This cross-sectional study included 31 patients with COPD subjected to PFTs (including the N2SBW test) and a cardiopulmonary exercise test (CPET). Patients were also evaluated using the following questionnaires: the COPD assessment test (CAT), the 36-Item Short Form Health Survey (SF36) and St. George's Respiratory Questionnaire (SGRQ). Peak oxygen uptake (peak VO2) was negatively correlated with the phase III slope of the N2SBW (SIIIN2)  and positively correlated with forced expiratory volume in one second . Breathing reserve was negatively correlated with SIIIN2, closing volume/vital capacity, and residual volume (RV)  and positively correlated with FEV1, forced vital capacity (FVC) and FEV1/FVC . SIIIN2 and CAT were independently predictive of VO2 and breathing reserve at peak exercise. RV, FVC, and FEV1 were independently predictive of the SF36-physical component summary, SF36-mental component summary, and breathing reserve, respectively. The SGRQ did not present any independent variables that could explain the model. In stable COPD patients, inhomogeneity of ventilation explains a large degree of exercise intolerance assessed by CPETs and, to a lesser extent, poor quality of life.Pulmonary function tests (PFTs) traditionally used in clinical practice do not accurately predict exercise intolerance in patients with chronic obstructive pulmonary disease (COPD). The aim of this study was to assess whether the nitrogen single-breath washout (N Chronic obstructive pulmonary disease (COPD) is a major public health problem worldwide that affects health-related quality of life (HRQoL) of individuals and causes disability and high morbidity and mortality . The HRQ1) is a marker of airflow obstruction in the proximal airways, whereas airflow obstruction in COPD occurs primarily in the distal airways (2SBW) test because it is a simple, non-invasive tool and can detect heterogeneity in the ventilation distribution and small airway disease in cases in which traditional PFTs indicate normal values (2SBW (SIIIN2) and closing volume/vital capacity (CV/VC) indicate heterogeneity in the ventilation distribution and small airway disease, respectively  are valuable tools for the diagnosis and assessment of the severity of COPD and have many clinical applications, such as evaluating prognosis and response to therapy . Spirome airways ,5. In re airways . There hl values . Increasectively . Two recith COPD ,9.2) and a low breathing reserve have been identified as the main contributors that limit exercise (The cardiopulmonary exercise test (CPET) can detect abnormalities in the functional capacity of the respiratory and cardiovascular systems earlier than other tests . In patiexercise –13. Diffexercise ,12. In fexercise ,13.2SBW test explain exercise intolerance in stable COPD patients. Secondly, we sought to evaluate whether parameters of the N2SBW test explain poor quality of life of these patients.As the outcomes of traditional PFTs do not correlate well with CPET parameters, it is important to investigate other pulmonary function methods that are better associated with exercise intolerance in patients with COPD. Considering that ventilatory limitation aggravated by exercise is a major contributor to the reduced functional capacity in COPD patients, we hypothesized that heterogeneity in the ventilation distribution and small airway disease may contribute to better understand exercise intolerance in these patients . Thus, tBetween April 2015 and January 2017, we conducted a cross-sectional study involving 38 consecutive patients with COPD recruited at the Policlínica Newton Bethlem, Rio de Janeiro, Brazil. The exclusion criteria were age less than 40 years, smoking load of fewer than 10 pack-years, previous pleuropulmonary disease not related to COPD, use of supplemental oxygen therapy, previous thoracic surgery, respiratory exacerbations or infections in the past 4 weeks and any comorbid condition that might reduce exercise capacity. The project was approved by the Research Ethics Committee of the Centro Universitário Augusto Motta under CAAE No. 52885116.6.0000.5235 and complied with the Declaration of Helsinki. All subjects signed a consent form.The m-MRC scale is a subjective tool that assesses limitations in ADLs and has been validated for use in Portuguese language . The m-MThe CAT is a specific questionnaire used to measure the HRQoL of patients with COPD. This one-dimensional measure contains 8 items related to the impairment of health status. At the end of the test, the scores of all the responses are summed, and the clinical impact of COPD is evaluated according to the stratification of CAT scores. The total score ranges from 0 to 40, and higher scores indicate poorer health status .vs ≥2 not leading to hospital admission or ≥1 leading to hospital admission), patients were classified as follows: A ; B ; C , and D   and a mental component summary (MCS). The scores range from 0 to 100, with higher scores indicating better HRQoL .The SGRQ is a validated, well-known self-administered tool that measures symptoms, ADLs, and HRQoL of patients with chronic respiratory diseases, particularly COPD. It consists of 50 items divided into 3 domains: symptoms (8 items), activity (16 items) and impact (26 items). Each item has a derived weight, and lower values indicate better HRQoL .1 values after salbutamol use as follows: GOLD 1 ; GOLD 2 ; GOLD 3 , and GOLD 4   in the Hospital Universitário Pedro Ernesto of the Universidade do Estado do Rio de Janeiro, Brazil, following the standardization of the American Thoracic Society . The ref2SBW test with a 3000 HDpft  according to the recommendations of the American Thoracic Society/European Respiratory Society . Then, patients exhaled slowly at a flow rate of approximately 0.3–0.5 L/s until reaching the RV. The concentration of exhaled N2 was recorded by a device located at the airway opening. Two indices derived from this procedure were reported as a percentage of the predicted values: SIIIN2, which is the change in N2 concentration between 25 and 75% of the expiratory volume, and the CV/VC ratio, which is the percentage of the VC that is exhaled after the beginning of airway closure , carbon dioxide output (VCO2), minute ventilation (VE) and related variables were calculated breath-by-breath  coupled with a metabolic analyzer  in the Laboratório de Movimento Humano at the Centro Universitário Augusto Motta according to previous recommendations . We usedy-breath . Breathiy-breath . Heart ry-breath . The equy-breath .2, breathing reserve, CAT, SF36 PCS, SF 36 MCS, and SGRQ with demographic, clinical and resting lung function variables were analyzed using Pearson correlation coefficients. Due to the small number of patients for each item on the m-MRC scale, the patients were categorized into 2 groups according to the m-MRC scale (<2 or ≥2); this cut-off separates ‘low breathlessness' from ‘high breathlessness' . The associations between the variables peak VOessness' . The ass2SBW test) and possible confounders  that were independently related to performance during the CPET (peak VO2 and breathing reserve) and HRQoL . The data that were tested as independent variables in the forward stepwise regression analysis were as follows: forced vital capacity (FVC), FEV1, FEV1/FVC, forced expiratory flow during the middle half of the FVC (FEF25–75%), TLC, RV, RV/TLC, SIIIN2, and CV/VC. These models were adjusted for relevant confounders .Forward stepwise regression analysis was used to identify the resting lung function parameters . The overall performance measures of the regression models was evaluated using the coefficient of determination (R2), adjusted for the number of variables retained in the proposed model (cumulative R2).In addition, the forward stepwise regression analysis was used to identify the variables provided by spirometry and body plethysmography that were independently related to parameters of the Nvs predicted outcomes, along with regression lines showing the slope and intercept) and the limits of agreement (LoA) plot for each of the proposed models . Differences were considered significant for P values less than 0.05.Among the 38 patients evaluated for inclusion in this study, 7 were excluded for the following reasons: respiratory exacerbations in the month before recruitment (n=3), history of pleuropulmonary disease not associated with COPD (n=3), and use of home oxygen therapy (n=1). The average age was 67.5±9.9 years, and the average smoking load was 50.9±26.2 pack-years. All but 4 patients were treated with inhaled bronchodilators and/or inhaled corticosteroids. The demographic and clinical data, resting lung function parameters, and CPET data are shown in 2 was negatively correlated with SIIIN2  and positively correlated with FEV1 and FEF25-75% . Breathing reserve was negatively correlated with SIIIN2, CV/VC, RV and RV/TLC  and positively correlated with FEV1, FVC, FEV1/FVC and FEF25-75% . We further assessed the correlations between parameters of the N2SBW test with resting lung function variables and possible confounders. SIIIN2 was negatively correlated with FEV1, FVC, FEV1/FVC and FEF25-75%  and positively correlated with RV and RV/TLC . CV/VC was negatively correlated with FEV1, FVC, FEV1/FVC and FEF25-75%  and positively correlated with age, TLC, RV and RV/TLC . Changes in the demographic, clinical, lung function and CPET variables according to the m-MRC scale (class <2 vs class ≥2) were also evaluated; the results of this analysis indicated no significant difference in the variables between the patients divided into the 2 classes of the m-MRC scale.The univariate correlations between the demographic, clinical, lung function and CPET data are shown in 2SBW test) could independently predict performance during the CPET and the HRQoL in these patients. The results of the forward stepwise regression analysis for the prediction of the CPET and HRQoL variables are shown in N2 was an independent predictor of the CAT score, peak VO2 and breathing reserve. RV, FVC and FEV1 were independent predictors of SF36 PCS, SF 36 MCS and breathing reserve, respectively. The SGRQ could not be predicted by any independent variables in the models. We further assessed whether the resting lung function variables would demonstrate an independent role in predicting the parameters of the N2SBW test. FEV1 and RV/TLC were the only independent predictors for SIIIN2 and CV/VC, respectively  and the observed correlation for the main outcome (peak VO2) and SIIIN2 (r=-0.681). For an alpha of 0.05, the power was 99.3%, showing the adequacy of the studied sample size to get significant results.To provide a context for interpreting the null findings, a 2SBW test were associated to a lesser extent with the CPET parameters and HRQoL. In addition, other lung function variables measured by spirometry and body plethysmography were associated with performance during exercise and HRQoL of patients with COPD. To our knowledge, this study is the first to demonstrate that the parameters of the N2SBW test partially explain the exercise intolerance and poor quality of life in stable COPD patients.The main finding of this study was that, in patients with COPD, a greater inhomogeneity in ventilation corresponded to a lower oxygen consumption and breathing reserve at peak exercise. The inhomogeneity of ventilation was also positively associated with poor HRQoL of these patients. Compared with the heterogeneity in ventilation, the changes in small airways measured by the NN2 (but not the CV/VC ratio) was the only independent predictor of peak VO2. Interestingly, Jones et al. (2 had a higher ventilatory inefficiency (higher ventilation/carbon dioxide output nadir) during the CPET. Therefore, we believe that increased SIIIN2 at rest indirectly indicates the ventilatory inefficiency detected during symptom-limited maximal exercise. Of note, our models were adjusted for relevant confounders . Furthermore, we used the predicted values for the lung function parameters, which adjust the absolute values for gender, age, weight and height . Breathing reserve depends on several factors that are responsible for ventilatory demand, including metabolic demand, body mass, dead space ventilation and neuroregulatory mechanisms  and that these abnormalities are associated with significant patient-centered outcomes, including shortness of breath and exercise intolerance (In our study, the strongest correlation