacock's tail, are particularly puzzling. Such preferences can evolve if directly favoured by natural selection or when the exaggerated trait, although maladaptive per se, indicates high overall quality of the male's genotype. Two recent analyses suggested that the advantage to mate choice based on genetic quality is too weak to explain extreme cases of exaggeration of display traits and the corresponding preferences. We studied coevolution of a female mate-preference function and a genotype-dependent male display function where mutation supplies variation in genotype quality and mate preference is costly. Preference readily evolves, often causing extreme exaggeration of the display. Mate choice and trait expression can approach an equilibrium, or a limit cycle, or exaggeration can proceed forever, eventually causing extinction.


Leaving on a jet plane: the trade in fraudulently obtained airline tickets.
Every day, hundreds of people fly on airline tickets that have been obtained fraudulently. This crime script analysis provides an overview of the trade in these tickets, drawing on interviews with industry and law enforcement, and an analysis of an online blackmarket. Tickets are purchased by complicit travellers or resellers from the online blackmarket. Victim travellers obtain tickets from fake travel agencies or malicious insiders. Compromised credit cards used to be the main method to purchase tickets illegitimately. However, as fraud detection systems improved, offenders displaced to other methods, including compromised loyalty point accounts, phishing, and compromised business accounts. In addition to complicit and victim travellers, fraudulently obtained tickets are used for transporting mules, and for trafficking and smuggling. This research details current prevention approaches, and identifies additional interventions, aimed at the act, the actor, and the marketplace.


Natural Xanthones from Garcinia mangostana with Multifunctional Activities for the Therapy of Alzheimer's Disease.
Natural xanthones have diversity pharmacological activities. Here, a series of xanthones isolated from the pericarps of Garcinia mangostana Linn, named α-Mangostin, 8-Deoxygartanin, Gartanin, Garciniafuran, Garcinone C, Garcinone D, and γ-Mangostin were investigated. Biological screening performed in vitro and in Escherichia coli cells indicated that most of the xanthones exhibited significant inhibition of self-induced β-amyloid (Aβ) aggregation and also β-site amyloid precursor protein-cleaving enzyme 1, acted as potential antioxidants and biometal chelators. Among these compounds, α-Mangostin, Gartanin, Garcinone C and γ-Mangostin showed better antioxidant properties to scavenge Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) free radical than Trolox, and potent neuroprotective effects against glutamate-induced HT22 cell death partly by up-regulating HO-1 protein level and then scavenging reactive oxygen species. Moreover, Gartanin, Garcinone C and γ-Mangostin could be able to penetrate the blood-brain barrier (BBB) in vitro. These findings suggest that the natural xanthones have multifunctional activities against Alzheimer's disease (AD) and could be promising compounds for the therapy of AD.


Inferring HIV Escape Rates from Multi-Locus Genotype Data.
Cytotoxic T-lymphocytes (CTLs) recognize viral protein fragments displayed by major histocompatibility complex molecules on the surface of virally infected cells and generate an anti-viral response that can kill the infected cells. Virus variants whose protein fragments are not efficiently presented on infected cells or whose fragments are presented but not recognized by CTLs therefore have a competitive advantage and spread rapidly through the population. We present a method that allows a more robust estimation of these escape rates from serially sampled sequence data. The proposed method accounts for competition between multiple escapes by explicitly modeling the accumulation of escape mutations and the stochastic effects of rare multiple mutants. Applying our method to serially sampled HIV sequence data, we estimate rates of HIV escape that are substantially larger than those previously reported. The method can be extended to complex escapes that require compensatory mutations. We expect our method to be applicable in other contexts such as cancer evolution where time series data is also available.


Exploring matrix effects and quantifying organic additives in hydraulic fracturing associated fluids using liquid chromatography electrospray ionization mass spectrometry.
Hydraulic fracturing (HF) operations utilize millions of gallons of water amended with chemical additives including biocides, corrosion inhibitors, and surfactants. Fluids injected into the subsurface return to the surface as wastewaters, which contain a complex mixture of additives, transformation products, and geogenic chemical constituents. Quantitative analytical methods are needed to evaluate wastewater disposal alternatives or to conduct adequate exposure assessments. However, our narrow understanding of how matrix effects change the ionization efficiency of target analytes limits the quantitative analysis of polar to semi-polar HF additives by means of liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). To address this limitation, we explored the ways in which matrix chemistry influences the ionization of seventeen priority HF additives with a modified standard addition approach. We then used the data to quantify HF additives in HF-associated fluids. Our results demonstrate that HF additives generally exhibit suppressed ionization in HF-associated fluids, though HF additives that predominantly form sodiated adducts exhibit significantly enhanced ionization in produced water samples, which is largely the result of adduct shifting. In a preliminary screening, we identified glutaraldehyde and 2-butoxyethanol along with homologues of benzalkonium chloride (ADBAC), polyethylene glycol (PEG), and polypropylene glycol (PPG) in HF-associated fluids. We then used matrix recovery factors to provide the first quantitative measurements of individual homologues of ADBAC, PEG, and PPG in HF-associated fluids ranging from mg L-1 levels in hydraulic fracturing fluid to low μg L-1 levels in PW samples. Our approach is generalizable across sample types and shale formations and yields important data to evaluate wastewater disposal alternatives or implement exposure assessments.