was observed between SIIIchanisms . A low bchanisms  evaluateolerance ,28. Empholerance ,29.N2, our results indicate that changes in the CV/VC may not be a major contributor to CPET measurements in patients with COPD because CV/VC was not retained in any of the regression models that were developed. In the N2SBW test, an increased CV/VC reflects air trapping because of small airway closure, and therefore, it can potentially provide information about early abnormalities in the small airways in COPD (In contrast to the variations in SIII in COPD . Althoug in COPD .2 values that explained less than 50% of the variance (N2 alone explained 35% of the variability of the CAT scores. The clinical variables and traditional PFTs were not retained in the regression models in our study, likely because of the strong association between CAT and SIIIN2. In contrast to CAT, we were unable to build a regression model for the m-MRC scale. This result was expected because the CAT is a multidimensional scale that includes seven other items, not only shortness of breath, whereas the m-MRC scale is one-dimensional and assesses only dyspnea (Although COPD is primarily a lung disease, it also causes significant systemic effects that can lead to a decline in health status and HRQoL . A recenvariance . Conside dyspnea .In this study, the SF36 PCS was associated with various physiological parameters at rest. However, the strongest correlation was observed between the increase in RV and the decrease in SF36 PCS, and RV was the only physiological parameter that was included in the SF36 PCS regression model. Air trapping caused by the combined effect of peripheral airway disease and increased ventilatory demand imposes critical mechanical restrictions and worsens dyspnea, which, in turn, leads to the decline of HRQoL of patients with COPD . Interes2 and the SGRQ score. Similarly, other researchers reported that peak VO2 is a poor indicator of health status (Our findings were similar to those of Morishita-Katsu et al.  who evalh status ,39, suggh status .2SBW test assessments in randomized controlled trials. In this context, the N2SBW test may contribute to the categorization of patients and the evaluation of responses to various clinical and surgical therapies for COPD.Our study is characterized by the following limitations. First, this study included a small number of subjects from a single center and employed a cross-sectional design; therefore, the temporal effect on the stability of the response of each evaluated tool is unknown. Due to the multiple statistical tests performed, some of the findings may be a result of chance. Therefore, there is the possibility of type I error, i.e., accepting an alternative hypothesis when the results can be attributed to chance . Second,2SBW test measurements and CPET parameters in stable COPD patients. In these patients, inhomogeneity of ventilation explains much of the exercise intolerance assessed by the CPET and, to a lesser extent, the poor HRQoL. Compared with the N2SBW test, the variables measured by spirometry and body plethysmography more strongly influenced the HRQoL and, to a lesser degree, oxygen consumption in patients with COPD. The contribution of the N2SBW test to COPD studies should be further explored, as this test has the potential to be used as a complementary tool in clinical practice to assess these patients.In conclusion, there was a relationship between N


Diethylnitrosamine (DEN) induces hepatocarcinogenesis, increasing mitotic hepatocytes and leading to chronic inflammation. In addition, type 1 diabetes mellitus (T1DM) is also characterized by a proinflammatory state and by requiring insulin exogenous treatment. Given the association of diabetes, insulin treatment, and cell proliferation, our specific goal was to determine whether the liver in the diabetic state presents a greater response to DEN-induced cell cycle alteration, which is essential for the malignant transformation. Male C57BL/6 mice (four-week-old) were divided into 4 groups: C, C + DEN, T1DM, and T1DM + DEN. Mice were euthanized ten weeks after DEN injection. DEN per se produced an increase in liver lipid peroxidation levels. Besides, in T1DM + DEN, we found a greater increase in the proliferation index, in comparison with C + DEN. These results are in agreement with the increased expression observed in cell cycle progression markers: cyclin D1 and E1. In addition, a proapoptotic factor, such as activated caspase-3, evidenced a decrease in T1DM + DEN, while the Vascular Endothelial Growth Factor (VEGF) and the protooncogene p53 showed a higher increase with respect to C + DEN. Overall, the results allow us to highlight a major DEN response in T1DM, which may explain in part the greater predisposition to the development of hepatocarcinoma (HCC) during the diabetic state. Liver cancer is one of the most common malignancies worldwide. Hepatocellular carcinoma (HCC) accounts for about 80%–90% of all liver cancers and is the fifth most common cause of cancer mortality , 2. MajoDiethylnitrosamine (DEN) is a well-known potent hepatocarcinogen agent present in tobacco smoke, water, cured and fried meals, cheddar cheese, agricultural chemicals, cosmetics, and pharmaceutical products , 10. CytDEN is known to induce damage in many enzymes involved in DNA repair and is normally used to induce liver cancer in experimental animal models –16. ExpoType 1 diabetes mellitus (T1DM) is a serious and growing health problem worldwide and is associated with severe acute and chronic complications that negatively influence both the quality of life and survival of affected individuals. Clinically, altered liver size is seen in both juvenile and adult diabetic patients, which can be the result of alteration in cell number, cell growth, and/or cell death (apoptosis) , 20. TheWe demonstrated that hyperglycemia increases ROS levels, specifically hydroxyl radicals, which initiate mitochondrial apoptosis either directly and/or by acting through Bcl2 family proteins, thus inducing the formation of the mitochondrial permeability transition pore (MPTP), the release of cytochrome c, and the activation of caspase-3 . In this Guide for the Care and Use of Laboratory Animals . All the experimental protocols were performed according to the Regulation for the Care and Use of Laboratory Animals (Expedient 6109/012 E.C. Resolution 267/02) and approved by the Institutional Animal Use Committee of the National University of Rosario, Argentina.Four-week-old male C57BL/6 mice were provided by the School of Medicine, National University of Rosario, and were maintained at the animal facilities of the School of Biochemistry of the National University of Rosario. Animals received humane care according to criteria outlined in then = 5 per group), housed in plastic cages, and kept on hardwood bedding in animal facilities with a 12-hour light/dark cycle and controlled temperature (23 ± 2°C) and ventilation. Water and a standard diet were provided ad libitum throughout the experiment. Mice of group 1  received a single intraperitoneal (i.p.) injection of both citrate buffer at week 6 and saline solution at week 7, vehicles of STZ (diabetogenic agent) and DEN (carcinogenic agent), respectively. Animals of group 2 (C + DEN group) were injected with a single dose of DEN  [Mice were randomly divided into 4 experimental groups   at week g, i.p.)  at week g, i.p.) , 27. MicLipid peroxidation (LPO) levels were determined as indirect measurements of ROS . A fractTo investigate modifications in proliferation activity among the experimental groups, liver slides were examined by immunohistochemical staining with anti-proliferating cell nuclear antigen (PCNA) antibody, determined by the method of Greenwell et al. . The PCNPCNA, as a naturally occurring cell marker of proliferating cells, offers an alternative method for investigating cell proliferation. Molecular studies indicate that the synthesis of PCNA is initiated in the nucleus in late G1 phase and continues during the S phase. The different staining patterns recognized in this study are believed to reflect individual phases of the cell cycle. Cells expressing no staining in the nucleus or cytoplasm are expected to be quiescent Go phase cells. Minimal nuclear staining would be consistent with G1 phase cells. Deep red nuclear staining is characteristic of S phase. G2 cells present speckled nuclear and cytoplasmic staining. In mitosis (M) the nucleoplasm and cytoplasm coalesce with the loss of nuclear boundaries. This could account for the diffuse speckled cytoplasmic staining specifically observed in all actively mitotic cells , 43.https://imagej.nih.gov/). Equal loading and protein transference were checked by detection in mitochondrial fraction by prohibitin, cytosolic fraction by GAPDH, and nuclear fraction by Ponceau S staining of the membranes. According to the bibliography, nuclear loading controls are modified in association with diabetes or HCC, and thus Ponceau S staining is the most reliable loading control in nucleus in STZ or DEN-treated mice livers [Tissue samples were homogenized in 300 mM sucrose with protease inhibitors. Cytosolic, mitochondrial, and nuclear extracts were prepared as described previously . Proteine livers , 47. t-test; one-way or two-way analysis of variance followed by Tukey's test was applied wherever necessary. Differences were considered to be statistically significant when p < 0.05.Data are expressed as means ± SEM. Significance was determined by Student'sDEN treatment induced an increase of 102% in the generation of TBARS in C + DEN as compared to control animals, highlighting the known oxidizing effect of DEN  Figure . BesidesAs cyclin E1 and cyclin D1 have an important role in the progression of cell cycle from phase G1 to phase S, we evaluated both proteins' nuclear levels by Western blot. Analyses revealed an increase of cyclin E1 and cyclin D1 nuclear localization in T1DM + DEN group when compared to T1DM group  Figures . Also, TBax : Bcl-xL could reflect accurately the status of apoptosis, since the fate of the cells  is largely dependent on the mitochondrial Bax : Bcl-xL ratio , 51. We In most hepatocarcinogenesis models, a very long latent period occurs, following initial exposure to the carcinogen. During this latent period, the liver cells are not quiescent. Preneoplastic hepatocytes differ from normal hepatocytes according to the expression of many mediators, some of which are overexpressed, while the expressions of others are decreased , 52. In The pathogenic equation for T1DM presents a complex interrelation of metabolic, genetic, and environmental factors, as well as inflammatory mediators. Among the latter, it is mostly unclear whether they reflect the disease process or are simply signs of systemic or local responses to the disease . The higIncreased generation of ROS and decreased antioxidant enzymes in liver tissues have been reported in many models of DEN-induced hepatocellular carcinoma , 56. DatProgrammed cell death plays an important role in the genesis of cancer. In this regard, several proteins that are structurally related to Bcl-2, an inhibitor of apoptosis, have been identified . DiverseVEGF is the main growth factor implicated in angiogenesis. It has been demonstrated by immunohistochemistry that VEGF expression is increased in 36% to 83% of HCC cells . Park etTaken together, our results show a greater liver sensitivity to DEN treatment in T1DM group, suggesting that the degree of chronic inflammation, increased ROS levels, and alterations in cell cycle balance derived from insulin treatment, among other factors that are hepatic hallmarks of these mice, could play a fundamental role in its sensitization.In conclusion, data from this study suggest for the first time that, in T1DM + DEN group, the liver cells are not quiescent and exhibit alterations in the levels of certain proteins involved in cell cycle progression. During a diabetic pathology accompanied by insulin treatment, the exposure to a carcinogen such as DEN could lead to an altered early stage in the hepatocarcinogenesis progression. In this regard, this precondition could represent a risk factor to an accelerated DEN-induced process. In this sense, it becomes highly relevant to understand the pathways involved in the development of the initial phases of HCC in T1DM and therefore find the main targets responsible for the alteration in the cell cycle progression.


Cognitive insight represents the ability to question and criticize the validity of one’s beliefs, to recognize when beliefs may be faulty, and to then rely on external feedback to make correct assessments of a situation. Cognitive insight is characteristically impaired in persons with schizophrenia and related psychoses. The Beck Cognitive Insight Scale (BCIS) is the most widely used tool to assess cognitive insight, yet there is no consensus regarding clinical cutoff values. Cognitive insight is predictive of better response to psychosocial treatment and the ability to accept critical feedback from treatment teams, thus cutoffs are an important next step needed to facilitate the clinical interpretation of the BCIS. Some studies have attempted to develop diagnostic cutoffs, yet no study has proposed clinical cutoffs to differentiate levels of cognitive insight between patients with schizophrenia.Three hundred and eighty-five English or French-speaking patients with a schizophrenia spectrum disorder (203 first-episode and 182 multiple-episode psychosis patients) and 185 healthy controls completed a battery of clinical and neuropsychological tests, including the BCIS. Patients and controls were matched on age, sex, level of education, and socio-economic-status. Correlations were calculated between the composite index and previously identified correlates of cognitive insight. Variables significantly correlating with the BCIS composite index were then included in a clustering analysis to classify patients according to their clinical profile. Two clinical profiles representing low and high cognitive insight were identified, and were based on global functioning and IQ. Composite index scores at the 33rd percentile in the low cognitive insight cluster and the 66th percentile in the high cognitive insight cluster were calculated.Functioning and IQ significantly correlated with the BCIS composite index and were included in a clustering analysis, using a pre-determined number of two clusters. Independent samples t-tests revealed that the 2 clusters differed significantly on the BCIS self-reflectiveness score (t(372) = -3.93, p < .001) and on the composite index (t(372) = -3.17, p = .002). There was no difference between clusters on self-certainty (t(372) = .31, p = .76). Patients in cluster A had a mean SR, SC, and composite index of 12.65 , 7.78  and 4.87 , respectively, while mean scores for patients in cluster B were 15.11 , 7.64  and 7.47 . In cluster A, the values of the 33rd and 66th percentiles were 2.6 and 7 respectfully. In cluster B, these values were 5 and 9. We are proposing that 33% of patients with the lowest composite index scores in cluster A represent those with low cognitive insight. Accordingly, 33% of patients with the highest composite index scores classified in cluster B represent those with high cognitive insight. Low cognitive insight is thus represented by a score of 3 or below, borderline scores range from 4 to 9, and high cognitive insight is represented by a score of 10 or above.We proposed clinical cutoffs for the BCIS with a theoretical basis anchored in patient clinical profiles (functioning and IQ). Clinical cutoffs will facilitate and better orient treatment teams in the clinical interpretation of the BCIS and ergo to patients’ level of cognitive insight. The development of such cutoffs will help to reduce heterogeneity in psychosocial group intervention, will facilitate interventions aimed at increasing cognitive insight, and improve communication between patients and their treatment teams.


However, they have been considered non-active for surface enhanced Raman spectroscopy (SERS). In this work, we explore the scattering contributions of Pd and Pt for the detection of organic molecules. The Pd and Pt nanostructures were synthesized on silicon substrate using a modified galvanic displacement method. The results show Pt nanoparticles and dendritic Pd nanostructures with controlled density and size. The influence of surfactants, including sodium dodecyl sulfate and cetyltrimethylammonium bromide, on the size and morphology of the nanostructures was investigated. The Pd and Pt nanostructures with a combination of large size and high density were then used to explore their applicability for the detection of 10 Galvanic displacement is one of the electrochemical methods used to fabricate metals on semiconductor surfaces ,2. GalvaIn galvanic displacement, the semiconductor substrate acts as an electron source via its surface oxidation. Likewise, metal ions in solution, taking up electrons supplied by the substrate, are reduced to metal atoms. Thus, the overall displacement process occurs when metals are deposited on the semiconductor substrate and the surface oxide is dissolved, as described by the following chemical equations ,3:(1)An−5 M thiocyanate [−2 M benzenethiol, with significant EFs for Pd and Pt [−6 M R6G and EF up to 105 for 4-mercaptopyridine have been achieved with Pd urchins [Platinum (Pt) and palladium (Pd) are noble transition metals, whose excellent catalytic and electrochemical properties have been widely used in modern technologies, including chemical and biosensing ,34,35, pocyanate  with an ocyanate . While td and Pt . Monodisd and Pt ,50, Pd fd and Pt ,52 and Pd and Pt ,53. So f urchins , respect−5 M and 10−2 M, as well as EFs up to 102 and 10, respectively. Additionally, the influence of surfactants, including sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB), on the size and morphology of the nanostructures was investigated.Few authors have reported successful fabrication of metallic nanostructures on Si without HF ,20. As dA range of characterization techniques were employed to assist the investigation. Scanning electron microscopy (SEM) was used to determine morphology, size, and density of the nanostructures. The chemical information of the nanostructures was obtained by energy dispersive X-ray spectroscopy (EDX). Finally, surface plasmon resonance (SPR) absorption and SERS properties of the nanostructures were recorded by UV-Vis spectroscopy and Raman spectroscopy, respectively.2PtCl4, ≥99.9%), sodium tetrachloropalladate(II) , and cetyltrimethylammonium bromide  purchased from Sigma–Aldrich , sodium dodecyl sulfate  from Fisher Scientific , which were used as received. Hydrofluoric acid  was purchased from Thermo Fisher Scientific . For SERS experiments, Rhodamine 6G dye  and paraoxon-ethyl  were purchased from Sigma–Aldrich . All solutions were prepared using deionized (DI) water. HF was contained in a polypropylene beaker for experimental use. The substrate was Si (100) wafer  purchased from El-Cat Inc. . Prior to deposition, Si wafer was cleaved into 1.5 cm × 1.5 cm2 pieces. The chemicals used for cleaning Si substrate were acetone and ethanol .The chemicals used for metallic deposition included potassium tetrachloroplatinate(II) (Kw/w) HF solution for 2 min to eliminate surface oxide and activate the surface. The substrate was then instantly immersed for 5 min in 0.3 mM K2PtCl4 or Na2PdCl4 for Pt or Pd deposition, respectively, followed by rinsing with deionized water. This deposition procedure will be denoted as 1 deposition cycle, which includes immersion in HF, treating with the metal precursor solution and rinsing with DI water. Multiple deposition cycles were also used in the present investigation. For experiments that were used to study the influence of surfactants on the final outcome, the metallic precursor solution was composed of 0.1 mM SDS or CTAB with 0.3 mM K2PtCl4 or Na2PdCl4. Finally, the Pt and Pd nanostructures were air dried and stored for further characterization.The Pd and Pt nanostructures were synthesized by the galvanic displacement method. The synthesis procedure includes cleaning the Si substrate with acetone, followed by ethanol and DI water. Each cleaning step was performed for 10 min, in an ultrasonic bath, to degrease and de-contaminate the substrate. For the deposition, the Si substrate was immersed in 10%  of the stock solution was drop casted and dried under ambient conditions. Prior to drop casting the analyte, the sample was treated with ambient air plasma for 1 min. under medium radio frequency power level (11 W) by a plasma cleaner .The size and morphology of the nanostructures were studied by scanning electron microscopy (SEM) using a FEI Quanta-250  SEM instrument at 10 kV accelerating voltage. The SEM instrument was equipped with an Oxford Aztec energy dispersive X-ray (EDX) analysis system, which was used to conduct elemental analyses of the deposited nanostructures. UV-Vis absorption spectroscopy was recorded by a Perkin Elmer Lambda 25  spectrophotometer. To prepare samples for UV-Vis measurements, each sample was immersed in 1 mL DI water and sonicated at the highest power for 1–3 min to detach the metallic nanostructures from the Si substrate and disperse them in DI water. SERS measurements were performed at room temperature on a Renishaw Dispersive Raman Spectrometer  with an Ar-ion laser with a wavelength of 488 nm, using 50× objective lens, with incident power of 5 mW for 4 accumulations, each of 30 s duration. SERS spectra were collected from several random regions on each sample to confirm homogeneity.Upon the completion of one deposition cycle, Pd nanostructures appeared to be quasi-spherical a. The laIn contrast, Pt nanostructures showed a steady increase in density, with respect to an increasing number of deposition cycles . This in42−, thus attract and concentrate Pd precursors toward the Pd nanostructures. Sodium dodecyl sulfate (SDS), an anionic surfactant, and cetyltrimethylammonium bromide (CTAB), a cationic surfactant, have been widely used as stabilizers ,58 and mChemical compositions of the as-synthesized samples were determined by EDX . The Pd In addition to EDX, UV-Vis data was also obtained. −5 M R6G dye, drop casted onto Pt and Pd nanostructures. For comparison, the same concentration of R6G was also drop cast onto bare Si substrate without metallic deposition. No noticeable Raman signal was detected using this bare Si substrate. On the other hand, several strong Raman modes were observed for the Pd nanostructure sample. Nearly all of these Raman modes were also visible for the Pt nanostructure sample. However, their intensities were significantly reduced.To evaluate the Pt and Pd nanostructures as possible platforms for SERS application, R6G was chosen as a probe molecule. R6G has been widely used for SERS due to their well-defined vibrational features. To obtain the best performance of SERS substrates, samples with a combination of high-density and large metallic nanostructures were used. These samples were achieved after ten deposition cycles see . Figure −2 M paraoxon drop casted on Pt and Pd nanostructures. The three strongest fingerprint peaks, representing different vibrational modes of the paraoxon molecule, were clearly seen in these spectra [In addition to R6G detection, paraoxon was used, which served as a real sample. Here, Pt and Pd nanostructure platforms were used to demonstrate