Does aquatic foraging impact head shape evolution in snakes?
Evolutionary trajectories are often biased by developmental and historical factors. However, environmental factors can also impose constraints on the evolutionary trajectories of organisms leading to convergence of morphology in similar ecological contexts. The physical properties of water impose strong constraints on aquatic feeding animals by generating pressure waves that can alert prey and potentially push them away from the mouth. These hydrodynamic constraints have resulted in the independent evolution of suction feeding in most groups of secondarily aquatic tetrapods. Despite the fact that snakes cannot use suction, they have invaded the aquatic milieu many times independently. Here, we test whether the aquatic environment has constrained head shape evolution in snakes and whether shape converges on that predicted by biomechanical models. To do so, we used three-dimensional geometric morphometrics and comparative, phylogenetically informed analyses on a large sample of aquatic snake species. Our results show that aquatic snakes partially conform to our predictions and have a narrower anterior part of the head and dorsally positioned eyes and nostrils. This morphology is observed, irrespective of the phylogenetic relationships among species, suggesting that the aquatic environment does indeed drive the evolution of head shape in snakes, thus biasing the evolutionary trajectory of this group of animals.


Identity-by-descent and association mapping of a recessive gene for Hirschsprung disease on human chromosome 13q22.
Hirschsprung disease (HSCR) is a congenital disorder of unknown etiology characterized by the absence of enteric ganglia in the distal colon. We have ascertained a large, inbred, Mennonite kindred which demonstrates a high incidence of Hirschsprung disease (HSCR). Genealogical analysis of all kinship relationships identified a single common ancestral couple for all parents of affected offspring. Segregation analysis yielded a segregation ratio of 10.67% for males and 5.45% for females. We searched for locations of the gene(s) responsible for HSCR in this pedigree by genotyping three small multicase families and locating genomic regions demonstrating identity-by-descent followed by linkage disequilibrium analysis of 28 additional nuclear families. Based on this novel strategy, we report the mapping of a new locus for HSCR to chromosome 13q22. Nine microsatellite markers spanning 10 cM in this region were genotyped on thirty-one nuclear families. Significant nonrandom association was detected with alleles at markers D13S162, D13S160, D13S170, and AFM240zg9. In addition, our studies reveal preliminary evidence for a genetic modifier of HSCR in this kindred on chromosome 21q22.


[Predictive factors of alcohol use problems among patients visiting an emergency room].
To assess the predictive effect of key individual-level characteristics to identify cases of alcohol use problems among patients visiting an emergency room. The study sample was composed of 371 people attending an emergency room in a public hospital in Lima, Peru, during a period of seven complete days in January, 2005. For data gathering, we used a questionnaire for demographic information, the SIDUC/CICAD for recent use (i.e., in the last 6 hours) of psychoactive substances before arriving to the emergency room, and the AUDIT, to identify alcohol use problems in the last year. Univariate and multivariate logistic regression models were used to estimate the predictive effect of age, sex, area of attention in the emergency room, presence of physical injuries and recent use of alcohol. The odds of being a case of alcohol use problem for males is 26 times the odds of having that problem for females (p-value<0,001). Stratified analysis by sex and adjusted by age and area of attention in the emergency room, showed that males who had recent use of alcohol were more likely of having alcohol use problems as compared to those who did not have recent alcohol use (OR=5.2; 95% CI: 2.4 - 11.5; p<0,001), while for females such an association did not exist. These results support screening initiatives for those males who have recent alcohol use before arriving to an emergency room in order to identify cases of alcohol-related problems and refer them for an appropriate counseling or medical treatment.


How does visual language affect crossmodal plasticity and cochlear implant success?
Cochlear implants (CI) are the most successful intervention for ameliorating hearing loss in severely or profoundly deaf children. Despite this, educational performance in children with CI continues to lag behind their hearing peers. From animal models and human neuroimaging studies it has been proposed the integrative functions of auditory cortex are compromised by crossmodal plasticity. This has been argued to result partly from the use of a visual language. Here we argue that 'cochlear implant sensitive periods' comprise both auditory and language sensitive periods, and thus cannot be fully described with animal models. Despite prevailing assumptions, there is no evidence to link the use of a visual language to poorer CI outcome. Crossmodal reorganisation of auditory cortex occurs regardless of compensatory strategies, such as sign language, used by the deaf person. In contrast, language deprivation during early sensitive periods has been repeatedly linked to poor language outcomes. Language sensitive periods have largely been ignored when considering variation in CI outcome, leading to ill-founded recommendations concerning visual language in CI habilitation.


Metabolic response of glioma to dichloroacetate measured in vivo by hyperpolarized (13)C magnetic resonance spectroscopic imaging.
The metabolic phenotype that derives disproportionate energy via glycolysis in solid tumors, including glioma, leads to elevated lactate labeling in metabolic imaging using hyperpolarized [1-(13)C]pyruvate. Although the pyruvate dehydrogenase (PDH)-mediated flux from pyruvate to acetyl coenzyme A can be indirectly measured through the detection of carbon-13 ((13)C)-labeled bicarbonate, it has proven difficult to visualize (13)C-bicarbonate at high enough levels from injected [1-(13)C]pyruvate for quantitative analysis in brain. The aim of this study is to improve the detection of (13)C-labeled metabolites, in particular bicarbonate, in glioma and normal brain in vivo and to measure the metabolic response to dichloroacetate, which upregulates PDH activity. An optimized protocol for chemical shift imaging and high concentration of hyperpolarized [1-(13)C]pyruvate were used to improve measurements of lactate and bicarbonate in C6 glioma-transplanted rat brains. Hyperpolarized [1-(13)C]pyruvate was injected before and 45 min after dichloroacetate infusion. Metabolite ratios of lactate to bicarbonate were calculated to provide improved metrics for characterizing tumor metabolism. Glioma and normal brain were well differentiated by lactate-to-bicarbonate ratio (P = .002, n = 5) as well as bicarbonate (P = .0002) and lactate (P = .001), and a stronger response to dichloroacetate was observed in glioma than in normal brain. Our results clearly demonstrate for the first time the feasibility of quantitatively detecting (13)C-bicarbonate in tumor-bearing rat brain in vivo, permitting the measurement of dichloroacetate-modulated changes in PDH flux. The simultaneous detection of lactate and bicarbonate provides a tool for a more comprehensive analysis of glioma metabolism and the assessment of metabolic agents as anti-brain cancer drugs.


Low telomerase activity in CD4+ regulatory T cells in patients with severe chronic GVHD after hematopoietic stem cell transplantation.
CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) play an important role in the control of chronic graft-versus-host disease (cGVHD). In this study, we examined telomere length and telomerase activity of Treg and conventional CD4(+) T cells (Tcon) in 61 patients who survived more than 2 years after allogeneic hematopoietic stem cell transplantation. Cell proliferation and expression of Bcl-2 were also measured in each subset. Treg telomere length was shorter and Treg telomerase activity was increased compared with Tcon (P < .0001). After transplantation, Treg were also more highly proliferative than Tcon (P < .0001). Treg number, telomerase activity, and expression of Bcl-2 were each inversely associated with severity of cGVHD. These data indicate that activation of telomerase is not sufficient to prevent telomere shortening in highly proliferative Treg. However, telomerase activation is associated with increased Bcl-2 expression and higher Treg numbers in patients with no or mild cGVHD. In contrast, patients with moderate or severe cGVHD have fewer Treg with lower levels of telomerase activity and Bcl-2 expression. These results suggest that failure to activate Treg telomerase may restrict proliferative capacity and increase apoptotic susceptibility, resulting in the loss of peripheral tolerance and the development of cGVHD.


Mass spectrometry-based metabolic profiling reveals different metabolite patterns in invasive ovarian carcinomas and ovarian borderline tumors.
Metabolites are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes. We have used a metabolite profiling approach to test the hypothesis that quantitative signatures of primary metabolites can be used to characterize molecular changes in ovarian tumor tissues. Sixty-six invasive ovarian carcinomas and nine borderline tumors of the ovary were analyzed by gas chromatography/time-of-flight mass spectrometry (GC-TOF MS) using a novel contamination-free injector system. After automated mass spectral deconvolution, 291 metabolites were detected, of which 114 (39.1%) were annotated as known compounds. By t test statistics with P < 0.01, 51 metabolites were significantly different between borderline tumors and carcinomas, with a false discovery rate of 7.8%, estimated with repeated permutation analysis. Principal component analysis (PCA) revealed four principal components that were significantly different between both groups, with the highest significance found for the second component (P = 0.00000009). PCA as well as additional supervised predictive models allowed a separation of 88% of the borderline tumors from the carcinomas. Our study shows for the first time that large-scale metabolic profiling using GC-TOF MS is suitable for analysis of fresh frozen human tumor samples, and that there is a consistent and significant change in primary metabolism of ovarian tumors, which can be detected using multivariate statistical approaches. We conclude that metabolomics is a promising high-throughput, automated approach in addition to functional genomics and proteomics for analyses of molecular changes in malignant tumors.


Estimating the total rate of DNA replication using branching processes.
Increasing the knowledge of various cell cycle kinetic parameters, such as the length of the cell cycle and its different phases, is of considerable importance for several purposes including tumor diagnostics and treatment in clinical health care and a deepened understanding of tumor growth mechanisms. Of particular interest as a prognostic factor in different cancer forms is the S phase, during which DNA is replicated. In the present paper, we estimate the DNA replication rate and the S phase length from bromodeoxyuridine-DNA flow cytometry data. The mathematical analysis is based on a branching process model, paired with an assumed gamma distribution for the S phase duration, with which the DNA distribution of S phase cells can be expressed in terms of the DNA replication rate. Flow cytometry data typically contains rather large measurement variations, however, and we employ nonparametric deconvolution to estimate the underlying DNA distribution of S phase cells; an estimate of the DNA replication rate is then provided by this distribution and the mathematical model.


Superficial moist heat's lack of influence on soleus function.
It is reported that thermotherapy decreases motoneuron-pool recruitment. Any decrease in recruitment might have a significant impact on an athlete's ability to return to competition. To determine whether moist heat application influences involuntary motoneuron-pool recruitment or voluntary plantar-flexion peak torque of the soleus muscle immediately or 30 min after application. A 3 x 3 crossover design. Biomechanics laboratory. Eighteen healthy subjects with no history of lower extremity surgery or injury 12 months before the study volunteered. A series of short-duration, high-intensity stimuli was delivered to the tibial nerve to find the H(max) and M(max) measures. Immediately after the H(max) and M(max) measures, subjects were positioned on an isokinetic dynamometer where they performed 5 submaximal warm-up repetitions. Immediately after the warm-up, 5 maximum-intensity peak plantar-flexion torque repetitions were performed. After the dynamometer measures, subjects returned to the table, where a moist heat pack, no heat pack, or a dry nonheated heat pack was applied. H(max), M(max), peak plantar-flexion torque, surface temperature (degrees C), and ambient temperature (degrees C). Moist heat did not influence the H:M(max) ratio or peak plantar-flexion torque. Temperature increased with moist heat pack. Ambient temperature remained constant. Moist heat did not influence involuntary motoneuron-pool recruitment or voluntary peak plantar-flexion torque of the soleus muscle immediately or 30 min after application.