their ability to detect a real sample like paraoxon. Paraoxon is an organophosphorus compound and is used to regulate pests in crops and plants. It is highly toxic, causing detrimental health problems if exposed to food and drinking water . SERS ca spectra . Raman sSERS and IRS are Raman intensities of SERS and non-SERS substrates, respectively, while cSERS and cRS are analyte concentrations used for SERS and non-SERS substrates, respectively [SERS to IRS. However, since the Raman signal obtained from bare Si substrate (IRS) was too weak to be detected by our instrument, it was not possible to obtain absolute values for EF. Instead, we roughly estimated EFs in a manner similar to those reported by Yamamoto et al. [2, which was ~three times higher than that for Pt. Similarly, EF values for paraoxon on Pd and Pt both were ~10.The enhancement factor (EF) was determined to quantitatively evaluate the effectiveness of the SERS substrate. The most widely used definition of the EF is described asectively . In the o et al. . Thus, tThe Raman enhancement can be attributed to the local electromagnetic enhancement ,74. For In summary, we report a modification of the galvanic displacement method, in which Pt and Pd nanostructures were deposited on an n-type Si substrate with limited exposure to HF. By performing multiple cycles of the metallic deposition steps, large dendritic Pd and Pt nanostructures with a high density and small size distribution were obtained. Further, the Pd and Pt nanostructures were successfully used as SERS platforms for detecting low concentrations of organic analytes. From the results, it is clearly evident that Pd and Pt nanostructures, prepared by the modified galvanic displacement method, can produce stable and reproducible nanostructures that can be used for SERS application. Additionally, SDS and CTAB surfactants were utilized, which produced an increase in anisotropic growth of the dendritic Pd nanostructures. A higher density of Pt nanostructures was observed when CTAB was utilized. Future investigation will mainly focus on Pt and Pd samples, prepared in the presence of surfactants.


Purpose: Nitric oxide (NO) bioavailability is reduced during acute altitude exposure, contributing toward the decline in physiological and cognitive function in this environment. This study evaluated the effects of nitrate : −1·min−1) rested and performed exercise twice at moderate  and twice at very-high  simulated altitude. Participants ingested either 140 ml concentrated 2), pulmonary oxygen uptake (Results: Pre-exercise plasma  and 4.2%  at 3,000 and 4,300 m, respectively (p = 0.019). Oxygenation of the gastrocnemius was elevated during the TT consequent to BRJ (p = 0.011). The number of false alarms during the Rapid Visual Information Processing Task tended to be lower with BRJ compared with PLA prior to altitude exposure (p = 0.056). Performance in all other cognitive tasks did not differ significantly between BRJ and PLA at any measurement point (p ≥ 0.141).Conclusion: This study suggests that BRJ improves physiological function and exercise performance, but not cognitive function, at simulated moderate and very-high altitude. Nevertheless, it remains to be established whether a higher dose of Several studies conducted at sea-level indicate that Against this background, the purpose of the present study was to evaluate the effects of −1·min−1 volunteered and provided fully informed written consent to participate in the present study. All participants were non-smokers, normotensive and not currently taking any medication. None of the participants had traveled to an altitude >1,500 m during the preceding 3 months and all were currently residing <500 m. The study received institutional ethical approval and adhered to the principles of the Declaration of Helsinki.Ten healthy men with a mean (SD) age of 23 (3) years, body mass of 78.0 (12.5) kg, stature of 180.3 (8.1) cm, and sea-level maximal rate of oxygen uptake  or very-high altitude . There were, in total, four different experimental conditions, which were conducted in a randomized order. These were: (1) 3,000 m simulated altitude with Participants attended the laboratory on five separate occasions within a 7 week period. Visits were separated by a minimum of five and maximum of 12 days, and conducted at the same time each day (±1 h) to minimize the influence of circadian variance. On the first laboratory visit, participants completed an incremental exercise test to volitional exhaustion in normoxia to elucidate  Conkin, , to simu−1 for the first stage, and increased by 1 km·h−1 each subsequent stage. The treadmill gradient was set to 10% for all submaximal stages, and participants were required to carry a 10 kg backpack. Gas data obtained during this first part of the test was used to calculate the A two-part incremental test was conducted on a motorized treadmill  at sea-level. During the first part of the test, participants completed four submaximal stages of 3 min duration, interspersed with 1 min recovery periods. The treadmill speed was set at 1 km·h−1, which replicated the low intensity nature of high altitude hiking . Blood was immediately centrifuged for 3 min at 5,000 rpm. Plasma was subsequently extracted into opaque cryotubes , which were pre-treated with 6.5 mM nethylmaleimide (NEM) and 0.1 mM diethylenetriaminepentaacetic acid (DTPA) to minimize the interchange between NO metabolites . Subsequent measures of SpO2 and HR were conducted at rest in hypoxia immediately prior to exercise, during the final 5 min of steady-state walking, immediately following the TT, and prior to exiting the hypoxic chamber.Following BP assessments, SpOCognitive function was assessed at rest prior to hypoxic exposure, at rest in hypoxia, during steady-state walking , and 5 min following TT completion. All cognitive assessments were conducted using the Cambridge Neuropsychological Test Automated Battery , and administered via a touch screen tablet computer . Three tests which have previously been applied to evaluate the efficacy of nutritional supplementation on cognitive function were administered  and incongruent stimuli . The total number of correct responses and response times to the test were analyzed.The RVP is a test of sustained attention. The participant is presented with a white box in the center of the computer screen. Single digits ranging from 2 to 9 are presented one at a time in a pseudo-random order inside the box, appearing at a rate of 100 digits per minute. The participant is required to detect specific 3-digit sequences, including 2-4-6, 4-6-8, and 3-5-7. As soon as a target sequence is detected, the participant is required to touch a button on the screen. The task administration time was ~10 min. The number target sequences correctly identified, response time for correctly identified sequences, and number of false alarms were analyzed.The SSP is a visuospatial analog of the Digit Span Task, and measures working memory capacity. The participant is initially presented with a black screen containing 9 white boxes. In a variable sequence, some of the boxes briefly change color. The participant is required to touch the boxes in the same order in which they changed color. Initially, two boxes change color. For every successful correct response, the number of boxes changing color is increased by one, up to a maximum of nine boxes changing color. The sequence and color change of boxes is varied throughout the task. The test terminates when the participant fails to correctly identify the sequence at a particular level on three occasions, or correctly identifies all nine boxes that change color during the final stage of the task. The task administration time was ~5 min. The longest sequence of correctly identified boxes and mean number of attempts to pass were analyzed.2, a small patch was shaved on the left lateral gastrocnemius. The portable NIRS device was secured to the leg via a combination of surgical tape and an elastic non-compressive bandage. The bandage also served the combined role of shielding the device from extraneous light. The device placement was replicated between trials with reference to local anatomical sites and the small shaved patch on the leg. The device uses the modified Beer-Lambert Law to monitor concentration changes in oxyhaemoglobin ([O2Hb]), deoxyhaemoglobin ([HHb]), and total hemoglobin ([tHb]). Values are expressed relative to the first datum, which was collected following 5 min of seated rest in normoxia. The [HHb] signal, which is regarded as being essentially blood volume insensitive during exercise  * 100. Data was recorded at 10 Hz, and a differential pathlength factor (DPF) of 4.94 was used, in accordance with the findings of Duncan et al.  is a popular non-invasive technique for evaluating tissue oxygenation Neary, . In the n et al. . Mean mu2Hb], [HHb], and [tHb] relative to the first datum, which was obtained following 5 min of seated rest in normoxia. Data were recorded at 10 Hz and averaged for each variable across all channels to provide an index of pre-frontal cortex oxygenation. The DPF was adjusted according to the participants age , and overall for the 3 km TT.Cerebral oxygenation was also monitored via means of a cerebral NIRS device. A dual wavelength (760 and 850 nm), light weight (230 g), portable cerebral NIRS device  was used for this purpose. The unit consisted of a headband with 8 light emitters and two light detectors, with an interoptode distance of 3.5 cm. The device placement was replicated between trials by positioning the bottom of the headband 1 cm above the eyebrows, and the middle of the headband in the center of the forehead. This device principally measures oxygenation of the medial prefrontal cortex. As with the muscle tissue NIRS device, the cerebral NIRS applies the modified Beer-Lambert Law to monitor changes in , muscle [HHb], muscle [tHb], and TT performance across experimental conditions. Cerebral [HbO2], cerebral [HHb], and cerebral [tHb] were compared during TT via two-way  ANOVA, and during each cognitive testing period via three-way  repeated measures ANOVA. Significant three-way interaction effects were explored using one-way ANOVA and two-way interactions were explored using paired t-tests. Statistical significance was set at p < 0.05 and Bonferroni corrections were applied to all post-hoc tests. Cohen's d is presented for information on effect sizes. Effect sizes were interpreted as ≤0.2 trivial, >0.2 small, >0.6 moderate, >1.2 large, >2 very large, and >4 extremely large  repeated measures ANOVA was employed to assess differences in plasma  was significantly lower during TT exercise, and tended to be lower during steady-state exercise (which involved simultaneous cognitive tasks) following n et al.  reportedn et al. . In hypon et al. . Thus, iThe effects of 2 observed in the present study following Several mechanisms have been suggested to explain the beneficial effects of +, Pi, and ADP (Vanhatalo et al., The effects of The present study makes an important contribution toward