A second gene for gonadotropin-releasing hormone: cDNA and expression pattern in the brain.
In vertebrates, the gonadotropin-releasing hormone (GnRH) decapeptide is secreted from hypothalamic nerve terminals to regulate reproduction via control of synthesis and release of pituitary gonadotropins. Only one GnRH peptide has been found in mammals, with one exception, although numerous other vertebrate species express more than one of the eight known decapeptide forms as shown by immunocytochemical labeling of distinct cell groups in the brain. However, neither the functional nor the evolutionary relationships among these GnRH forms are clear, because only one preprohormone gene sequence from any species has been reported. The most ubiquitous alternative form of GnRH is [His5,Trp7,Tyr8]GnRH (also referred to as chicken-II), which differs from the mammalian sequence at amino acids 5, 7, and 8. This peptide has been shown to have the most potent releasing-hormone activity, although immunocytochemical staining has suggested it is synthesized only in the mesencephalon. Here we report the cloning and expression pattern of the gene for the precursor of this form from the teleost fish Haplochromis burtoni. This is the second GnRH-encoding gene to be characterized in this species. The newly discovered preprohormone gene differs from that previously reported in two ways. First, whereas the original gene predicts only a single associated peptide, this one predicts two associated peptides, both of which appear to be unique. Second, the gene for [His5,Trp7,Tyr8]GnRH is expressed in only one cell group in the mesencephalon. In contrast, the previously reported gene is expressed only in the terminal nerve. The striking differences between the preprohormone structure and localization suggest that the genes coding for the two known GnRH forms in H. burtoni did not arise from a recent duplication event. Interestingly, neither of the two genes found to date in this species is expressed in cells which project from the hypothalamus to the pituitary, suggesting that yet a third gene coding for GnRH may exist.


Gastric dysplasia and gastric cancer: Helicobacter pylori, serum vitamin C, and other risk factors.
Gastric cancer is generally thought to arise through a series of gastric mucosal changes, but the determinants of the precancerous lesions are not well understood. To identify such determinants, we launched a follow-up study in 1989-1990 among 3433 adults in Linqu County, China, a region with very high rates of gastric cancer. Data on cigarette smoking, alcohol consumption, and other characteristics of the participants were obtained by interview in 1989-1990, when an initial endoscopy was taken. At study entry, antibodies to Helicobacter pylori were assayed in 2646 adults (77% of people screened), and levels of serum micronutrients were measured in approximately 450 adults. Follow-up endoscopic and histopathologic examinations were conducted in 1994. Antibodies to H. pylori, levels of serum micronutrients, and other baseline characteristics were compared between subjects whose condition showed progression to dysplasia or gastric cancer from study entry to 1994 and subjects with no change or with regression of their lesions over the same time frame. All P: values are two-sided. The presence of H. pylori at baseline was associated with an increased risk of progression to dysplasia or gastric cancer (odds ratio [OR] = 1.8; 95% confidence interval [CI] = 1.2-2.6). The risk of progression to dysplasia or gastric cancer also was moderately increased with the number of years of cigarette smoking. In contrast, the risk of progression was decreased by 80% (OR = 0.2; 95% CI = 0.1-0.7) among subjects with baseline ascorbic acid levels in the highest tertile compared with those in the lowest tertile, and there was a slightly elevated risk in those individuals with higher levels of alpha-tocopherol. H. pylori infection, cigarette smoking, and low levels of dietary vitamin C may contribute to the progression of precancerous lesions to gastric cancer in this high-risk population.


Do Neural Network Weights Account for Classes Centers?
The exploitation of deep neural networks (DNNs) as descriptors in feature learning challenges enjoys apparent popularity over the past few years. The above tendency focuses on the development of effective loss functions that ensure both high feature discrimination among different classes, as well as low geodesic distance between the feature vectors of a given class. The vast majority of the contemporary works rely their formulation on an empirical assumption about the feature space of a network's last hidden layer, claiming that the weight vector of a class accounts for its geometrical center in the studied space. This article at hand follows a theoretical approach and indicates that the aforementioned hypothesis is not exclusively met. This fact raises stability issues regarding the training procedure of a DNN, as shown in our experimental study. Consequently, a specific symmetry is proposed and studied both analytically and empirically that satisfies the above assumption, addressing the established convergence issues. More specifically, the aforementioned symmetry suggests that all weight vectors are unit, coplanar, and their vector summation equals zero. Such a layout is proven to ensure a more stable learning curve compared against the corresponding ones succeeded by popular models in the field of feature learning.


A Foreign Body Through the Shoe of a Person With Diabetic Peripheral Neuropathy Alters Contralateral Biomechanics: Captured Through Innovative Plantar Pressure Technology.
High plantar pressure as a result of diabetic peripheral neuropathy is often reported as a major risk factor for ulceration. However, previous studies are confined to laboratories with equipment limited by cables, reducing the validity of measurements to daily life. The participant concerned in this case report was wearing an innovative plantar pressure feedback system as part of a wider study. The system allows for continuous plantar pressure monitoring and provides feedback throughout all activities of daily living. The participant concerned was a 59-year-old male with type 2 diabetes who presented with severe peripheral neuropathy. In addition, the right ankle had previously undergone fusion. Between monthly study appointments, the participant unknowingly had a screw embedded in his right shoe, while pressure was being recorded. Although no significant differences in pressure were present for the right foot with the embedded screw, the contralateral foot showed significantly higher pressure when the screw was embedded, compared with pre and post time periods. The increase in pressure on the contralateral foot is expected to result from the protrusion of the screw in the right shoe, causing a perturbation to balance and a shift in the center of pressure toward the contralateral side. This compensatory effect is likely to have been magnified by the limited mobility of the fused right ankle. These findings highlight the importance of checking both feet for ulcer risk, in the event of receiving high-pressure feedback. This innovative technology may improve our understanding of diabetic plantar foot ulcer development.


Parental misinterpretations of over-the-counter pediatric cough and cold medication labels.
Concerns about the safety and efficacy of over-the-counter cold medications have led to a recent US Food and Drug Administration public health advisory against their use in children <2 years of age. Our goal was to examine caregiver understanding of the age indication of over-the-counter cold medication labels and identify factors, associated with caregiver understanding. Caregivers of infant children (< or =1 year old) were recruited from clinics at 3 institutions. Questions were administered regarding the use of 4 previously common "infant" over-the-counter cold and cough medicines labeled to consult a physician if used in children <2 years of age. Literacy and numeracy skills were assessed with validated instruments. A total of 182 caregivers were recruited; 87% were the infants' mothers. Mean education level was 12.5 years, and 99% had adequate literacy skills, but only 17% had >9th-grade numeracy skills. When examining the front of the product label, 86% of the time parents thought these products were appropriate for use in children <2 years of age. More than 50% of the time, parents stated they would give these over-the-counter products to a 13-month-old child with cold symptoms. Common factors that influenced parental decisions included label saying "infant," graphics (eg, infants, teddy bears, droppers), and dosing directions. Caregivers were influenced by the dosing directions only 47% of the time. Caregivers with lower numeracy skills were more likely to provide inappropriate reasons for giving an over-the-counter medication. Misunderstanding of over-the-counter cold products is common and could result in harm if medications are given inappropriately. Label language and graphics seem to influence inappropriate interpretation of over-the-counter product age indications. Poorer parental numeracy skills may increase the misinterpretation of these products. Opportunities exist for the Food and Drug Administration and manufacturers to revise existing labels to improve parental comprehension and enhance child safety.


Ring Finger 149-Related Is an FGF/MAPK-Independent Regulator of Pharyngeal Muscle Fate Specification.
During embryonic development, cell-fate specification gives rise to dedicated lineages that underlie tissue formation. In olfactores, which comprise tunicates and vertebrates, the cardiopharyngeal field is formed by multipotent progenitors of both cardiac and branchiomeric muscles. The ascidian Ciona is a powerful model to study cardiopharyngeal fate specification with cellular resolution, as only two bilateral pairs of multipotent cardiopharyngeal progenitors give rise to the heart and to the pharyngeal muscles (also known as atrial siphon muscles, ASM). These progenitors are multilineage primed, in as much as they express a combination of early ASM- and heart-specific transcripts that become restricted to their corresponding precursors, following oriented and asymmetric divisions. Here, we identify the primed gene ring finger 149 related (Rnf149-r), which later becomes restricted to the heart progenitors, but appears to regulate pharyngeal muscle fate specification in the cardiopharyngeal lineage. CRISPR/Cas9-mediated loss of Rnf149-r function impairs atrial siphon muscle morphogenesis, and downregulates Tbx1/10 and Ebf, two key determinants of pharyngeal muscle fate, while upregulating heart-specific gene expression. These phenotypes are reminiscent of the loss of FGF/MAPK signaling in the cardiopharyngeal lineage, and an integrated analysis of lineage-specific bulk RNA-seq profiling of loss-of-function perturbations has identified a significant overlap between candidate FGF/MAPK and Rnf149-r target genes. However, functional interaction assays suggest that Rnf149-r does not directly modulate the activity of the FGF/MAPK/Ets1/2 pathway. Instead, we propose that Rnf149-r acts both in parallel to the FGF/MAPK signaling on shared targets, as well as on FGF/MAPK-independent targets through (a) separate pathway(s).


Anti-Adhesive Activity of Cranberry Phenolic Compounds and Their Microbial-Derived Metabolites against Uropathogenic Escherichia coli in Bladder Epithelial Cell Cultures.
Cranberry consumption has shown prophylactic effects against urinary tract infections (UTI), although the mechanisms involved are not completely understood. In this paper, cranberry phenolic compounds and their potential microbial-derived metabolites (such as simple phenols and benzoic, phenylacetic and phenylpropionic acids) were tested for their capacity to inhibit the adherence of uropathogenic Escherichia coli (UPEC) ATCC®53503™ to T24 epithelial bladder cells. Catechol, benzoic acid, vanillic acid, phenylacetic acid and 3,4-dihydroxyphenylacetic acid showed anti-adhesive activity against UPEC in a concentration-dependent manner from 100-500 µM, whereas procyanidin A2, widely reported as an inhibitor of UPEC adherence on uroepithelium, was only statistically significant (p < 0.05) at 500 µM (51.3% inhibition). The results proved for the first time the anti-adhesive activity of some cranberry-derived phenolic metabolites against UPEC in vitro, suggesting that their presence in the urine could reduce bacterial colonization and progression of UTI.