our understanding of the applications of 2 of altitude by reducing the FIO2. This exercise environment allowed greater control over potentially confounding variables vs. testing at terrestrial altitude, and helped minimize participant burden. Recent work has also reported broadly similar responses to normobaric hypoxia and terrestrial altitude (Woods et al., Firstly, our research was conducted in a normobaric hypoxic chamber which simulated the low PORelative to placebo, This study was carried out in accordance with the recommendations of the Leeds Beckett University Research Ethics Committee, and adhered to the principles set out in the Declaration of Helsinki. The protocol was approved by the Leeds Beckett University Research Ethics committee. All subjects gave written informed consent prior to participation.OS conceived the study. OS, LD, MB, KD, JM, EW, DW, LX, BS, MS, and JO designed the study. OS and JM performed the data collection. LX and MS performed chemiluminescent analysis. OS and KD conducted the statistical analysis. OS, JM, and KD drafted the manuscript. OS, LD, MB, KD, JM, EW, DW, LX, BS, MS, and JO contributed in the revision of the manuscript. All authors approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Pregnancy at early age is the most significant modifiable factor which consistently decreases lifetime breast cancer risk. However, the underlying mechanisms haven't been conclusively identified. Studies in mice suggest a reduction in progesterone-receptor (PR) sensitive epithelial cells as well as a downregulation of the Wnt signaling pathway as being one of the main mechanisms for the protective effect of early pregnancy. The aim of our study was to validate these findings in humans.We collected benign breast tissue of 125 women who had been stratified according to age at first pregnancy and the occurrence of subsequent breast cancer, and performed immunohistochemistry for PR, Wnt4 and the Wnt-target Versican.p = 0.0135). In women with early pregnancy, expression of Versican and Wnt4 was significantly lower compared to nulliparous women (p = 0.0036 and p = 0.0241 respectively), and Versican expression was also significant lower compared to women with late pregnancy (p < 0.0001).The number of PR positive epithelial cells was significantly lower in the group of women with early pregnancy and no subsequent breast cancer compared to the group of nulliparous women with subsequent invasive breast cancer (Our results confirm prior observations in mice and suggest a role of downregulation of epithelial Wnt signaling in the protective effect of early pregnancy in humans. This results in a decreased proliferation of stem/progenitor cells; therefore, the Wnt signaling pathway may represent a potential target for breast cancer prevention in humans. With an estimated 1.7 million cases per year worldwide, breast cancer is the most frequent cancer in women. Despite substantial progress in early detection and treatment, 521.900 women die of their disease each year .The protective effect of a full-term pregnancy is most pronounced for a pregnancy occurring before the age of 20, resulting in a lifetime breast cancer risk reduction of >50% in comparison to nulliparous women . In addialterations in circulating hormones [decrease of hormone responsiveness of the mammary gland [alterations in the composition of the mammary extracellular matrix (ECM) [terminal epithelial cell differentiation occurring during pregnancy [The underlying mechanisms of this protective effect have not been conclusively identified and may not be explainable by a solitary mechanism, but by a combination of mechanisms  such as hormones , 8, a pary gland , 8, longix (ECM) , 8, 10, regnancy , 11–13.alteration of cell fate of specific mammary epithelial cells including mammary stem/progenitor cells represents another promising explanation. Persistent changes in transduction pathways, growth and transcription factors and/or cell cycle regulatory molecules during pregnancy may allow breast epithelial cells to become less proliferative and more resistant to carcinogenesis [Parity induced ogenesis , 14. Of ogenesis , 16.Meier-Abt et al.  analyzedThe results of their study suggested that in mice, early pregnancy leads to a reduced response of the mammary gland to progesterone, caused primarily by a decrease in the proportion of PR positive luminal cells. Consecutively, a downregulation of Wnt4 expression in the luminal epithelial cells, and a reduction of the Wnt signaling pathway in basal stem/progenitor cells was identified . Wnt ligIn a second study, the authors demonstrated that the reduction of PR expressing cells and downregulation of Wnt signaling pathway in parous mice are of life-long duration , and thaIn humans, only a few studies have analyzed the effect of pregnancy on breast cancer cells. A decrease of PR positive luminal cells as well as Wnt signaling can also be observed in parous compared to nulliparous women , 25. HowOf the 125 patients identified, 35 (28%) were nulliparous, 52 (42%) had full-term pregnancy before the age of 30 and 38 (30%) after the age of 30   .To determine if the pattern of expression of PR, Versican, Wnt4, CK15 and β-Catenin in benign breast tissues differed between the groups, we performed an immunohistochemical analysis for these markers. Representative staining for Versican, Wnt4 and PR are shown in Figures ) Figure . Additio) Figure . There wp = 0.0449)   compared to the group of women with early pregnancy  . Interes) Figure .p < 0.0001, Fisher's exact test)  and compared to nulliparous women   compared to the women with late pregnancy  Figure . Within ct test) .p >0.05).No association between PR  and Versican  in any of the individual groups could be found  with immunohistochemistry against PR, Versican, Wnt4, CD15 and β-Catenin. The number of cases was too small for a statistical analysis, but looking at the results there were no differences between the cancer cases. 3 breast cancers were completely negative for PR, and all but one were negative for Wnt-4 and Versican. It thus seems as if in these few available breast cancer cases, there was no particular activation of the Wnt-pathway.In the present study, we performed immunohistochemistry for PR, Wnt4, Versican, β-catenin and CK-15 on breast tissue from different subpopulations of women, stratified according to age at first pregnancy and occurrence of breast cancer. In accordance to correspondent mouse studies , 23, we th and 23rd week of pregnancy, the reduction of PR positive epithelial cells was distinct when compared to nulliparous women. In their study, a possible association with subsequent breast cancer was not analyzed [In accordance with the results of our study, PR positive cells are reduced not only in parous rodents, but also in parous humans , 26, wheanalyzed . The resanalyzed , 23, whoanalyzed .Meier-Abt et al. , 23 examOur analysis of Wnt4 expression showed similarities to the results of PR. Additionally, expression of Wnt4 was significantly higher in nulliparous compared to parous women. However, since the number of cases evaluable for Wnt4 in our study was small (12 cases), further experiments to test this hypothesis are warranted.By analyzing β-catenin, Versican and CK15, we examined important targets of the Wnt pathway. Although no significant difference was observed in the expression of β-catenin and CK15, a Wnt target and an epithelial stem cell marker, respectively, between the various groups, we found a clear impact of pregnancy and, interestingly, early pregnancy, on the expression of Versican, with significantly lower expression in women with early pregnancy compared to those with late pregnancy and to nulliparous women.Our findings thus support the hypothesis that early pregnancy, possibly through a decrease of PR positive luminal cells, leads to a decreased activation of the Wnt- signaling pathway and subsequent lower expression of the Wnt target Versican, which, by inhibiting the proliferation of mammary stem/progenitor cells, may be the reason for the lower incidence of subsequent breast cancer in these women.Nevertheless, the limiting factors of our study need to be addressed. First, the number of patients in each group is rather small and heterogeneous. Specifically, the number of cases that could be analyzed for Wnt4 was small. Despite the small sample size, we demonstrated a relationship between a higher expression of certain components of the Wnt pathway, such as PR, Wnt4 and Versican, and the occurrence of subsequent breast cancer.This is the first study examining the effect of early pregnancy on PR expression and the activation of the canonical Wnt/β-catenin pathway in human breast tissue. We demonstrated a lower expression of PR, Wnt4 and Versican in women with an early pregnancy compared to women with a late pregnancy and compared to nulliparous women. Our results suggest, in accordance with previous studies in mice, that the downregulation of epithelial Wnt signaling is associated with the protective effect of early pregnancy against breast cancer in humans. The reduction in Wnt signaling may occur through a decrease of PR-positive luminal cells, which results in a decreased expression of the PR target Wnt4 and the Wnt4 target Versican, and consequently in a decreased proliferation of stem/progenitor cells. The Wnt signaling cascade might thus represent a potential target for breast cancer prevention strategies and further studies to define the underlying molecular mechanisms are needed.Based on the database of the Institute of Pathology at the University Hospital Basel and associated clinical information, a total of 125 women who underwent biopsy of benign breast tissue were identified. The cohort was divided into six groups based on age at first pregnancy and the occurrence of subsequent breast cancer , Versican , Cytokeratin(CK)15 , β-Catenin  and Wnt4 . For all immunohistochemical stainings, DAB was used as a chromogen. All antibodies were extensively tested and validated using normal breast tissue (progesterone), normal thyroid tissue (Wnt4), normal brain tissue (Versican), normal esophagus (CK15) and desmoid tumors (β-Catenin).Immunohistochemical analyses were performed by R.M and S.M., blinded for the subgroups of the patients. For PR, Wnt4, CK15 and β-Catenin, 200 breast epithelial cells were counted and the number of positive cells for each marker was recorded. For Versican, the staining was scored as: 0 = absent staining, 1 = only nuclear staining, 2 = weak cytoplasmic staining, and 3 = strong cytoplasmic staining of epithelial cells. For statistical analyses, 0 andd were considered negative for Versican, and 2 and 3 (cytoplasmic staining) were considered positive. Examples of staining for Versican and Wnt4 are given in Figures P-values < 0.05 were considered statistically significant. All analyses were performed using Graphpad Prism 6.0  or SPSS v.20 .Statistical analyses for categorical and non-categorical variables were performed using Fisher's Exact and Mann-Whitney U tests. All tests were two-sided.