Surface Modification of Polyacrylonitrile Membrane by Chemical Reaction and Physical Coating: Comparison between Static and Pore-Flowing Procedures.
The influences of static and pore-flowing procedures on the surface modification of a polyacrylonitrile (PAN) ultrafiltration membrane through chemical reaction and physical coating were investigated in detail. For chemical modification by ethanolamine, a membrane modified by the pore-flowing procedure showed a higher flux and different morphology. The reasons were explained by two effects: the pore-flowing resistance to the random thermal motion of PAN at high temperatures and different reaction kinetics related to the reactant concentration profile on the interface between the membrane and reaction solution and the kinetic property of the fluid (driving force and miscibility) and reaction (time and rate). For physical coating modification, a dense and flat layer via a loose and random layer was formed during the pore-flowing process and static process, which changed the flux and antifouling property of the membrane. The membrane prepared by dead-end filtration showed the best trade-off between the flux and antifouling property. Overall, the procedure kinetics plays an important role in the optimization of membrane modification.


Handheld Co-Axial Bioprinting: Application to in situ surgical cartilage repair.
Three-dimensional (3D) bioprinting is driving major innovations in the area of cartilage tissue engineering. Extrusion-based 3D bioprinting necessitates a phase change from a liquid bioink to a semi-solid crosslinked network achieved by a photo-initiated free radical polymerization reaction that is known to be cytotoxic. Therefore, the choice of the photocuring conditions has to be carefully addressed to generate a structure stiff enough to withstand the forces phisiologically applied on articular cartilage, while ensuring adequate cell survival for functional chondral repair. We recently developed a handheld 3D printer called "Biopen". To progress towards translating this freeform biofabrication tool into clinical practice, we aimed to define the ideal bioprinting conditions that would deliver a scaffold with high cell viability and structural stiffness relevant for chondral repair. To fulfill those criteria, free radical cytotoxicity was confined by a co-axial Core/Shell separation. This system allowed the generation of Core/Shell GelMa/HAMa bioscaffolds with stiffness of 200KPa, achieved after only 10 seconds of exposure to 700 mW/cm2 of 365 nm UV-A, containing >90% viable stem cells that retained proliferative capacity. Overall, the Core/Shell handheld 3D bioprinting strategy enabled rapid generation of high modulus bioscaffolds with high cell viability, with potential for in situ surgical cartilage engineering.


Canonical wnt signaling is required for commissural axon guidance.
Morphogens have been identified as guidance cues for postcrossing commissural axons in the spinal cord. Shh has a dual effect on postcrossing commissural axons: a direct repellent effect mediated by Hhip as a receptor, and an indirect effect by shaping a Wnt activity gradient. Wnts were shown to be attractants for postcrossing commissural axons in both chicken and mouse embryos. In mouse, the effects of Wnts on axon guidance were concluded to depend on the planar cell polarity (PCP) pathway. Canonical Wnt signaling was excluded based on the absence of axon guidance defects in mice lacking Lrp6 which is an obligatory coreceptor for Fzd in canonical Wnt signaling. In the loss-of-function studies reported here, we confirmed a role for the PCP pathway in postcrossing commissural axon guidance also in the chicken embryo. However, taking advantage of the precise temporal control of gene silencing provided by in ovo RNAi, we demonstrate that canonical Wnt signaling is also required for proper guidance of postcrossing commissural axons in the developing spinal cord. Thus, axon guidance does not seem to depend on any one of the classical Wnt signaling pathways but rather involve a network of Wnt receptors and downstream components.


Nickel hypersensitivity and orthodontic treatment: a systematic review and meta-analysis.
Nickel-containing alloys are widely used in orthodontic appliances, even though nickel is by far the most common contact allergen. However, the scientific evidence concerning allergic reactions to nickel in orthodontic patients has not been evaluated systematically. The objective of this study was to investigate whether the prevalence of nickel hypersensitivity is affected by orthodontic treatment. Unrestricted electronic and manual searches were performed until July 2013 for human clinical studies assessing orthodontic treatment and nickel hypersensitivity. Methodological limitations were evaluated with the Downs and Black tool. Crude and adjusted odds ratios (ORs) with their 95% confidence intervals (CIs) were calculated from random-effects meta-analyses, followed by subgroup and sensitivity analyses. Thirty studies were included in the review, and 24 datasets with 10 184 patients in the meta-analyses. Orthodontic treatment had no significant effect on nickel hypersensitivity (n = 11; crude OR 0.99; 95%CI: 0.78-1.25; p = 0.914). However, when confounding from factors such as sex and piercings was taken into account, orthodontic treatment was associated with a lower risk of hypersensitivity (n = 1; adjusted OR 0.60; 95%CI: 0.40-0.80; p < 0.001). This was even more pronounced when orthodontic treatment was performed prior to piercing (n = 7; crude OR 0.35; 95%CI: 0.24-0.50; p < 0.001). Orthodontic treatment seems to have a protective role against nickel hypersensitivity, especially when it precedes piercings.