The cellular membrane of eukaryotes consists of phospholipids, sphingolipids, cholesterol and membrane proteins. Among them, cholesterol is crucial for various cellular events  in addition to its essential role as an ingredient of steroid hormones, vitamin D, and bile acids. From a micro-perspective, at the plasma membrane, recent emerging evidence strongly suggests the existence of lipid nanodomains formed with cholesterol and phospholipids . Thus, it is important to elucidate how cholesterol behaves in membranes and how the behavior of cholesterol is regulated at the molecular level. To elucidate the complexed characteristics of cholesterol in cellular membranes, a couple of useful biosensors that enable us to visualize cholesterol in cellular membranes have been recently developed by utilizing domain 4 (D4) of Perfringolysin O , a cholesterol-binding toxin. This review highlights the current progress on development of novel cholesterol biosensors that uncover new insights of cholesterol in cellular membranes. Cells produce cholesterol through the de novo pathway in the endoplasmic reticulum (ER) and this lipid is transported to cellular compartments via either vesicular or non-vesicular routes . A part One of the hallmarks of cholesterol is that this lipid has a rigid four-ring sterol backbone that is hydrophobic and possesses only a hydroxyl group as a polar head group . The larClostridium perfringens and binds to cholesterol in the exofacial leaflets of the PM resulting in pore formation and cell lysis [To elucidate the characteristics of cholesterol in cellular membrane in great details, visualization of cholesterol in a single cell is required and, until now, a variety of cholesterol biosensors have been developed . Dehydroll lysis . PFO strll lysis ,46; Figull lysis ,48. By ill lysis ; Figure 125I-PFO*) using a scintillation counter  is a domain of PFO which size is 13 kDa and D4 is the smallest segment that is sufficient to bind to cholesterol with no cytotoxicity ,48,63,64A. D4 assD434S mutant, named D4H (D4 with higher affinity), is applicable to visualize cholesterol in the cytosolic leaflets of the PM by expression in cytosol [A previous study found that a point mutation, D434S in domain 4 decreases the threshold for cholesterol-binding to PFO  and it w cytosol . Cytosol cytosol ,67,74; F cytosol , suggest cytosol . PtdSer  cytosol . A hallm cytosol ,76. By d cytosol . Importa cytosol . In vitr cytosol , suggest cytosol . EM anal cytosol . Taken t cytosol ,74. Furt cytosol .D434S), other point mutations were introduced in D4 in addition to D434 [To decrease still high threshold for cholesterol-binding to D4H  ,51,60,61


Recent findings on hearing loss (HL) and healthy aging have highlighted important clinical and policy implications concerning quality of life in older adults. Our objective was to explore the impact of HL on quality of life in older Americans, independent of survival, using the 2000 Medical Expenditure Panel Survey (MEPS) and 2010 Census data.A retrospective cohort of 2,567 subjects aged 60–90 in the MEPS who provided information on self-reported HL, health-related quality of life and chronic conditions. The EQ-5D visual analog scale (VAS) transformation was used to estimate marginal utility decrements for 5-year age categories and conditions, including HL, hypertension, diabetes, angina, joint pain, asthma, emphysema, or blindness. The modeled decrements were applied to the 2010 US census population to estimate annual quality-adjusted life years (QALYs) lost.Of the respondents, 15.4% had mild HL and 1.1% had moderate/severe hearing loss. Other conditions (utility decrement) included: joint pain 53% (.0643), hypertension 47.2% (.0292), diabetes 15.6% (.0577), angina 9.8% (.0352), asthma 7.9% (.0288), emphysema 4.5% (.1186), blindness 0.8% (.0836), and average age 71.0 with decrements .0033 per year. The decrement from hearing loss ranked 4th at 174,689 in the US population.The substantial impact of hearing loss on healthy aging may not be obvious when quality of life decrements include survival or when diluted with younger populations. Careful consideration of clinical interventions for age-related HL is warranted and further research is needed on the effect of HL on quality of life in otherwise healthy older adults. Patient-centered outcomes research (PCOR) is an important US policy initiative that is aimed at “promoting science that sees through the eyes of patients” . The PCOPresbyacusis, or age-related HL, is among the most common chronic conditions of aging, with 26.7 million individuals age 50 and older reporting hearing loss; however, only approximately 14% of older adults use hearing aids [HL can affect many aspects of medical care and may be correlated with other health problems. Hearing loss has been associated with perceived problems in access to medical care , negativThis study examined data for 2,567 subjects, ages 60–90, in the 2000 Medical Expenditure Panel Survey (MEPS). Self-reported HL, HRQoL, and chronic conditions present were extracted from the publicly available 2000 national MEPS survey. The MEPS sample was limited to individuals 60–90 years of age in order to represent the aging population. HL was defined as “mild” if the respondent indicated they “can hear most conversations”, and “moderate/severe” if they could hear “some conversations”. The EQ-5D VAS response was used as the indicator of HRQoL . We compThe MEPS data sample was extracted from a public use data set that contains no subject identifiers, and was therefore classified as non-human research.Age, gender, and race distributions were examined using means and proportions, as appropriate. The EQ-5D VAS responses in the MEPS data were converted to utilities using an approach developed by Dolan  and applAll statistical analyses were conducted using SAS version 9.3 statistical software . Population estimates were performed using Microsoft Excel.Respondents in the MEPS national survey data set had a mean age of 71.0 years ; 45.7% were male, and 10.5% of black race; 15.4% reported mild HL and 1.0% reported moderate/severe hearing loss; 49.0% reported a diagnosis of one or more of: myocardial infarction, asthma, angina, diabetes, joint pain, emphysema, cancer, hypertension, kidney disease, osteoporosis, stroke, blindness, or memory problems. The mean EQ-5D VAS scores observed were 72.6  and the mean utility score based on the reported VAS score was 0.81 , which is representative of the US population over the age of 65 . SubjectModerate-to-severe HL  was associated with decrements double that of blindness  and 30% worse than emphysema . Health-related quality of life utilities and p-values comparing quality of life decrements due to HL and other health conditions are shown in Applying the utility loss for a condition to the total US population integrates the effects of prevalence of a condition with the utility decrement associated with the specific disease. The greatest number of QALYs lost in 2010 for the US population over the age of 60 is due to the effects of diabetes, hypertension, and joint pain. However, approximately 175,000 QALYs were lost due to HL , which sThese results add to a growing body of evidence that HL significantly affects patients’ evaluation of their quality of life. Mild HL appears to have a small effect on HRQoL, as indicated by a utility score of −0.01, indicating a 1% decrement in HRQoL . For theth when estimating the impact on healthy aging in the US geriatric population. It is important that we begin to examine ways to close this knowledge gap before the large cohort of aging baby boomers begin to experience the loss of HRQoL that is evident with presbyacusis.These exploratory findings warrant careful consideration of age-related HL in clinical practice and further research to examine the effect of impaired hearing and of the need for hearing loss interventions. Significant differences in some models related to age, gender, and race emphasize the importance of providing rehabilitation approaches that can be adapted to patient needs and preferences, and adjusted according to the etiologies of age-related HL. Given that a large proportion of patients with HL do not use hearing aids, our findings that HL is clearly associated with decrements in HRQoL on par with common chronic conditions make it clear that a pervasive void in understanding the need for intervention may exist, both among patients and medical practitioners. HL may be expected to have a substantial impact on healthy aging, which is not obvious when quality of life decrements are calculated to include survival, or when its effects are diluted by including younger populations as is commonly done in studies of national and international burden of illness. From a public health perspective HL ranks 31 in the global burden of disease (GBD) , 19, wit


P < 0.0001). More specifically, compared to the arch, the rear of the foot and front of the foot were significantly more conducive to the generation of melanomas . The distribution pattern was not associated with differences in age, gender, right/left foot involvement, ulceration, mitosis, lymph node metastasis, tumor thickness, or stage. The overall, distant metastasis-free, and recurrence-free survival rates did not differ significantly between the stress-bearing and non-stress-bearing areas. Furthermore, while acral melanomas tended to develop on stress-bearing areas, the distribution pattern was not associated with the prognostic index or survival.The pathogenesis of melanomas emerging in plantar surfaces remains unclear; however, mechanical stress has been reported to increase the formation of melanomas. In this study, we conducted a multicenter retrospective analysis of 153 acral melanomas diagnosed between 2000 and 2015 in Taiwan. The male-to-female ratio of the patients in question was 1:1.28, and the mean age at diagnosis was 68 years. We examined whether melanomas which developed in different areas of the patients’ soles differed in their associations with various clinicopathological characteristics and survival. Testing by goodness of fit indicated that stress-bearing areas were significantly more conducive to the generation of melanomas than non-stress-bearing areas ( Melanomas on sun-exposed areas are seen less frequently in Asians than in Caucasians. Relatedly, ultraviolet radiation does not appear to be associated with melanomas in Asian populations4. Other typical risk factors for melanoma, such as a personal or family history of previous melanoma, fair skin, and preexisting melanocytic nevi, seem to be less applicable in acral melanomas5. Meanwhile, the relevance of mechanical stress to acral melanomas has previously been reported with respect to Scots, Japanese, and Koreans patients7. To our knowledge, however, no studies regarding the relationship between mechanical stress and acral melanomas in Taiwanese patients over a given time period have previously been reported. Therefore, we established a clinical