The Determination of Finger-Flexor Critical Force in Rock Climbers.
Purpose: To determine if the mathematical model used for the estimation of critical force (CF) and the energy store component W' are applicable to intermittent isometric muscle actions of the finger flexors of rock climbers, using a multisession test. As a secondary aim, the agreement of estimates of CF and W' from a single-session test was also determined. The CF was defined as the slope coefficient, and W' was the intercept of the linear relationship between total "isometric work" (Wlim) and time to exhaustion (Tlim). Methods: Subjects performed 3 (separated by either 20 min or >24 h) tests to failure using intermittent isometric finger-flexor contractions at 45%, 60%, and 80% of their maximum voluntary contraction. Results: Force plotted against Tlim displayed a hyperbolic relationship; correlation coefficients of the parameter estimates from the work-time CF model were consistently very high (R2 > .94). Climbers' mean CF was 425.7 (82.8) N (41.0% [6.2%] maximum voluntary contraction) and W' was 30,882 (11,820) N·s. Good agreement was found between the single-session and multisession protocol for CF (intraclass correlation coefficient [ICC3,1] = .900; 95% confidence interval, .616-.979), but not for W' (ICC3,1 = .768; 95% confidence interval, .190-.949). Conclusions: The results demonstrated the sensitivity of a simple test for the determination of CF and W', using equipment readily available in most climbing gyms. Although further work is still necessary, the test of CF described is of value for understanding exercise tolerance and to determine optimal training prescription to monitor improvements in the performance of the finger flexors.


AQP4 Antibody Assay Sensitivity Comparison in the Era of the 2015 Diagnostic Criteria for NMOSD.
We have compared five different assays for antibodies to aquaporin-4 in 181 cases of suspected Neuromyelitis optica spectrum disorders (NMOSD) and 253 controls to assess their relative utility. As part of a clinically-based survey of NMOSD in Australia and New Zealand, cases of suspected NMOSD were referred from 23 centers. Clinical details and magnetic imaging were reviewed and used to apply the 2015 IPND diagnostic criteria. In addition, 101 age- and sex-matched patients with multiple sclerosis were referred. Other inflammatory disease (n = 49) and healthy controls (n = 103) were also recruited. Samples from all participants were tested using tissue-based indirect immunofluorescence assays and a subset were tested using four additional ELISA and cell-based assays. Antibodies to myelin oligodendrocyte glycoprotein (MOG) were also assayed. All aquaporin-4 antibody assays proved to be highly specific. Sensitivities ranged from 60 to 94%, with cell-based assays having the highest sensitivity. Antibodies to MOG were detected in 8/79 (10%) of the residual suspected cases of NMOSD. Under the 2015 IPND diagnostic criteria for NMOSD, cell-based assays for aquaporin-4 are sensitive and highly specific, performing better than tissue-based and ELISA assays. A fixed cell-based assay showed near-identical results to a live-cell based assay. Antibodies to MOG account for only a small number of suspected cases.


Comprehensive immunotherapy combined with intratumoral injection of zoledronate-pulsed dendritic cells, intravenous adoptive activated T lymphocyte and gemcitabine in unresectable locally advanced pancreatic carcinoma: a phase I/II trial.
Dendritic cell (DC)-based vaccines prepared using various antigen loading methods have been studied for cancer immunotherapy. The in vivo provocation of immunity by the direct injection of DCs without using tumor-specific antigens into tumors after apoptosis-inducing chemotherapy is more applicable. We previously reported that zoledronate-pulsed DCs (Zol-DCs) may induce tumor-antigen-specific CD8+ T cells by activating Vγ9γδT cells. In this report, we studied the feasibility, safety, and efficacy of a comprehensive immunotherapy involving the combined intratumoral injection of Zol-DC, gemcitabine (GEM) and αβT cells in locally advanced pancreatic carcinoma. Seven of 15 patients showed a stable disease (SD) and most of the patients showed long-term clinical responses. The FACT-BRM score was significantly higher in the patients with SD. Additionally the CD8+/Treg ratio significantly increased in SD patients after treatment. The median over-all survival and progression-free-survival of 15 patients were 12.0 months and 5.5 months, respectively. Patients with a pretreatment neutrophil/lymphocyte ratio (NLR) lower than 5.0 showed significantly longer survival. Even in an analysis limited to the patients with an NLR lower than 5.0, the patients whose CD8+/Treg ratio increased more than twofold tended to survive longer. In conclusion, the comprehensive immunotherapy using Zol-DCs, systemic αβT cells, and GEM may synergistically show a therapeutic effect on locally advanced pancreatic carcinoma. By using appropriate and precise biomarkers, such as NLR and CD8+/Treg ratio, the present comprehensive immunotherapy could be more beneficial for patients with pancreatic carcinoma.


Community challenges when using large plastic bottles for Solar Energy Disinfection of Water (SODIS).
Communities living in developing countries as well as populations affected by natural or man-made disasters can be left at great risk from water related diseases, especially those spread through the faecal-oral route. Conventional water treatments such as boiling and chlorination can be effective but may prove costly for impoverished communities. Solar water disinfection (SODIS) has been shown to be a cheap and effective way for communities to treat their water. The exposure to sunlight is typically carried out in small volume plastic beverage bottles (up to 2 l). Given the water requirements of consumption and basic personal hygiene, this may not always meet the needs of communities. Recent work has shown 19-L plastic water dispenser containers to be effective SODIS reactors, comparable in efficacy to PET bottles. In this paper we outline the need for studying SODIS in large volumes and discuss 4 main associated challenges. Apart from clean water needed for consumption, access to adequate water is essential for sanitation and hygiene. Contamination of treated water through unwashed hands or vessels contributes heavily to the spread of water borne pathogens in communities. Traditional water treatments such as boiling and chlorination can be effective but may prove financially burdensome for low income communities. SODIS in large vessels could be used as a simple method to meet water requirements in low income and disaster affected populations. However, there have been some concerns associated with the conventional SODIS method; we identify the main ones to be: (1) cold or cloudy weather; (2) the fear of leaching in plastic bottles; (3) water turbidity, and; (4) community acceptance. The application of SODIS in large bottles like WDCs has the potential to be an efficient and cost effective method of disinfecting water, either for consumption until more rigorous water treatments can be put in place, or for sanitation and hygiene to curb the spread of fecal contamination. Further research is needed that can address some of the limitations and challenges associated with the use of large bottles for SODIS.