database of acral melanomas in Taiwanese patients and analyzed whether any differences in frequency and clinicopathological characteristics existed among melanomas that developed in different areas of the sole. In this study, the associations between the distribution pattern of the melanomas and the overall survival (OS), distant metastasis-free survival (DMFS), and recurrence-free survival (RFS) rates were evaluated11.In Asia, over half of all melanomas are acral melanomasin situ in 21 cases, 1.00 mm or less in 23 cases, 1.01 to 2.00 mm in 37 cases, 2.01 to 4.00 mm in 33 cases, and 4.01 mm or more in 39 cases. The mean Breslow thickness of invasive melanoma was 4.1 mm. At presentation, 13.1% of the cases presented melanoma in situ, 68.6% of the cases were at stage I or II, and 18.3% of the cases were at stage III or IV. After diagnosis, the mean duration of patient follow-up was 4.1 years.Summarized data regarding 153 melanoma patients are listed in Supplementary Table 2 pixels  based on the same previous study6. For the 114 lesions located on the soles of the patients (excluding those on the borders), the locations of the melanomas were as follows: 52 lesions (0.9 lesions per square centimeter) were located on the rear of the foot, 37 (0.82 per square centimeter) were located on the front of the foot, 16 (0.46 per square centimeter) were located on the midfoot, and 9 (0.21 per square centimeter) were located on the arch. The density of tumors on the entire plantar surface was 0.46 lesions per square centimeter. The front of the foot, midfoot, and rear of the foot are considered to be stress-bearing regions, while the arch is regarded as the non-stress-bearing portion. Testing by goodness of fit indicated that the stress-bearing areas were significantly more conducive to the growth of melanomas than non-stress-bearing areas (P < 0.0001) , but the midfoot was not significantly more conductive to the generation of melanomas (P = 0.059). Moreover, the distribution pattern of the melanomas was not associated with age, gender, right/left side, ulcer, mitosis, lymph node metastasis, tumor thickness, or stage  . The results of Cox univariate and multivariate analyses of the OS, DMFS, and RFS rates are shown in Table The 5-year OS rate of the 153 melanoma patients was 57.8%. When stratified by stress-bearing (n = 105) and non-stress-bearing (n = 9) melanomas, the 5-year OS rates of the melanoma patients were 55.8% and 65.6%, respectively. Kaplan-Meier curves revealed no significant OS, DMFS, and RFS differences between melanomas developed in stress-bearing and non-stress-bearing areas (st) Fig. . There w13. Acral melanomas may present a greater diagnostic challenge because the lesions often are presented and diagnosed late, adversely affecting outcomes12. The present study constitutes one of the largest studies of acral melanomas to date, including data on 153 foot melanomas included in a multi-center database for Taiwan. The mean age of the melanoma patients in question was 68 years, which was similar to the mean ages of patients in previous Japanese and Chinese studies but older than the mean age of patients in a Korean study of acral melanomas7. The mean Breslow thickness in this study was 4.1 mm, which was similar to the mean thickness reported for the aforementioned Chinese data but much thicker than the mean thickness values of other previous studies regarding cutaneous melanomas, suggesting that acral melanomas had a more advanced thickness1. There was a slight female predominance among the melanoma patients in this study , consistent with previous reports14. Compared with those of Korean and Chinese patients, the acral melanomas of the Taiwanese patients in this study had a lower rate of ulceration (32.7% vs. 42.3% and 47.9%) and a lower proportion of melanomas thicker than 4 mm (25.5% vs. 32.9% and 40.8%), findings which might explain the higher 5-year survival rate of the patients in this study (57.8% vs. 49.3% and 53.3%)5. Compared to those previously seen in Caucasians, meanwhile, the acral melanomas of the Taiwanese patients in this study were usually diagnosed at a late stage and resulted in a lower survival rate14.Acral melanomas are the most commonly occurring subtype of melanoma in Asians and have many unique characteristics compared with the cutaneous melanomas seen elsewhere in terms of presentation and prognosis17. A recent study showed that acral melanomas have distinct genetic features, with a lower frequency of ultraviolet-induced characteristics18. Due to their anatomical locations, it has been hypothesized that trauma might be a predisposing factor for acral melanomas5. Supporting this hypothesis, the subsite distribution of acral melanomas is noticeable19. Mechanical stresses such as plantar pressure and shear stress are higher on the front and rear areas than on other areas of the sole20. This may be an important feature in determining where skin breakdown develops more frequently. One more crucial factor may be that volar skin is hypopigmented even in black skin16. This hypopigmentation could be a considerable cause of melanomas in volar skin, given that melanin molecules function as a blocker of the free radicals generated locally by inflammation16.Melanomas on volar sites are of interest because they arise on skin that receives little or no sun exposure, unlike cutaneous melanomas on other anatomic sites7. The relevance of mechanical stress to melanomas was also later reported in Koreans patients, but that study only conducted anatomic mapping of the distribution of melanomas and did not include data on the RFS and DMFS rates5. Minagawa et al. established an analysis method to quantitatively evaluate the association between mechanical stress points and melanomas6. However, their sample size was small and they did not analyze clinicopathological factors other than thickness6. Furthermore, a survival analysis was not included in their study14. Using a measure of goodness of fit, we found that stress-bearing areas, especially the rear and front of the foot, were significantly more conducive to the development of melanomas than non-stress-bearing areas6. As in previous reports, there were no significant differences between the genders regarding the distinct sites on the sole on which melanomas were found, as well as no differences between left and right soles in our study7. In addition, the prognostic indexes including ulceration, thickness, mitosis, positive lymph node, and stage did not differ significantly between the stress- and non-stress-bearing portions5. Although the acral melanomas tended to develop on stress-bearing-areas, their distribution pattern was not associated with death, metastasis, or recurrence5. The focus of this study, as well as previous studies, was on investigating the link between mechanical stress and acral melanomas on the soles, hence whether the research findings can be applied to acral melanomas found on the hand is still a matter of uncertainty21. Further research involving the collection of enough data on palmar melanoma cases is required. The etiology of acral melanomas remains unclear, but several risk factors have been identified, including penetrative injury, heavy exposure to agricultural chemicals, and ethnicity15. Future molecular studies are needed to determine the mechanism through which mechanical stress is associated with higher incidences of melanomas.In a previous study of 47 cases, plantar melanomas were noted to be more common on the weight-bearing areas of the soles in a Caucasian population6. In addition, cases for which clinical photographs and clinical data  were lacking were excluded (n = 74) from the study. Since the relevant clinical photographs were not available, these cases could not be appropriately categorized into groups for further analysis. Cases involving lesions located on the hands were excluded (n = 20) due to the relatively small sample size for this type of lesion6. After excluding the cases above, 153 acral melanomas remained  and it was performed according to the principles of the Declaration of Helsinki. Five hundred and seventeen consecutive pathology reports regarding melanomas diagnosed between January 1, 2000, and December 31, 2015, at National Taiwan University Hospital and Kaohsiung Chang Gung Memorial Hospital were obtained using a computer-assisted search. All the cases were evaluated by at least 2 pathology faculty members at the time of diagnosis. Pathology reports from the study period were evaluated, and biopsies from specimens that were not adequate to check the lesion precisely were omitted (n = 63). Cases involving the local recurrence or metastasis of melanomas (n = 130) were likewise excluded from the study, as were melanomas not classified as acral melanomas (n = 77)22. The influence of each factor on survival was calculated using univariate analysis by the Cox proportional hazard models. In light of the fact that thickness, ulceration, and lymph node metastasis are features considered in staging, the stage factor was not examined in the Cox regression models. Because of missing values in many cases, mitosis was also excluded from the Cox regression models. All tests were two-sided. P values of less than 0.05 were regarded as statistically significant. Analysis were performed using SAS 9.4 .Descriptive statistics were used to summarize the data. Associations between the distribution of lesions and clinicopathological factors were evaluated with the Fisher exact test, Chi-square test, or goodness of fit test, when indicated. Survival probabilities were assayed using the Kaplan-Meier method and were evaluated by log-rank testsSupplementary information


Bifidobacterium, Megasphaera, Haemophilus and Streptococcus; and five genera enriched in infants with eczema: Escherichia/Shigella, Veillonella, Faecalibacterium, Lachnospiraceae incertae sedis and Clostridium XlVa. Several species, such as Faecalibacterium prausnitzii and Ruminococcus gnavus, that are known to be associated with atopy or inflammation, were found to be significantly enriched in infants with eczema. Higher abundance of Akkermansia muciniphila in eczematous infants might reduce the integrity of intestinal barrier function and therefore increase the risk of developing eczema. On the other hand, Bacteroides fragilis and Streptococcus salivarius, which are known for their anti-inflammatory properties, were less abundant in infants with eczema. The observed differences in genera and species between cases and controls in this study may provide insight into the link between the microbiome and eczema risk.Eczema is frequently the first manifestation of an atopic diathesis and alteration in the diversity of gut microbiota has been reported in infants with eczema. To identify specific bacterial communities associated with eczema, we conducted a case-control study of 50 infants with eczema (cases) and 51 healthy infants (controls). We performed high-throughput sequencing for V3–V4 hypervariable regions of the 16S rRNA genes from the gut fecal material. A total of 12,386 OTUs  at a 97% similarity level were obtained from the two groups, and we observed a difference in taxa abundance, but not the taxonomic composition, of gut microbiota between the two groups. We identified four genera enriched in healthy infants:  Eczema is a chronic inflammatory disorder of the skin, which commonly starts during infancy. Its prevalence has been increasing worldwide, and it is more prevalent in affluent societies. It afflicts children mainly in the first year of life 60%) and affects up to 30% of infants , 2, whil0% and afLactobacillus and Bifidobacterium [Bifidobacterium dominate the large-bowel bacterial community of human infants during the first months of life [After the term “probiotic” was defined by Lilly DM in 1965 , severalacterium . The queacterium . MembersBifidobacterium and Lactobacillus are present at a significantly greater abundance in non-eczematous infants at several time points  [Enterococcus and Shigella had a greater abundance in eczematous infants during the early stages of infancy [The diversity of the gut microbiota was reported to be significantly reduced in infants with eczema , 12. Mose years) , 13, 14.We conducted this study to compare the structures of gut commensal bacteria between healthy infants and infants who had experienced eczema during the first year of their life through a stool sample collected at 12–13 months of age, and to identify specific bacterial communities that may be associated with eczema. We performed high-throughput sequencing for V3–V4 hypervariable regions of the 16S rRNA gene from gut fecal material to characterize and compare the difference in the gut microbiota between healthy and eczematous infants.The study was reviewed and approved by the ethics committee board of Shanghai Institute of Planned Parenthood Research (IRB00008297). All participants gave their written informed consent before participation in the study.We conducted a case-control study to examine the association between microbiome and the risk of eczema among infants. Study subjects were selected from a birth cohort, in which 1,400 pregnant women were recruited in their first trimester in the Maternity and Child Care Hospital, Minhang District of Shanghai. Of the followed women, 1,048 delivered live births. We conducted home visits for all live-born infants at 12 months after obtaining their mothers’ consent. During the visit, the parents were asked to report their children’s history of physician-diagnosed eczema since birth using a structured questionnaire. Among them, 150 infants were invited to participate in a study examining the microbiota of infants. A total of 101 mothers agreed to participate and infants’ stool samples were collected at 12–13 months of age using a self-collection kit following a standard collection procedure. Among the 101 fecal samples, 51 samples were from case infants who had developed eczema within the first 12 months, while 50 were from healthy control infants who had not developed eczema during the first year of life.® 2.0 Fluorometer ; the integrity and size were checked by 1.5% agarose gel electrophoresis containing 0.5 mg/ml ethidium bromide. All DNA was stored at -20°C until further analysis.DNA was extracted from 300 mg of feces using a QIAamp DNA stool mini kit  according to the manufacturer’s instructions. The amount of DNA was determined using a Qubit5’- TACGGRAGGCAGCAG-3’) and 798R (5’- AGGGTATCTAATCCT -3’) primers specific for the V3-V4 hypervariable regions of the 16S rRNA gene [The bacterial genomic DNA was amplified with 343F (RNA gene . Barcode® 2.0 Fluorometer (Life technologies).Each PCR reaction was performed in a 50 μl system, and the products were extracted with the QIAquick gel extraction kit (Qiagen) and quantified on a NanoDrop 2000C spectrophotometer and QubitThe samples were pooled in an equal amount and sequenced on a Roche 454 GS FLX+ System  according to the manufacturer’s recommendations. The sequencing was started from the the 454 FLX Titanium adapter.Raw pyrosequencing reads obtained from the sequencer were first treated using the 454 SOP of Mothur (version 1.32.0) , in a prThe high-quality sequences were assigned to samples according to barcodes. After the removal of primers, operational taxonomic units (OTUs) were calculated by comparing to the SILVA  referencThe online software RDP classifier  was usedThe authors confirm that all data underlying the findings are fully available without restriction. The sequence data from this study have been deposited in the GenBank Sequence Read Archive under accession number SRS1054294.A total of 972,520 high quality sequences were generated from 50 eczema cases and 51 controls whose characteristics are presented in Since the sample with the fewest sequences had 6,134 reads, we normalized each sample to 6,134 reads using command "sub.sample” of Mothur. After data normalization, a total of 12,386 OTUs at a 97% similarity level were obtained from the two groups, comprising 7,418 species level OTUs in healthy control infants and 8,566 OTUs in infants with eczema. The Good’s coverage was estimated to be 98.82% for healthy controls and 98.69% for eczematous infants, indicating a sufficient sequencing depth for an eczema-associated fecal microbiota investigation in infants. This was also supported by the rarefaction curve of 101 samples, which tended to be asymptote after the rarest OTUs (only one observation) were removed according to the former method , suggestNeither the bacterial richness nor diversity  was significantly different between the two groups  and prinTo compare the microbiota composition of two groups, a Venn diagram of OTUs at a 97% similarity level was generated . AlthougThrough the rank abundance curves of the two groups, we revealed that the majority of the abundance came from a few OTUs . From thFirmicutes, Proteobacteria, Bacteriodetes and Actinobacteria)  . These pEnterobacteriaceae, Ruminococcaceae, Veillonellaceae, Bacteroidaceae, Lachnospiraceae, Bifidobacteriaceae, Verrucomicrobiaceae, Streptococcaceae, Pasteurellaceae and Lactobacillaceae.At the family level, 83 families were identified. Two groups shared 57 families. The healthy and eczematous groups had 14 and 12 unique families, respectively. The top ten shared families, which accounted for more than 92.7% of the total sequences, were Escherichia/Shigella, Bacteroides, Bifidobacterium, Veillonella, Faecalibacterium, Megasphaera, Haemophilus, Lachnospiraceae incertae sedis, Streptococcus, Clostridium XlVa and Megamonas. The genera Bifidobacterium and Bacteroides belonged to the phylum Actinobacteria and Bacteroidetes, respectively; and Escherichia/Shigella and Haemophilus were members of the phylum Proteobacteria; the other seven genera were classified to the phylum Firmicutes.In total, the sequences from fecal microbiota represented 170 bacterial genera, with 143 genera in infants with eczema and 136 genera in healthy infants. The 11 most abundant genera   accounteEnterobacteriaceae and Bacteroidaceae.To investigate the difference in the taxa abundance of fecal microbiota between healthy infants and infants with eczema, Metastats was used to identify the differentially abundant phylotypes  including phylum, family and genus  Tables. Megamonas and Bacteroides) and 41 less abundant genera  in healthy infants; the other five genera, Escherichia/Shigella, Veillonella, Faecalibacterium, Lachnospiraceae incertae sedis and Clostridium XlVa, were enriched  in infants with eczema. There were 27 and 34 unique genera in the microbiota of healthy infants and infants with eczema, respectively  between the two groups, including nine out of the 11 most abundant genera  between the two groups. In infants with eczema, 170 OTUs were enriched versus 154 in healthy infants. Here, we focused solely on the abundant OTUs (proportion>0.1%), and 18 of them could be assigned into 13 species based on the Silva and NCBI databases . Among tPorphyromonadaceae and Desulfovibrionaceae and genus Clostridium XlVa. This was consistent with the Metastats-derived results  or type of feeding (infant formula or breast-fed), the following genera were consistently abundant in infants with eczema: ae sedis . The genr eczema . Meanwhi allergy , and incuman gut ; and the infants .Microbial composition analysis using several bioinformatics methods revealed that, though the taxonomic composition of the intestinal microbiota showed only slight difference between cases and controls, the abundance of OTUs in infants with eczema was significantly different from that of healthy infants.Enterobacteriaceae is usually abundant in infants with eczema [Escherichia/Shigella, a member of the family Enterobacteriaceae, was increased in infants with eczema. The most abundant OTU belonging to genus Escherichia/Shigella occupied 8.8% and 8.4% of all OTUs in infants with and without eczema, respectively, and these were enriched in infants with eczema .The family h eczema , 27. In Faecalibacterium prausnitzii accounted for 11.78% of eczematous infants compared with 5.70% of healthy infants , consistent with an earlier report [The second most abundant OTU and two other abundant OTUs all belonging to the species  infants . The resr report .Bifidobacterium were enriched in healthy infants, B. bifidum and B. longum  might indicate an imbalanced Th1/Th2 and partly contribute to allergy-like eczema.In the genus  infants . It was one year , and thethe host –35. BecaVerrucomicrobia, 128 were classified into the genus Akkermansia, which accounted for 1.78% of gut microbiota in eczematous infants but only 0.92% in healthy infants. Two abundant OTUs of the genus Akkermansia were identified as belonging to the species Akkermansia muciniphila, which occupied 94.70% of the genus Akkermansia in eczematous infants. In the animal model, higher abundance of A. muciniphila was associated with allergic dermatitis, wherein A. muciniphila, being a mucin-degrading bacterium in the intestine, reduced the integrity of intestinal barrier function causing increased uptake of allergenic proteins [A. muciniphila also contribute to eczema in infants.Out of 129 OTUs that were found to belong to the phylum proteins . Thus, wLactobacillus are well known as probiotics. For the components of 16S rRNA gene, the region V3/V4 was not suitable for identifying Lactobacillus species, so no Lactobacillus sp. was identified in the gut microbiota at the species level. However, at the genus level, we found that Lactobacillus was indeed increased in healthy infants. One member of the genus Streptococcus, S. salivarius, was enriched in healthy infants (1.88%), emphasizing its anti-inflammatory property [Species of the genus property .Ruminococcus gnavus was more abundant in eczematous infants Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file.S1 Table(XLS)Click here for additional data file.S2 Table(XLS)Click here for additional data file.S3 Table(XLS)Click here for additional data file.