Process-Induced Changes in the Quantity and Characteristics of Grain Dietary Fiber.
Daily use of wholegrain foods is generally recommended due to strong epidemiological evidence of reduced risk of chronic diseases. Cereal grains, especially the bran part, have a high content of dietary fiber (DF). Cereal DF is an umbrella concept of heterogeneous polysaccharides of variable chemical composition and molecular weight, which are combined in a complex network in cereal cell walls. Cereal DF and its distinct components influence food digestion throughout the gastrointestinal tract and influence nutrient absorption and other physiological reactions. After repeated consumption of especially whole grain cereal foods, these effects manifest in well-demonstrated health benefits. As cereal DF is always consumed in the form of processed cereal food, it is important to know the effects of processing on DF to understand, safeguard and maximize these health effects. Endogenous and microbial enzymes, heat and mechanical energy during germination, fermentation, baking and extrusion destructurize the food and DF matrix and affect the quantity and properties of grain DF components: arabinoxylans (AX), beta-glucans, fructans and resistant starch (RS). Depolymerization is the most common change, leading to solubilization and loss of viscosity of DF polymers, which influences postprandial responses to food. Extensive hydrolysis may also remove oligosaccharides and change the colonic fermentability of DF. On the other hand, aggregation may also occur, leading to an increased amount of insoluble DF and the formation of RS. To understand the structure-function relationship of DF and to develop foods with targeted physiological benefits, it is important to invest in thorough characterization of DF present in processed cereal foods. Such understanding also demands collaborative work between food and nutritional sciences.


Adenosine receptor antagonism attenuates angiotensin II effects on adrenergic neurotransmission in the rabbit isolated vas deferens.
Angiotensin II augments adrenergic neurotransmission in the rabbit isolated vas deferens and suppresses purinergic neurotransmission. This study tests the hypothesis that angiotensin II augments adrenergic neurotransmission by depressing the neuronal release of ATP, resulting in suppressed formation of the inhibitory neuromodulator, adenosine or a related purine. Exogenous ATP added to the vasa deferentia increased adenosine formation and depressed adrenergic neurotransmission thus providing indirect support for the hypothesis. The adenosine receptor antagonist, 8-(sulfophenyl)theophylline (10 and 100 microM) depressed responses to exogenous adenosine and ATP but did not alter contractile responses to nerve stimulation or exogenously administered norepinephrine thus indicating that endogenous adenosine had no basal influence upon neurotransmission. However, the 8-(sulfophenyl)theophylline reduced angiotensin II effects on both adrenergic neurogenic contractions and evoked norepinephrine release. Additionally, the augmentation of adrenergic neurogenic contractions by angiotensin II was enhanced in the presence of ATP. These results are consistent with an ATP involvement in angiotensin effects on adrenergic neurotransmission and contrary to the initial hypothesis, suggest that purines enhance adrenergic neurotransmission in the presence of angiotensin II.


Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro.
Despite clinical success with anti-spike vaccines, the effectiveness of neutralizing antibodies and vaccines has been compromised by rapidly spreading SARS-CoV-2 variants. Viruses can hijack the glycosylation machinery of host cells to shield themselves from the host's immune response and attenuate antibody efficiency. However, it remains unclear if targeting glycosylation on viral spike protein can impair infectivity of SARS-CoV-2 and its variants. We adopted flow cytometry, ELISA, and BioLayer interferometry approaches to assess binding of glycosylated or deglycosylated spike with ACE2. Viral entry was determined by luciferase, immunoblotting, and immunofluorescence assays. Genome-wide association study (GWAS) revealed a significant relationship between STT3A and COVID-19 severity. NF-κB/STT3A-regulated N-glycosylation was investigated by gene knockdown, chromatin immunoprecipitation, and promoter assay. We developed an antibody-drug conjugate (ADC) that couples non-neutralization anti-spike antibody with NGI-1 (4G10-ADC) to specifically target SARS-CoV-2-infected cells. The receptor binding domain and three distinct SARS-CoV-2 surface N-glycosylation sites among 57,311 spike proteins retrieved from the NCBI-Virus-database are highly evolutionarily conserved (99.67%) and are involved in ACE2 interaction. STT3A is a key glycosyltransferase catalyzing spike glycosylation and is positively correlated with COVID-19 severity. We found that inhibiting STT3A using N-linked glycosylation inhibitor-1 (NGI-1) impaired SARS-CoV-2 infectivity and that of its variants [Alpha (B.1.1.7) and Beta (B.1.351)]. Most importantly, 4G10-ADC enters SARS-CoV-2-infected cells and NGI-1 is subsequently released to deglycosylate spike protein, thereby reinforcing the neutralizing abilities of antibodies, vaccines, or convalescent sera and reducing SARS-CoV-2 variant infectivity. Our results indicate that targeting evolutionarily-conserved STT3A-mediated glycosylation via an ADC can exert profound impacts on SARS-CoV-2 variant infectivity. Thus, we have identified a novel deglycosylation method suitable for eradicating SARS-CoV-2 variant infection in vitro. A full list of funding bodies that contributed to this study can be found in the Acknowledgements section.